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    05 April 2020, Volume 40 Issue 4
    Special Issues of Novel Coronavirus Pneumonia(1)
    The novel coronavirus SARS-CoV-2: high pathogenecity and its prevention and therapy
    LIU Li
    2020, 40(4):  433-439. 
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    Coronavirus has been regarded as safe pathogen that causes no diseases or only induces milder syndromes. However, At the first 20 years as human society enters into the 21st century, the world has experienced three coronaviral epidemics(SARS-CoV in 2002—2003, MERS-CoV in 2012 and SARS-CoV-2 in 2019—2020) with the most severe in the latest one. By March 12, 2020, this outbreak has caused more than 126 000 confirmed cases with over 4 600 deaths. Similar with the previous two outbreaks, SARS-CoV-2 infection was also caused by cross species transmission that was believed to be derived from bats to human, although the intermediate hosts are still remained unknown. As the same as the previous two coronaviruses, SARS-CoV-2 also belongs to the pathogen that can infect lower respiratory tracts. Patients with underlying diseases such as diabetes and hypertension are more susceptible to this infection. The severe patients manifest not only different degrees of dyspnea, but also can sometimes develop functional damages in heart and/or kidney. The results of pathological autopsy indicated that the obstructive pulmonary ventilation failure in critical ill patients might be mainly due to the complete filling of excessive amount of viscous liquids in alveolar cavities and tracheas. Patients' conditions could be even worse when they develop the followings: lymphopenia, the increased level of CRP, cytokine storm caused by the large secretion of pro-inflammatory cytokines. Because of the quite differences in syndromes from patient to patient, diagnosis and treatment should be carefully evaluated and fully based on the illness as well as the patient’s condition so as to avoid improper treatments that might aggravate disease progression and even cause the deadweight loss in lives. This review attempts to summarize the current knowledge about the typical features of this novel coronavirus such as the structure of the viral genome, the origin of the virus, life cycle, morbidity, disease progression and the strategies of clinical prevention and treatments.
    Thoughts on biosafety education of experimental course of pathogenic biology under COVID-19 epidemic situation
    ZHANG Zhuo-yang, YUAN Zhen-dong, YANG Yang, YANG He-ping, XIAO Jia-qi, ZHAO Wei
    2020, 40(4):  440-443. 
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    The current COVID-19 epidemic situation is grim, and the government has realized the importance of biosafety. This paper summarizes the situation and experience of biosafety management and education in pathogenic biology teaching laboratory from the aspects of management system, hardware construction and software construction. And through the questionnaire survey of the eight-year clinical students in Shanghai Jiao Tong University School of Medicine, we have a preliminary understanding of the students' views on the way of learning biosafety knowledge and the situation of mastering biosafety knowledge. It is expected that through the consideration of biosafety education, the curriculum design of biosafety education of pathogenic biology will be discussed to improve the knowledge level of biosafety of medical students.
    Occurrence and prevention of novel coronavirus pneumonia from the theory of plague
    MA Ya-hui, MA Shi-cai, LIAN Jian-wei, ZHENG Hong
    2020, 40(4):  444-447. 
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    Starting from the related theory of plague, this thesis reveals the relationship between plague and novel coronavirus pneumonia, it also provides integrated Chinese and western medicine treatment ideas for the occurrence and defense of 2019-nCoV pneumonia. According to the characteristics of this disease, it can be classified in the category of “plague” in Traditional Chinese Medicine (TCM). This thesis expounds the meaning and development of the plague, discusses the stage differentiation and treatment of 2019-nCoV from view of TCM. New insights into the occurrence, treatment and defense of 2019-nCoV pneumonia are helpful to cure patients with 2019- nCoV pneumonia more effectively.
    Nursing management of 2019-nCoV pneumonia patients in Emergency Department
    ZHANG Jing-min, GAO Yong-li
    2020, 40(4):  448-450. 
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    The 2019-nCoV infected pneumonia is an acute respiratory infectious disease with strong infectivity. The Emergency Department of West China Hospital of Sichuan University is the key department of screening and diagnosis of novel coronavirus pneumonia. Starting from the nursing management, this paper expounds the nursing management methods of the patients admitted to the emergency department and the effective measures, and has achieved remarkable results.
    Original Articles
    Association between SHMT1 gene promoter methylation and ischemic stroke
    HU Jing-cen, LI Lian, WANG Shu-yu, XU Lei, YANG De-lun, HAN Li-yuan, XU Guo-dong
    2020, 40(4):  451-455. 
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    Objective To explore the relationship between serine hydroxymethyltransferase (SHMT1) gene methylation and ischemic stroke patients. Methods The quantitative methylation-specific PCR was used to measure the level of SHMT1 promoter methylation in 290 healthy control individuals and 141 ischemic stroke patients. Results The results showed that the SHMT1 methylation level in the stroke group was 24.87(16.97~35.46), higher than that in the control group 6.58(2.43~15.14) (P<0.05). After adjusting for relevant risk factors, SHMT1 methylation was a risk factor for stroke(OR=1.051, 95% CI= 1.034~1.068). Area under the curve was 0.804, 95% CI=0.760~0.849(P<0.01). The level of uric acid was associated with SHMT1 methylation in the control group(rs=0.17, P<0.01), and the level of TG was associated with SHMT1 methylation in the stroke group(rs=0.18, P<0.05). SHMT1 methylation expression was negatively correlated with mRNA expression(r=-0.472, P<0.01). Conclusions The SHMT1 gene is more methylated and results in low expression in ischemic stroke patients, so SHMT1 hypermethylation is a risk factor for stroke.
