Objective:To investigate the expression of potassium calcium-activated channel subfamily N member 4(KCNN4)in pancreatic cancer tissues and its impact on tumor progression,and to explore its role in clinical diagnosis and prognosis evaluation of pancreatic cancer.Methods:Using the GEPIA2 data analysis platform,the expression of KCNN4 in pancreatic cancer tissues and its correlation with patient prognosis were analyzed by integrating data from the TCGA and GTEx databases.Cancerous and adjacent non-cancerous tissues from 24 patients with pancreatic cancer who underwent surgical resection at ChangHai Hospital of the Naval Medical University were collected.The expression of KCNN4 in pancreatic cancer tissues was validated using qPCR,Western blotting,and immunohistochemical staining.The expression of KCNN4 in human pancreatic cancer cell lines BXPC3 and PANC-1 was knocked down using shRNA.CCK-8 and colony formation assays were performed to detect tumor cell proliferation and growth.A murine KPC cell pancreatic cancer model was established to investigate the effects of KCNN4 knockdown on the growth of orthotopic pancreatic tumor and overall survival(OS)in mice.Results:Analysis of TCGA and GTEx data revealed that KCNN4 was highly expressed in pancreatic cancer tissues(P<0.05)and was associated with shortened OS and disease-free survival(DFS)in patients(both P<0.05).The expression levels of KCNN4 mRNA and protein were significantly elevated in pancreatic cancer tissues compared with those in adjacent non-cancerous tissues(all P<0.01).Knockdown of KCNN4 led to significantly reduced growth rates and fewer colony formations in pancreatic cancer cells(both P<0.01).The murine orthotopic pancreatic tumor experiment revealed that KCNN4 knockdown inhibited tumor progression and prolonged the OS of mice.Conclusion:KCNN4,highly expressed in pancreatic cancer tissues,promotes pancreatic cancer cell proliferation and tumor progression,and its expression is closely associated with patient prognosis,suggesting that KCNN4 may serve as a promising target for clinical diagnosis and prognosis evaluation of pancreatic cancer.

[摘 要] 目的：通过生物信息学分析以及细胞生物学实验研究角蛋白6A（KRT6A）对胰腺导管腺癌（PDAC）诊断、预后判断、免疫微环境以及PDAC细胞PANC1增殖、凋亡等生物学行为的影响。方法：通过GEPIA平台整合TCGA（The Cancer Genome Atlas）数据库与GTEx（Genotype-Tissue）数据库中的数据，分析KTRT6A在PDAC组织中的表达情况，并通过CIBERSORT工具分析KRT6A表达与PDAC组织中免疫细胞浸润的关系，然后通过GSEA方法研究与KRT6A基因表达相关的肿瘤信号通路。选取长海医院病理科保存的60例PDAC组织与癌旁组织标本进行免疫组化分析，验证KRT6A在肿瘤组织中表达情况；通过干扰RNA敲减PANC1细胞中KRT6A的表达，采用CCK-8实验以及流式细胞术检测敲减KRT6A对细胞的增殖、凋亡的影响。结果：利用TCGA与GTEx数据库数据分析发现，KRT6A在人PDAC组织中呈高表达，且与患者较差的生存期存在关联（P=0.015）。利用CIBERSORT软件以及GSEA分析发现，KRT6A高表达的PDAC组织中M2型巨噬细胞浸润性升高（P=0.034），且与Wnt通路（NES:1.7359272，P<0.05）、磷酸戊糖途径（PPP）（NES:1.5613053，P<0.05）等信号通路上调有关联（P<0.05或P<0.01）；免疫组化结果进一步验证了KRT6A在PDAC组织中呈高表达（P<0.001）。增殖和凋亡实验发现，干扰KRT6A能够显著抑制PANC1细胞的增殖（P<0.05）以及凋亡（P<0.001）。结论：KRT6A在人PDAC组织中呈高表达，敲减其表达能够抑制PANC1细胞的增殖和凋亡，具有作为PDAC诊断与预后判断新靶标的潜力。

