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Table of Content

    05 January 2022, Volume 42 Issue 1
    Invited Reviews:Nanotechnology, Drug Delivery and Regeneration Medicine
    Brain-targeted nanoparticle drug delivery systems: research advances
    CAO Yi, JIANG Chen
    2022, 42(1):  2-14.  doi:10.16352/j.issn.1001-6325.2022.01.007
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    The blood-brain barrier (BBB) is a special structure in the central nervous system (CNS), whose delicate properties can protect the brain from the attack by harmful large molecules and organisms within blood circulation. However, the very barrier also prevents the brain-targeted drugs from taking effects, posing great challenge to the treatment of neurodegenerative diseases, glioma and other brain diseases. With the development of nanotechnology, more and more nanoparticles (NPs) have been developed as drug carriers, which can facilitate the drug with various mechanisms to get across the BBB. This article starts with introduction of the BBB, explaining the complexity of BBB structure and its unique features to form barriers, so that we can understand the difficulty in transporting drug into the BBB and the potential methods of targeting and permeate the barrier. Next, we focus on three main types of NPs that have been applied in the brain-targeted drug delivery system: polymeric-based NPs; biomimetic-based NPs and inorganic-based NPs. Furthermore, we deal with the specific NPs drug delivery strategies, which include absorption-mediated, carrier-mediated and receptor-mediated transcytosis. At the end of the article, we talk about the leading trend and future development of the brain-targeted nanoparticle drug delivery systems.
    Design and biomedical applications of core-shell tecto dendrimers
    SONG Cong, WANG Da-yuan, SHEN Ming-wu, SHI Xiang-yang
    2022, 42(1):  15-25.  doi:10.16352/j.issn.1001-6325.2022.01.010
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    Core-shell tecto dendrimers (CSTDs) are a kind of superstructured dendrimeric nanoconstructs with relatively high-generation dendrimers as the cores and low-generation dendrimers as the shells. Recently, there is growing evidence that the performance of CSTDs constructed with poly(amidoamine) (PAMAM) dendrimers of different generations as reactive modules is not only superior to that of single-generation dendrimers, but also overcomes some biomedical limitations (e.g., restricted drug loading capacity, limited tumor passive targeting based on enhanced permeability and retention effect) of single-generation dendrimers. Herein, the design of CSTDs for biological imaging, chemotherapy, gene therapy, and combination therapy are reviewed. The current challenges and future development prospects of CSTDs in the field of biomedicine are also analyzed.
    Research progress of nanomaterials for the female reproductive system tissue engineering
    HU Xue-chun, XU Hai-yan
    2022, 42(1):  26-32.  doi:10.16352/j.issn.1001-6325.2022.01.003
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    The health of the reproductive system is closely associated with females' fertility ability and the quality of life. However, the incidence of diseases related to the female reproductive system has been increasing and the age of onset is becoming younger, generating numerous negative consequences on the female reproductive health. Therefore, there are urgent demands to develop effective therapeutic technologies and strategies. In recent years, the progress of tissue engineering and regenerative medicine has shown promising potentials of novel approaches for the treatment of related diseases by guiding tissue regeneration to restore the organs' functions. In this article, we introduced the advances of scaffolds construction inspired by nanostructures and biomaterials as well as the applications in the premature ovarian failure, follicle culture in vitro, intrauterine adhesions, and the regeneration of defected uterine and pelvic organ prolapse. In addition, existing challenges and efforts that should be considered in the future were discussed.
    Application of nanomaterials in medical detection and disease diagnosis
    XU Ying, ZHANG Yu
    2022, 42(1):  33-40.  doi:10.16352/j.issn.1001-6325.2022.01.005
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    Nanomaterials have the characteristics of small size and high specific surface area. These characteristics make it unique from other traditional materials in the properties of light, sound, electricity, heat, and magnetism. With the continuous development of nanomaterials and the needs of clinical medicine, more and more functionalized nanomaterials and related devices have been widely used in medical detection with the advantages of high sensitivity, high specificity, accuracy, and speed. The rapid development of nanomaterials can provide new methods for clinical diagnosis and provide more scientific basis for disease prevention and treatment. Nanomaterials have great development potential and broad application prospects in the diagnosis of a variety of diseases. In this article, the progress in the medical detection and diagnosis of infection, tumor, diabetes, neurodegenerative disease and cardiovascular diseases is reviewed; future perspectives are discussed as well.
    Progress of nucleic acid drugs and non-viral carriers in gene therapy
    LIU Jian
    2022, 42(1):  41-50.  doi:10.16352/j.issn.1001-6325.2022.01.002
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    Gene therapy based on nucleic acid drugs is the ultimate treatment technology for gene abnormality related diseases. However, nucleic acid drugs need the help of carriers to enter target cells and reach specific intracellular locations, therefore, the development of safe and efficient delivery systems for nucleic acid drugs is one of the fundamental technologies for gene therapy. Compared with viral vectors, non-viral vectors have higher safety, but are still facing the challenge of relative low transfection efficiency. With the advances of nanotechnology, the efficiency of non-viral carriers has been gained significantly improvements. In this paper, some representative nucleic acid-based drugs and vaccines are introduced, and the progresses of non-viral delivery carriers for uses in gene therapy have been reviewed.
