Table of Content

05 April 2021, Volume 41 Issue 4

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Original Articles

Identification of a novel BTK variant in a Chinese family with X-linked agammaglobulinemia

ZHANG Han, SUN Yang, WANG Rong-rong, ZHANG Wen, ZHANG Xue

2021, 41(4):  467-471. 

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Objective To identify the pathogenic variant in a Chinese family with X-linked agammaglobulinemia (XLA). Methods A trio family with suspected X-linked agammaglobulinemia was recruited. The pathogenic variant was detected by the whole exome sequencing, and then confirmed by Sanger sequencing. cDNA sequencing was performed to find the abnormal splicing of the BTK variant. Quantitative real-time PCR was conducted to evaluate the mRNA expression of BTK in the patient. Results A novel hemizygous splicing variant (c.240+3A＞C) was identified in the BTK gene from this patient. The variant co-segregated with the phenotype of the family members was not listed in the public databases, such as dbSNP153,ExAC, gnomAD or the Human Gene Mutation Database. Further Sanger sequencing demonstrated that the BTK c.240+3A＞C variant led to a 106 bp from intron 3 of BTK insertion between exon 3 and exon 4 of the BTK transcript. The mRNA expression of BTK in the patient was significantly reduced as compared to a control individual. Conclusions The novel c.240+3A＞C splicing variant in BTK likely results in X-linked agammaglobulinemia in this family.

Association of genetic variant rs3662C＞A in the gene of p21-activated kinase 2 with risk and prognosis of stroke as well as its effect on transcription activity

SI Bin, ZHANG Shu-yuan, YANG Yun-yun, CHEN Yu, ZHANG Wei-li

2021, 41(4):  472-478. 

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Objective To identify the relationship between variant rs3662C＞A in the gene of p21-activated kinase 2 (PAK2) and risk of stroke as well as prognosis, and to explore the effect of rs3662C＞A on the transcriptional activity of PAK2 gene. Methods In a multi-center case-control study including 733 cerebral atherothrombotic stroke, 488 lacunar infarction, 435 intracerebral hemorrhage and 1 727 healthy controls, the relationship between variant and risk of three stroke subtypes was examined using the multivariate logistic regression model. The stroke patients were prospectively followed up for a median of 4.5 years, and the relationship between rs3662 and cardiovascular events as well as mortality was analyzed using the Cox proportional-hazards model. The effect of rs3662C＞A on the transcriptional activity of PAK2 gene was examined using the luciferase reporter assays. Results The rs3662A allele was associated with a decreased risk of atherosclerotic stroke as compared with rs3662CC genotype, and odds ratio was 0.67 (95% CI: 0.47~0.95,P＜0.05) in a dominant genetic model. This relationship had sex-specific difference and was significant incline to male. During the follow-up, atherosclerotic stroke patients carrying with rs3662A allele had a significantly higher risk of cardio-cerebrovascular events and cardiovascular mortality; hazards ratio was respectively 1.84 (95% CI: 1.16~2.92,P＜0.05) and 2.06 (95% CI:1.05~4.04,P＜0.05). The rs3662C significantly reduced the luciferase activity by 79% (P＜0.01). Conclusions The carriers with rs3662A allele of PAK2 gene have a reduced risk for atherosclerotic stroke but a worse prognosis. It is found that rs3662A affects the PAK2 expression level.

Effects of X-ray radiation on expression of DNA damage repair related genes in C.elegans

CHEN Li-na, LIU Zhi, ZHANG Chao

2021, 41(4):  479-483. 

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Objective To study the effects of different high-dose X-ray on the expression of DNA damage repair related genes, the mortality and fertility in C.elegans. Methods Wild-type C.elegans were irradiated with 200 Gray (Gy) or 400 Gy of X-ray, then the expression of 14 DNA damage-repair related genes were quantified by RT-qPCR. STRING was adopted to analyze the protein-protein interaction of the differential genes.C.elegans survival was counted to assess the effect of X-ray on nematodes mortality and fertility. Results X-ray radiation caused abnormal expression of multiple DNA damage repair related genes in C.elegans as compared to the control group. Mortality of C.elegans increased(P＜0.05)and fecundity decreased(P＜0.05)after radiation. Conclusions X-ray radiation promotes apoptosis and inhibits fertility of C.elegans by changing the expression of multiple DNA damage and oxidation damage related genes. The age-1,cep-1,ced-4,rad-51 and daf-2 are sensitive to radiation, which are potential targets for radiotherapy.

Annexin A1 alleviates microvascular complications in type 1 diabetic mice

BIAN Fang, LI Ning, LI Qi, LI Kang, JIN Wen

2021, 41(4):  484-489. 

