Table of Content

05 May 2019, Volume 39 Issue 5

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Hepatitis B virus X protein inhibits the proliferation and promotes the apoptosis of mouse foot cell line MPC5

2019, 39(5):  617-622. 

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Objective To investigate the effect of hepatitis b virus X (HBx) on the proliferation and apoptosis of condition immortalized mouse foot cell line(MPC5).Methods We used pEX plasmid with HBx gene transfected MPC5 cells，and quantitative Real-time PCR(qRT-PCR) tested the transfection efficiency. The experiment was divided into three groups: HBx transfection group (MPC5-pEX-HBx group):liposome+HBx-plasmid; Negative control group (MPC5-pEX-neo group) : liposome+empty plasmid; Blank control group (MPC5 group) : liposome. Survival rate of each group was determined by MTT method. The cell apotosis was analyzed by flow cytometry. Western blot was employed to detect the Nerhrin、STAT3、JAK2 proteins level. Results There was the highest expression of HBx in 48 hours after transfection with MPC5 cells. The expression level of Nephrin protein in HBx transfection group was lower than that in blank and negative control( P＜0.01). After transfection of HBx gene, the podocyte proliferation was inhibited, and the apoptosis rate was increased significantly (P<0.01), what’s more, STAT3 and JAK2 expression were all increased than control groups(P<0.01). Conclusion HBx can reduce the nephrin expression in podocyte, and inhibit it proliferation and promote which apoptosis, this mechanism may be related to the activation of STAT3/JAK2 signaling pathway.

Mechanism of rL-RVG-induced apoptosis of gastric cancer cell line SGC via alpha 7-nAChR

2019, 39(5):  623-629. 

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Objcetive To explore the possible mechanism of the recombinant avirulent NDV LaSota strain expressing the rabies virus glycoprotein (rL-RVG) inducing the apoptosis of gastric cancer cells by the acetylcholine receptor pathway.Methods The gastric cancer cell lines SGC at logarithmic proliferation stage were randomly divided into the rL-RVG, newcastle disease virus（NDV）and PBS groups.Furthermore,the acetylcholine receptor （α7-nAChR）inhibitor groups and the agonist pretreatment groups were set up, respectitively. At 24 hours after infected by viruses,CCK8 assay was used to determine the inhibitory effect of virus on the growth of gastric cancer cell lines.western blot detected the protein expression of alpha 7-nAChR, RVG, NDV and apoptosis-related proteins including cleved-caspase3 and BAX/BCL-2 as well as the expression of P-ERK and ERK. Immunofluorescence was also used to examine the expression of P–ERK. Results CCK8 showed that the concentrations of the recombinant and wild virus inhibited the growth of gastric cancer cell line SGC were greater than 10-2 /mL and 10-3 /mL, and the rL-RVG group was significantly more inhibited than NDV and PBS group(P<0.05).Western blot showed apoptosis-related proteins cleved-caspase 3, bax/bcl-2 in rL-RVG group and NDV group increased significantly compared with PBS group, the protein expression of P-ERK/ERK in the virus group was lower than the control group significantly. The apoptosis-related protein expression levels in the rL-RVG/NDV/PBS groups pretreated with choline receptor inhibitor significantly more increased than that in the groups without pretreatment.Besides,the groups pretreated with choline receptor agonist had obviously decreased apoptosis protein expression level compared with the groups without pretreating.Western blot and immunofluorescence showed the expression of P-ERK decreased in the virus group, and the recombinant virus group decreased significantly compared with the NDV and PBS group(P<0.05).Furthermore, the P-ERK protein expression in groups pretreated with acetylcholine receptor inhibitor descended.On the other hand, the level of P-ERK ascended while pretreated with choline receptor agonist(P<0.05).Conclusion One of the mechanism that rL-RVG induced apoptosis of gastric cancer cell line SGC by alpha 7-nAChR is suppressing the ERK signaling pathway.

Optimization of alginate-chitosan porous composite scaffold for in vitro proliferation of human umbilical cord-derived mesenchymal stem cells

2019, 39(5):  630-635. 