    Original Articles
    Compound heterozygous variation of PATL2 causes oocyte maturation defect
    HUO Ming-zhu, ZHANG Yi-le, XU Jia-wei, SHI Hao, LIU Yi-dong, NIU Wen-bin
    2022, 42(1):  51-55.  doi:10.16352/j.issn.1001-6325.2022.01.004
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    Objective To investigate the genetic etiology of a patient with primary infertility and oocyte maturation defect. Methods Whole exome sequencing (WES) was carried out on patient's DNA to detect potential pathologic variants. The positive sites detected by WES were verified by Sanger sequencing, and the bioinformatics analysis of the mutation was analyzed. Results The patient was found to harbor compound heterozygous variants of the PATL2 in exon 14 and 13 c.1 374A>G (p.Ile458Met) and c.1 289_1 291delTCC (p.Leu430del), which were respectively inherited from her father and mother. Conclusions The compound heterozygous variation of PATL2 may be the genetic mechanism to explain infertility resulted from the disturbance of oocyte maturation.
    Effects of 5-lipoxygenase activating protein inhibitor MK886 on proliferation and apoptosis of human esophageal cancer cell lines
    SHI Tie-wei, LI Tian-zhu, ZHOU Jing, NA Ri-su, SUN Qi, XU Li-yan, BAI Chun-ying
    2022, 42(1):  56-61.  doi:10.16352/j.issn.1001-6325.2022.01.006
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    Objective To investigate the effects of 5-lipoxygenase activating protein (FLAP) inhibitor MK886 on the proliferation and apoptosis of human esophageal carcinoma cell lines (KYSE-150 and TE-3) and to explore the potential mechanism. Methods KYSE-150 and TE-3 cells were treated with MK886(2.5,5,10,20,40 and 80 μmol/L). xCELLigence RTCA system was used to to detect the half maximal inhibitory concentration(IC50)of MK886. The expressions of proteins with apoptosis and autophagy were detected by Western blot and cell cycles were detected by flow cytometry. Results MK886 decreased the proliferation of human esophageal carcinoma cells(KYSE-150 and TE-3) (both P<0.05).IC50 of MK886 in KYSE-150 group was 29.11 μmol/L and in TE-3 group of 27.47 μmol/L.The proportion of G0/G1 phase was increased(both P<0.001)However, in the group treated with concentrations of MK886 50 μmol/L,G2/M phase was increased(P<0.05). The expression of cleaved-caspase-3 and LC-3A/B-Ⅱwere increased with MK886 (both P<0.001). Conclusions MK886 inhibits proliferation of human esophageal carcinoma cell lines and induces apoptosis.
    Balanophora polysaccharide decreases the expression of BDNF and TrkB in liver tissues of rat with experimental liver injury
    XIE Ya, CHEN Ying, XIA De-yao, TANG Xian-e, WANG Feng-jie, CHEN Xian-bing
    2022, 42(1):  62-67.  doi:10.16352/j.issn.1001-6325.2022.01.009
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    Objective To study the effects of Balanophora polysaccharides (BPS) on the expression of BDNF, TrkB and other apoptosis-related proteins in rats with experimental liver injury induced by D-gal. Methods Rats were randomly divided into control group, D-gal group (200 mg/kg of hypodermic D- galactose), groups with BPS of low dosage (BPS-L), medium dosage (BPS-M) and high dosage (BPS-H), which were treated with intragastric administration by 50, 100 and 200 mg/kg BPS per day, respectively. After six weeks, serum alanine transaminase(ALT) and aspartate aminotransferase(AST) were measured. HE staining was used to observe liver tissue morphology. The localization of BDNF and TrkB was determined by immuno-histochemical staining. Western blot was used to detect BDNF, TrkB, Bcl-2, caspase-3 and Bax protein expression. ELISA was used to determine the malondialdehyde(MDA) content and superoxide dismutase(SOD) activity in liver tissues. Results Compared with the control group, the hepatocytes showed edema and punctate necrosis in D-gal group, with increased serum level of ALT and AST(P<0.05). The results also showed decreased SOD activity and increased MDA content (P<0.05), increased Bax, caspase-3, BDNF, TrkB expression (P<0.05) and decreased Bcl-2 expression (P<0.05). Compared to D-gal group, the hepatocyte degeneration and necrosis were obviously alleviated in BPS groups with each dose accompanied by decreased serum ALT and AST levels (P<0.05), increased SOD activity and decreased MDA content (P<0.05), reduced Bax and caspase-3 expression of pro-apoptotic proteins, increased expression Bcl-2 anti-apoptotic proteins (P<0.05) and decreased expression of BDNF and TrkB (P<0.05). Conclusions Balanophora polysaccharide had a protective effect on experimental liver injury caused by D-gal. The mechanism may be related to the decrease of the expression of BDNF and TrkB followed by inhibition of hepatocyte apoptosis.
    LncRNA expression profile in human malignant embryonic rhabdomyosarcoma cell line infected with Coxsackie virus group B type 5
    TENG Pei-ying, LI Jing, SHI Yu-lu, YANG Fan, OU Xia, CHEN Wei
    2022, 42(1):  68-74.  doi:10.16352/j.issn.1001-6325.2022.01.011
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    Objective To explore the molecular mechanism of the interaction between Coxsackie virus group B type 5 (CVB5) and the host, The long non-coding RNA(lncRNA) expression profile in human malignant embryonic rhabdomyosarcoma cell line (RD) infected by CVB5 was investigated. Methods After 24 hours of infection with 1MOI CVB5 RNA was sampled and RNA-sequencing was used to identify the differential expression profile of lncRNA in the cells. Differentially expressed transcripts were analyzed by cluster analysis, GO analysis and KEGG pathway enrichment. At the same time, RNAfold was used to predict the secondary structure of lncRNA. Results Compared to the uninfected group, 1 754 mRNAs and 508 lncRNAs were up-regulated and 3 106 mRNAs/760 lncRNAs were down-regulated after CVB5 infection. Co-expressed genes that differentially express lncRNA were mainly enriched in biological processes such as molecular structure activity, protein molecular binding, and humoral immune response; lncRNA targets were mainly involved in olfactory transduction pathway, cytokine-cytokine receptors interaction, neuroactive ligand-receptor interaction and other pathways. In addition, the seven differen-tially expressed lncRNAs verified by quantitative real-time PCR (RT-qPCR) were consistent with the sequencing results. Conclusions lncRNA is mainly involved in the regulation of the immune processes, which lays the foundation for a full understanding of the regulatory role of lncRNA in CVB5 infection.