    Function of molecular chaperones in the secretion of two-partner-secretion to the periplasm in Gram-negative bacteria
    ZHANG Yan-juan, WANG Zhe, CAI Kun, Huang Pi-ying, CHU Yin-di, FAN En-guo
    2020, 40(4):  456-459. 
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    Objective To investigate the function of general chaperones in the secretion of two-partner-secretion (TPS) protein across the inner membrane to the periplasm of Gram-negative bacteria. Methods The general chaperones were knocked-out and the strains were used to prepare spheroplast to see the effects on the secretion of FhaB*, which is a substrate of the representative model TPS substrates FhaB/FhaC system. Results Single knock-out of chaperones Skp,DegP,PpiD,YfgM and double knock-out of PpiD/YfgM has no or little effect on the secretion of FhaB* into the periplasm, while single knock-out of SurA or double knock-out of Skp/DegP severely affect the secretion of FhaB*. Conclusions There may be two molecular chaperone pathways mediated by SurA or by both SKP and DegP in Gram-negative bacteria, which can mediate the substrate protein of the two-partner-secretion system secretes into the peripheral space.
    Construction of three strains of recombinant oncolytic vaccinia virus and their anti-tumor activity
    LI Dan-yang, FANG Jing-jing, TANG Hui
    2020, 40(4):  460-468. 
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    Objective The aim of this study is to construct a recombinant oncolytic vaccinia virus with CCL5 and SSTR2 genes and to detect its antitumor activity. Methods Recombinant vaccinia virus eukaryotic expression plasmid pSC65-CCL5-SSTR2-Luc+ was constructed. The purified recombinant oncolytic vaccinia virus rVV-CCL5-SSTR2-Luc+ was obtained after homologous recombination with WR vaccinia virus and 23 rounds of screen- ing and purification. The concentrated recombinant oncolytic vaccinia virus was abtained by expanded culture, concentration and purification. The expression changes of CCL5 and SSTR2 in HeLa cells after oncolytic vaccinia virus infection were determined by ELISA and Western blot respectively. The killing activity of rVV-CCL5-SSTR2-Luc+ on two tumor cell lines was detected by CCK-8. Results The recombinant vaccinia eukaryotic expression plasmid pSC65-CCL5-SSTR2-Luc+ was constructed successfully; Purified recombinant oncolytic vaccinia virus rVV-CCL5-SSTR2-Luc+ with titer as 9.4×108 PFU/mL was obtained after homologous recombination, screening, purification and concentration; ELISA and Western blot results showed that after HeLa cells were infected with rVV-CCL5-SSTR2-Luc+, the expression level of CCL5 and SSTR2 protein in HeLa cells was significantly higher than those in the control group rVV-Luc+ (P<0.05 and P<0.01, respectively); The results of CCK8 indicated that the killing activity of rVV-CCL5-SSTR2-Luc+ on human embryonic renal epithelial cells HEK293T was very low, while the killing activity of rVV-CCL5-SSTR2-Luc+ on two tumor cell lines increased gradually with the infection time. In addition, the killing activity of oncolytic vaccinia virus rVV-CCL5-SSTR2-Luc+ on tumor cells was significantly increased as compared with that of the control group rVV-Luc+ (HCT116: 48 h after infection P<0.05; Hela: at 48 h, P<0.05; at 72 h, P<0.01). Conclusions Purified recombinant oncolytic vaccinia virus rVV-CCL5-SSTR2-Luc+ is constructed and purified successfully, its tumoricidal activity is verified preliminarily, so this conclusion lays out an experimental foundation for subsequent in vivo and in vitro researches.
    miR-451 negatively regulates EMT of the diabetic nephropathy by Akt
    GUO Hong-bao, LIU You-xia, JIA Jun-ya, YAN Tie-kun
    2020, 40(4):  469-472. 
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    Objective To study microRNA-451 (miR-451)'s influence on diabetic nephropathy epithelial mesenchymal transdifferentiation (EMT). Methods Taking human healthy kidney tissues and diabetic nephropathy kidney tissues, real-time PCR was used to detect the expression of miR-451, Akt and EMT related genes (E-cadhering, vimentin and alpha smooth muscle actin)in different groups; Western blot and immuno-histochemical staining (IHC) were used to detect the expression of Akt and EMT related proteins in different groups. Results The expressions of miR-451 and E-cadherin mRNA significantly reduced in diabetic nephropathy kidney tissues as compared with healthy kidney tissues, and the expressions of Akt, vimentin and alpha-SMA mRNA significantly increased. The expressions of Akt, vimentin and alpha-SMA protein significantly increased, and the expression of E-cadherin protein decreased significantly (P<0.05). Conclusions The miR-451 is likely to negatively influence diabetic nephropathy epithelial mesenchymal trans differentiation by Akt.
    Shc3 over-expression inhibits apoptosis of HCC cells and is involved in drug resistance
    ZHANG Xin-ran, ZHANG Yi, GAO Hui-er, QIANG Zhao-yan, LI Yong-mei
    2020, 40(4):  473-478. 