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Objective To investigate the potential value of annexin A1 (ANXA1) in preventing and alleviating microvascular complications in type 1 diabetes (T1D). Methods Plasma ANXA1 level was detected in T1D patients and healthy people. T1D mice were treated with human recombinant ANXA1 (rANXA1) for 8 weeks. After the administration, the mice were subjected to the oral glucose tolerance test (OGTT). The left ventricular ejection fraction (EF) and left ventricular fraction shortening (FS) of the mice were evaluated by echocardiography. Serum urea and creatinine of mice in each group were detected by Au400. The concentration of insulin, urinary albumin and ANXA1 were measured by ELISA. The heart and kidney tissues of mice were stained with PAS or Sirius red. Western blot was used to detect protein kinase B (Akt)/p-Akt, p38/p-p38, c-Jun amino terminal kinase (JNK)/p-JNK, and extracellular signal-regulated kinase (ERK1/2)/p-ERK1/2 expression level. Results Comparing with healthy people, the plasma ANXA1 level of T1D patients decreased, while the level of C-reactive protein (CRP) increased (P＜0.05). hrANXA1 reduced the area under the curve (AUC) of the T1D mouse glucose tolerance test (OGTT) and increased serum insulin level(P＜0.05). hrANXA1 increased the left ventricular ejection fraction (EF) and left ventricular short axis shortening rate (FS) of mice (P＜0.05); hrANXA1 reduced the serum urinary albumin to creatinine ratio (ACR) and urea level of mice (P＜0.05). hrANXA1 promoted the phosphorylation of Akt in mouse heart and kidney tissues, and inhibited the phosphorylation of p38, JNK and ERK1/2 (P＜0.05). Conclusions The protective effect of ANXA1 on the heart and kidney of T1D mice is attributed to the inhibition of MAPK signaling pathway and activation of Akt survival pathway.

miR-30a inhibits autophagy of rat cardiomyocyte line H9c2 induced by angiotensin 2

YU Tian-le, MA Han, WU Xin-hua, CHEN Zhang-rong, SUN Biao, PENG Yuan, LIU Hong

2021, 41(4):  490-494. 

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Objective To explore the effect of miR-30a on autophagy of rat cardiomyocytes induced by angiotensin 2(Ang2). Methods 1)H9c2 cells were treated with Ang2 2×10^－6 mol/L and NE 5×10^－6 mol/L respectively for 48 h,miR-30a expression was measured by RT-qPCR, LC3 and Beclin-1 protein expression were detected by Western blot. 2)miR-30a mimics, miR-30a inhibitor and miR-30a NC were transfected into H9c2 cells by Lipofectamine2000 and incubated for 24 h, the transfection efficiency was detected by fluorescence microscope.RT-qPCR and Western blot were applied to detect the effect of miR-30a on autophagy of H9c2 induced by angiotensin 2. Results 1)Comparing with the control group, the expression of miR-30a mRNA in Ang2 treatment group and NE group was significantly decreased (P＜0.05). Protein expression of LC3 and Beclin-1 in Ang2 group and NE group was significantly increased (P＜0.001). 2)Comparing with Ang2 treatment group, the expression of LC3 and Beclin-1 protein in Ang2+ miR-30a mimics group significantly decreased (P＜0.001). Conclusions Ang2 reduces the expression of miR-30a and induces H9c2 cardiomyocyte autophagy, miR-30a mimics can inhibit Ang2 induced H9c2 autophagy.

RNF43 over-expression inhibits the proliferation,migration and invasion of clear cell renal cell carcinoma cell line 786-O

SUN Zhi-chao, WANG Xiu-zhi, YANG Shu-hui, PU Yang

2021, 41(4):  495-500. 

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Objective To assess RNF43 on the proliferation,migration,and invasion of clear cell renal cell carcinoma(ccRCC) cell line 786-O. Methods GEPIA website was visited to analyze RNF43 expression and perform prognosis analysis of the patients with clear cell renal cell carcinoma. RNF43 over-expressing 786-O was established by cationic transfection. Inhibitory efficiency of RNF43 over-expression and Wnt inhibitor LGK974 on 786-O cells were checked by Western blot or qPCR. The effects of RNF43 over-expression and LGK974 on cell proliferation,migration and invasion of 786-O were analyzed by CCK8,clone formation,wound healing assay and Transwell assay respectively. Results RNF43 was reduced in ccRCC tissues. Low expression was associated with poor prognosis. RNF43 over-expression or LGK974 treatment led to decreased expression of β-catenin and non-phospho (active) β-catenin in 786-O cells(P＜0.05),along with significantly reduced cell proliferation,migration,and invasion(P＜0.001). Conclusions RNF43 inhibits proliferation,migration and invasion of 786-O cells by suppressing Wnt/β-catenin pathway.

Expression and function of Met-tRNAi^Met carrier proteins in acute myeloid leukemia

SU Peng-zhong, HE Jia-huan, YU Shan, WANG Xiao-shuang, YU Jia

2021, 41(4):  501-507. 