    PF-127-LV-NTFs three-dimensional composite scaffold in culturing rat neural stem cells
    LIANG Guo-xin, LIANG Yin-ru, LIU Shu-xian, CHEN Kang-zhen, LIN Yong, QIN Jiang-xia, WU Hong-fu
    2022, 42(1):  75-81.  doi:10.16352/j.issn.1001-6325.2022.01.008
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    Objective To investigate the effect of LINGO-1 shRNA (LV) and neurotrophic factors cocktail (NTFs,NTFcocktail) encapsulated by Pluronic F-127(PF-127)hydrogel (PF-127-LV-NTFs)three-dimensional(3D) composite scaffolds on the proliferation and differentiation of rat neural stem cells. Methods The LV was constructed and infected with NSCs at different MOI. RT-qPCR and Western blot were used to detect the expression of LINGO-1 in NSCs. Brain derived neurotropic factor(BDNF), neurotrophin-3(NT-3), platelet-derived growth factor(PDGF),insulin-like growth factor 1(IGF-1), epidermat growth factor(EGF), basic fibroblast growth factor(bFGF) and glial cell derived neurotrophic factor(GDNF) were prepared as NTFs. The LV and NTFs were mixed with PF-127 and divided into different groups to culture NSCs. CCK-8 was used to detect the cell viability of NSCs, the survival rate of NSCs was detected by LIVE/DEAD staining, and immunofluorescence staining was used to detect the proliferation and differentiation of NSCs. Results The optimal MOI for LV was 5. CCK-8 assay results showed that there was no significant difference in the survival rate of NSCs. The LIVE/DEAD and Nestin immune-fluorescence staining results showed that compared with control, the survival rate of NSCs and the proportion of neuron differentiation positive were the highest in PF-127+LV+NTF group(P<0.01). Conclusions PF-127-LV-NTFs 3D composite scaffold is favouring to support the proliferation and the directional differentiation of NSCs of rat.
    CircRNA_23113 inhibits the proliferation and migration of lung adenocarcinoma cells
    WANG Ling-xia, YAN Yu-lan, YUAN Hai-tao, CAI Ye-wei, LIU De-hui, GU Wen-lu, CAO Bi-yue
    2022, 42(1):  82-88.  doi:10.16352/j.issn.1001-6325.2022.01.012
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    Objective To explore the expression of circRNA_23113 in lung adenocarcinoma and its influence on the proliferation/migration of the adenocarcinoma cells. Methods Screening 5 cases of lung adenocarcinoma and adjacent tissues by high-throughput sequencing from which screened out the differentially expressed circular RNA circRNA_23113. Real-time fluorescence quantitative PCR was used to detect the expression of circRNA_23113 in the adenocarcinoma tissues, serum and cells; the over-expression plasmid of circRNA_23113 was constructed, and its effect on cell proliferation and migration was measured by CCK-8 method, clone formation experiment, cell scratching and Transwell chamber method; Western blot was used to detect the expression of β-catenin, cyclin D1 and c-myc protein. Results CircRNA_23113 was significantly less expressed in tissues, serum and cells of patients with lung cancer (P<0.05); Over-expression of circRNA_23113 inhibited the proliferation and migration of A549 and H1299 cells(P<0.05); CircRNA_23113 inhibited the expression of β-catenin, cyclin D1 and c-myc(P<0.05). Conclusions CircRNA_23113 is poorly expressed in lung adenocarcinoma, and over-expression may inhibit the proliferation and migration of lung adenocarcinoma cells.
    Regulating effects of GSK650394 on behavior and hippocampal neurotrophy in chronic stress model of depression in rats
    LIU Dan, LIN Xiao-yuan, YANG Hui, LIU Tong-tong, LIU Ping-an, MENG Pan
    2022, 42(1):  89-93.  doi:10.16352/j.issn.1001-6325.2022.01.013
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    Objective To investigate the protective effect of serum and glucocorticoid-regulated kinase 1(SGK1) inhibitor GSK650394 on depression-like behaviors and hippocampus neurotrophy in depressive model rats. Methods SD rats were randomly divided into control group, depression model group and GSK650394 group. Depression model was established by chronic mild unpredictable stress. The behavioral changes of the SD rats were observed by forced swimming, 1% sugar consumption test and Morris water maze. The changes of corticosterone(CORT) and monoamine neurotransmitters in plasma and serum were observed by ELISA. The changes of BDNF, NT-3 and NGF in hippocampus were detected by Western blot. Results Compared with the control group, the model group showed that sucrose preference decreased and swimming immobility time increased significantly(P<0.01). Escape latency(EL) and latency time(Lat.T) prolonged significantly (P<0.05 or P<0.01) and the content of CORT in plasma increased significantly (P<0.01). The content of 5-HT, NE in serum and the expression of BDNF, NT-3, NGF in hippocampus significant increased in the model group. Compared with model group, GSK650394 significantly increased sucrose preference, decreased immobility time(P<0.05 or P<0.01), shortened EL and Lat.T(P<0.05), decreased the content of CORT in plasma (P<0.01) and enhanced the level of 5-HT and NE in serum (P<0.01) and also promoted the expression of NT-3, BDNF and NGF in hippocampus. Conclusions SGK1 inhibitor GSK650394 can significantly relieve depression like behavior potentially through improving the nutrition profile of neurotrophin in hippocampus.