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    Objective To investigate the apoptosis and drug resistance mechanism of Src homolog and collagen homolog 3 (Shc3) in human hepatocellular carcinoma (HCC). Methods Human hepatocellular carcinoma cell lines HCCLM3 and HepG2 with stable Shc3 downregulation and their scramble cell control groups were selected, Huh7 and HepG2 cells with stable Shc3 up-regulation and their pCDH control groups were selected. Protein expression levels of Shc3,MEK,p-MEK,ERK and p-ERK were determined by Western blot analysis. The mRNA level of Shc3 were determined by real-time PCR. Apoptotic experiment was used to observe cell apoptosis. CCK-8 assays were used to observe cell proliferation. Results Shc3 down-regulation and up-regulation cell lines were proved. Compared with scramble cells, Shc3 down-regulation significantly decreased activity of MEK/ERK pathway (P<0.05), promoted cell apoptosis significantly (P<0.05). Compared with pCDH cells, Shc3 up-regulation increased cell tolerance to sorafenib (P<0.05), Moreover, significantly enhanced activity of MEK/ERK pathway (P<0.05). Conclusions Shc3 may increase the resistance of HCC cells to sorafenib by interfering with apoptosis of HCC cells and activating MEK/ERK pathway. The results of this study provide theoretical basis and laboratory basis for targeting at Shc3 in the treatment of hepatocellular carcinoma.
    Significance of abnormal expression of fatty acid metabolic enzymes in the diagnosis of autism
    CHEN Jian-ling, FU Ying-mei, YU Shun-ying, CHEN Jing, DU Ya-song
    2020, 40(4):  479-482. 
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    Objective To investigate the significance of expression level of fatty acid metabolic enzymes in the diagnosis of autism. Methods The protein content of fatty acid synthase (FASN), acetyl-CoA carboxylase (ACC), acetyl-CoA synthase 1 (AceCS1) and long chain acyl-CoA synthase 1 (ACSL1) in freshly frozen cerebellum tissues of autistic children were determined by Western blot. The levels of FASN, ACC, AceCS1 and ACSL1 in the peripheral blood of autistic children in China were detected by enzyme-linked immunosorbent assay (ELISA). Results The levels of ACC and FASN in cerebellum and peripheral blood of autistic patients were higher than that of control groups (P<0.05 or P<0.01). The expressions of FASN, ACC, ACSL1 and AceCS1 in cerebellum tissue and peripheral blood of patients with autism were positively correlated with the severity of autism (all P<0.01). Conclusions The severity of autistic symptoms in children is related to the expression changes of fatty acid metabolic enzymes, and the abnormal expression of fatty acid metabolic enzymes may be a group of biomarkers for the diagnosis of autism in children.
    Construction of HEK293 and C2C12 cell models transfected by LMNA mutant and related intracellular sublocalization of lamin A/C
    TAN Dan-dan, CHAI Jing-yan, LIU Jian-yun, NIE Hong-bing, XIONG Hui, WU Xiang-bin
    2020, 40(4):  483-489. 
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    Objective To construct an expression vector of fusion protein from wild type LMNA and mutant and the lentivirus vector of LMNA mutant, to study the expression and sublocalization of lamin A/C and the change of nucleus in HEK293 and C2C12 cells. Methods The wild type LMNA was cloned into pEGFP-N1 plasmid to construct pEGFP-N1-LMNA, then pEGFP-N1-LMNA-I373V plasmid for c.1117A>G site-directed mutant was constructed. The pEGFP-N1-LMNA and pEGFP-N1-LMNA-I373V were transfected respectively into HEK293 and C2C12 cells. Then C2C12 cells were selected with G418 and the sublocalization of GFP-labeled lamin A/C and nuclear morphology were observed under fluorescent microscope. pHBLV-h-LMNA-I373V-3*flag-GFP-PURO was constructed by using pEGFP-N1-LMNA-I373V plasmid as template. After packaging and detecting the titer of pHBLV-GFP-PURO and pHBLV-LMNA-C1117-3*flag-GFP-PURO, C2C12 cells were transfected by the lentivirus. The sublocalization of lamin A/C and abnormal nuclear morphology were examined by immunofluorescence staining. Results The sequences of pEGFP-N1-LMNA, pEGFP-N1-LMNA-I373V and pHBLV-LMNA-C1117-3*flag-GFP-PURO were the same as the target gene. Lamin A/C distributed uniformly at the inner nuclear membrane in HEK293 and C2C12 cells transfected by pEGFP-N1-LMNA, but distributed abnormally, forming distinct aggregates in the cells with transfection of the mutant construct. C2C12 exhibited lower transfection efficiency of plasmid than HEK293. However, transfection efficiency of lentivirus in C2C12 was improved significantly. Lamin A/C forming distinct aggregates in nucleus and abnormal nuclear morphology were also showed in C2C12 after transfection of mutant lentivirus. Conclusions Two kinds of eukaryotic expression vectors of LMNA mutant, and HEK293 and C2C12 cell models transfected by LMNA mutant are successfully constructed, which provides scientific basis for the pathogenesis of diseases caused by LMNA mutant.
    Silencing NLRP3 inflammasome suppresses apoptosis of human pulmonary artery endothelial cells induced by cigarette smoke extract
    XUE Hong, LIN Chang-jian, XIE Bao-song, XU Neng-luan, YAO Xiu-juan, WANG Hong, XIE Wei-ping
    2020, 40(4):  490-495. 
