Objective:To investigate the effects of the psychological stress on peripheral blood lymphocytes apoptosis and Bax,Bcl-2 gene expression in rats after being given different intensity of noise stress.Methods:All 24 SD rats were divided into three groups:control,high intensity of the noise stress,middle intensity of the noise stress. The apoptosis in peripheral blood lymphocytes(PBL)of rats was demonstrated with the flow cytometer to determine the percentages of apoptosis in rat PBL,and the expression changes of related Bcl-2,Bax genes were observed by us- ing immunohistochemical method.Results:1)After high intensity of the noise stress for three weeks,the apoptosis in rat PBL increased significantly compared with the control.2)After high intensity of the noise stress for three weeks, Bcl-2 expression as down-regulation in PBL,but the B ax expression remained obviously high.Conclusion:It indi- cates that apoptosis exists in the immunocytes of rats in the course of high intensity of the psychological stress and it consists with the expression of the related regulatory genes.

Acute Disease ; Female ; Humans ; Intubation, Intratracheal ; Male ; Organophosphate Poisoning ; Pesticides ; poisoning ; Respiration, Artificial ; Tracheotomy ; methods

Objective To observe the expression level of HOXB6 gene on the differentiation and proliferation of hematopoietic stem cells(HSC) to colony forming unit-granulocyte-monocyte(CFU-GM),erythroid progenitor(CFU-E) and colony forming unite-T-lymphocyt(CFU-TL).And the proliferation procress was affected by all-transretinoic acid(ATRA).Methods 1.By the colony culture in vitro,the impact of ATRA on the CFU-GM,CFU-E,CFU-TL colony formation were surveyed;the expressions of HOXB6 gene were observed on the differentiation procress of HSC to CFU-GM,CFU-E and CFU-TL affected by ATRA on the 3rd,7th,12th.2.Real-time fluorogenic quantitative reserve transcription-polymerize chain reaction(FQ-RT-PCR) method was used to explore the possible mechanism of ATRA up-regulated to human cord blood CFU-GM,CFU-E and CFU-TL in gentic level.Results 1.HOXB6 gene had a regulatory function in the differentiation procress of hematopoiesis.During the differentiation and proliferation of HSC to colony forming CFU-GM or CFU-E,CFU-TL in vitro,the expressions of HOXB6 gene were significant positive.2.Compared with the expressions of CFU-GM and CFU-TL HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 7 and lower on day 12,respectively in each group.Compared with the expression of CFU-E HOXB6 mRNA on day 3,the quantity of HOXB6 mRNA was obviously higher on day 12 in each group.3.Compared with the HOXB6 gene of control group,the expressions of HOXB6 gene of group ATRA were up-regulated remarkable.Conclusions 1.HOXB6 gene has a regulatory function in the differentiation procress of hematopoiesis.HOXB6 gene plays an important role in the progress which can be associated with the regulating effect of HOXB6 gene on the differentiation and proliferation of CFU-GM,CFU-E and CFU-TL.2.During the differentiation and proliferation of HSC,the expression of HOXB6 gene is regular in time.3.Low concentration of ATRA(60 ?mol?L-1) can up-regulate the expression of HOXB6 gene,which confirms the theory that the normal hematopoie-tic lineage determination and maturation rely on the stable and consistent expression of HOXB6 gene.