    Specific necroptosis inhibitor-1(Nec-1) attenuates glial scar formation in rat models with spinal cord injury
    CHEN Sheng, WANG Chun-ming, YAN Xue-fei, MAO Yuan-qing
    2022, 42(1):  94-99.  doi:10.16352/j.issn.1001-6325.2022.01.014
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    Objective To observe the effect of necroptosis inhibitor-1(Nec-1) on glial scar formation in rats with spinal cord injury and its mechanism. Methods The rats were randomly divided into sham group, model group (falling method) and RIPK1 inhibitor Nec-1 group (10 μmol/L Nec-1 10μL was injected into lateral ventricle). BBB method was used to measure the motor function of hind limbs, immunofluorescence was used to detect the formation of glial scar, and Western blot was used to detect the expression of cathepsin B, caspase-3, GFAP and vimentin. Results On the 14th day after operation, compared with the sham group, the BBB scores of the model group and Nec-1 group were significantly lower (P<0.05), compared with the model group, the BBB score of Nec-1 group was significantly higher (P<0.05). NF-200 fluorescence was not found in the sham operation group, but in the model group and Nec-1 group, the intensity of NF-200 fluorescence labeling in Nec-1 group decreased significantly(P<0.05). On the 7th and 14th day after operation, compared with the sham operation group, the protein level of cathepsin B, caspase-3, GFAP and vimentin in the model group and Nec-1 group was significantly higher(P<0.05),while the protein level of cathepsin B, caspase-3, GFAP and vimentin in Nec-1 group decreased significantly (P<0.05). Conclusions Nec-1 may regulate expression level of cathepsin B and caspase-3, reduce the expression of GFAP and vimentin proteins, reduce the formation of glial scar after spinal cord injury and promote the recovery of neural function.
    Effects of furosemide on PERK/eIF2α/CHOP pathway and secondary brain injury in rat models with intracerebral hemorrhage
    ZHANG Yu, GAO Jian-zhou, HAN Shao
    2022, 42(1):  100-105.  doi:10.16352/j.issn.1001-6325.2022.01.016
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    Objective To investigate the effects of furosemide(FUR) on PERK/eIF2α/CHOP pathway and secondary brain injury in rats with intracerebral hemorrhage (ICH). Methods The rats were divided into sham operation group, model group(intracerebral injection of collagenase type Ⅳ in ICH), furosemide low dose (FUR-L) groups, medium dose (FUR-M) groups,high dose (FUR-H) groups and ganglioside group, with eighteen rats in each. After treatment, the neurological function was evaluated and scored by Longa and the water content of brain tissue was measured; Evans blue extravasation test was used to detect the blood-brain barrier injury; HE staining microscopy was used to detect the damage of hippocampus neurons; enzyme-linked immunosorbent assay (ELISA) was used to detect the serum level of interferon-γ (IFN-γ) and interleukin-6 (IL-6); Western blot was used to detect the expression of p-PERK/PERK, p-eIF2α/eIF2α and CHOP. Results Compared with those in the sham operation group, the hippocampal neurons in the model group were degenerated and necrotic, the shrinkage was smaller, the number was significantly decreased, severe pathological damage was found, Longa score, Evans blue exudation, brain water content, level of serum IFN-γ and IL-6,the expression of p-PERK/PERK, p-eIF2α/eIF2α and CHOP were all significantly increased (P<0.05). Compared with those in the model group, The morphological changes of hippocampal neurons in the FUR-L, FUR-M and FUR-H groups and ganglioside group were restored to different degrees and the pathological damage was alleviated, Longa score, brain water content, Evans blue exudation, level of serum IFN-γ and IL-6, expression of p-PERK/PERK, p-eIF2α/eIF2α and CHOP were decreased (P<0.05). Conclusions Furosemide can down-regulate the expression of PERK/eIF2α/CHOP pathway protein, relieve brain edema and hippocampal neuron damage and repair the neural function in ICH animal.model.
    Silencing eEF2K combined with sodium tanshinone ⅡA sulfonate synergistically inhibit proliferation of human lung adenocarcinoma cell line A549
    WANG Bu, YUAN Sheng-fang, ZHANG Chang-hong, YUAN Cheng, HUANG Pan-deng, ZHANG Zhi-hua, ZHAO Jian-qing
    2022, 42(1):  106-113.  doi:10.16352/j.issn.1001-6325.2022.01.017
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    Objective To investigate the effects of eukaryotic elongation factor-2 kinase (eEF2K) gene transfection and sodium tanshinone Ⅱ A sulfonate (STS) on proliferation, invasion and migration of lung adenocarcinoma cell line A549 and its mechanism. Methods The optimal concentration of STS was screened out by preliminary experiment. A549 cells were divided into siRNA-NC group (transfected with siRNA-NC), siRNA-eEF2K group (transfected with siRNA-eEF2K), siRNA-NC+STS group (treated with 10 μg/mL STS after siRNA-NC) and siRNA-eEF2K+STS group (treated with 10 μg/mL STS after siRNA-eEF2K).The survival rate, clone formation, cell counting of transmembrane cells, migration and apoptosis of A549 cells were detected by CCK-8 method, clone formation test, Transwell chamber test, scratch test and flow cytometry. The protein expression levels of protein kinase B (AKT), phosphorylation (p)-AKT, Ki67, matrix metalloproteinase-2 (MMP-2) and B-cell lymphoma-2 genes (Bcl-2) were detected by Western blot. Results Compared with those in siRNA-NC group, the cell survival rate, clone formation rate, number of transmembrane cells, migration rate and p-AKT, Ki67, MMP-2 and Bcl-2 in proteins siRNA-eEF2K+STS group were significantly lower, while the apoptosis rate was significantly higher (P<0.05). The siRNA-eEF2K+STS group had the largest range of changes in the above indexes. Conclusions Silencing eEF2K combined with STS can synergistically inhibit the proliferation, invasion and migration of A549 cells and promote cell apoptosis. The mechanism is potentially related to the inhibition of protein expression of p-AKT, Ki67, MMP-2 and Bcl-2.