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Objective Alginate-chitosan porous composite scaffold prepared and optimized was used to culture human umbilical cord-derived mesenchymal stem cells in order to provide the application of stem cells in tissue engineering and disease treatment. Methods We prepared porous composite scaffold, and which was optimized by grafting polylysine on its surface. Next, mesenchymal stem cells were isolated from umbilical cord, seeded and serially cultured in the scaffolds. Cell growth and proliferation were monitored, and cell proliferation curves were made using cytometry and CCK-8 kit techniques. Moreover, stem cell ageing was evaluated by senescence β-Galactosidase staining kit. Results This assay successfully prepared and optimized the alginate-chitosan composite scaffold. We found that the mesenchymal stem cells could easily attach to and cover the surface of the scaffold grafted with polylysine. Moreover, the stem cells grew in agglomerates rapidly. Conclusions This result proves that the alginate-chitosan scaffold provides a suitable environment for cell growth and has the feasibility for cell culture in vitro. This scaffold can be used as potential material for tissue engineering applications.

Effect of Kawasaki disease on intestinal microbiota in children

2019, 39(5):  636-640. 

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Objective To explore the effect of Kawasaki disease on intestinal microbiota in children. Methods The V1~V2 variable region in 16S rRNA gene was amplified by PCR from fecal samples collected from affected with Kawasaki disease and matched healthy children, and analyzed for bacterial taxonomic content following high-throughput sequencing. Microbiota in affected and healthy children were compared by alpha diversity, beta diversity, linear discriminant analysis and LDA effect size analysis. Results The richness (Ace index, P<0.001) and structure (ANOSIM based on unweighted UniFrac distances，P<0.01) of the intestinal microbiota in the Kawasaki disease group had changed significantly. In particular, Lactobacillus (P<0.01), Veillonella (P<0.01), and Clostridium (P<0.001) were significantly diminished in affected children, while Bacteroides (P<0.05), Enterococcus (P<0.001), and Parabacteroides (P<0.001) were significantly expanded. Conclusions Dysbiosis of intestinal microbiota occurs in children with Kawasaki disease. This result provides a partial basis for the later Kawasaki disease microecology research.

Effect of adenomatous polyposis coli on mitochondria during fat metabolism in human hepatoma cell line BEL-7402

2019, 39(5):  641-645. 

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Objective To observe the effect of adenomatous polyposis coli (APC) on cell mitochondria in the process of fat metabolism in human hepatoma cell lines. Methods Human hepatoma cells (BEL-7402) cells were transfected with APC gene knockdown plasmid, and oleic acid (OA) was used to stimulate knockdown cells and control cells to produce lipid accumulation. Oil red O staining was used to detect the lipid accumulation in each group. ATP content detection kit was used to detect the ATP content of each group. The content of oxygen free radicals (ROS) in each group was detected by fluorescent probe labeling. Results After transfection of the plasmid, the mRNA expression of APC gene decreased (P<0.05) and protein expression decreased (P<0.05). Reduced cell lipid accumulation after stimulation with oleic acid in APC knockdown group compared with control cells (P<0.05), ATP content increased (P<0.05), and the number of intracellular oxygen free radicals decreased (P<0.01). Conclusions APC affects the physiological function of mitochondria during lipid metabolism in human hepatoma cell lines.

Berberine promotes M1 proinflammatory phenotype to M2 anti-inflammatory phenotype polarization in macrophage cell line RAW264.7

2019, 39(5):  646-651. 

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Objective To test the effect of berberine (BBR) on polarization of RAW264.7 mouse macrophages. Methods RAW264.7 cells were divided into control group, high lipid- and inflammatory-model group (stimulated with 100μg/L lipopolysaccharide and aggregated LDL), BBR 5 μmol/L group, BBR 10 μmol/L group and BBR 20 μmol/L group. ELISA and flow cytometry were used to assess specific cell markers (TNF-α, MCP-1, CD86, IL-4, IL-13, TGF-β1 and CD206) to define M1 and M2 polarization. Protein expression of apolipoprotein E (apoE) and very-low-density lipoprotein receptor (VLDL-R) or mRNA expression of apoE receptor-2 (apoER2) were detected by Western blot and RT-qPCR,respetively. Results BBR decreased M1 macrophage-specific cell markers TNF-α and MCP-1 levels (P<0.05)and downregulated the expressions of CD86. In contrast, M2 macrophage-specific cell markers IL-4, TGF-β1 and CD206were upregulated by BBR. BBR could promote the protein expression of apoE and the mRNA expression of VLDL (P<0.001). Conclusions BBR treatment may shift RAW264.7 mouse macrophage to antiinflammatory M2 phenotype by promoting the binding of apoE to VLDLR.

Increased expression of FoxO3a and MMP2 in vascular smooth muscle cells of patients with chronic thromboembolic pulmonary hypertension

2019, 39(5):  652-656. 

