Objective To optimize multiplicity of infection ( MOI) and antibiotics ( blasticidin) concentration selecting BSD gene in construction of monoclonal stable cell line by lentivirus vector-mediated RNA interence silenced gene SGMS2.Methods The INS-1 cells were transfected by fluorescence labeled negative control SGMS2-siRNA lentivirus at MOI of 0, 10, 30, 60 and 120 TU number/cell.The cells were photographed under fluorescent microscopy after 72 h cultivation, then fluorescence ratio and apoptosis rate were calculated to determine optimal MOI.The INS-1 cells were treated by blasticidin with different concentrations of 0, 1, 2, and 3 μg/mL, and the apoptosis rate was observed to acquire optimal concentration of antibiotics.The INS-1 cells were transfected by negative control SGMS2-siRNA lentivirus and SGMS2-siRNA lentivirus (virus titer:1 ×108TU/mL) at optimal MOI and positive-transfected cells were selected by blasticidin at optimal concentration, then mixed cell lines were acquired.The monoclonal cell line was constructed at fluorescence ratio of 90%.Results The optimal MOI was 60 with 100% fluorescence ratio, less than 0.5% apoptosis rate and keep original cellular morphology.The optimal concentration of blasticidin was 2 μg/mL with cell adherence disappear and all cells apoptosis.The Ct value of INS-1-SEMS2 cells detected at the second time was 28.21, which was greater than 27.58 at the first time.The interfering efficiency of siRNA was 77.78% which indicated a successful expression of siRNA and construction of monoclonal stable cell line ( INS-1-SEMS2 ).Conclusion The monoclonal stable cell line was successfully constructed by lentivirus vector-mediated RNA interence silenced gene SGMS2.

Tissue factor pathway inhibitor-2(TFPI-2) is a matrix-associated Kunitz-type serine proteinase inhibitor,and is considered to play an important role in some pathophysiological processes,including artherosclerosis,tumor metastasis and angiogenesis.Extracellular signals can regulate TFPI-2 gene expression through modulating promoter or signaling pathway or other factors.The mechanism,more over,has become one of the focus in recent years.

Objective To investigate the contamination of carbapenem-resistant Acinetobacter baumannii (CRAB) from object surface of key departments in a hospital,and identify whether these CRAB were homologous. Methods Environmental hygienic monitoring in intensive care unit (ICU),emergency intensive care unit(EICU), hemodialysis room and operating room was conducted.Acinetobacter baumannii (A.baumannii)isolated from ICU and EICU environmental specimens were amplified and typed by enterobacterial repetitive intergenic consensus-poly-merase chain reaction (ERIC-PCR).Results Except hand hygiene of health care workers in EICU was qualified, bacterial count of object surface of ICU and EICU were all unqualified;detection results of specimens from hemodi-alysis room and operating room were all qualified.A total of 53 specimens were taken from object surface of ICU and EICU,7 (13.21 %)A.baumannii isolates were isolated,and all were CRAB isolates,6 of which were of the same genotype and were identical with A.baumannii from patients’sputum.Conclusion CRAB isolated from object surface in key departments is homologous,cleaning and disinfection of environmental object surface should be inten-sified to reduce the occurrence of healthcare-associated infection.

Adult ; Heat Stroke ; diagnosis ; Humans ; Male ; Middle Aged ; Occupational Diseases ; diagnosis

Adult ; Benzene ; poisoning ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; chemically induced ; diagnosis ; Male ; Occupational Diseases ; diagnosis ; Physical Examination

Proanthocyanidins is an effective antioxidants and free radical scavenger derived from kinds of plants in nature. It has some advantages that high absorptivity and bioactivity, low toxicity, which contain broad pharmacological functions such as dropping high blood pressure, reducing blood fat, anti-inflammatory, anti-cancer, anti - aging, anti - fatigue and preventing cardiovascular and cerebrovascular diseases. In this paper, we summarized its physicochemical properties, pharmacologic action and the ophthalmological studies which mainly in keratopathy, cataract, glaucoma, uveitis, retinopathy, traumatic optic neuropathy and asthenopia. With the deeper research and more unequivocal in the pharmacological mechanism of proanthocyanidins, it should make the most of its values in medicine. And some more economical and effective drugs are expected to be provided for the clinical ophthalmology in the near future.

The authors analyzed the negative impacts on hospital public welfare performance,caused by such organizational elements imbalances of public hospitals,as objective structure,allocation of rights and responsibilities,and budget resources.In view of these setbacks,they proposed countermeasures for timely solution of such imbalances and maxim optimal combination,for the purpose of more effective reform of such hospitals.

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.

Type 1 diabetes mellitus and autoimmune thyroid disorders are the most common combination of autoimmune polyendocrine syndrome type Ⅲ(APS Ⅲ). However, APS Ⅲ combined with myasthenia gravis is rare. We described a male patient with myasthenia gravis, type 1 diabetes mellitus, and Hashimoto thyroiditis, who was diagnosed as APS Ⅲ. The human leukocyte antigen (HLA)type was analyzed in this patient. We subsequently reviewed 11 cases of APS Ⅲ combined with myasthenia gravis. This review revealed that HLA-DR9/DQ9 might be a specific HLA subtype associated with APS Ⅲ and complicated with myasthenia gravis .

Adolescent ; Ammonia ; poisoning ; Female ; Humans