·AIM: To investigate the effect of the dipeptide Arg-Glnon retinal neovascularization of retinopathy of prematurity(ROP) in the oxygen-induced retinopathy (OIR) animalmodel.·METHODS.- Forty-eight 7-day-old C57BL/6J mice were exposed to 750mL/L oxygen for 5 days and then to normal situation to produce the murine model of oxygen-induced retinopathy (OIR). All mice received twice daily intra- peritoneal injections of PBS or the dipeptide Arg-Gln(1.0, 3.0, 5.0g/kg per day), starting on postnatal day 12 and continuing till postnatal day 17. Experimental groups (36 mice, 12 in each group) received Arg-Gln, while the control group (12 mice) received PBS. All mice were executed at postnatal day 17. The changes of retinal vessels of mice were observed by ADPase histochemical technique and HE staining was used to count preretinal neovascular nuclei. RNA was isolated from retinas of 28 mice (7 in each group) selected at random and VEGF mRNA level of each group was measured by real-time RT-PCR.·RESULTS; Neovascularization reduced in retinas of the dipeptide Arg-Gln treated group in a dose-dependent manner. Compared with control group, experimental group had diminished non-perfusion area and neovascular tufts in retinal flatmount. The number of the endo-theliocyte nuclei of new vessels extending from retina to vitreous was significantly less in the eyes of the experimental group than in control group. Arg-Gln at 5g/kg per day reduced preretinal neovascularization by about 75% (P＜ 0.01). There was a significant reduction in VEGF mRNA at the 17th day in Arg-Gln treated group compared with control group(P＜0.01).·CONCLUSION; Arg-Gln dramatically inhibits retinal angiogenesis in OIR and this effect is associated with a reduction in retinal VEGF mRNA level. It appears to be a safe way to prevent and treat some neovascular retinal diseases including retinopathy of prematurity.

Abstract: Objective　To evaluate the effects of sunitinib on Japanese encephalitis virus (JEV) infection in vitro and vivo. Methods　The 4-week-old BALB/c mice infected with JEV by intraperitoneal injection. The infected mice were treated with sunitinib for 5 days and 10 days respectively. After that, the change of weight and survival rate were evaluated continuously. The viral load variation in mice brain were detected by qRT-PCR. Indirect immunohistochemical staining assay (IFA) was used to detect the number and distribution of CD4+/CD8+T cells in mouse brain. IFA was also used to observe the expression of virus E protein in the brain of mice. Vero cells were infected with JEV in vitro and given a certain concentration of sunitinib to observe the cell survival status. The expression of virus E protein in cells was detected by IFA. Results　Continuous administration of sunitinib significantly improved the survival rate of infected mice. Survival rate and body weight changes showed that the 5-day's administration strategy was significantly better than the 10-day's administration strategy. The treatment of sunitinib decreased the infiltration of CD4+/CD8+T cells in the brain and reduced the changes of vascular sleeve. However, the variation of viral load and E protein expression in brain were not obvious. The cytopathic effect (CPE) of infected Vero cells were slightly relieved after giving sunitinib, and the expression of E protein was also slightly changed. Conclusion　Sunitinib can significantly reduce the mortality of infected mice, and the 5-day's administration strategy is significantly better than the 10-day's administration strategy. Sunitinib decrease T lymphocyte infiltration in brain of mice, relieve the encephalitis symptoms ,and prolonged the life of mice.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Electrophoresis, Capillary ; Esters ; analysis ; Phthalic Acids ; analysis ; Sensitivity and Specificity

Adenoma, Villous ; metabolism ; pathology ; surgery ; Aged ; Diagnosis, Differential ; Female ; Humans ; Keratin-20 ; metabolism ; Keratin-7 ; metabolism ; Mullerian Ducts ; pathology ; Papilloma ; pathology ; Vaginal Neoplasms ; metabolism ; pathology ; surgery

Objective To evaluate the role of early cleavage embryo in combination with embryo growth rate and morphology scoring in embryo selection in in vitro fertilization/intracytoplasmic sperm injection (IVF/ICSI) cycles. Methods Six hundred and ten IVF/ICSI cycles were randomly assigned to group A(269 cycles) and group B(341 cycles). In group A, transferred embryos were chosen according to embryo growth rate and morphology scoring by 72 h(D3) after fertilization, while early cleavage embryo was added to the selecting system in group B. The pregnancy rate and implantation rate were compared between two groups, and the clinic outcomes were compared between transfers with early cleavage embryos and without early cleavage embryos in group B. Results The pregnancy rate and implantation rate in group B were significantly higher than those in group A (P < 0.05). Transfers with early cleavage embryos also achieved much higher pregnancy rate and implantation rate in group B (P < 0.01). Conclusion Compared with embryo growth rate and morphology scoring, early cleavage embryo in combination with embryo growth rate and morphology scoring can improve the clinical outcomes in IVF/ICSI cycles.