    Propofol inhibits proliferation of human endometrial cancer cell line RL95-2
    LI Yong-xiao, SUN Yan, ZHANG Rui-sheng
    2022, 42(1):  114-119.  doi:10.16352/j.issn.1001-6325.2022.01.015
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    Objective To investigate the influence of propofol on the proliferation and apoptosis of endometrial cancer cell line RL95-2 and the effect on mTOR/S6K1 signaling pathway. Methods Human endometrial cancer cell line RL95-2 was divided into control group and propofol group (different concentrations of propofol were added). CCK-8 method was used to detect cell proliferation; Clone formation assay was used to detect the ability of cell clone formation; Flow cytometry was used to measure cell apoptosis; Colorimetric was used to measure the activities of caspase-3 and caspase-9; Western blot was used to detect the phosphorylation levels of protein kinase B (Akt), mammalian target of rapamycin (mTOR) and ribosomal protein S6 kinase 1 (S6K1). Results Compared with the control group, the proliferation rate and colony forming ability of RL95-2 cells in propofol groups were significantly lower (P<0.05), the apoptosis rate and the activities of caspase-3 and caspase-9 were significantly higher(P<0.05), the phosphorylation levels of p-Akt, p-mTOR and p-S6K1 were significantly lower(P<0.05). Conclusions Propofol may inhibit the proliferation and promote the apoptosis of endometrial cancer cell line by regulating mTOR/S6K1 pathway.
    Recombinant Treponema pallidum protein Tp47 increases permeability of vascular endothelial cells through inducing uPA
    YU Bao-dan, CHEN Cong, KE Wu-jian, LYU Ping
    2022, 42(1):  120-125.  doi:10.16352/j.issn.1001-6325.2022.01.018
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    Objective To investigate the effect of recombinant Treponema pallidum protein Tp47 (rTpp47) on the expression of uPA by THP-1 macrophages and on the permeability of human umbilical vein/vascular endothelial cells(HUVECs). Methods THP-1 cells were stimulated with rTpp47 for 24 hours and then the culture supernatant and THP-1 cells were collected respectively. The expression of urokinase-type plasminogen activator (uPA) in THP-1 cells was detected by ELISA and Western blot. After stimulation of HUVECs monolayers by THP-1 cell culture supernatant, FITC-dextran was used to evaluate the permeability of HUVECs. Western blot method was used to observe the effect of uPA on the expression of tight junction protein claudin-5 of HUVECs and the protein kinase C(PKC) signaling pathway of THP-1 cells stimulated by rTpp47. Results The synthesis and secretion of uPA of THP-1 cells stimulated by rTpp47 were significantly higher than those in the control group(P<0.05, P<0.001). When HUVECs were stimulated by the THP-1 cell supernatant for 12 hours and 24 hours, the relative permeability of vascular endothelial cells in the experimental group was significantly higher than the control group (P<0.05 for 12 hours and P<0.000 1 for 24 hours). Amiloride, the synthetic inhibitor of uPA, significantly inhibited the uPA expression and secretion of THP-1 cells stimulated by rTpp47 (P<0.0001), alleviated the inhibition effect of rTpp47 on claudin-5 expression of HUVECs (P<0.05), and significantly suppressed PKC phosphorylation(P<0.05) in THP-1 cells induced by rTpp47. Conclusions The rTpp47 can stimulate the synthesis and secretion of uPA in THP-1 cells through a potential mechanism of activating PKC signaling pathway and uPA may increase the permeability of vascular endothelial cells.
    An investigation of Helicobacter pylori infection and potential risk factors of colonization found in 10 661 cases from routine physical examination
    LEI Rong, YANG Dan, YUAN Fang-tao, ZHOU Xiao-yan, JIANG Yi-min, LIU Mi
    2022, 42(1):  126-130.  doi:10.16352/j.issn.1001-6325.2022.01.019
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    Objective To investigate the situation of Helicobacter pylori(H.pylori) infection in healthy people and analyze the related risk factors. Methods Retrospective analysis was conducted on 10 661 healthy subjects who underwent carbon 13-urea breath test (13C-UBT)on January 1, 2019 to December 30, 2020 in affiliated hospital of North Sichuan Medical College. According to the infection status of Helicobacter pylori (H.pylori), the subjects were divided into H.pylori positive group and H.pylori negative group. H.pylori infection and platelet (PLT), triglyceride(TG), total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), fasting blood glucose (FBG), T3, T4, TSH and pepsinogen Ⅰ (pepsinogen Ⅰ) were analyzed. PepsinogenⅠ, pepsinogen Ⅱ(pepsinogen), gastrin 17(G-17), homocysteine (Hcy), uric acid (UA) and total bilirubin (TBIL)were also examined. Results 1)The total number of positive infections: 4 382, the total positive rate: 41.10%, including 2 717 male positive cases with morbidity as 25.49%. There were 1 665 cases of positive infection in females, and the morbidity was 15.62%, which was lower than that in males (P<0.05). 2)Compared with the negative group, the age of the positive group was slightly lower than that of the negative group (P<0.05), and the HDL-C, pepsinogen Ⅰ/Ⅱ, Hcy and TBil were all lower than those of the negative group (P<0.05). BMI, SBP, DBP, TG, FBG, pepsinogen Ⅰ, pepsinogen and G-17 in H.pylori positive group were all higher than those in negative group (P<0.05). 3)DOB value was correlated with age, FBG, pepsinogen Ⅰ, pepsinogen, G-17, pepsinogen Ano/Cha and TBil (P<0.001). 4) The parameters P<0.05 were further included in binary Logistic regression analysis, and male, BMI, FBG, pepsinogen Ⅱ, pepsinogen branch/root were risk factors for H.pylori infection (P<0.05). Conclusions H.pylori infection is more common in male population, BMI, FBG, pepsinogen Ⅱ, pepsinogen L/N are the risk factors for H.pylori infection. Early intervention of these risk factors is expected to reduce the infection of H.pylori and provide theoretical basis for the primary prevention of this gastrointestinal diseases.