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Objective To investigate the expression of Met-tRNAi^Metcarrier proteins eIF2A, eIF2D and MCTS1 in normal hematopoietic stem cells (HSCs) and acute myeloid leukemia (AML) cells, and to study the effect of carrier proteins on the functions of AML cell lines. Methods The expression of carrier proteins among multiple stages of mouse HSCs differentiation was analyzed by public datasets. The expression of carrier proteins among multiple stages of human HSCs differentiation was analyzed in single cell RNA-sequencing datasets. The expression of carrier pro- teins among AML patients and normal was analyzed in public datasets. eIF2A or eIF2D expression was inhibited by lentivirus-mediated gene transduction in AML cell line MOLM13, cell proliferation and cell cycle was analyzed.AML cell line was treated with salubrinal (10 μmol/L) and serum starvation, which make cells in stress, and the expression of eIF2A and eIF2D were detected. Results The expression of eIF2A and eIF2D in hematopoietic stem and progenitor cells (HSPCs) was higher than mature blood cells, and in AML patients was higher than normal. On the contrary, the expression of MCTS1 had no difference in normal hematopoietic differentiation process, as well as AML vs normal. eIF2A or eIF2D knock down significantly suppressed cell proliferation (P＜0.0001), as well as caused cell cycle arrest in G₂/M (P＜0.001). After MOLM13 was treated with salubrinal and serum starvation, the expression of eIF2A and eIF2D was upregulated. Conclusions eIF2A and eIF2D may regulate AML cell growth through breaking cell cycle arrest.

Down-regulation of Notch1 gene promotes autophagy of human liver cancer cell line HepG2

LI Chang-an, LIU Chun-xiu, XIE Xiao-juan, SUN Ai-ping

2021, 41(4):  508-513. 

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Objective To investigate the effect of Notch1 gene on the proliferation, apoptosis, autophagy and Akt/mTOR signaling pathway of HepG2 liver cancer cells. Methods Thirty-two cases of liver cancer tissues from patients undergoing liver cancer surgery in the Third Affiliated Hospital of Xinxiang Medical College were selected, and the expression of Notch1 mRNA was detected by fluorescence quantitative PCR. Notch1 siRNA eukaryotic expression vector was constructed, transfected into HepG2 cells by liposome, RT-qPCR and Western blot were applied to check the interference effect of Notch1, MTT method was used to detect cell proliferation and flow cyto- metry was used to detect cell apoptosis, Western blot was also applied in the detection of apoptosis-related proteins BCL-2, BAX, cleared-caspase-3, autophagy-related proteins LC3B, Beelinl, ATG7, ATG5 and Akt/mTOR signaling pathway related proteins Akt, p-Akt, p-mTOR, p-P70S6K. Results The relative expression of Notch1 gene in primary hepatocellular carcinoma tissue was significantly higher than that in adjacent tissues (1.865±0.791 vs 0.709±0.414) (P＜0.001). After Notch1 siRNA was transfected into HepG2 cells, the expression of Notch1 mRNA and protein in Notch1 siRNA group was significantly lower than that in NC siRNA group and in control group (P＜0.001). The cell proliferation rate of Notch1 siRNA group at 24, 48, 72 and 96 h was significantly lower than that of NC siRNA group and control group (P＜0.001). Forty-eight hours after transfection, the apoptosis rate and Bax level of Notch1 shRNA group were significantly higher than that of NC siRNA and control group, and the expression of Bcl-2 and cleaved caspase-3 were significantly lower than that of NC siRNA group and control group (P＜0.001). The expression of autophagy-related proteins LC3B, Beelinl, ATG7 and ATG5 in Notch1 siRNA group was significantly higher than that of NC siRNA group and control group (P＜0.001). The expression level of p-Akt, p-mTOR, and p-P70S6K in Notch1 siRNA group was significantly lower than those in NC siRNA group and control group (P＜0.001). Conclusions Down-regulation of expression of Notch1 gene can significantly inhibit the proliferation of liver cancer HepG2 cells, induce cell apoptosis, and increase cell autophagy. The mechanism may be related to the inhibition of Akt/mTOR signaling pathway.

Mechanism of in vitro sensitization of temozolomide to MGMT⁺ glioma cell line by FM19G11

FANG Huang-yi, YOU Chao-guo, PANG Chen, ZHANG Zhong-ding, ZHANG Zhe, KUANG Tong-shuai, SHENG Han-song

2021, 41(4):  514-520. 

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Objective To investigate whether FM19G11 affects the sensitivity of human-derived malignant glioma cell line T98G to temozolomide (TMZ) and underlying molecular mechanisms. Methods T98G cells were treated with 0, 0.5, 1 and 2 μmol/L of FM19G11 and TMZ separately and jointly. The CCK-8 assay and cell clone formation assay were used to detect the proliferation of T98G cells. The Hoechst 33258 staining assay was used to detect the morphological changes of apoptosis in T98G cells. RT-PCR was used to detect the mRNA expression of HIF-1α and its downstream genes VEGF and EPO. Western blot was used to detect the protein level of O⁶-methylguanine-DNA methyltransferase (MGMT), NF-κB P65, HIF-1α,VEGF and EPO. Results Compared with the treatment with TMZ alone, the combined treatment of FM19G11 plus TMZ had a more pronounced effect on the cell proliferation inhibition and morphological changes of apoptosis in T98G cells(P＜0.05). The mRNA and protein of HIF-1α,VEGF and EPO and the protein expression of MGMT, NF-κB P65 were all decreased.(P＜0.05). Conclusions FM19G11 mainly inhibits the expression of HIF-1α, and then down-regulates the expression of MGMT protein, thereby increasing the sensitivity of glioblastoma T98G cells to TMZ. At the same time, FM19G11 also down-regulates genes relating to tumor proliferation and metastasis downstream of HIF-1α and inhibit the NF-κB signaling pathway and then inhibits tumor growth.

