1.Effects of H2 O2 on endometrial stromal cells in the decidual mouse
Jinchun QIN ; Lingyun JIANG ; Yufu JIN ; Aiping QIN
Journal of Medical Postgraduates 2017;30(2):122-126
Objective Among the factors affecting embryo implantation, oxidative stress is one that receives much medical attention. The purpose of the study was to explore the effect of H2 O2 on the endometrial stromal cells ( ESCs) in the decidual mouse model of oxidative stress of ESCs. Methods Primary ESCs cultured by enzymatic digestion with a mesh filter were divided into a control, an experimental, and a model group. Decidualization was induced in the mice of the experimental and model groups by inter-vention with 10 nM E2 and 1μmol/L P4 for 72 h in vitro. Then, the animals of the model group were treated with different concentra-tions of H2 O2 for 4 hours. The primary ESCs were identified by immunohistochemistry, the expression of dPRP mRNA determined byRT-PCR, the proliferation of the decidual ESCs treated with H2 O2 analyzed by CCK-8, and the level of ROS detected by flow cytometry. Results Primary mouse ESCs were successfully isolated, cultured, and identified, which were shaped like spindles and polygons, radial-ly aligned under the microscope. Immunofluorescence analysis showed positive expression of vimentin and negative expression of cytokeratin. The purity of the primary mouse ESCs was ( 96. 3 ± 0. 49 )%. Theexpression of dPRP mRNA was significantly higher in the ESCs treated with E2 and P4 than in the control (0.0002±0.0000 vs 1.0010±0.0011, P<0.01). H2O2 at ≥150μmol/L suppressed the proliferation of the decidual ESCs by 6.9% (P<0.05), and in a concen-tration-dependent manner, reaching the maximum inhibition rate of 70.6% at the concentration of 300μmol/L. The level of intracellular ROS was markedly increased in the ESCs treated with H2O2 at 50μmol/L (27.77±4.20) and 100μmol/L (43.57±6.58), with statisti-cally significant difference from that in the control group (17.47±0.61) (P=0.001). Conclusion H2O2 at 100μmol/L can signifi-cantly elevate the intracellar ROS level without affecting the proliferation of decidual primary ESCs, and therefore can be used for estab-lishing the model of oxidative stress of ESCs in decidual mice.
2.Clinicopathologic analysis of small cell neuroendocrine carcinoma of the cervix
Jinchun QIN ; Li LI ; Zhijun YANG ; Desheng YAO ; Fei LI ; Jieqing ZHANG
Chinese Journal of Obstetrics and Gynecology 2011;46(5):360-363
Objective To analyse the clinico-pathologic characteristics,diagnosis,therapy and prognostic of small cell neuroendocrine carcinoma of the cervix(SCNCC).Methods The clinic-pathological features of 12 patients with SCNCC treated in Tumor Hospital of Guangxi Medical University,admitted during March 2006 to July 2010,were analyzed retrospectively.Results Of 12 patients,the mean age was 38.7 years(rang 28-57 years),6 had stages Ⅰ b1-Ⅱa,6 had stagesⅡb-Ⅳ.Among 8 patients(Ⅰ b1-Ⅲb)underwent surgery,4 of them received neoadjuvant chemotherapy,8 of them received adjuvant chemotherapy and(or)radiotherapy.All had greater than one-half stromal invasion,4 patients had positive pelvic lymph nodes metastases.The positive ratio of the chromogranin(CgA),synaptophysin,neuronspecific enolase(NSE),cytokeratins(CK),CD56 tested by immunohistochemical staining were 8/12,9/10,4/4,4/4,4/4,respectively.Median follow-up period was 3 months(1-22 months).Among 8 patients underwent surgery,2 patients developed lung metastases,1 patient developed liver and lung metastases,1 patient developed liver metastases concurrently with bone metastases,disease-free survival (DFS)were 3 months(Ⅰ b2 with positive lymph nodes),4.6 months(Ⅱ a),7 months(Ⅰ b1),17 months (Ⅰ b2);2 patient died(8.5 and 11.3 months,respectively)after surgery;4 patients are alive and show no evidence of disease.Among 4 patients untreated,1 patients received concurrent chemoradiation and are alive for 10.1 months.Two patient untreated(Ⅲb,Ⅳ)died after 0.6 and 1.3 months final diagnosis,respectively.One patient Was lost follow-up.Conclusions SCNCC is a highly malignant tumor with rare morbility,propensity for distant spread and dismal prognosis.Final diagnosis of SCNCC depends on pathomorphology and immunohistochemical analysis.Combined therapeutic modalities may in favor of survival in some patients.
3.Application of high efficiency promoters in microbial production of 4-hydroxybutyric acid.
Qin ZHOU ; Jinchun CHEN ; Guoqiang CHEN
Chinese Journal of Biotechnology 2012;28(1):48-55
4-Hydroxybutyric acid (4HB) is a psychotropic drug used for polymer synthesis such as poly (4-hydroxybutyric acid) (P4HB) and poly (3-hydroxybutyric acid-co-4-hydroxybutyric acid) (P3HB-co-4HB). 1,4-butanediol (BD) can be converted to 4-hydroxybutyric acid by alcohol dehydrogenase (DhaT) and aldehyde dehydrogenase (AldD). In this study, high efficiency promoters including T7 promoter and P(Re) promoter were cloned to increase expression of dhaT and aldD, and thus accelerate the conversion from BD to 4HB. A. hydrophila 4AK4 (pZQ01), the recombinant strain under the control of T7 promoter, produced 6.00 g/L 4HB from 10 g/L BD with the productivity increased by 43.20%. While A. hydrophila 4AK4 (pZQ04), the strain under the control of T7 promoter, produced 4.87 g/L 4HB from 10 g/L BD, and the productivity was increased by 16.23%. Thus, the gene expression was increased by T7 and P(Re) promoters, leading to an accelerated biosynthesis of 4HB.
Aeromonas hydrophila
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genetics
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metabolism
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Genetic Engineering
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Hydroxybutyrates
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metabolism
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Promoter Regions, Genetic
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genetics
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Recombination, Genetic