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    Objective To investigate the mechanism of cigarette smoke extract (CSE) inducing apoptosis of pulmonary artery endothelial cells (HPAECs), especially focus on the role of NLRP3(NLR family pyrin domain protein 3) inflammasome. Methods After transfection with NLRP3 siRNA and NC siRNA, HPAECs from experimental group and control group were stimulated by 10% CSE for 12 h, annexin V/PI flow method was used to detect the apoptosis of the two groups of cells before and after CSE stimulation. Cultured HPAECs were further divided into four groups:control group, CSE group (10% CSE solution was added), NAC+CSE group (10 mmol/L NAC pretreatment for 1 h and then 10% CSE culture solution) and NAC group (10 mmol/L NAC treatment for 1 h). After 12 h culture, intracellular reactive oxygen species(ROS) were detected by DCFH-DA fluorescent probe. The expression of NLRP3 and caspase-1 was determined by Western blot. Results After CSE stimulation, the survival rate of HPAECs in siNC group decreased, and the apoptosis was significantly increased as compared with that before CSE stimulation (P<0.05). After CSE stimulation, the apoptosis rate of siNLRP3 group decreased compared with that after the same CSE stimulation in siNC group (P<0.05).The intracellular ROS level in the CSE group was significantly increased as compared with that in the control group, which in the NAC+CSE group was decreased compared with that in the CSE group (P<0.05). In the CSE group, NLRP3 and caspase-1 were significantly increased compared with the control group, while the expression of NLRP3 and caspase-1 in NAC+CSE group was lower than that in CSE group (P<0.05). Conclusions CSE could induce apoptosis of HPAECs accompanied by increased NLRP3 inflammatory corpuscle expression. Silencing of NLRP3 protein expression can inhibit the apoptotic effect of CSE on HPAEC.
    Intrauterine infusion of human chorionic gonadotrophin improves endometrial receptivity in patients with polycystic ovarian syndrome
    SUN Jia-zhen, PAN Rui-yang, SHEN Su-fang
    2020, 40(4):  496-500. 
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    Objective To investigate the effect of intrauterine infusion of human chorionic gonadotrophin(hCG) on endometrial receptivity, estrogen and progesterone and their receptors in patients with polycystic ovary syndrome(PCOS). Methods The control group(n=23) consisted of healthy females, the study group consisted of PCOS patients. The study group 1(n=25) was given letrozole, and the study group 2(n=20) was given letrozole. Two days before ovulation and on the ovulation day, hCG 500 IU/mL was intraperitoneally administered. Endometrium was taken from D6 to D8 after ovulation in each group. Immuno-histochemical staining was used to detect the expression of estrogen receptor(ER) and progesterone receptor(PR). Scanning electron microscopy(SEM) was used to observe endometrial pinopode. The serum estrogen(E2) and progesterone(P) were examined by through chemi- luminescence on the endometrial day. Sixteen days after ovulation, the pregnancy rate of each group was calculated. Results The expression rate of the mature stage pinopode in the study group 1 was lower than that in the control group. The expression rate of the mature stage pinopode in the study group 1 was higher than that of study group 1(P<0.05). The P level of the implantation window and the positive expression rate of PR in the study group 1 was lower than that in the control group. The P level of the implantation window and the positive expression rate of PR in the study group 1 was higher than that in the study group 1(P<0.05).The pregnancy rate of study group 1 was lower than that of the control group, and the pregnancy rate of study group 2 was higher than that of study group 1(P<0.05). Conclusions PCOS patients are stunted during the implantaqtion window. Intrauterine perfusion of hCG may increase the number of mature pinopode and improve endometrial receptivity. This is potentially related to increased P level and PR expression.
    Effects of ARID1A loss on transcriptional regulation in hepatocellular carcinoma cell line PLC/PRF/5
    WANG Ding-ding, LIANG Jun-bo, WANG Xiao-yue
    2020, 40(4):  501-505. 
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    Objective To explore the transcriptional alterations regulated by ARID1A in hepatocellular carcinoma cell line PLC/PRF/5. Methods CRISPR/Cas9 technology was utilized to generate ARID1A knockout cells. RNA-seq was then performed in both ARID1A intact and ARID1A null PLC/PRF/5 cells to generate gene expression profiles. Differentially expressed genes were identified using DESeq2. Functional enrichment analysis of differentially expressed genes was performed using metascape database. Results Successful generation of the ARID1A knockout PLC/PRF/5 cells was confirmed by Western blot and Sanger sequencing. A total of 978 differentially expressed genes were identified by RNA-seq, among which the expression of 480 genes was up-regulated and that of 498 genes was down-regulated. These differentially expressed genes were mainly enriched in several biological processes including cell adhesion, regulation of hormone levels, hormone metabolism and regulation of MAP kinase activity. Conclusions The results indicate that ARID1A loss induces alterations in transcription in PLC/PRF/5, which may provide novo clues for elucidating the underlying mechanism of ARID1A in tumorigenesis.
    Irbesartan attenuates renal injury with diabetic nephropathy rats
    ZHANG Ya, HUANG Wen-hui, HOU Zhi-min, LIU Ju-xiang, LIU Jing
    2020, 40(4):  506-511. 
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    Objective To observe the effect of Irbesartan on renal injury in diabetic nephropathy (DN) rats. Methods The rats were randomly divided into control group, DN model group and Irbesartan treatment group. At the age of 22 weeks, body weight, random blood sugar, serum creatinine,urea nitrogen,and 24-hour urinary microalbumin were measured.The morphological changes of kidneys were observed. The expression of microRNA-192 in rat kidney was detected by fluorescence quantitative PCR,the expression of TGF-beta 1, Zeb1 and collagen Ⅰ in rat kidney were detected by Western blot and immuno-histochemistry. Results Compared with the control group, the body weight, random blood sugar, serum creatinine,urea nitrogen,and 24-hour urinary micro-albumin of the model group rats increased significantly (P< 0.05); Compared with the DN group, the body weight, random blood sugar, serum creatinine,urea nitrogen,and 24-hour urinary micro-albumin of the Irbesartan treatment group rats decreased significantly(P<0.05). Irbesartan treatment can improve the general condition and renal pathological damage of DN rats. Compared with the normal control group, the expression of microRNA-192 in the kidney of DN rats in the DN model group was down-regulated (P<0.05), while that in the Irbesartan group was up-regulated (P<0.05). WB and immuno-histochemical results showed that the expression of TGF-beta 1, ZEB1 and collagen Ⅰ protein in kidney tissue of DN model group was significantly increased as compared with that of normal group (P<0.05); Compared with DN group, Irbesartan treatment reduced expression of TGF-beta 1, ZEB1 and collagen Ⅰ protein, with statistical significance (P< 0.05). Conclusions Irbesartan attenuates renal damage in diabetic nephropathy rats.