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To study the role of FoxO3a and MMP2 in chronic thromboembolic pulmonary hypertension (CTEPH). Methods Twenty patients with CTEPH admitted to the China-Japan Friendship Hospital during 2018.1-2018.12 were enrolled in this study. Pulmonary vascular smooth muscle cells (VSMCs) were obtained from 5 cases of 20 surgical specimens. Pulmonary artery trunk of donor was used as a control. The expression of FoxO3a and MMP2 in the surgical specimens and VSMCs from the normal group and disease group were observed by immunohistochemistry and Western Blot. Results The integral absorbance value (IA) of FoxO3a in the CTEPH group was: 3269±338, which was significantly increased compare with normal group 420±46 (P<0.01); the IA of MMP2 respectively was 1799±139, 4936±521 (P<0.01); The expression of MMP2 and FoxO3a in the CTEPH group was significantly increased compare with normal group (P<0.05). Conclusions Overexpression of FoxO3a/MMP2 may be involved in the vascular remodeling, thus accelerating the occurrence and development of CTEPH.

Changes of vascular smooth muscle cells in patients with carotid atherosclerosis

2019, 39(5):  657-662. 

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Objective To explore the vascular smooth muscle cells (VSMCs) number and distribution in carotid atherosclerotic plaque and provide theoretical basis for clinical analysis of carotid atherosclerotic plaque stability. Methods 68 patients with carotid artery plaque after CEA were collected. All the surgical specimens were stained with Movat and immunochemistry to detect VSMCs, and the MA, MA/mm2 and the thickness of fibrous cap was counted under the microscope. Results The stable and unstable plaques were 87.5% and 12.5% in the asymptomatic group, 25.0% and 75.0% in the symptomatic group, respectively (P<0.001). MA of VSMCs in asymptomatic and symptomatic groups were 1650 +58, 1343 +54 (P<0.001); 1506 +59, 1312 +58 in stable and unstable plaques, respectively (P<0.05). In the distribution, the asymptomatic group and the symptomatic group were (1664±73) MA/mm2, (1112±69) MA/mm2 in basal part (P<0.001), (1697±76) MA/mm2, (1412±81) MA/mm2 in the shoulder part (P<0.05), (1620±65) MA/mm2, (1321±66) MA/mm2 in the fibrous cap (P<0.01), respectively. With the increase of fibrous cap, MA/mm2 of VSMCs also increased in both groups, showing a significant positive correlation (P<0.001). Conclusions The number and density of VSMCs in symptomatic and unstable plaques are less than those in asymptomatic and stable plaques. The thickness of fibrous cap is positively correlated with the density of VSMCs.

Dynamic change of autophagy in mouse spermatogenesis identified by RNA-Seq

2019, 39(5):  663-667. 

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Objective To reveal the dynamic expression patterns of autophagy-related genes during spermatogenesis according to the RNA sequencing data sets of 3 population cells related to mouse spermatogenesis. Methods RNA-Seq data during spermatogenesis were downloaded from the GEO database, KEGG(Kyoto Encyclopedia of Genes and Genomes) pathway enrichment analysis of differentially expressed genes were conducted to detect significantly altered pathways between adjacent stages. 783 autophagy-related genes were obtained from the Autophagy Database, and the expression levels of 636 autophagy-related genes(the average FKPM>1) in spermatogenesis were displayed by heatmap. Results Autophagy and autophagy related pathways were significantly enriched between spermatogonia and pachytene but not significantly enriched between pachytene and round spermatid. In addition, 636 autophagy-related genes have three major expression patterns during spermatogenesis. Conclusions Autophagy had undergone significant changes in initiation of meiosis at the RNA level and autophagy-related genes were specifically expressed during spermatogenesis.

Expression of DNA methyltransferases in prostate cancer and the effects on cell proliferation and invasion in vitro

2019, 39(5):  668-672. 

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Objective To investigate the expression of DNA methyltransferases (DNMTs) in prostate cancer tissues and prostate cancer cell lines and the effects on cell proliferation, apoptosis, migration and invasion of prostate cancer cells in vitro. Methods The prostatic tissue samples were collected from 24 patients, including 12 prostate cancer and 12 benign prostatic hyperplasia. Immunohistochemical staining was used to detect the expression of DNMT1 and DNMT3b protein. The prostate cancer cell lines (DU145, PC-3) and human normal prostate epithelial cell line (RWPE-1) were cultured. The expression of DNMT1 and DNMT3b was detected by Western blot and RT-qPCR. SiRNA was used to down regulate the expression of DNMT1 and DNMT3b in DU145 cells, and the changes in biological behaviors of DU145 cells were detected. Results The expression of DNMT1 and DNMT3b in prostate cancer tissues was higher than that in benign prostatic hyperplasia (P<0.05). The expression of DNMT1 and DNMT3b in DU145 and PC-3 cells was higher than that in RWPE-1 cells (P<0.05), and the expression of DNMT1 and DNMT3b in DU145 cells was higher than that in PC-3 cells (P<0.05). Down regulation of the expression of DNMT1 and DNMT3b in DU145 cells could significantly inhibit cell proliferation, promote cell apoptosis and reduce cell migration and invasion (P<0.05), in which the DNMT1 inhibition group had the most significant effect. Conclusions The expression of DNA methyltransferases (DNMT1 and DNMT3b) in prostate cancer is higher than that in normal prostate, and DNMT1 has the most significant effect on cell biological behaviors.

