Objective:Through gene cloning,expression and activity analysis of hBcl 2 to provide enough proteins for the development of new drugs on the basis of 3 D structue of hBcl 2 protein.Methods:Gene cloning using PCR amplification,identification with Western Blot and MALDI TOF MS.Results:hBcl 2 gene was cloned,inserted into pET28a(+) and solublely expressed in E.Coli.Its molecule weight was confirmed through MALDI TOF MS,which fits exactly with its theoretical value.After purification to reach electrophoresis homogeneity,it showed the ability of combining specifically with BH 3 domain of Bak,and then provide the basis for the further research on small chemical compounds which could specifically bind hBcl 2 protein.Conclusion:In this work hBcl 2 was successfully expressed and recovered its binding activity in a soluble form. [

Objective To clone and express the ns5a gene of hepatitis C virus(HCV) 1b strain DY.Methods By using the prokaryotic cell gene engineering,HCV ns5a gene was amplified with nested PCR from the plasmid HCV17 of HCV 1b strain DY full-length gene and inserted into the cloning pMD18-T vector.The cloned HCV ns5a gene was separated and subcloned into expression vector pET-28a and induced by IPTG in E.coli.BL21.The expressed product was identified by SDS-PAGE and Western-blot methods.Results Recombinant expression plasmid pet-28a-ns5a was constructed and expressed successfully.Conclusion HCV ns5a gene was cloned and expressed.This might be helpful for further studies on the nature and biological properties of the ns5a gene.

This article reviewed the measures taken by the hospital against a catastrophic fire hazard and exploration in its response and work organization of nursing care of the wounded.Proposed in this paper are development of such five systems as the pre-plan,exercises,personnel,quality control and incentives,which are expected to improve the nursing capacity of the hospital in emergency rescue,for sustainable development of nursing emergency rescue work.

Objective:To discuss the effect of pharmacist intervention on the use of antimicrobial agents in the clinical pathway of community-acquired pneumonia ( CAP) in our hospital to standardize the rational medication and promote the rational use of antimicro-bial agents. Methods:Totally 100 bacterial CAP patients in 2013 ( before the intervention) and 2014 ( after the intervention) in the pneumology department were studied. The antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospi-talization time, therapeutic effects and so on in the two groups were observed during the treatment. Results: There were significant differences between the two groups in antibacterial drug cost, total hospitalization cost, use intensity of antimicrobial drugs, hospitaliza-tion time and so on, whereas there was no significant difference in the curative effect. Conclusion:After the pharmacist intervention, the application of antibiotics is more rational, the antibiotics use density and per capita cost are reduced, the hospitalization day is shortened and the value of pharmacists is also improved.

The growth of M-07e human megakaryocytic leukemia cells is strictly dependent on GM-CSF. In M-07e cells, the GM-CSF receptor (GM-CSF R) is composed of two subunits: a low affinity alpha subunit and a phosphorylated beta subunit, which is constitutively linked to lyn(53/56) protein tyrosine kinase. In this study, The role of lyn kinase in regulating TGF-beta 1-induced apoptosis in M-07e cells was examined. The removal of rhGM-CSF from the culture medium resulted in down-regulation of lyn kinase activity, followed by growth inhibition and programmed cell death. Apoptosis of M-07e cells was accompanied with a massive cleavage of Bcl-2 and Bax proteins into shortened fragments with molecular mass of 22 kD and 18 kD, respectively. Using specific inhibitors, the cleavage of Bcl-2, but not Bax, was found to be processed through activated caspase-3 (CPP32), which is abundantly expressed in M-07e cells. TGF-beta 1 inhibited rhGM-CSF-stimulated cell growth and promoted apoptosis in M-07e cells with a pattern identical to that induced by rhGM-CSF depletion, which included massive cleavage of both Bcl-2 and Bax proteins and inactivation of lyn kinase activity. TGF-beta 1 did not affect the levels of lyn protein or the beta-subunit, neither did it block the interaction between these two components. Also, TGF-beta 1 treatment did not diminish the expression of the alpha subunit in M-07e cells. Our results showed that TGF-beta 1 inhibits cell proliferation and promotes apoptosis in M-07e cells by inactivating the GM-CSF R-associated lyn kinase activity. Further, This study showed that Bcl-2 cleavage by activated CPP32 is a naturally occurring event associated with apoptosis, which is under the regulation of lyn kinase activation.