    Identification of potential target genes for isocitrate dehydrogenase mutations in cholangiocarcinoma
    WANG Di, LI Qing, TONG Wei-min, NIU Ya-mei
    2020, 40(4):  512-517. 
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    Objective To identify potential target genes for isocitrate dehydrogenase (IDH) mutations in cholangiocarcinoma. Methods DNA methylation 450K data and mRNA expression data of the TCGA CHOL project were downloaded. Probe-wise differential methylation analysis and differential expression analysis were performed through limma and DESeq2 in the IDH-mutant vs IDH-WT comparison. Then, Spearman correlation analysis was performed between the normalized expression levels of differentially expressed genes and the methylation levels of corresponding differentially methylated sites. Genes with high methylation, low expression, and strong negative correlation were screened for enrichment analysis to explore their functions. A protein interaction network was constructed and the core modules were found by MCODE. Results The analysis revealed 11 605 differentially methylated sites, of which 10 427 were hypermethylated; and 651 of the 735 differentially expressed genes were down-regulated. Among them, 143 down-regulated genes and 328 hypermethylation sites formed 330 strongly negatively correlated combinations. The above 143 genes are enriched in the regulation of epidermal growth factor receptor signaling pathway, cell extravasation and cell division. By constructing a protein-protein interaction (PPI) network, 10 genes of its core module were identified, which were involved in biological processes such as epithelial cell differentiation and development, and cellular protein localization. Conclusions By integrating the TCGA cholangiocarcinoma transcriptome and DNA methylation data, the potential target genes for IDH mutations are obtained and the core modules of their PPI network are identified, providing new clues for the role of IDH mutations in the development of cholangiocarcinoma.
    Recombinant globular adiponectin relieves kidney injury in diabetic mice
    YANG Li-li, GUO Zhi-xin, FENG Jin-yi, SONG Ying-lun, SHI Wen-jiao
    2020, 40(4):  518-522. 
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    Objective To investigate whether recombinant globular adiponectin can reduce kidney injury in diabetic mice by regulating autophagy and endoplasmic reticulum stress. Methods Mice were randomly divided into control group,diabetic model group, low and high dose of adiponectin group (5 and 15 μg/kg dose of recombinant globular adiponectin was injected). There were 8 mice in each group. After eight weeks, blood samples were collected to determine the level of blood glucose, urea nitrogen and creatinine by biochemical method and the level of insulin was measured by enzyme-linked immunosorbent assay. Bilateral kidneys were taken to calculate the ratio of the kidney mass to body mass. The renal cortex tissue samples were stained with HE, and the rest were kept at -70 ℃. The mRNA expression of endoplasmic reticulum stress indicators (GRP78, CHOP and caspase-12) and autophagy indicators (LC3, Beclin1 and p62) was determined by real-time quantitative PCR. Results Compared with control group, the level of blood glucose, creatinine and urea nitrogen, and the ratio of the kidney/body mass were significantly increased (P<0.05), and the level of blood insulin was significantly reduced (P<0.05) in model group. There were significant pathological changes in kidney in diabetic mice in model group compared with control group. The mRNA expression of GRP78, CHOP, caspase-12 and p62 was significantly increased (P<0.05), and the mRNA expression of LC3 and Beclin1 was significantly reduced(P<0.05) in kidney in model group compared with control group. Compared with model group, adiponectin can significantly relieve the changes in model group (P<0.05). Conclusions Recombinant globular adiponectin may alleviate the renal injury in diabetic mice by regulating autophagy and endoplasmic reticulum stress.
    Comparison of liquid-phase chip and real-time fluorescent quantitative PCR for the detection of infectious diarrhoeal pathogens in children
    HAN Cong-hui, CAO Guang-jin, ZHANG Fu-zhen, HU Liang-shan, CAO Dong-lin, LI Ling
    2020, 40(4):  523-527. 
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    Objective To investigate the efficiency of liquid-phase chip on the detection of infectious diarrhea pathogens in children, and provide reference for rapid clinical diagnosis of infectious diarrhea diseases. Methods Stool samples of 96 infants and children with diarrhea were collected and detected by xTAG®Gastrointestinal Pathogen Panel (GPP), and the results were compared with the real-time PCR method. Results Total of 7 pathogens were detected by liquid-phase chip technology, with a positive detection rate of 65.63%(63/96), and the Salmonella detection rate was highest as 50%(48/96). The positive rate of single pathogen infection was 39.58%(38/96), and the positive rate of mixed infection was 26.04%(25/96). Overall, compared with the results of real-time PCR, the sensitivity and specificity of the liquid-phase chip in the detection of diarrhea pathogens were 100% and 71.74%, respectively, and negative predictive value and positive predictive value were 100.00% and 79.37%, respectively(P<0.05). Conclusions Compared with real-time PCR, liquid-phase chip is a character-ized by high throughput, rapid and reliable so support laboratory diagnosis of contagious diarrhea
    Melatonin attenuates the oxidative injury of smoking rat carotid arteries
    YANG Gen-huan, WANG Yan, LIAO Peng-zhi, JIA Yu-long
    2020, 40(4):  528-531. 