PAK1 correlation with the subcellular localization and protein level of survivin in mouse oocyte meiosis

2019, 39(5):  673-676. 

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Objective To study the relationship of activated PAK1 (pPAK1Thr423) with the subcellular localization and protein level maintenance of survivin in the meiotic progression of mouse oocytes. Methods Immunofluorescent staining was employed to analyze the spatial-temporal correlation between pPAK1Thr423 and survivin during oocyte meiosis; Western blot was applied to analyze the change of survivin level in oocytes treated with PF3758309. Results In mouse oocytes at metaphase I stage (MI), pPAK1Thr423 was co-localized with survivin between chromosome arms and on centromeres of homologous chromosomes, the centromeric co-lozalization remained at metaphase II stage (MII), while that across chromosome axis was confined in the local space between sister centromeres. The protein level of survivin was significantly decreased in oocytes treated with PF3758309. Conclusions pPAK1Thr423 may regulate the subcellular localization and protein maintenance of survivin, so as to participate in CPC regulating chromosome separation in mouse oocyte meiosis.

miR-138 expression is increased in rat cardiomyocyte cell line H9C2 with myocardial oxidative stress

2019, 39(5):  677-681. 

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Objective To observe the changes of miR-138 on Sirt1 expression in ethanol induced Rat H9C2 myocardial cell lines and its downstream oxidative stress and mitochondrial apoptosis-related protein expression. Methods Cells were divided into control group, ethanol group (E group) and miR-138 inhibitor group (I group), every groups were cultured with different concentrations of alcohol culture media for 24h, cell viability was detected by MTT assay and 200mmol/L alcohol culture media was selected as the optimal concentration for following experiment. Group I was cultured with miR-138 inhibitor transfection media. After 12-hour-incubation, total superoxide dismutase (T-SOD) activity was analysed by hydroxylamine method and the content of malondialdehyde (MDA) was detected by thiobarbituric acid method. The expression of mRNA of miR-138 and Sirt1 were detected by RT-qPCR. The expression of Sirt1, Bax, Bcl-2 and caspase-3 protein was detected by Western blot. Results 1.Cell viability could be suppressed by alcohol. 2.Compared with control group, the content of MDA, the expression of miR-138 and both the protein expressions of Bax and caspase-3 were increased significantly (P<0.05) in ethanol group, the T-SOD activity, the mRNA expression of Sirt1, protein expressions of Sirt1 and Bcl-2 decreased significantly (P<0.05) in ethanol group. Compared with ethanol group, the content of MDA, the expression of miR-138 and both the protein expressions of Bax and caspase-3 decreased significantly (P<0.05) in miR-138 inhibitor group, the T-SOD activity, the mRNA expression of Sirt1, protein expressions of Sirt1 and Bcl-2 increased significantly (P<0.05) in miR-138 inhibitor group. Conclusions miR-138 increases oxidative stress of rat H9C2 cardiomyocytes induced by ethanol, Sirt1 may be the target of miR-138.

Effects of SDF1/CXCR4 axis on the angiogenesis induced by degenerative nucleus pulposus cells

2019, 39(5):  682-689. 