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

To construct a recombinant attenuated Salmonella strain carrying the p57 and BLID genes and to evaluate its inhibitory effect on breast cancer cells and apoptosis induction in vitro,the recombinant attenuated Salmonella LH430/pEGFP-p57-BLID was constructed.RT-PCR and Western blot were used to detect the transcription and expression of the recombinant attenuated Salmonella in cancer cells.The CCK-8 method was used to assess the safety of the recombinant at-tenuated Salmonella and its impact on the activity of breast cancer cells.The cell scratch assay was used to analyze the influence of the recombinant attenuated Salmonella on the migratory ability of cancer cells.Flow cytometry and Western blot were used to analyze the effect of the recombinant attenuated Salmonella on the apoptosis of MDA-MB-231 cells.The bands of target genes carried by the recombinant attenuated Salmonella could be amplified by PCR,and these target genes could be transcribed and expressed in MDA-MB-231 cells.The recombinant attenuated Salmonella showed good growth characteristics and genetic stability.Compared with the PBS group,recombi-nant attenuated Salmonella LH430/pEGFP-p57-BLID significantly reduced(P＜0.01)the activity of MDA-MB-231 cells and significantly inhibited(P＜0.01)the migration of MDA-MB-231 cells.Recombinant attenuated Salmonella LH430/pEGFP-p57-BLID can obviously increase the number of apoptosis in MDA-MB-231 cells,significantly upregulate(P＜0.01)the expression of apoptotic protein Bax,and significantly downregulate(P＜0.01)the expression of apoptotic protein Bcl-2.The results showed that the recombinant attenuated Salmonella had an inhibitory effect on breast cancer MDA-MB-231 cells and could induce apoptosis,which laid a foundation for the subsequent study of anti-tumor in vivo.

OBJECTIVE: The CYP4V2 gene of two pedigrees affected with Bietti crystalline corneoretinal dystrophy was analyzed to indentify the cause of the disease and provide a basis for clinical diagnosis. METHODS: The probands were subjected to next generation sequencing (NGS). Suspected variants were verified by Sanger sequencing. Pathogenicity of the variants were searched through relevant databases and PubMed by following the ACMG guidelines. RESULTS: A homozygous variant in the CYP4V2 gene c. (802-8) _810delTCATACAGGTCATCGCTinsGC was detected in proband from pedigree 1, parents did not detect; CYP4V2 genes c. (802-8)_810delTCATACAGGTCATCGCTinsGC and c. 958 C>T (p.Arg320X) compound heterozygous variants existed in the proband of pedigree 2,both parents were variant carriers. The results of Sanger sequencing showed that the variant of CYP4V2 gene in the two families was consistent with the NGS sequencing. The c. (802-8)_810delTCATACAGGTCATCGCTinsGC of CYP4V2 gene was splicing variant, and both splicing variant and nonsense variant could produce truncated nonfunctional protein products. Based on standards and guidelines by American College of Medical Genetics and Genomics, the CYP4V2 genes c. (802-8)_810del TCATACAGGTCATCGCTinsGC and c. 958 C>T (p.Arg320X) were predicted to be pathogenic variants (PVS1+PS1+PM2+PM3). CONCLUSION The homozygous variant c. (802-8) _810delTCATACAGGTCATCGCTinsGC and the complex heterozygous variants c. (802-8) _810delTCATACAGGTCATCGCTinsGC and c.958C>T (p.Arg320X) in CYP4V2 gene are the cause of the disease in the probands of two pedigrees , respectively.
Corneal Dystrophies, Hereditary/pathology* ; Cytochrome P450 Family 4/genetics* ; Genetic Variation ; Humans ; Mutation ; Pedigree ; Phenotype ; Retinal Diseases/pathology*