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    Objective To investigate the effect of melatonin on oxidative injury of rat carotid arteries after balloon injury. Methods Twenty male sprague-dawley rats were randomized into control, melatonin, smoking, and smoking plus melatonin. An established protocol of smoking was carried out. The carotid arteries were collected 7 days later. The average content of ROS was measured. And Western blot was used to detected the expression of NF-κB, MCP-1, VCAM-1 and ICAM-1. Results Smoking significantly induced the increase of ROS and the expression of NF-κB, MCP-1, VCAM-1 and ICAM-1 in rat carotid arteries. Moreover, cigarette smoke exposure inhibited the expression of eNOS. Melatonin minimized the effect resulted from smoking. Conclusions Melatonin may attenuate the the oxidative injury and the expression of inflammatory mediators in rat carotid arteries resulted from smoking.
    Silencing HOXB7 inhibits metastasis and increases the sensitivity to cisplatin in human cervical cancer CASKI cells
    SUN Hui-xia, GUO Zhe, XU Jing
    2020, 40(4):  532-537. 
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    Objective To explore the effect of HOXB7 on migration, invasion and the sensitivity to cisplatin in cervical cancer CASKI cells. Methods The expression level of HOXB7 in chemotherapy sensitive tissues and chemotherapy resistant tissues was detected by immunohistochemistry and Western blot. The expression level of HOXB7 in CASKI cells and cisplatin-resistant CASKI/DDP cells was detected by immunofluorescence and Western blot. sh-HOXB7 was transfected into CASKI cells and the cell invasion and migration were analyzed by transwell analysis, the sensitivity to cisplatin was detected by CCK-8 assay and flow cytometry. Results High expression of HOXB7 was found in chemoresistant tissues and CASKI/DDP cells (P<0.001); Silencing HOXB7 inhibited migration and invasion (P<0.001) and increased the sensitivity to cisplatin in CASKI cells (P<0.001). Conclusions Silencing HOXB7 inhibits metastasis and increases the sensitivity to cisplatin in cervical cancer CASKI cells.
    Clinical Sciences
    Clinical value of ultrasonography in the evaluation of multiple organs in patients with hyperuricemia
    GAO Ze-yan, LIU Xiao-yue, CHEN Li-meng, TAN Li, ZHANG Yi-xiu, LI Jian-chu, LYU Ke
    2020, 40(4):  538-542. 
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    Objective To explore and evaluate the clinical value of color Doppler ultrasound in the evaluation of multiple organs in patients with hyperuricemia. Methods The research included 50 patients with newly diagnosed hyperuricemia who were admitted to Peking Union Medical College Hospital from July 2015 to May 2017. Serum uric acid (sUA) was recorded and color Doppler ultrasound was used to examine the bilateral first metatarsophalangeal joint, knee joint, kidney, carotid artery and liver in each patient. The potential correlation between sUA level and organ lesions were searched. The subjects were further divided into gouty arthritis (GA) group and asymptomatic chyperuricemia (AH) group. The multiple organ diseases were analyzed and compared. Statistical methods used in cluded chi-square test and Spearman correlation analysis. Results In the 50 patients with newly diagnosed hyperuricemia, the rate of joint lesions diagnosed by ultrasound was 56.0%; the rate of renal morphological changes was 72.0%; the rate of nonalcoholic fatty liver disease (NAFLD) was 64.0%, of which 59.4% was moderate to severe fatty liver, and the sUA level was positively correlated with the detection rate of NAFLD(P<0.05),the detection rate of carotid atherosclerosis was 36.0%, and the level of sUA was positively correlated with the thickness of the common carotid artery (P<0.05).The detection rate of two or more organ lesions in joint, kidney, liver and carotid artery was 80.0%, and the level of sUA was positively correlated with the detection rate of multiple organ lesions (P<0.05). There was a significant difference in the detection rate of joint lesions and carotid atherosclerosis between the GA group and the AH group (P<0.05). Conclusions Color Doppler ultrasound has certain clinical application value for early diagnosis, comprehensive evaluation of lesion progression and follow-up treatment of multiple organ lesions such as joint lesions and renal damage, nonalcoholic fatty liver and carotid atherosclerosis in patients with hyperuricemia.
    Enhanced recovery after surgery promotes postoperative recovery of patients receiving robot assisted laparoscopic insulinoma enucleation
    MA Man-jiao, ZHANG Xiu-hua, LIU Zi-jia, WU Wen-ming, ZHAO Yu-pei
    2020, 40(4):  543-546. 
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    Objective To compare the postoperative recovery of patients receiving robotic laparoscopic insulinoma enucleation before and after implementing enhanced recovery after surgery (ERAS) protocol. Methods Anesthetic data of patients who received robotic laparoscopic insulinoma enucleation from January 2013 to March 2016 in Peking Union Medical College Hospital were collected and analyzed. All patients were be divided into routine group and ERAS group. Results The study enrolled 68 patients, whose obesity rate was 69.1%. The intraoperative fluid volume of ERAS group was lower than that of the routine group (P<0.05), while the difference of urine output was not significant. The maximum peak airway pressure of ERAS group was also lower than that in the routine group(P<0.05). The postoperative hospital stay length of ERAS group was shorter (P<0.05). The postoperative complications were uncommon in the ERAS group. Conclusions ERAS protocol can shorten the hospital stay length of patients receiving robotic laparoscopic insulinoma enuleation and postoperative complications are likely to be reduced.