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Objective To investigate the influence and relative mechannism of stromal cell derived factor 1 (SDF1)secreted by nucleus pulposus cells(NPCs) on the vascularization of degenerated intervertebral disc. Methods NPCs of degenerated intervertebral disc were obtained and cultured. The expression of SDF1 in NPCs was up regulated and down regulated by transfected virus, then NPCs were classified as DOWN(down regulation),D(degeneration) and UP(up regulation) groups according to the expression of SDF1. Human umbilical vein endothelial cells(VECs) were cultured. Different groups of NPCs or conditional medium from NPCs were respectively co-cultured with VECs, during the period, AMD3100, the CXCR4 receptor inhibitor was also added to the co-culture system. The CCK8, cell migration assay and tube formation assay were used to detect the angiogenesis ability of VECs. Result Each group of NPCs was successfully constructed. After co-cultured with VECs, the CCK8, cell migration assay and tube formation assay showed that VECs cell vitality, migration ability and tube formation ability were enhanced as the expression of SDF1 was increased(P<0.05). After adding AMD3100, the cell vitality, migration ability and tube formation ability of VECs were significantly suppressed in D and UP groups(P<0.05). Conclusion SDF1 is secreted by NPCs of degenerated intervertebral disc, the SDF1/CXCR4 axis could influence the ability of vitality, migration and tube formation of VECs, and may play an important role in vascularization of human degenerated intervertebral disc.

Over-expression miR-181a aggravates H2O2-induced HUVECs injury

2019, 39(5):  690-695. 

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Objective To investigate the expression of miR-181a in hydrogen peroxide (H2O2)-injured human umbilical vein endothelial cells (HUVECs) by H2O2, and the effect and mechanism of miR-181a on cell viability and apoptosis. Methods The viability and apoptosis of HUVECs was measured by MTT and flow cytometry assays. miR-181a and X-linked inhibitor of apoptosis (XIAP) expressions were determined by qRT-qCR and Western blot analyses. The binding sites of miR-181a in XIAP 3'UTR were predicted using Targetscan software and verified by Dual-Luciferase and Western blot assays. Results 1) H2O2 inhibited cell activity while promoted apoptosis of HUVECs in a dose-dependent manner (P<0.05). 2) miR-181a was significantly elevated in H2O2-treated HUVECs (P<0.05). 3) Knockdown of miR-181a promoted H2O2-induced cell activity and inhibited apoptosis (P<0.05). 4) XIAP was identified to be a target for miR-181a. 5) Overexpression of miR-181a inhibited H2O2-induced cell activity and promoted apoptosis, which was reversed by XIAP restoration. Conclusions These findings suggested that miR-181a stimulated H2O2-induced cell activity while inhibited cell apoptosis by targeting XIAP, leading to the aggravation of HUVECs injury.

Dexmedetomidine alleviates acute lung injury induced by lipopolysaccharide in rats

2019, 39(5):  696-700. 

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Objective To explore the role and mechanism of demedetomidine in mitigation of lipopolysaccharide(LPS) induced acute lung injury(ALI) in rats. Methods SD rats were randomly divided: control group, lipopolysaccharide group (LPS group, intraperitoneal injection LPS 5 mg/kg), dexmedetomidine+LPS group (LD group, intraperitoneal injection demedetomidine 25 μg/kg and LPS 5 mg/kg). After LPS injection 6h later, broncho-alveolar lavage fluid (BALF) in left lung was collected for determination of concentrations of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor (TNF-α) by enzyme linked immunosorbent assay (ELISA). And the right lung was removed to observe the pathological changes by HE staining and measure the lung injury score(LIS). The wet to dry lung weight (W/D) ration was calculated. The expression of HGMB1, TLR4 and TLR2 were detected by western blot. Results Compared with control group, the ALI, W/D ration, IL-1β, IL-6 and TNF-α in BALF, and the expression of HMGB1, TLR4 and TLR2 were significantly increased in LPS group (P<0.05). Compared with LPS group, the changes of each index in LD group were significantly relieved (P<0.05). Conclusion Dexmedetomidine relieves LPS-induced ALI in rats through inhibiting the expression of HGMB1 and TLRs.

Gender difference of GABAAα3 expression in the ventrolateral periaqueductal gray of rats with neuropathic pain

2019, 39(5):  701-704. 

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Objective To investigate the gender difference of GABAAα3 receptor expression in the ventrolateral periaqueductal gray (vLPAG) of rats with neuropathic pain. Methods Twenty-four SD rats were divided into 2 groups: 12 females in group F and 12 males in group M. Neuropathic pain model was established by spared nerve injury (SNI) of sciatic nerve branches. The mechanical pain threshold and cold pain threshold were recorded at each time point (1 day before operation and 1, 3, 7, 14 days after operation). Western blotting was used to detect the expression level of GABAAα3 receptor at vLPAG. Results The mechanical and cold allodynia were significantly higher in both group F and group M after operation. The mechanical pain threshold of group F was lower than that of group M (P<0.05), and the frequency of cold foot contraction in group F was higher than that in group M (P<0.05). 14 days after the model establishment, the expression level of GABAAα3 receptor protein increased significantly at vLPAG, and it was higher in group F than that in group M (P<0.05). Conclusions Female rats are more sensitive to neuropathic pain than male rats and the up-regulation of GABAAα3 receptor expression at vLPAG may be related to the decrease of pain threshold in rats.