    Clinical Sciences
    Comparative analysis of supraclavicular area mobility in locally advanced esophageal cancer radiation therapy with two different fixed ways
    WAN Bao, HUAN Fu-kui, ZHAO Yu, CHEN Huan, GENG Song-song, YAN Hui, ZHANG Yan-xin
    2022, 42(1):  131-135.  doi:10.16352/j.issn.1001-6325.2022.01.023
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    Objective To compare the setup errors provided by the chest and abdomen flat frame fixation device and integral cervico-thoracic fixation device in supraclavicular regions of locally advanced esophageal cancer patients. Methods The cone beam computed tomography (CBCT) images of 50 patients with locally advanced esophageal cancer and received radiotherapy in supraclavicular region were retrospectively analyzed. The chest and abdomen flat frame fixation device (group A) or the integral cervicothoracic fixation device (group B) were used respectively with 25 in each.There were 175 CBCT images in group A and 230 CBCT images in group B. The setup errors of the two groups were analyzed, and the movement and displacement of acromioclavicular joint in three-dimensions were measured. The differences were compared by independent sample t test and rank sum test. Results The setup errors were significantly different between group A and group B. The P value in X axis (left-right) and Z axis (anterior-posterior) were both less than 0.001 (P<0.001);The P value in pitch was less than 0.001 (P<0.001)and roll was less than 0.05 (P<0.05). The movement amplitude of acromioclavicular joint (ΔX, ΔY, ΔZ) of groups A and B were (0.10±0.10)cm and (0.08±0.08)cm in X axis, (0.14±0.13)cm and (0.11±0.11)cm in Y axis, (0.16±0.12)cm and (0.11±0.12)cm in Z axis. The three-dimensional displacement of groups A and B were (0.27±0.14)cm and (0.20±0.14)cm, (P<0.001). Conclusions If the influence of rotation angle is well controlled, the application of the integral cervicothoracic fixation device is better to immobilize the acromioclavicular joint and reproduce the setup position for locally advanced esophageal cancer patients. Special attention should be paid for the influence of rotation angle on the location accuracy of target.
    Antibiotic sensitivity of Rhodococcus equi in clinical specimens is correlated with efficacy of clinical treatment
    LI Jing
    2022, 42(1):  136-138.  doi:10.16352/j.issn.1001-6325.2022.01.021
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    Objective To isolate and identify Rhodococcus equi(R.equi) from clinical specimens for potential relationship between antibiotic sensitivity and efficacy of antibiotic chemotherapy. Methods From April 2019 to April 2020, 17 cases patients with positive isolation of R. equi were grouped as test one and and 17 cases of patients underwent empirical treatment only were collected as the control group. Results The efficacy of antibiotic chemotherapy with sensitivity test (test group) showed significantly better than that of the control group. Conclusions Rational use of antibiotics chemotherapy supported by sensitivity test can significantly improve the effect of chemotherapy.
    Direct detection of Plasmodium female gametocyte-specific transcript s25 mRNA in blood by CLIP-PCR
    YANG Jun-yuan, PIAN Hong-ru, YANG Ming-zhu, ZHENG Zhi
    2022, 42(1):  139-144.  doi:10.16352/j.issn.1001-6325.2022.01.024
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    Objective To establish a molecular detection method for the Plasmodium female gametocyte. Methods Probes for capture and ligation probe-PCR (CLIP-PCR) were specific designed based on Plasmodium female gametocyte special RNA targets, which were the transcripts of the ookinetes surface protein (s25). After the whole blood cells were lysed, the RNA targets were directly released and captured on the 96-well plate by “sandwich” hybridization without extraction. Washing away the unbound probes, the ligation probes which also hybridized to the RNA target were ligated to form a single-strand amplification template with specific-designed sequence at both ends. The SYBR Green qPCR was carried out with universal primers. Or with TaqMan probe binding sequence was designed at the end of the templates, the TaqMan qPCR was carried out with universal primers and universal fluorescent probes. The sensitivity, specificity and repeatability of CLIP-PCR were evaluated and compared with the RT-qPCR. And it was also used to detect a small number of clinical samples. Results This method was successfully applied to detect s25 mRNA with high sensitivity and specificity. Both RT-qPCR and CLIP-PCR could detect s25 mRNA targets as low as 11 copies. However, CLIP-PCR was simpler compared to RT-qPCR. And it could accurately detect gametocytes in blood of malaria patients. The detection of 96 samples could be completed within 3 h. Conclusions SYBR Green CLIP-PCR and TaqMan CLIP-PCR for the female gametocyte of Plasmodium detection provides a sensitive and efficient method for large-scale screening of gametocytes and control of malaria transmission.
