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.

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

·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.

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

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

Objective:To express mycobacterium tuberculosis protein MPB 64 in baculovirus insect cell expression system ,and identify its immunogenicity.Methods:The target gene MPB64 was connected to pFastBac vector ,then the pFastBac-MPB64 plasmid which was harvested would transformed to DH10Bac competent,and the target gene was transposition into Bacmid by Tn7 transposase fragment,therefore Bacmid-MPB64 Shuttle vector was obtained.The shuttle vector was packaged by liposomes and transfected Sf 9 cells to harvest P1-generation virus ,then high titers of P4 generation virus was harvested by repeat transfected Sf 9 cells three times.The target protein MPB64 was purified from the supernatant by Q Sepharose FF and Ni affine chromatography ,which were used to immunize BALB/c mice.Antibody changes in serum would be detected ,and the proliferation of immunized mice spleen cells would be detected by MTT,detected the IFN-γsecretion by MPB64 stimulated spleen cells by ELISA method.Results: MPB64 successfully expressed in insect cells.The purity of target protein was over 90% and yield up to 35 mg/L after purification.Purified protein can effectively stimulate BALB/c mice to produce antibodies , increase the content of IFN-γmedium in mice spleen cells ,and significantly promoting proliferation in spleen cells between 0.2-100 μg/ml.Conclusion: MPB64 which has immunogenicity was successfully expressed in baculovirus insect cell expression system ,that open a new avenue for tuberculosis vaccine production.

Prolactin is a polypeptide hormone which mainly acts on the reproductive system and plays an important role in penile erection and ejaculation. Prolactin receptors have a variety of short forms apart from the classic long form, which are widely expressed in male reproductive glands. High levels of prolactin can induce erectile dysfunction and results in secondary male infertility, which are mainly associated with the inhibition of dopaminergic activity, reduction of the testosterone level, and contraction of the cavernous smooth muscle. Moreover, low levels of prolactin can result in ejaculatory dysfunction. This article updates the views on the expressions of prolactin receptors in the male reproductive system, the effects of prolactin on penile erection and ejaculation, and its action mechanisms.
Ejaculation ; physiology ; Erectile Dysfunction ; Humans ; Infertility, Male ; Male ; Muscle, Smooth ; physiopathology ; Penile Erection ; physiology ; Prolactin ; physiology ; Receptors, Prolactin ; physiology ; Reproduction