Silencing PTEN gene inhibits cardiomyocyte injury induced by H2O2 in rats

2019, 39(5):  705-709. 

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Objective To investigate the effect and mechanism of phosphatase and tensin homolog deleted on chromosome ten (PTEN) silencing on cardiomyocyte induced by H2O2 injury in rats. Methods The isolated rat cardiomyocytes were divided into blank control group and H2O2 treated group. (50, 100 and 200 μmol/L), and the LDH release of cardiomyocytes was detected by kit, apoptosis of cardiomyocytes was detected by flow cytometry, and the expression of PTEN was detected by RT-qPCR and Western blot. The expression of PTEN were detected by RT-qPCR and Western blot after transfection of siR-PTEN. The effect of siR-PTEN on LDH release of cardiomyocytes induced by oxidative stress was detected by kit, the effect of siR-PTEN on cardiomyocyte apoptosis induced by oxidative stress was detected by flow cytometry. Results Compared with the control group, the LDH release and apoptosis rate of myocardial cells increased significantly (P < 0.05) and the expression of PTEN mRNA and protein increased significantly (P < 0.05) after H2O2 treatment. After transfection of siR-PTEN, the LDH release rate and apoptosis rate of cardiomyocytes were significantly decreased (P<0.05). Conclusion PTEN silencing could inhibit the injury of cardiomyocytes induced by oxidative stress.

Distribution of oral microbiota in patients with gastric cardia cancer

2019, 39(5):  710-713. 

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Objective To describe and compare the distribution and differences of oral microbiota between gastric cardia carcinoma and healthy controls, and provide clues and insights to explore the etiology of gastric cardia carcinoma (GCC). Methods A total of 33 GCC from October 2015 to January 2016 and 33 healthy controls from August 2017 to January 2018 were enrolled in the study, from Linzhou Cancer Hospital, Henan province. Oral saliva specimens from participants were collected and microbial DNA extraction were conducted. The 16S rRNA V4 region was sequenced by Illumina MiniSeq sequencing platform. Alpha-, beta- diversity and relative abundances were used to compare the differences between GCC and healthy control groups. Results There was a decreased observed Operational Taxonomic Units in GCC compared with the healthy controls (175 vs 198, P < 0.05), while the Shannon index did not show significant difference. The beta diversity was significantly different between two groups (P < 0.001). The relative abundance of Prevotella and Veillonella in the saliva of GCC was significantly higher than those of healthy controls (P < 0.001). Conclusions The oral saliva microbiota of GCC is significantly different from that of healthy controls.

Impact of air pollution on blood pressure in middle-aged and elderly patients with hypertension in Beijing

2019, 39(5):  714-719. 

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Objective To investigate the associations of air pollution with blood pressure in middle and old-aged hypertensive patients in Beijing. Methods A total of 85 local residents of hypertension aged 45 to 85 years old and with good physical activity in Beijing was recruited. Exclude patients with other serious cardio-pulmonary diseases, infections, or allergic diseases. We collected the 24 hourly concentration of PM2.5, PM10, SO2, NO2, O3 and CO from the nearest Beijing monitoring sites for 5 days. Linear mixed effect model with a random intercept for each participant to account the dependent repeated measures was used to estimate the associations between 6 air pollutants and blood pressure. Results In the single-pollutant model, short-term exposure to 6 pollutants except O3 resulted in an increase in SBP（P < 0.05）. PP decreased with O3 exposure（P < 0.05）and increased with SO2 exposure（P < 0.05）. Conclusions Short-term exposure to air pollution may significantly elevate the SBP and PP of hypertensive patients.

Losartan potassium down-regulates the expression of uric acid transporter -1 in renal tubular epithelial cells of rats with uric acid renal fibrosis

2019, 39(5):  720-725. 