Down-regulation of miR-199a-5p inhibits lipopolysaccharide- induced apoptosis of rat type Ⅱ alveolar epithelial cell line AECⅡ

WANG Wei, ZHAO Si-lin, FAN Fu-yuan, JIN Zhao-hui, LI Da, FU Yan

2021, 41(4):  521-527. 

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Objective To investigate the expression of miR-199a-5p in the serum of patients with chronic obstructive pulmonary disease (COPD) and its effect on lipopolysaccharide (LPS)-induced apoptosis of rat type Ⅱ alveolar epithelial cells (AECⅡ) and possible mechanism. Methods Forty-five patients with COPD and 45 controls were enrolled in the first affiliated hospital of Hunan University of Traditional Chinese Medicine from April 2017 to December 2018 . AEC Ⅱ cells were cultured in vitro and were divided into control group , LPS group, anti-miR- 199a-5p+ LPS group, anti-miR-NC+LP group,anti-miR-199a-5p+si-FZD4+Lgroup and anti-miR-199a-5p+si-NC+LPS group. RT-qPCR was used to detect the expression of miR-199a-5p and FZD4 mRNA in serum and cells, flow cytometry was used to detect the apoptosis rate and Western blot was used to detect FZD4, B-lymphoma-2 (Bcl-2) and B-lymphoma-2 related proteins (Bax) in cells. The dual luciferase reporter gene experiment verified the targeting relationship between miR-199a-5p and FZD4. Results Compared with the control, the expression of miR-199a-5p in serum of COPD patients significantly increased (P＜0.05), and the expression of FZD4 mRNA significantly reduced (P＜0.05). Compared with the control group, the expression of miR-199a-5p in the LPS group significantly increased(P＜0.05), the mRNA and protein expression of FZD4 significantly reduced (P＜0.05), the apoptosis rate and the expression level of Bax protein significantly increased (P＜0.05), and the expression of Bcl-2 protein significantly reduced (P＜0.05). Compared with anti-miR-NC+LPS group, cell apoptosis and the expression of Bax protein in the anti-miR-199a-5p+LPS group significantly reduced (P＜0.05), while the expression of Bcl-2 protein significantly increased (P＜ 0.05). There was no significant change in the detection indexes between the LPS group and the anti-miR-NC+LPS group (P＞0.05). miR-199a-5p targets FZD4, compared with anti-miR-199a-5p+si-NC+LPS group, cell apoptosis rate and the expression level of Bax protein in the anti-miR-199a-5p+ si-FZD4+LPS group all significantly increased (P＜0.05), while the expression of Bcl-2 protein significantly reduced (P＜0.05). Conclusions The expression of miR-199a-5p is increased in serum of COPD patients and in LPS-induced AECⅡcells. Down-regulating miR-199a-5p expression may inhibit LPS-induced AECⅡ cell apoptosis by up-regulating FZD4 expression.

Screening aging related proteins in urine from healthy adults

ZHENG Shu-xin, ZHAO Min-di, SHAO Chen, SUN Hai-dan, LIU Xiao-yan, GUO Zheng-guang, SUN Wei

2021, 41(4):  528-532. 

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Objective To screen the differential proteins related to aging in human urine and to find out the functional changes of proteins related to aging. Methods Fourteen healthy men and 15 healthy women were enrolled and divided them into three groups according to their ages (20-30, 40-50 and over 60 years old). The differential proteins were screened by label-free quantification and 2DLC-MS/MS. Results In male group, there were 100 proteins differentially expressed in three groups, 66 were over-expressed and 34 were under-expressed. In female group, there were 94 proteins differentially expressed in three groups, 79 were overexpressed and 15 were under-expressed. There were 12 differential expressed proteins related to aging in both male and female groups. These protein pathways mainly focused on coagulation system, protein ubiquitination pathway, LXR/RXR activation and lactose degradation III, etc. Conclusions Proteins related to aging can be found in urine, which may be used as biomarkers of aging and providing basis for subsequent discovery of disease biomarkers.

Differential expression of piRNAs in brain from patients with Alzheimer's disease

SUN Tian-yi, LIU Fan, QIU Wen-ying, MA Chao

2021, 41(4):  533-538. 

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Objective To investigate the differential expression of piRNAs in brain tissues from patients with Alzheimer's disease (AD), and to study the roles of these piRNAs related to AD. Methods With age and sex matching, 6 AD (pathological AD) and 7 control subjects were selected. RNAs were converted into piRNAs after extracting from the prefrontal lobe brain tissues, then piRNAs were reversed transcription into cDNA. Based on preliminary results, 10 piRNAs expressed in brain tissues were selected: DQ571030, DQ597973, DQ576872, DQ597479, DQ586404, DQ590835, DQ600318, DQ581610, DQ586113 and DQ571669. The expression of piRNAs was detected by real-time PCR. Those mRNA related with piRNAs of significant differences were further studied. Results A trend of decrease was found in the expression of all the 10 piRNAs in the AD group. The expression of 4 piRNAs DQ597973, DQ576872, DQ597479 and DQ600318 were significantly decreased in the AD group(P＜0.05). The related mRNA LRRC37A3 of DQ600318 was further studied. DQ600318 showed the most significant difference in expression. The expression of LRRC37A3 was also significantly decreased in the AD group (P＜0.01). Conclusions The decreasing expression of DQ600318 and LRRC37A3 is potentially related to AD, and the detection of DQ600318 may function as a marker of AD pathogenesis.