    Mini Reviews
    Research progress on mechanism of Schwann cells promoting peripheral nerve regeneration
    YANG Yi-duo, GUO Hai-dong, SHAO Shui-jin, LIU Yu-pu
    2022, 42(1):  145-148.  doi:10.16352/j.issn.1001-6325.2022.01.020
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    Schwann cells (SCs) have a strong plasticity. After peripheral nerve injury, Schwann cells dedifferentiate into repair phenotype and dominate the repair process. C-Jun N-terminal kinase, mitogen activated protein kinase(MAPK) and other signaling pathways and transcriptional regulators are involved in regulating Schwann cell-mediated repair process. This review discusses the main signaling pathways and transcriptional regulators of Schwann cell promoting peripheral nerve regeneration.
    Research progress in colitis-induced hepatic barrier dysfunction
    SHU Meng-xuan, YUAN Fang-chen, ZHOU Meng-qi, TU Shi-yan, XU Yue, QIN Hong-yan
    2022, 42(1):  149-153.  doi:10.16352/j.issn.1001-6325.2022.01.022
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    Liver barrier functions as a main structure to protect the liver from exogenous harmful substances and play an important role in maintaining secretion function, regulating bile transport and intercellular information transmission. Liver barrier protein alteration and barrier dysfunction can be induced by endotoxin, inflammatory cytokines and bile acids, which may be involved in the occurrence of colitis-related liver diseases. To explore the outcomes and underlying mechanism of intestinal inflammation on liver barrier function may support the understanding of pathogenesis of colitis-related liver diseases and thus provide clues and basic information for the prevention and treatment of colitis-related liver diseases.
    Methodology and application of human physiological reference intervals estimation
    HE Hui-jing, SHAN Guang-liang
    2022, 42(1):  154-158.  doi:10.16352/j.issn.1001-6325.2022.01.030
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    Establishment of physiological reference interval (RI) is important for health status evaluation, disease screening and diagnosis, and the effect assessment of clinical therapy. Currently, there is no review on methodology of RI establishment in China. Therefore, by reviewing of literatures on methodology of RI establishment, this article comprehensively described the purpose and significance of RI establishment,its statistical methods of calculation and its application. Three methods are commonly used for RI calculation: the nonparametric, parametric and robust estimate. Researchers may use a specific method based on research purpose and the characteristics of the data. This review provides methodological reference for RI establishment in China and also some guidelines to better understand the meaning of physiological constant RI in health practices.
    Research progress on IL-6/JAK/STAT3 in the development and treatment of prostate cancer
    CHEN Hao, CHEN Tao, WU Jing
    2022, 42(1):  159-162.  doi:10.16352/j.issn.1001-6325.2022.01.025
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    IL-6/JAK/STAT3 is one of the most significant signaling pathways activated by IL-6.IL-6 and p-STAT3 which are highly expressed in prostate cancer (PCa) tissue and metastatic tumors So this pathway may play a role in the occurrence of prostate cancer, promotion of malignant cell proliferation, invasion and metastasis of the tumor. It may also function in the development of castration resistance and drug resistance through activation androgen receptor(AR). The multiple roles of IL-6/JAK/STAT3 and its activated downstream factors in tumor progression provide a good basis for the development of chemotherapy drugs. Currently, many targeted inhibitors have been proved to be effective in inhibiting tumor progression and more effective drugs are expected to be developed.
    Progress of exosomes in diagnosis and treatment of prostate cancer
    LI Ying-jie, YANG Ting-kai, JI Zhi-gang, WEN Jin
    2022, 42(1):  163-167.  doi:10.16352/j.issn.1001-6325.2022.01.026
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    Prostate cancer(PCa) is a common malignancy in males. Exosomes are lipid bilayer vesicles containing protein, mRNA and miRNA, which are active in intercellular communications. Exosomes promotes the development of prostate cancer, support immune evading and chemotherapy resistance, stimulate angiogenesis, induce epithelial-mesenchymal transition(EMT) of tumor cells, and contribute to the formation of pre-metastasis niche, thus promoting the invasion and metastasis of prostate cancer. Based on the role in tumor biological behavior and heterogeneity in different cells, exosomes hold great potentials as biomarker for diagnosis and prognosis of prostate cancer, and are expected to become a favorable drug carrier. It has a promising prospect for cancer therapy.
    Research progress on biomarkers for embryonic development
    XIE Cong-cong, WANG Xue-ying, AN Zhuo, YAO Sheng-jie, MA Jing, WANG Shu-song
    2022, 42(1):  168-172.  doi:10.16352/j.issn.1001-6325.2022.01.027
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    Screenings high-quality gametes and high developmental potential embryos are focus of assisted reproductive technology. In recent years, studies have found that the embryo culture medium contains proteins secreted by the embryo, free DNA fragments, cytokines and so on, and the nutrients needed for embryo development are obtained from the culture medium. Through using non-invasive methods, for example embryo metabonomics, proteomics, to analyze the composition of the embryo culture medium. These biomarkers are helpful for rapid and accurate screening of high-quality embryos with the most developmental potential. This paper summarizes the effects of amino acids, carbohydrates, cytokines and proteins on embryonic development.
    Immune microenvironment and immunotherapy of pituitary adenomas
    GUO Xiao-peng, XING Bing, MA Wen-bin
    2022, 42(1):  173-178.  doi:10.16352/j.issn.1001-6325.2022.01.034
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    Pituitary adenomas (PAs) are the second most common primary central nervous system tumors. Although most PAs can be cured by surgical operation, medicine treatment or radiotherapy, some PAs may recur, invade adjacent structures or even metastasize to other organs after traditional treatments, so decreaseing patient overall survival and deteriorate their life quality. In recent years, studies on the immune microenvironment of brain tumors emerged, especially in the field of glioma, providing potential effective treatment targets for immunotherapy and setting the foundation for improving patient survival. However, previous studies have only conducted preliminary analyses on immune cells, immune molecules, and cytokines in the microenvironment of PAs. The research on immunotherapy of PAs with animal model and humans is only at an early phase.