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Objective To observe the effect of losartan potassium on the expression of Uric acid transporter1(URAT1) in renal epithelial cells of hyperuricemia rats with renal interstitial fibrosis. Methods The rats were randomly divided intonormal group,model group(it was administrated with adenine 100 mg/(kg?d) and Ethambutol 250 mg/(kg?d) by gastric gavage)，losartan potassium low ,in and high (25、50 and75 mg/kg?d) dose treatment group.Serum uric acid, serum creatinine, urea nitrogen, urinary uric acid, urinary creatinine and uric acid excretion fraction were detected at 1, 2, 3and 4W after modeling. Pathological changes of uric acid renal fibrosis rats were observed by HE and Masson staining methods,and the protein expression of URAT-1 was detected in renal tubular epithelial cells by Western blot. Results With the extension of time, serum uric acid level of model group was significantly higher than that of normal group(P<0.01), uric acid level was lower than normal group(P<0.01), serum uric acid level decreased and uric acid level increased in the treatment group, The high dose group was the most obvious(P<0.05).The tubulointerstitial injury index in the model group was significantly higher than that in the normal group (P<0.05), and the tubulointerstitial injury index was significantly decreased after losartan potassium intervention (P<0.05) . The expression of URAT1 protein were up-regulated in the model group at each time period (P<0.05), and with the progress of pathological damage showed an upward trend, but decreased significantly in losartan potassium intervention groups(P<0.05). Conclusions　 Losartan potassium may reduce the high expression of URAT1 and significantly reduce the level of serum uric acid in hyperuricemia rats.

QKI5~miR-124~c-Myc pathway promotes leukemia cell proliferation

Gui-hua Yang

2019, 39(5):  726-730. 

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Objective To investigate the function and mechanism of RNA binding protein QKI5 in human acute myeloid leukemia (AML). Methods The clinically diagnosed acute leukemia patients were collected as the experimental groups. The expression level of QKI mRNA were detected by RT-qPCR in experimental groups and normal controls. Lentivirus expressing QKI5 was used to transduce into HL-60 cells. CCK-8 analysis was performed to detect the influence of QKI5 on HL-60 cell proliferation. PI staining and FACS assay were used to assess the effects of QKI5 on HL-60 cell cycle progression. RT-qPCR were performed to investigate the changes of miRNA. Results QKI5 mRNA was down-regulated in AML compared to the normal control（p<0.05）. Overexpression of QKI5 repressed HL-60 cell proliferation and cell cycle progression. QKI5 specifically regulated the miR-124 biogenesis. miR-124 inhibits HL-60 cell proliferation via targeting c-Myc expression. Conclusions The QKI5~miR-124~c-Myc regulatory axis has important roles in acute myeloid leukemogenesis.

Clinical characteristics of biopsy-positive giant cell arteritis and the relevance with the positive pathology results

2019, 39(5):  731-734. 

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Objective To summarize the characteristics of biopsy-positive giant cell arteritis patients and the relevance with positive biopsy results. Methods The data of the giant cell arteritis in-patients were retrospectively analyzed who underwent the temporal artery biopsy from January 1997 to December 2017. The clinical characteristics of the patients with a positive were compared with those with a negative biopsy result. Results Twenty-six of 46 biopsies were positive for giant cell artery and 19 negative. The biopsy-positive group has higher age and shorter course. Percentage of scalp tenderness、myalgia and non-specific manifestations are higher, while hearing loss is lower in positive group. The patients in positive group had less smoking history and tumor history, and lower lymphocytes and hemoglobin. Only lymphocyte count (odds ratio 0.205) was independent predictor of the biopsy results. Conclusions The group with positive temporal artery biopsy result has significantly different characteristics from negative group. The number of lymphocytes is an independent influencing factor to the pathologic result of temporal artery biopsy.

Clinical analysis of laparoscopic nephron sparing surgery for treatment of complex renal cyst

2019, 39(5):  735-737. 

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Objective To analyses the clinical experience of laparoscopic nephron sparing surgery for treatment of complex renal cyst. Methods 19 cases of complex renal cyst treated with laparoscopic partial nephrectomy were analyzed retrospectively. There were 7 males and 12 females, with the mean age of 43.3 years (28 to 59 years). The locations were 9 cases in the middle of the kidney, 4 cases in the upper pole and 6 cases in the lower pole of the kidney. The mean diameter of the renal cyst mean was 3.32cm (2.5 to 5.1cm). There were 6 cases of type II, 7 cases of type IIF, 4 cases of type III and 2 cases of type IV according to the Bosniak classification. Results 19 patients were performed laparoscopic nephron sparing surgery successfully. The mean operation time was (76±22.4) min (60 to 130 min), and the mean intraoperative blood loss was 38ml (20 to 100 ml). The mean renal artery occlusion time was 17.0min (14 to 28min), and there was no significant change in serum creatinine before and after operation. The pathological results were 18 cases of renal clear cell carcinoma and 1 case of simple renal cyst, with negative margins. Conclusions Laparoscopic nephron sparing surgery can resect the possible malignant renal cyst completely no matter it is benign or malignant, and can preserve the kidney while avoiding some medical risks.

