1.Azithromycin up-regulated class Ⅰ integron-integrase gene expression in biofilm-forming Pseudomonas aeruginosa
Jiansen CHEN ; Yuhong PAN ; Xiangyan WU
Chinese Journal of Infection Control 2015;(9):597-600
Objective To evaluate the effect of azithromycin on class Ⅰ integron-integrase gene (intI 1 )mRNA expression in biofilm-forming (BF)Pseudomonas aeruginosa (PA).Methods intI 1 of 10 PA strains isolated from a hospital were detected,1 strain with positive BF+intI 1 was selected for culture,blank control group and three az-ithromycin trial groups (divided according to 3 concentrations:16 mg/L,32 mg/L,and 64 mg/L)were set,experi-ments were repeated 5 times,expression of intI 1 mRNA were detected by RT-PCR.Results Relative expression of intI 1 mRNA in azithromycin groups of 16 mg/L,32 mg/L,64 mg/L,and control group were (1 .15 ±0.04), (12.47±3.10),(19.71 ±0.78 ),and (1 .00 ±0.00),respectively,there were significant difference among four groups(F =163.82,P <0.001 );intI 1 mRNA expression between 16mg/L azithromycin group and control group was not significantly different (P >0.05),but among other groups were significantly different (P <0.05 ),intI 1 mRNA expression in azithromycin groups increased with the enhancing concentration of azithromycin in culture so-lution .Conclusion Expression of intI 1 gene mRNA in BF PA can be up-regulated by the present of azithromycin, which may improve the probability of drug-resistant genes,and promote drug-resistant gene recombination.
2.Amplification of?-globin gene to assess the quality of genitourinary tract samples for PCR detection
Jiansen CHEN ; Zhixin CHEN ; Wenjuan ZHOU ; Bin ZHU
Chinese Journal of Clinical Laboratory Science 2006;0(06):-
Objective To investigate whether amplification of?-globin gene can be used to monitor the quality of samples for PCR detection of DNA of Chlamydia trachomatis(CT).Methods First-void urines(FVU),endocervical swabs(ECS),and urethral swabs (UTS) were collected for PCR detection of CT-DNA and cellular?-globin gene.Samples negative for?-globin gene were retested after 10-fold dilution.Results The positive rates of?-globin gene in FVU and ECS were 95.6%(255/264),and 91.7%(413/450) respectively, both higher than that(77.8%,172/221) in UTS(P
3.Construction and immunological protection analysis of attenuated Salmonella typhimurium harbouring Helicobacter pylori oipA DNA vaccine
Miaoduan LIN ; Jiansen CHEN ; Feifei SHE ; Hao CHEN
Chinese Journal of Immunology 2010;26(4):335-340
Objective: To optimize Helicobacter pylori(H.pylori)oipA gene and develop strain of vaccine in Attenuated Salmonella typhimurium SL7207 haxboring recombinant plasmid pVAX1-oipA and pVAX1-optioipA, then to study its protection against H.pylori in C57BL/6 mice.Methods: oipA gene was modified by codon optimization,inserting the Kozak sequence and engineering CpG motifs.The optioipA gene and oipA gene was inserted into the eukaryotic expressing vector pVAX1 respectively to develop recombinant plasmid pVAX1-optioipA and pVAX1-oipA.The AGS cells were transfected by pVAX1-oipA,pVAX1-optioipA and pVAX1 respectively.Then the OipA protein expression was confirmed by Western blot.pVAX1-oipA,and pVAX1-optioipA were converted to LB5000 for methylation and then the modified eukaryoticvector was electrotransfered to final host SL7207 respectively, SL7207/pVAX1-oipA, SL7207/pVAX1-optioipA was identified with PCR and enzyme digestion.Specific serum IgG antibody was detected by indirect ELISA after oral inoculation with vaccine strains.Mice were challenged with live Sydney strain(SS1)three times at 0,2,4 d(5 × 10~8CFU/each mouse).Fonr weeks after challenge, the mice were sacrificed.The colonization of H.pylori in gastric mucosa were detected by rapid urease test and quantitative culture of H.pylori.Results: The AGS cells transfected with pVAX1-oipA and pVAX1-optioipA had expressed corresponding production, moreover, the expression level of pVAX1-optioipA was higher than those of pVAX1-oipA,SL7207/pVAX1-oipA and SL7207/pVAX1-optioipA confirmed with PCR,enzyme digestion;Two weeks after last immunization,immunized mice.were induced to produce anti-OipA IgG antibodies, and the levels of anti-OipA IgG antibody in pVAX1-optioipA group was obviously higher than in pVAX1-oipA group(P<0.01); After challenged with SS1, the infection rate of pVAX1-oipA and pVAX1-optioipA group was 60%(9/15)and 26.67%(4/15), which was significantly lower than 100%(10/10)in the control group(P<0.01).Conclusion:Attenuated Salmonella typhimurium SL7207 containing pVAX1-oipA, pVAX1-optioipA is successfully constructed.It can protect mice from H pylori infection and optimizing oipA gene can enhance the protection efficiency.