Background Perfluorinated compounds (PFCs) are persistent organic pollutants. Dietary exposure to PFCs among pregnant women may lead to elevated risks of adverse events during pregnancy and postpartum depression. Objective To estimate potential risk of dietary exposure to PFCs among pregnant women in Xuhui District, Shanghai, and the relationship between dietary PFCs exposure and risks of adverse events during pregnancy and postpartum depression. Methods This study was a small cohort study which recruited women residing in Xuhui District from July 2017 to September 2018. All information was collected through questionnaires by trained investigators. Basic information and dietary information were collected at the time of inclusion, where the dietary information was obtained through the Food Frequency Questionnaire (FFQ). A follow-up visit was conducted within 42 d after delivery to collect the occurrence of adverse events during pregnancy and postpartum depression. According to the results of dietary survey, sampling and PFCs level testing were conducted on commercially available staple foods, poultry and livestock meat, aquatic products, vegetables, milk and dairy products, eggs, etc. The food sampling points covered all subdistricts in Xuhui District, including farmers' markets, supermarkets, convenience stores, and shops. Daily exposure level and hazard ratio (HR) of PFCs per capita were calculated according to the dietary survey and laboratory testing. Logistic regression was used to estimate the relationship between PFCs exposure and the risks of adverse events during pregnancy and postpartum depression. The levels of PFCs exposure were further divided into 4 groups according to interquartile range, and OR and P_trend value were calculated respectively. Results A total of 345 pregnant women were recruited in this study, whose average age was (29.61±4.92) years. Among them, 26.37% subjects reported at least one adverse event during pregnancy, and 30.14% subjects reported an Edinburgh Postnatal Depression Scale (EPDS) score ≥12. A total of 132 food samples were collected and tested, and PFCs were positive mainly in milk and dairy products, poultry meat, livestock meat, eggs, fresh water products, and sea water products. The PFCs positive rate in fresh water products was the highest, reaching 100%. The average concentration of PFCs in the positive samples was 0.018-2.10 μg·kg⁻¹. The HR of PFCs was 4.44. A higher risk of postpartum depression was found along with a higher perfluorooctane sulfonate (PFOS) exposure level (P_trend=0.02). Conclusion Dietary PFCs exposure may pose a health risk to pregnant women. The study findings suggest that decreasing the intake of freshwater products might help reduce the risk of PFCs exposure among pregnant women in Xuhui district, Shanghai.

Objective To explore the regulatory effect and mechanism of Zhongfeng Xinglou Tongfu Capsule on the brain inflammatory response of mice with cerebral hemorrhage through cholecystokinin octapeptide(CCK-8)mediated the interleukin 10(IL-10)/signal transduction and transcriptional activator 3(STAT3)signaling pathway.Methods A total of 110 C57BL/6 mice were divided into sham operation group,cerebral hemorrhage group,Zhongfeng Xinglou Tongfu Capsule low-,medium-,and high-dose groups according to the random number table method,with 22 mice in each group.The intracerebral hemorrhage model was established by collagenase method.At 2 hours after the establishment of the model,Zhongfeng Xinglou Tongfu Capsule suspension was intragastrically administered at a dose of 10 mL·kg-1.The sham operation group and the cerebral hemorrhage group were intragastrically administered with the same dose of normal saline.The neurofunctional score and left-turn rate of the mice were recorded.The dry and wet weight method was used to detect brain water content on the 72 h after surgery,and the Western Blot method was used to detect tumor necrosis factor-α(TNF-α),interleukin 1β(IL-1β),interleukin 6(IL-6),interleukin 10(IL-10),and the relative expression levels of JAK-1,STAT3,and p-STAT3 pathway proteins around the hematoma of mice at 72 h after surgery.On the 3rd and 5th day after surgery,Elisa method was used to detect the relative expression level of CCK-8,qPCR method was used to detect the expression levels of IL-10 and STAT3 mRNA.Results Compared with the cerebral hemorrhage group,the NDS scores of mice in the low-,medium-,and high-dose groups of Zhongfeng Xinglou Tongfu Capsules were significantly reduced on the 3rd and 5th days after intervention treatment(P＜0.01).And the left-turn rate in the low-dose group on the 3rd and 5th days after intervention treatment was reduced(P＜0.05),the left-turn rate in the medium-dose group was reduced on the 3rd day(P＜0.05)and the 5th day(P＜0.01).Furthermore,the left-turn rate was obviously reduced on the 3rd and 5th day in the high-dose group(P＜0.01).The brain water content of mice in the low-dose group was reduced(P＜0.05),and the brain water content of mice in the medium-and high-dose groups was significantly reduced(P＜0.01).In the low-dose group,the expressions of pro-inflammatory factors TNF-α,IL-6 and JAK-1,STAT3,and p-STAT3 pathway proteins were significantly reduced(P＜0.01),while the expression of IL-1β was reduced(P＜0.05),the expression of IL-10 increased significantly(P＜0.01).The expression of pro-inflammatory factors TNF-α,IL-1β,IL-6,as well as JAK-1,STAT3,and p-STAT3 pathway proteins in the medium-and high-dose groups was significantly reduced(P＜0.01),the expression of anti-inflammatory factor IL-10 increased significantly(P＜0.01).The content of CCK-8 in the low-dose group was increased on the 3rd day after treatment(P＜0.05),and the content increased significantly on the 5th day(P＜0.01).The content of CCK-8 in the medium-and high-dose groups obviously increased on the 3rd and the 5th day after treatment(P＜0.01).The relative expression level of IL-10 mRNA was significantly increased after treatment(P＜0.01),but the relative expression level of STAT3 mRNA was significantly decreased after treatment(P＜0.01).Conclusion Zhongfeng Xinglou Tongfu Capsule can target the content of brain-gut peptide CCK-8 and regulate the IL-10/STAT3 signaling pathway to reduce the secondary inflammatory reaction and cytotoxic brain edema after cerebral hemorrhage.