    Mini Reviews
    Research progress of nucleostemin signaling pathway in tumorigenesis and development
    HE Zhen-hua, SHEN Fu-hui, ZENG Fan-gang, LIU Qian-lin, YUAN Jing-min
    2020, 40(4):  547-551. 
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    Silencing nucleostemin (NS) can activate the p53 pathway and p53 inhibit proliferation of the tumor cell through the phosphatidylinositol 3-kinase (PI3K)/ protein kinase B (PKB or AKT)/ mammalian target of rapamycin (mTOR) signal pathway. Low expression of NS also inhibits the PI3K/AKT/mTOR signal pathway through a non-p53 pathway. Inhibition of p53 activates the Wnt/β-catenin signal pathway. High expression of NS also activates the Wnt/β-catenin signal pathway. Over-expression of c-Myc increases NS expression and inhibits p53 expression,which may promote cell proliferation.
    Ultrasound targeted microbubble destruction technology and its application in the treatment of breast cancer
    ZHENG Wen-yi, WU Yu-ming, HAO Yi
    2020, 40(4):  552-556. 
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    UTMD which uses UCAs as carriers and combines genes or drugs to treat a variety of diseases has shown great potential.The UCAs under the low-frequency ultrasound irradiation produce sound hole and cavitation effect, which effectively increases the concentration of genes or drugs in the targeting cells that enhance the lethargy to breast cancer cells. Although UTMD technology has been proved to support clinical performance, it still faces some challenges in terms of the practical application.
    Advances in the correlation of uromodulin and UMOD gene with chronic kidney disease
    YOU Rui-lian, XU Lu-bin, CHEN Li-meng
    2020, 40(4):  557-561. 
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    Uromodulin, also known as Tamm-Horsfal protein, is the most abundant protein in normal urine. It is exclusively secreted by renal tubular epithelial cells in thick ascending limb (TAL) of the Henle loop. Past studies showed that uromodu in had various biological functions, including regulating water and salt metabolism, immunomodulation, protection against renal calculus and urinary tract infection. UMOD is the gene encoding uromodulin. Recently, with the development of genome-wide association study (GWAS), it is found that the UMOD gene is significantly associated with the progression and prognosis of chronic kidney disease (CKD). Further studies using Mendelian randomization methods verified the causality between uromodulin and CKD.
    Medical Education
    Application of flipped classroom combined with case-based teaching model in regional anatomy
    GAO Xiang-hong, LIU Qing-hua, WANG Qing, WANG Xiao-cui, ZHENG Jie
    2020, 40(4):  562-565. 
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    Objective To analyze the effectiveness of flipped classroom combined with case-based teaching model in regional anatomy teaching. Methods Undergraduates of clinical medicine major from Weifang medical University were randomly divided into experimental group and control group. In the experimental group, teaching was conducted using the mode of flipped classroom combined with case-based teaching, whereas traditional teaching was conducted in the control group. Teaching effectiveness was evaluated through examination and questionnaire at the end of course. Results Students of the experimental group scored higher in theoretical examination, mid-term test, peacetime test and final academic performance than those of control group. The questionnaire results showed that students from the experimental group were significantly better in learning initiative, self-learning capacity, clinical reasoning, problem analysis and solving capacity compared with the control group. Conclusions Combination of flipped classroom and case-based learning can improve the teaching effectiveness and be of great inspiration for the teaching reform of topographic anatomy.
    Exploration of curriculum development of statistics for medical students
    LIU Yun-yi, FENG Tie-nan, ZHU Rong-jia, TAN Jing-yu, WENG Lyu-kan
    2020, 40(4):  566-569. 
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    Objective Investigating the perspectives and collecting suggestions from medical students on the course of Medical Statistics, as well as their own career development plan and needs, so to provide reference for educational reform. Methods The investigation was implemented with self-made paper questionnaires which contained three parts: personal information, perspectives of Medical Statistics and self-learning condition with career plan. Results Eighty-two percent students (477) are satisfied with the courses of Medical Statistics. Most students believe it as an important course. However, they all think their application skills are relatively weak. Conclusions The cultivation of statistical thinking should be underlined together with a targeted curriculum system suitable for all needs and the implementation of diversified and effective teaching methods.
    Standardized training mode and evaluation system in critical care medicine for the training intensivtists
    HE Huai-wu, YUAN Si-yi, ZHOU Xiang, ZHOU Yuan-kai, TANG Bo, SU Long-xiang, Liu Da-wei, LONG Yun
    2020, 40(4):  570-573. 
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    Objective To explore the standardized training model and assessment system for training intensivtists. Methods The training doctors in ICU of Peking Union Medical College Hospital from 2015—2017, the implementation of “basic training, infection control, critical thinking and critical care medicine research” module training,and set up to 3 months for a cycle of “theory + operation + subjective assessment” of comprehensive clinical ability assessment system. Results A total of 161 training doctors from all over the country participated in the intensive clinical standardized training. The initial pass rate of the theoretical examination was 34%: after 3 months' of basic training, the passing rate of theoretical examination was significantly increased up to 75.5% (P<0.05). The first pass rate of aseptic training: 25%, through the enhanced training a significant increase to 95% (P<0.05). In the first assessment cycle (the first 3 months), the passing rate of clinical competence assessment was 44%, and the passing rate of clinical competence assessment was 91% in second assessment cycles (sixth months). Conclusions The critical knowledge, sense of control and critical thinking are relatively weak in intensive care medicine training doctors, and they can be strengthened after clinical standardized training. The comprehensive evaluation system of “theoretical assessment + operation assessment + subjective evaluation” is helpful to evaluate the critical clinical ability of the training intensivtists.