4.Clinical efficacy of high-selective laparoscopic high ligation of spermatic vein
International Journal of Surgery 2020;47(9):624-627
Objective:To evaluate the clinical efficacy and safety of high-selective laparoscopic high ligation of spermatic vein for the treatment of varicocele.Methods:A retrospective analysis was performed about 419 cases clinical data which have received high-selective laparoscopic high ligation of spermatic vein in Nanyang Second General Hospital from January 2010 to December 2018. The patients were divided into endoscopy group ( n=289) and open group ( n=130). the patients of endoscopy group were treated with laparoscopic high ligation of spermatic vein, the patients of open group were treated with high ligation of spermatic vein according inguinal incision. The operation time, hospital stay, intraoperative blood loss, postoperative analgesic application, postoperative complications, recurrence, and postoperative semen quality improvement were compared between the two groups. Measurement data were presented as mean±standard deviation ( Mean± SD), and comparison between groups was performed by t-test; count data groups using Chi-square test. Results:This group of 419 patients were all successful operation, all patients with intraoperative and postoperative stable vital signs. The operation time [(26.78±5.14) min vs (61.45±9.75) min], hospitalization time [(4.14±0.39) d vs (10.62±1.19) d] of the patients in the endoscopy group was better than that of the open group, and the difference were statistically significant ( P<0.01). Comparison of intraoperative blood loss between the two groups [(18.32±3.05) mL to (29.14±2.41) mL], the difference was not statistically significant ( P=0.053). The postoperative analgesic application rate, complication rate, and recurrence rate of patients in the endoscopic group were 12.5% (36/289), 2.1% (6/289), and 0.7% (2/289), respectively. The control group was 13.8%(18/130), 7.7% (10/130), 5.4% (7/130), the difference between the two groups were statistically significant ( P<0.05). The postoperative semen quality of the two groups of patients improved to varying degrees. There were 357 cases with definite improvement in postoperative semen quality, including 231 cases in the endoscopic group and 126 cases in the open group, but the difference between the two groups was not statistically significant ( P>0.05). Conclusions:High-selective laparoscopic high ligation of the spermatic vein for the treatment of varicocele, the operation is quick, the high ligation of the varicocele does not damage the blood supply of the testicular artery, the complication rate is low, and the recurrence rate is low. It is worthy of clinical application.
5.Progress of lipoxygenase in liver ischemia reperfusion injury
Fei WANG ; Wen HOU ; Shuang YANG ; Jiansen LU ; Decheng CHEN ; Wenjing XU ; Lei LIU
Chinese Journal of Hepatobiliary Surgery 2023;29(3):230-234
Liver ischemia reperfusion injury (IRI) is one of the main causes of liver dysfunction or functional failure after liver transplantation or liver resection. As the main organ of lipid metabolism, liver is closely related to lipid metabolic balance. Lipoxygenase is a non-heme iron-containing oxidases that oxidizes polyunsaturated fatty acids to produce hydroxy-eicosanotetraenoic acid. Lipoxygenase is excessively expressed during liver ischemia, causing lipid metabolic disorders. High expression of several proinflammatory cytokines induced by lipoxygenase during liver reperfusion. Lipid peroxidation induced by lipoxygenase leads to the production of lipid oxygen free radicals, which induces iron death mainly characterized by lipid peroxidation, thus affecting apoptosis and tissue damage. This review mainly introduces the latest progress of lipoxygenase in liver IRI.
6.Mechanism of mesenchymal stem cell-derived exosome for alleviating hepatic ischemia-reperfusion injury
Decheng CHEN ; Wen HOU ; Shuang YANG ; Jiansen LU ; Fei WANG ; Wenjing XU ; Lei LIU
Chinese Journal of Organ Transplantation 2023;44(5):317-321
The review focuses upon the mechanism of exosome derived from mesenchymal stem cells in hepatic ischemia-reperfusion injury(IRI)to provide references for clinical application of exosomes in alleviating hepatic IRI.