Hypoxia microenvironment promotes invasion and proliferation of human breast cancer cell line MCF-7

WANG Hai-yan, HUANG Ming-quan, PAN Guang-rui, WANG Gui-lin, LIANG Bin, QUAN Yi

2021, 41(4):  539-544. 

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Objective To investigate the expression of HIF-1 α, PAR-1 and VEGF in breast cancer tissues from patients with type 2 diabetes and the effect of hypoxia microenvironment on the invasion and proliferation of human breast cancer cell line (MCF-7). Methods Thirty-seven specimens of breast cancer from patients with complicated with type 2 diabetes mellitus were examined by surgical resection. The expression of HIF-1α, PAR-1, VEGF and microvessel density (MVD) in breast cancer and paracancerous tissues were examined by immunohistochemistry. MCF-7 cells were divided into control group (5.6 mmol/L glucose), high glucose group (30 mmol/L glucose), hypoxia group (1% O₂ 5% CO₂ 94% N₂) and high glucose plus hypoxia group. The cell morphology was observed by microscoped, the cell proliferation was detected by CCK-8 method, the invasive ability of cells was detected by Transwell chamber test, and the expression of HIF-1α, PAR-1, VEGF mRNA and protein were detected by RT-qPCR and Western blot. Results The expression of HIF-1α, PAR-1, VEGF and MVD count in breast cancer tissues were higher than those in periaracancerous tissues(P＜0.05); the number of cells passing through the basement membrane and the cell survival rate of MCF-7 in the high glucose hypoxia group were significantly higher than those in the control group (P＜0.01); and the expressions of HIF-1α, PAR-1, VEGF mRNA and protein in the hypoxia group and hypoxia high glucose group were significantly higher than those in the control group(P＜0.05). Conclusions Hypoxia and high glucose can promote the proliferation and invasion and the expression of HIF-1α, PAR-1, VEGF of MCF-7 cells, which is a potential target for clinical detection and treatment of breast cancer of patients with type 2 diabetes.

Effects of levetiracetam on neuroprotection and oxidative stress in neonatal rats with hypoxic-ischemic brain damage

WANG Hai-bin, MEN Qing-ling, CAI Yan-li, ZHOU Wen-long, GUO Hai-zhi, CHEN Rui-xiang

2021, 41(4):  545-550. 

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Objective To study the neuroprotective effect of levetiracetam (LEV) with hypoxia ischemia brain-damage (HIBD) model of neonatal rats. Methods The rats were randomly divided into three groups: model group, LEV low, medium and high concentration groups [gavage with 10, 30, 50 mg/(kg·d) LEV], with 10 rats in each, and another 10 rats in sham operation group. Twenty four hours after the last administration, HE staining was used to observe the pathological changes of rat brain, TUNEL method was used to detect the apoptosis of neurons, the expression of inflammatory factors interleukin-6 (IL-6), tumor necrosis factor α(TNF-α), and oxidative stress factors malondialdehyde(MDA), the content and activity of superoxide dismutas (SOD) in serum were detected by ELISA, and the protein expression of caspase-3, B-cell lymphoma-2(Bcl-2), nuclear factor erythroid 2-related factor 2(Nrf2), heme oxygenase(HO-1) in rat brain tissue were detected by Western blot. Results Compared with the sham operation group, the morphology of brain tissue cell in the model group were disordered, showed diffuse distribution, increased vacuolation and inflammatory infiltration, the apoptosis rate of neurons, the expression of IL-6, TNF-α, MDA in serum and caspase-3 in brain tissue were significantly higher(P＜0.05), and the expression of SOD in serum, Bcl-2, Nrf2 and HO-1 in brain tissue were significantly lower(P＜0.05); Compared with the model group, the above changes of LEV in low, medium and high concentration groups were significantly reduced(P＜0.05). Conclusions LEV can alleviate inflammatory reaction and oxidative stress, and play a neuroprotective role in HIBD newborn rats.

Angelica polysaccharide promotes the proliferation of rat cardiomyocyte line H9c2

YANG Ping, DAI Tian, ZHANG Su-chuan

2021, 41(4):  551-557. 

