1.Hsa-mir-202 inhibit the proliferation of lung cancer A549 cells by reducing expression of IL-10
Xiaobin FAN ; Qiuxiong LIN ; Yanfei LOU
Chinese Journal of Clinical Oncology 2014;(23):1512-1516
Objective:To study the effects of the overexpression of hsa-miR-202 on the proliferation and molecular mechanism of lung cancer A549 cells. Methods:A sequence of hsa-miR-202 with ppG/miR/eGFP/Blasitidin pasmid was directionally connected and a eukaryotic expression vector pmiR-202 of the target hsa-miR-202 gene was constructed. pmiR-202 was transtected to A549 cell with Lipofectamine 2000. The WST assay was used to detect the cell proliferation rate, and RT-PCR was used to detect the relative gene expression levels. Western blot analysis was used to detect the IL-10 protein expression levels. The interaction between miR-202 and IL-10 was examined using a luciferase reporter assay. Results:The design from the DNA sequencing results shows that a eukaryot-ic expression vector of miR-202 was successfully constructed. The proliferation inhibition rates of A549 cells by Pmir-202 were 12%, 38%, and 52%. The differences in the treatment group compared with the blank control and negative control groups were statistically significant. The RT-PCR results showed that the relative expression levels of miR-202 increased after transfection with pmiR-202. Over-expression of miR-202 can downregulate the relative gene and protein expression levels of IL-10, and the relative levels were 25%and 0.75, respectively. Compared with the blank control and the negative control groups, the difference was statistically significant (P<0.05). The fluorescent activity was reduced when transfection was performed with miR-202 mimics, and IL-10-3'UTR plasmid was cloned. Conclusion: pmiR-202 effectively inhibited the proliferation of A549 cells and exhibited a time-effect relationship with miR-202 by targeted combination with IL-10 3'UTR to downregulate IL-10 expression in A549 cells.
2.Drug resistance and molecular epidemiological analysis of KPC-producing Escherichia coli strains isolated in Hangzhou
Yuying WANG ; Yan QI ; Xuejing YANG ; Yanfei WANG ; Yan JIANG ; Zhengqing LOU ; Yunsong YU
Chinese Journal of Microbiology and Immunology 2016;36(5):359-363
Objective To analyze the epidemiological characteristics of Klebsiella pneumoniae car-bapenemase(KPC)-producing Escherichia coli(E. coli)strains isolated in Hangzhou,China. Methods A total of 25 KPC-producing Escherichia coli strains were collected from four hospitals in Hangzhou from July 2012 to January 2014. Antibiotic susceptibility of the isolates to 22 common antimicrobial agents was deter-mined by using Kirby-Bauer(K-B)disk diffusion method. PCR analysis and gene sequencing were used for bla KPC gene screening. The modified Hodge test was performed to detect the production of carbapenemase. Pulsed-field gel electrophoresis(PFGE)and multi-locus sequence typing(MLST)were used for homology analysis. Results All of the 25 clinical isolates were confirmed to be KPC-producing E. coli strains,harbo-ring the blaKPC-2 gene. These KPC-producing isolates showed high drug resistance rates and were resistant to almost all β-lactam antibiotics. PFGE typing classified the 25 isolates into three main homologous clone groups,including clone group A(4 isolates),clone group B(5 isolates)and clone group C(2 isolates), and some single clones(14 isolates). MLST typing classified the isolates into eight ST types,including ST131(14 isolates),ST167(3 isolates),ST2003(3 isolates),ST410(1 isolate),ST457(1 isolate), ST1463(1 isolate),STnew1(1 isolate)and STnew2(1 isolate). The typing results of PFGE and MLST were consistent with each other. Conclusion The prevalent KPC-producing E. coli strains in Hangzhou, China were ST131 type,which were resistant to multiple antibiotics and had been detected in several hospi-tals. The epidemic of KPC-producing E. coli strain often occurred at some special wards,such as Intensive Care Unit(ICU)and emergency ICU.
3.Characteristics of peripheral blood CD45RA+and CD45RO+T lymphocyte subsets in patients with cGVHD and its clinical significance
Ying LOU ; Chunmiao WANG ; Ting LIN ; Yanfei LUO ; Huiquan GAN ; Maohua ZHUO
Chinese Journal of Immunology 2014;(8):1117-1120
To characterize the CD45RA+and CD45RO+T lymphocyte subsets in the peripheral blood of patients with cGVHD induced by allogeneic haematopoietic stem cell transplantation ( allo-HSCT ) and to explore their relations with the disease.Methods:The peripheral blood was collected from 64 patients after allo-HSCT,including 21 non-cGVHD patients,15 light grade cGVHD patients,18 mild grade cGVHD patients and 10 severe grade cGVHD patients,then CD4+CD45RA+,CD4+CD45RO+, CD8+CD45RA+and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry ( FCM).Results: Compared with the control,the percent of CD4+CD45RA+T lymphocyte in patients with light,mild and severe grade cGVHD decreased markedly (P<0.05),the percent of CD4+CD45RO+T lymphocyte increased markedly (P<0.05).But there were not obviously change in the patient with different grade cGVHD.The percent of CD8+CD45RA+,CD8+CD45RO+T lymphocyte did not change obviously.Conclusion:CD4+CD45RA+and CD4+CD45RO+may play an important role in the pathogenesis of cGVHD.
4.Current status of the diagnosis and treatment of pancreatic exocrine insufficiency after pancreatectomy
Ning PU ; Yanfei AN ; Wenhui LOU
Journal of Clinical Hepatology 2023;39(12):2763-2769
Dyspepsia due to pancreatic exocrine insufficiency is a common complication after pancreatectomy; however, due to the lack of simple, efficient, and unified diagnostic methods, standardized treatment standards, and awareness of this disease among surgeons, it is difficult to get adequate diagnosis and treatment. Therefore, this article summarizes the research advances in the definition and pathogenesis of pancreatic exocrine insufficiency, the incidence rate of pancreatic exocrine insufficiency after different surgical procedures, and current diagnostic methods and treatment strategies, in order to provide a reference for further improving the diagnosis and treatment of pancreatic exocrine insufficiency after pancreatectomy.