7.Clinical effect of iliac artery interventional chemotherapy combination with bladder perfusion for high level of bladder urothelial carcinoma
Jiansen CHEN ; Ming LI ; Yueqing WU ; Mao TANG ; Zetian TANG ; Xinwu GUO ; Jinqian LI ; Kesong QIN
International Journal of Surgery 2017;44(9):583-586,后插1
Objective To evaluate the clinical effect of iliac artery interventional chemotherapy combination with bladder perfusion for high level of bladder urothelial carcinoma after transurethral bladder tumor electricity cut operation for those who intend to retain the bladder.Methods From February 2010 to December 2016,a total of 74 cases high-grade urothelial carcinoma of the bladder,giving chemotherapy and suitted support were retrospectively analyzed.The patients were divided into two groups according to different chemotherapy methods.Artery perfusion chemotherapy group:43 cases treated by transurethral bladder tumor cutting + bilateral iliac artery interventional perfusion chemotherapy + bladder perfusion in combination therapy;Intravenous chemotherapy group:31 cases underwent transurethral bladder tumor cut method + cisplatin,vein gemcitabine + bladder perfusion chemotherapy combined therapy.According to the ((x) ± s) between the groups.The method of count data is represented by chi-square test.Kaplan-meier method was used to describe the survival time of two groups of patients after operation.Results Two groups of patients with postoperative there was no statistically significant difference in disease-free survival time.In the bone marrow suppression (x2 =4.956,P =0.029);gastrointestinal tract reaction (x2 =5.912,P =0.012);dermatitis,mucositis (x2 =4.276,P =0.013),etc.,has the advantages in the iliac artery interventional perfusion chemotherapy (P < 0.05).Conclusion Via efficient iliac arterial infusion chemotherapy in patients with venous chemotherapy patients,to the retention of the bladder has a certain value,and the incidence of adverse reactions after surgery is low,have certain advantages.
8.The chemical reprogramming of unipotent adult germ cells towards authentic pluripotency and de novo establishment of imprinting.
Yuhan CHEN ; Jiansen LU ; Yanwen XU ; Yaping HUANG ; Dazhuang WANG ; Peiling LIANG ; Shaofang REN ; Xuesong HU ; Yewen QIN ; Wei KE ; Ralf JAUCH ; Andrew Paul HUTCHINS ; Mei WANG ; Fuchou TANG ; Xiao-Yang ZHAO
Protein & Cell 2023;14(7):477-496
Although somatic cells can be reprogrammed to pluripotent stem cells (PSCs) with pure chemicals, authentic pluripotency of chemically induced pluripotent stem cells (CiPSCs) has never been achieved through tetraploid complementation assay. Spontaneous reprogramming of spermatogonial stem cells (SSCs) was another non-transgenic way to obtain PSCs, but this process lacks mechanistic explanation. Here, we reconstructed the trajectory of mouse SSC reprogramming and developed a five-chemical combination, boosting the reprogramming efficiency by nearly 80- to 100-folds. More importantly, chemical induced germline-derived PSCs (5C-gPSCs), but not gPSCs and chemical induced pluripotent stem cells, had authentic pluripotency, as determined by tetraploid complementation. Mechanistically, SSCs traversed through an inverted pathway of in vivo germ cell development, exhibiting the expression signatures and DNA methylation dynamics from spermatogonia to primordial germ cells and further to epiblasts. Besides, SSC-specific imprinting control regions switched from biallelic methylated states to monoallelic methylated states by imprinting demethylation and then re-methylation on one of the two alleles in 5C-gPSCs, which was apparently distinct with the imprinting reprogramming in vivo as DNA methylation simultaneously occurred on both alleles. Our work sheds light on the unique regulatory network underpinning SSC reprogramming, providing insights to understand generic mechanisms for cell-fate decision and epigenetic-related disorders in regenerative medicine.
Male
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Mice
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Animals
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Cellular Reprogramming/genetics*
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Tetraploidy
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Pluripotent Stem Cells/metabolism*
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Induced Pluripotent Stem Cells/metabolism*
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DNA Methylation
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Spermatogonia/metabolism*
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Germ Cells/metabolism*