Background Diabetic retinopathy (DR) is one of the leading causes that result in adult irreversible blindness in many countries.Recent researches suggest that neurodegeneration is an important component of DR.To realize the disease process of retinal neutron is very important for prevention and treatment on DR.Objective This study was to investigate the change of retinal nerve fiber layer thickness in patients with type 2 diabetes mellitus.Methods Ninety-six eyes of 48 patients with type 2 diabetes mellitus were enrolled in Peking Union Medical College Hospital.The patients were assigned into non-diabetic retinopathy (NDR) group,background diabetic retinopathy(BDR) group,proliferative diabetic retinopathy (PDR) group and panretinal photocoagulation (PRP) group based on the fundus finding and fundus fluorescein angiography(FFA),and 24 normal subjects with matched age were included as control group.RNFL thickness was measured by GDxVCC system,including temporal,superior,nasal,inferior,total,(TSNIT) average,superior average,inferior average,TSNIT standard deviation and nerve fiber indication.The datas of the RNFL thickness were analyzed and comparison among different groups by one-way analysis of variance and Student Newman Keuls test.Results The TSNIT averages of the NDR group,BDR group,PDR group and PRP group were(56.54±5.28),(56.92±6.49),(53.04±6.14) and(53.17±9.30) μm,respectively,while that of the control group was (59.04±4.37) μm.The TSNIT average,superior average,inferior average,TSNIT standard deviation of the PDR group and PRP group compared with control group were significantly decreased,and the nerve fiber indication of the PDR group and PRP group was significantly increased (P =0.002,0.000,0.002,0.000,0.001 ;P =0.002,0.000,0.001,0.000,0.000).Compared with the control group,the TSNIT average,superior average,inferior average,TSNIT standard deviation were insignificantly decreased,and the nerve fiber indication was insignificantly increased in the NDR group and BDR group (P =0.187,0.235,0.333,0.106,0.202 ;P=0.262,0.063,0.072,0.098,0.062).Conclusions The decline of the RNFL thickness appears prior to DR findings.The RNFL thinning of PDR and PRP patients suggests the degeneration of neurons and atrophy of axonal.The neurodegeneration is an important component of DR.

OBJECTIVETo establish and evaluate a BA-ELISA method for the quantitative detection of cystic fibrosis transmembrane conductance regulator (CFTR) protein.

METHODSWe deliberately selected three tables of CFTR and made the synthetic peptide be expressed in E. coli, then used the antigen to immunize rabbits to obtain the anti-CFTR polyclonal serum. After that, 96 well plates were coated with the purified antibody against CFTR. The antigen CFTR which was extracted from human sperm was detected by anti-CFTR antibody labeled with biotin, horseradish peroxidase conjugated avidin, and the substrate. The concentrations of two kinds of antibodies and the experiment parameters were optimized. Thereby, the double antibody sandwich BA-ELISA method for the quantitative detection of CFTR protein was established. Furthermore, the reproducibility, specificity and so on were evaluated by clinical specimens of sperm.

RESULTSThe optimal concentration of coated anti-CFTR IgG was 4 µg/ml, while the biotin labeled anti-CFTR IgG was 10 µg/ml; the optimal blocking buffer was 1% BSA-PBST, the optimal time of the reaction between antigen and antibody was 60 min, the optimal chromogenic time was 15 min, the intra-assay and inter-assay coefficient were 2.16%-9.23% and 2.29%-11.71% respectively; The lowest detectable limit was 0.15 ng/ml; the standard curve had a good linear correlation of R2 = 0.962.

CONCLUSIONThe BA-ELISA method for the quantitative detection of CTFR protein is successfully established, and it is demonstrated that the method has strong specificity, high sensitivity and good reproducibility. It provides the basis and evidence of the further application of the method.

Animals ; Antibodies ; Cystic Fibrosis Transmembrane Conductance Regulator ; analysis ; Enzyme-Linked Immunosorbent Assay ; methods ; Escherichia coli ; Humans ; Peptides ; Rabbits ; Reproducibility of Results ; Sensitivity and Specificity