    Research progress of circular RNAs in prostate cancer
    FENG Tian-rui, YAN Wei-gang
    2022, 42(1):  179-183.  doi:10.16352/j.issn.1001-6325.2022.01.028
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    Circular RNAs (circRNAs) are non-coding RNAs with unique covalently closed circular structures and are widely found in eukaryotic cells. Recent studies have shown that circRNAs are involved in the patho-physiological processes of various diseases including malignant tumors through regulating microRNAs, binding to proteins and regulating transcription and splicing process. The special structure and complex functions of circRNAs make them have great research potential in the field of cancer. Prostate cancer(PCa) is one of the most common malignancies among the world. The the pathogenesis and treatment of PCa still need to be further studied.
    Medical Education
    Practice and consideration of Peking Union Medical College Hospital aiding Tibet in radiological images
    Bianbaciren, WANG Feng-dan, YIN Wu, JIN Zheng-yu
    2022, 42(1):  184-187.  doi:10.16352/j.issn.1001-6325.2022.01.031
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    Since the startup of the “group type” aid work for Tibet, five groups of radiologists from Peking Union Medical College Hospital have consecutively been sent to work in the Radiology Department of People's Hospital of Tibet Autonomous Region (PHTAR), with one-year shift. Because of their continuous work, remarkable changes have been achieved in PHTAR during the past five years in trms of upgrading the equipment and software, training local staff, launching research programs, publishing scientific papers and establishing management system and documentation of working guidelines and regulations. In this paper, all of these efforts and progresses are summarized, and the discussion with suggestion may facilitate the healthcare service in the future.
    Application of 3D printed eye model in direct ophthalmoscopy training for medical students
    WU Chan, LIU Yu-tong, DAI Rong-ping, ZHANG Mei-fen, ZHONG Yong, CHEN You-xin
    2022, 42(1):  188-192.  doi:10.16352/j.issn.1001-6325.2022.01.033
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    Objective To design an eye model that can simulate the fundus for teaching direct ophthalmoscopy and to evaluate its effectiveness. Methods Using 3D(3-dimensional) printing materials to creat an eye model and then randomly organized 92 eight-year medical students into two groups: group A (model training group) and group B (traditional training group). After training, real patients were invited to test the student for maximum time of 120 seconds for each student. That recorded the ability of the students to clearly see the optic disk and the time spend to determine the cup-to-disk ratio and whether the students were correct. Results Fourty-three students in group A (93.48%) successfully saw the fundus while 21 students in group B(45.65%) saw the fundus. The difference between the two groups was 47.83% and the 95% confidence interval was 29.59%-66.07% (P<0.001). The median time to see the fundus was 29 seconds in group A; the 95% confidence interval was 23-45 seconds. Based on the lower limit of the 95% confidence interval, it was estimated that at least 80 seconds was required in group B. The P value of the log-rank test was <0.001. It proved that the operation in group A is significantly faster than that of group B. Conclusions This 3D-printed eye model significantly improved the students' learning interest, learning efficiency. This method should be shared by other medical institutions to train students in ophthalmoscopy.
    Application of mind mapping in the teaching of human parasitology
    WEI Chun-yan, WANG Zhen-sheng, YANG Yin, WANG Heng
    2022, 42(1):  193-196.  doi:10.16352/j.issn.1001-6325.2022.01.036
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    Objective To explore the teaching effect of mind mapping to implement the concept of “ student-centered learning (SCL)”. Methods One hundred and eighteen undergraduates of four-year program in Nursing School, Peking Union Medical College enrolled in 2019 were selected as objects. Before the learning of the human parasitology, the students were investigated to find current situation of “students' autonomous learning”;Then they were guided to make up a mind map of what they need to learn in the course of human parasitology and the effect of mind mapping was evaluated at the end of the course. Results This method guided students to make up a mind map and so effectively stimulated their enthusiasm of active learning. This method significantly strengthened students' self-confidence,supported their deep understanding of knowledge and build their capacity of logic attitude of learning. This method may contribute to encourage and orient students' autonomous learning. Conclusions The application of mind mapping is an effective and feasible method to practice education concept of “student-centered learning” concept.
    Application of diversified teaching mode in ultrasound-guided interventional procedures for treatment of pain
    JIA Dong-lin, LI Shui-qing, JIANG Jie, JIANG Ling
    2022, 42(1):  197-200.  doi:10.16352/j.issn.1001-6325.2022.01.032
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    Objective To explore the application effect of diversified teaching mode in ultrasound-guided interventional procedures for treatment of pain. Methods Thirty learners who studied in the Department of Pain, Peking University Third Hospital from January 2018 to December 2020 were enrolled as participants, and were divided into control group and observation group. The control group adopted conventional teaching mode, while the observation group adopted diversified teaching mode. The clinical skill assessments, teaching quality and satisfaction scores of the two groups were compared. Results The excellent and good rate of clinical skill assessments in the observation group was 93.3%, which was higher than that in the control group, which was 73.3%(P<0.05). Scores of mastering basic theoretical knowledge, improving clinical thinking ability, motivating learning interests and improving disease diagnosis and treatment ability in observation group were higher than those in control group (P<0.01). Conclusions Diversified teaching mode improves ultrasound-guided interventional pain management skills, and supports the capacity building of clinical treatment.The method may function as a innovative clinical training pathway for other medical schools and healthcare institutions.