    Application of network-based training in standardized training of radiological residents in Beijing
    XUE Hua-dan, LI Juan, ZHANG Da-ming, SUN Hao, SONG Lan, LI Qi, JIN Zheng-yu
    2020, 40(4):  574-577. 
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    Objective To evaluate the application of network training platform in standardized training of residents in radiology department in Beijing. Methods The radiology residents who took the Beijing phase I skill examination in 2018 were given lectures about one year before the examination. The data of the online training platform visits, number of visitors, watch time and audience were analyzed and organized. The results of the questionnaire survey on the use of the training platform were also analyzed. The questions include whether the candidates liked the online training method and whether the online training platform can supplement the case types of daily work. Results For video courses and PPT courses, the number of course visits was significantly higher than the number of visitors(P<0.001). There was no significant difference between the duration of the course and the average actual viewing time. Visitors from departments other than radiology could be found in almost all courses. Generally, students like the online training platform and think that the training platform improves their learning efficiency and are helpful for their examination. In 2018, the scores and passing rate of the phase I examination for the standardized training of residents in the Beijing radiology department were 86% and 87.8% respectively, which were higher than that of 2017(84.5%,85.3%). Conclusions The network training platform is highly recognized in the standardized training of residents in radiology department and is helpful for residents to pass the phase I examination, which can be promoted in the future standardized training of radiology residents.
    Evaluation of curriculum setting and outcome of intensive training for clinical endocrinologists
    YUAN Tao, LI Qi, ZHANG Hua-bing, MAO Jiang-feng, ZHAO Jun, ZHU Hui-juan
    2020, 40(4):  578-581. 
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    Objective In order to meet the needs of endocrinology and metabolism specialists and so to improve the level of diagnosis and treatment, short-term intensive training was organized and its outcome was evaluated. Methods A short-term intensive training course for endocrinologists was implemented and the course evaluation form was designed by vote of WeChat.The learning results were statistically analyzed. Results The short-term intensive training course for endocrinologists were divided into 6 categories and covered by courses, including thyroid diseases (4), metabolic bone diseases (4), glucose, lipid and uric acid metabolic diseases (8), hypothalamic-pituitary diseases (9), adrenal diseases (5), and clinical reasoning training courses (5). All courses were completed in 6 days. The results were presented in both categories and ratings. The results of the category included “very satisfied”, “average” and “unsatisfied”, and the percentage of the number of voters was counted respectively. The five-points system was adopted for ratings, and the average score of all voters was calculated. Participants generally had high satisfaction with the course (average score: >4.5), and the evaluation results were 78.2%~100% in “very satisfied” and 0~21.8% in “average” category. Higher requirements for the teaching quality of common diseases (dissatisfaction rate of the trainees: 1.19%~2.30%), higher satisfaction for difficult and rare diseases (average score of 7 courses: 5). Conclusions The curriculum of endocrinologist clinical training should be developed to meet the needs of clinical work. It is necessary to develop a stratified training syste, divrsified teaching forms and quantitative course evaluation methods.
    Compilation of PBL plus CBL cases in the course of clinical biochemistry test
    ZHANG Xue-yan, YAO Hai-yan, HAO Chun-yan, CHEN Wei
    2020, 40(4):  582-584. 
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    This course covers commonly used methods in laboratory of clinical biochemistry. The teaching plan reported here is aiming for encourage students to learn the technology and methodology based on clinical needs. So the teaching plan is characterized by combination of clinical information and pathogenesis of illness with biochemical results and thus to support students' learning in an active pathway. An example is calcium concentration in circulation as a parameter to support clinical diagnosis of various illnesses.
    Practice and exploration of modularized teaching in the clinical postdoctoral training
    SHI Yu-xi, WEI Yi-zhen, ZHU Hui-juan, XUE Hua-dan, LUO Lin-zhi, PAN Hui, ZHANG Shu-yang
    2020, 40(4):  585-588. 
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    This study introduces the practice of modularized teaching in “core competency” training of the Clinical Postdoctoral Program of the Peking Union Medical College Hospital; moreover, taking the example of medical humanities model course, to compare the 2016 and 2017 module courses outcome by questionnaire and interview. The result shows that set up the “medical humanities model course” and the continuous optimization have greatly improved the course satisfaction. The Clinical Postdoctoral trainees believe module course is helpful to the accomplishing the “core competency”, to identifying the goal of training and enhancing the process management. However, the improvement of course content, format and its long-term effect, needs further exploration and discussion.
    Problems and countermeasures of general practitioners standardized training in the field of endocrine metabolic diseases
    WU Nan-nan, ZHAO Dong
    2020, 40(4):  589-592. 
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    The standardized training of general practitioners is the key to cultivate qualified general practitioners. Endocrine metabolic disease is an emerging discipline. Improving the clinical teaching quality of standardized training of general practitioners in the field of endocrine metabolic diseases is worthy of attention. In this paper, though analyzing teaching requirements and objectives,and based on our experience in clinical teaching practice, it summarized the problema in training of general practitioners in the field of endocrine metabolic diseases, such as the teaching content method, the teachers level, and student's own questions. Then, it puts forward countermeasures to improve and advance the training quality.