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Objective To study the effect of angelica polysaccharide on proliferation and apoptosis of rat cardiomyocyte line H9c2. Methods Rat cardiomyocyte H9c2 was divided into a control group (Control group), doxorubicin (ADR) induced H9c2 cell replication dilated cardiomyopathy cardiomyocyte group (ADR group), APS intervention (APS group), transfection anti-miR-NC (APS+anti-miR-NC group) and transfected anti-miR-4701-3p(APS+anti-miR-4701-3p group). Flow cytometry was applied to determine apoptosis, Western blot was employed to test B cell lymphoma/leukemia-2(Bcl-2), Bcl-2 related X protein (Bax), nuclear related antigen Ki-67(Ki-67) and proliferation cell nuclear antigen (PCNA) protein expression, enzyme-linked immunoassay (ELISA) method was used to detect B-type brain natriuretic peptide (BNP) level. Cell proliferation was detected by commercially available kit 8(CCK-8), and real-time quantitative PCR (RT-qPCR) was used to analyze the microRNA-4701-3p(miR-4701-3p) expression. Results Adriamycin significantly increased BNP content, apoptosis rate, and Bax protein level in H9c2 cells, and greatly reduced Bcl-2 protein expression(P＜0.05). Angelica polysac-charide obviously increased H9c2 cell viability, Ki-67, PCNA, Bcl-2 protein expression and miR-4701-3p expression after doxorubicin induction, and remarkably reduced cell apoptosis and Bax protein expression(P＜0.05). Knockdown of miR-4701-3p significantly reduced the cell viability, Ki-67, PCNA, and Bcl-2 protein levels of dilated cardiomyopathy H9c2 cells treated with angelica polysaccharide, while significantly increased its apoptosis rate and Bax protein expression(P＜ 0.05). Conclusions Angelica polysaccharide up-regulates the expression of miR-4701-3p, promotes the proliferation of adriamycin-induced rat cardiomyocyte line H9c2.

Clinical Sciences

Analysis of Traditional Chinese Medicine syndrome types in 52 COVID-19 patients with positive nucleic acid retest after discharge

CHEN Yun-kun, LIU Huang, ZHANG Wen-bin, WANG Jie

2021, 41(4):  558-561. 

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Objective To explore the four diagnostic information of Traditional Chinese Medicine and the distribution profile of syndrome types of common COVID-19 patients with positive nucleic acid retest after discharge in Xiaogan area, so as to provide basis for collecting experience and formulating scientific and efficient treatment regimen. Methods The clinical data of 52 cases of common COVID-19 patients with positive nucleic acid re-examination after discharge were collected from the First People's Hospital of Xiaogan City, Hubei Province with the basic information of patients, under lying diseases, four diagnosis information of Traditional Chinese Medicine and the distribution of syndrome types of Traditional Chinese Medicine. Results There were 22 cases (42.31%) showed abnormal clinical manifestations in 52 cases of common COVID-19 positive nucleic acid recheck after discharge, including 17 cases of cough (32.69%), followed by constipation, chest tightness, diarrhea, poor appetite, weakness, shortness of breath and other symptoms. The tongue was mainly dark red (50.00%), followed by light red and red. The tongue coating was mainly yellow greasy (53.85%), followed by white greasy, thin white, thin yellow and less coating. Clinical syndrome types were classified as 24 cases (46.15%) of dampness toxin stagnation lung syndrome, followed by cold dampness obstruction lung syndrome, lung spleen Qi deficiency syndrome and Qi Yin deficiency syndrome. Conclusions The syndrome differentiation of common COVID-19 patients with positive nucleic acid re-examination after discharge is mainly damp toxin stagnation of lung syndrome. The main pathogenic factors are damp and toxin, so the treatment should focus on dispelling dampness.

Squamous cell carcinoma of the thyroid gland: the analysis of primary and secondary disease

JIN Chao, LIU Gang, XU Xie-qun

2021, 41(4):  562-567. 

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Objective To discuss the clinical characteristics of patients with primary squamous cell carcinoma of the thyroid(PSCCT) and secondary squamous cell carcinoma of the thyroid(SSCCT). Methods The medical records of 17 PSCCT patients and 6 SSCCT patients admitted to Peking Union Medical College Hospital were systematically reviewed.Clinical symptoms, ultrasound, and pathological characteristics were compared between the two groups. Results The percentage of the female was higher than male and the mean age of diagnosis was about 56 years old in each group. The most common chief complaint of the patients was neck mass in both groups. The percentage of patients with cough(P＜0.01) was higher in SSCCT group than PSCCT group. The mean size of the tumor in PSCCT group was larger than SSCCT group(P＜0.05). Papillary thyroid carcinoma (PTC) could be complicated with or recurrent as PSCCT. The mean survival time after diagnosis was 17.1 months in the PSCCT patients and 13.5 months in the SSCCT patients. Predictors of overall survival (OS) in PSCCT patients included age, enlarged cervical lymph node, radical operation, and PTC. Conclusions SCCT is aggressive with neck mass as the most common chief complaint. PTC could be combined with or recurrent as PSCCT. Age, enlarged cervical lymph node, radical operation, and PTC were the predictors of OS in PSCCT patients.

Treatment of posterior short-segmental percutaneous pedicle screw fixation combined with unilateral percutaneous vertebroplasty for Kümmell disease stage Ⅲ without neurological symptoms

ZHU Wen-xiao, CUI Hong-xun, ZHAO Qing-an

2021, 41(4):  568-572. 