Optimization of plasmid DNA extraction method for improvement of transfection efficiency

2019, 39(5):  738-744. 

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Objective To optimize an extraction method for isolation of plasmid DNA to improve extraction and transfection efficiency of plasmid DNA. Methods The plasmid was extracted at room temperature and 4°C, and the extraction process was splitted to bacterial lysis, column and elution at two temperatures. Plasmid concentration and quality were detected by UV spectrophotometer and agarose gel electrophoresis，respectively. The working efficiency of plasmid was detected by fluorescence microscopy and Western blot after transfecting into mammalian cells. Results Plasmid extraction at low temperature significantly improved the plasmid yield and increased its working efficiency in mammalian cells. The three steps of plasmid extraction were sensitive to temperature, including bacterial lysis, absorption and elution, which all contributed to the yield and quality of extracted plasmid. Conclusions A simple and convenient plasmid extraction method is successfully obtained to increase plasmid yield and working efficiency in mammalian cells.

Progress on natural orifice transluminal endoscopic surgery

2019, 39(5):  745-749. 

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Natural orifice transluminal endoscopicsurgery (NOTES) is a surgical operation that is performed through the natural orifice of the human body. It has become a research hotspot due to its advantages such as less trauma, quicker recovery and beautiful appearance. With the development of technical equipment, the selection and operation methods are continuously enriched.

Research progress of fumarate hydratase gene defect in uterine leiomyoma

2019, 39(5):  750-754. 

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Recent studies have shown that the defect of fumarate hydratase (FH) may be related to the formation of uterine leiomyoma. This article reviews the biological characteristics of FH, the possible reasons for the formation of uterine leiomyoma caused by FH defect and the expression of different types of uterine leiomyoma. The aim is to provide new ideas for clinical diagnosis, treatment and prognosis of uterine leiomyoma.

Role of calcium signal in the synthesis of aldosterone and its regulation mechanism

2019, 39(5):  755-758. 

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Angiotensin II, adrenocorticotrophic hormone and potassium are the main physiological stimulus that promote aldosterone secretion. All of the three kinds of stimulus could activate voltage-gated calcium channel in adrenal cortical cells. Calcium influx through voltage-gated calcium channel is the common path in the synthesis of aldosterone. Calcium concentration in ATP1A1, ATP2B3, CACNA1D and KCNJ5 mutated aldosterone producing adenomas are also elevated. Calcium signal will promote aldosterone secretion though Ca2+-CaM-CaMK pathway.

Constructing a medical genetics course evaluation system for building self-learning capacity

2019, 39(5):  759-762. 

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An online teaching tool "rain classroom" was applied to the reform of the evaluation system of the medical genetics course. The formative assessment was strengthened while the summative assessment was well completed. Multiple methods were used, including preview, attendance, answering questions in class, stage quiz, case-base leaning, homework, final exam and so on. Quantitative evaluation of teaching in class and autonomous learning after class covered from learning attitude through learning result. The post-class questionnaire showed that the degree of satisfaction was high and the comprehensive evaluation system could effectively promote autonomous learning.

Analysis of the ability and affected factors on self-regulated learning in 8-year program medical students at Peking Union Medical College

2019, 39(5):  763-768. 

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Objective This study investigated the factors affecting self-regulated learning ability by exploring the current situation of self-regulated learning in eight-year medical students, and provides a basis for further improvement in this respect. Methods Questionnaire method was used to evaluate the time，the resources, the methods and the ability of self-regulated learning of the medical students in the 5th, 6th and 7th years, and to explore the relevant factors affecting self-learning ability. Results A total of 170 questionnaires were collected, 74 males and 96 females. The average time spent on self-regulated learning was (106.3+35.7) minutes per day. Textbooks, teachers, senior schoolmates are the most commonly used pathway to self-learning resources. PubMed is the most commonly used database, followed by UpToDate clinical consultant and then Wanfang/Weipu and CNKI. The score of learning anxiety in females are significantly higher than that of males. The analysis of the influencing factors of self-regulated learning shows that the students with better family economic conditions have higher score of self-regulated learning ability. According to grade classification, the total and strategy score of self-regulated learning ability in the fifth year are significantly higher than those in the sixth year. The self-regulated learning ability of excellent students is significantly higher than that of poor students in motivation, strategy and the total score. Conclusions The ability to self-regulate learning strategies is affected by many different factors. Students with the better the family economic condition, longer self-regulated learning time, better academic record have higher self-regulated learning ability.