Objective: To investigate the relationship between physical activity (PA) and the risk of incident hypertension among population in rural areas of China. Methods: The Community Intervention of Metabolic Syndrome in China & Chinese Family Health Study (CIMIC) was conducted in 2007-2008. Data on PA, smoking, drinking, blood pressure and other variables were obtained at baseline. Then the follow-up study of incident hypertension was performed during 2012-2015. A total of 41 457 participants aged ≥18 years and free from hypertension at baseline were included in the final analyses. PA was calculated as metabolic equivalent (MET) for each participant. Cox proportional hazard models were used to explore the relationship of PA with incident hypertension according to the quartiles of PA. Results: A total of 6 780 participants developed hypertension during an average follow up of 5.8 years. The annual incidence of hypertension was 2.80%. Compared to participants in the first quartile of PA, HR (95%CI) of incident hypertension decreased with the level of PA of 0.92 (0.86, 0.99), 0.72 (0.67, 0.77) and 0.70 (0.65, 0.75) for the 2^nd, 3^rd and 4^th quartile, respectively (P_trend<0.001). In subgroup analyses, compared to the first quartile, hazards of hypertension among normotensive participants (systolic blood pressure less than 120 mmHg (1 mmHg=0.133 kPa) and diastolic blood pressure less than 80 mm Hg) in the 2^nd, 3^rd and 4^th quartile were 0.82 (0.70, 0.95), 0.73 (0.63, 0.85) and 0.78 (0.67, 0.90), respectively (P_trend=0.002). Among participants with prehypertension (systolic blood pressure from 120 to 139 mmHg and/or diastolic blood pressure from 80 to 89 mmHg), similar trend for the relationship of PA and incident hypertension was also found with HR (95%CI) of 0.94 (0.87, 1.01), 0.71 (0.65, 0.77) and 0.66 (0.61, 0.71) for the 2^nd, 3^rd and 4^th quartile, respectively (P_trend<0.001). Conclusion There was linear trend association between PA and incident hypertension. Increased PA in daily life may be a protective factor against hypertension.

Objective To analyze the retrieval of master and doctorial dissertations entitledsystematic review/meta analysis in domestic universities of traditional Chinese medicine. Methods Master and doctorial dissertations entitledsystematic review/meta analysis in domestic universities of traditional Chinese medicine retrieved from China Dissertation Full-text Database were analyzed using Excel and SPSS 17. 0,respectively. Results Of the 423 master and doctorial dissertations included in this study,71. 9% were retrieved from both Chinese and English databases, 27. 7% were retrieved from Chinese databases,82. 8% were retrieved from two or more Chinese databases,54. 1% were retrieved from two or more English databases. The most frequently retrieved Chinese database was CKNI, followed by VIP, CBM and Wanfang. The most frequently retrieved English database was PubMed, followed by Cochrane Library, EMBASE and OVID. Conclusion The low use of traditional Chinese medicine databases is the main prob-lem in retrieval of master and doctorial dissertations entitledsystematic review/meta analysis in domestic univer-sities of traditional Chinese medicine. No significant difference is found in retrieved doctorial dissertations and mas-ter dissertations between 211and non-211 universities of traditional Chinese medicine.

Carcinoma, Hepatocellular ; diagnosis ; surgery ; therapy ; Hepatectomy ; Humans ; Liver Neoplasms ; diagnosis ; surgery ; therapy

Objective To observe the effect of p53 protein on smooth muscle cell(VSMC)in rabbit artery balloon injury.Methods Restenosis model of carotid artery after balloon injury was established in rabbits.30 rabbits were divided into 2 groups,the sham group(n = 6)and the vascular injury group(n = 24).With H.E.staining and automatic image analysis system,we investigated artery morphology alteration and measured the area of arterial intima and media.The expressions of p53 protein were detected by immunohistochemical analysis.Results With H.E.staining and automatic image analysis and immunohistochemistry,the results showed that the expression of p53 was significantly reduced and the intima area was increased in model group compared with the sham group(P ＜0.01).But the expression of p53 in media was remarkably reduced compared with intima(P ＜ 0.01).Conclusions The possible mechanism of preventing arterial restenosis in the balloon injury might be related with p53,which may be through inhibiting neointimal proliferation in arterial restenosis .