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Objective To study the clinical outcome of short segment percutaneous pedicle screw(PPS) combined with unilateral percutaneous vertebroplasty(PVP) for Kümmell disease stage Ⅲ without neurological symptoms. Methods Retrospective analysis of 11 patients with Kümmell disease stage Ⅲ hospitalized in the department of spine surgery of Luoyang Orthopedic-Traumatological Hospital of Henan Province from May 2018 to August 2019(5 males and 6 females),who were received posterior short-segment percutaneous pedicle screw fixation combined with unilateral percutaneous vertebroplasty. The mean age was 72.2 years old, with an average disease course of 5.5 months. All the patients had osteoporosis. The lesion segments included T11, T12 and L1. According to the follow up results,including the operation time,blood loss,the volume of injected bone cement,kyphosis Cobb angle,Oswesay disability index(ODI),height restoration of the vertebral, the incidence of adjacent vertebral fracture and stability of implant were assessed. Results The operations were successfully performed on the 11 patients, and all the patients were followed up for 12 to 15 months, average (14.27±1.01) months. Compared with preoperation, the observational targets (ODI, VAS score, Cobb angle and the height of anterior edge of injured vertebra) in last follow-up and a week after surgery were improved significantly(P＜0.05). Two cases had slight displacement of internal fixation during the follow-up, and 1 case had cement leakage without symptom to the disc during operation. Conclusions Short segment percutaneous pedicle screw fixation combined with unilateral percutaneous vertebroplasty is safe and effective for treatment of Kümmell disease stage Ⅲ without neurological symptoms, alleviate clinical symptoms and reduce the complication of long-term bed rest.

Mini Reviews

Advances in adeno-associated virus mediated gene therapy in tumors

YANG Xiao-juan, LI Zhen-hao, GOU Yuan-feng, HUO Xia-qin, PEI Ya-ping, LI Na, LIU Hui-ling

2021, 41(4):  573-577. 

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Adeno-associated virus (AAV) is a human parvovirus with defects and no disease.AAV vector, as an ideal carrier for the transduction of target genes, has certain advantages in long-term gene correction and gene therapy by transferring foreign bases into cells by using the virus's capacity to infect cells.Due to its advantages of extensive host, strong safety and continuous expression of foreign genes, AAV has been used in gene therapy for a variety of tumor.

SUMO modification and its regulation of signaling pathways

XIAO Yun-fei, WANG Jia-bin, GENG Hai-gang, LIU Qing-juan

2021, 41(4):  578-583. 

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SUMO modification is a kind of post-translational modification of proteins, which plays an important role in regulating protein stability, protein-protein interaction and subcellular localization. SUMO modification of protein is the main mechanism for the formation of diabetes, tumors and other diseases.It has become the key target for treatment. In this article, we have reviewed the latest advance in the relationship between protein SUMO modification and these pathways which are nuclear factor-κB (NF-κB) signaling pathway, mitogen-activated protein kinase (MAPK) signaling pathway and transforming growth factor-β (TGF-β).

Research progress on clinical characteristics and molecular genetic mechanism of pseudohypoparathyroidism

ZHAO Ping-ping, LIU Jing-fang

2021, 41(4):  584-588. 

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Pseudohypoparathyroidism (PHP) is a rare genetic disease characterized by parathyroid hormone (PTH) resistance, hypocalcemia and hyperphosphatemia. The pathogenesis is mainly related to the decrease of Gsα expression or impaired function caused by the mutation of GNAS gene encoding Gsα protein, which leads to the damage of Gsα/cAMP/PKA signal pathway. The purpose of this study is to explore the clinical features and molecular geneticmechanism of different types of PHP.

Role of mitochondrial dysfunction in the transition of acute renal injury to chronic kidney disease

ZHANG Xin, LIAO Xiao-hui

2021, 41(4):  589-592. 

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Acute kidney injury (AKI) is a clinical syndrome with a high incidence, which will significantly increase the incidence of chronic kidney disease (CKD). The mechanism of AKI-CKD transition is not clear. It may include cell cycle arrest, fewer capillaries around the tubule epithelium, inflammatory cell infiltration and so on. Mitochondrial dysfunction consequently leads to oxidative stress, inflammation response and renal tubular cell apoptosis resulting in AKI-CKD transition conclusively.

Research progress on the role of intestinal microbial imbalance in chronic constipation

YANG Jing-jing, CHEN Jia-yu

2021, 41(4):  593-597. 

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Chronic constipation is the prototype of functional constipation, and its morbidity increases gradually with aging. Intestinal microorganisms are involved in the occurrence, development, pathophysiology and other processes of chronic constipation. They can regulate the motor function of the gastrointestinal tract, and the destruction of the dynamic balance of intestinal microorganisms can lead to the occurrence of chronic constipation.

Medical Education

Preliminary exploration of standardized training of imaging residents based on mixed teaching method

CUI Cao-zhe, MA Yan-yun, WU Hui-hui, ZHANG Hui, NING Yan-yun, ZHANG Peng-li

2021, 41(4):  598-601. 

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Objective To explore a mixed teaching method that can effectively improve the clinical capacity of students in the department of imaging and residence training. Methods Sixty-six students were enrolled in the First Hospital of Shanxi Medical University in past recent 2 years and were randomly divided into traditional group, modern group and comprehensive group. Each group adopted corresponding teaching methods. Recording the OSCE scores before and after the standardized training and teaching work, and using questionnaires to obtain students' satisfaction with teaching work and self-evaluation. Results The results of the three groups before and after the standardized training were statistically different (P＜0.05). There was no significant difference in OSCE scores among the three groups before standardized training. After the standardized training of the three groups of students, there was a statistical difference between the OSCE performance groups(P＜0.05),and the comprehensive group was better than the traditional group and the modern group. The comprehensive teaching group had the highest average satisfaction (AS) scores with the teaching mode(4.36). The effective use of time AS scores (4.23 and 4.27) in traditional education group and comprehensive education group were higher than those in modern education group (3.55). The AS scores (3.64 and 3.78) of the traditional group in the application of self-learning ability and clinical skills were lower than those of the modern group and the comprehensive group. Conclusions The quadruple mixed teaching method can effectively improve the clinical knowledge and skills of resident trainees in the imaging department in the residential training.

Exploration on experimental teaching online of medical microbiology under COVID-19 pandemic situation

YU Zhi-jun, LU Li-li, WANG Yu-bin, JIANG Zhen-yu, DENG Hai-ying

2021, 41(4):  602-606. 

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Due to the COVID-19 pandemic, the routine performance of all universities was disrupted. It is a common problem faced by many colleges and universities that experimental teaching cannot be effectively carried out. The online experimental teaching course of medical microbiology on MOEC platform was applied in undergraduates in School of Medicine, Wuhan University of Science and Technology. This paper summarized the curriculum of designing, running, and the rules of evaluation of the training course.

Application of “making a nursing plan” in the teaching of human parasitology

WEI Chun-yan, DONG Yan, WANG Zhen-sheng, YANG Yin

2021, 41(4):  607-610. 

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Objective To improve the learing outcomes of parasitology by asking undergraduates of nursing school to make a nursing plan for patients with parasitic diseases during the process of teaching human parasitology. Methods Undergraduates of four-year curriculum in School of Nursing, Peking Union Medical College enrolled in 2019 (n=125) and in 2018 (n=150) were selected as objects in experiment group and control group respectively. The education in the experiment group was supplemented by writing a simulating nursing plan while in the control group only implemented the traditional teaching performance. The learning outcome was evaluated in forms of both test of key knowledge points and a questionnaire survey at the end of the last class undertaken by the author in each semester. Results The performance of the students in the experiment group was statistically better than the control group in terms of the test of key knowledge points, as well as the capacity building of self-learning , problem-analyzing and solving, nursing cognition for patients with parasitic diseases,understanding of related theory to practice, communication skill and team spirit, enthusiasm for learning parasitology, and the overall satisfaction to the teaching design and methods (P＜0.05). Conclusions The teaching of parasitology supplemented with creating simulating nursing plans for patients with parasitic diseases is guided by the concept of “workplace-based learning” in the current teaching system, and is proved to be a smart pathway of education in nursing school as a supplement to routine teaching process.

Blending teaching model of medical immunology and microbiology based on Zoom Meetings mixed with room classroom

PENG Gui-ying, HAO Yu, LI Jie

2021, 41(4):  611-614. 

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Considering the limitation of COVID-19 so that undergraduates can't return to the campus to continue classroom learning, on-line teaching based on Zoom Meetings mixed with rain classroom is being adopted to teach medical immunology and microbiology in students majoring in Chinese Medicine. At the same time, flipped classroom is also applied to this on-line teaching. In this article, it is described in detail how this blending teaching is constructed. What's more, personal teaching reflection and summary is also analyzed.

Application of KSO training system in laparoscopic skills training for urological residents

DONG Jie, XU Wei-feng, WU Xing-cheng, JI Zhi-gang

2021, 41(4):  615-618. 

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Objective To compare the training effect of KSO (Knowledge, Skill, Operation) training system with traditional clinical teaching methods on urological residents' surgical operation skills. Methods Twelve urological residents (average age 31 years old) with less than 10 laparoscopic operation experience were included in this study and were divided into two groups. Besides routine clinical work, the experimental group received KSO basic training for 6 weeks, twice a week, 3 hours each time, while the doctors in the control group received laparoscopic skills training by two operations every week for 6 weeks with traditional teaching methods. At the beginning and the end of the study, the differences of the basic operation ability of the two groups of doctors were compared, including basic clamping, advanced clamping, pile clamping, hooping, suture and knot tying. Results After the training, the operation time of KSO experimental group was significantly shorter than that of the traditional teaching control group in the four training items, including basic clamping, advanced clamping, pile clamping and hooping (P＜0.05). Conclusions Compared with the traditional clinical teaching, KSO training system has significant advantages in improving the basic operation ability of urological residents. It is a practical and effective training system.

Transformative learning theory for medical education from UK and USA and its implications to China

HOU Jian-lin, LI Ji-yun, LUO You-hui, WANG Wei-min

2021, 41(4):  619-622. 

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Transformative learning in medical education has been adapted in developed countries like UK and USA,and has achieved confirmed effects, such as medical students'attitude of learning and professional values acknowledgement during clinical management of diseases and patients, perception of medicine service in undeveloped areas and continues quality improvement of healthcare services. It is suggested that China may adapt the concept of transformative learning in medical education, pay special attention to the cultivation of medical students' competencies in critical thinking,systems-based practice, and appropriately introduce disorienting environment to challeng and stimulate studens'learning in order to optimize teaching effects.