G protein-coupled receptors(GPCRs) are the largest and most diverse group of trans-membrane proteins involved in signal transduction. They have been playing key roles in drug discover-y. Increasing orphan GPCRs (oGPCRs) whose endogenous ligands and functions are still to be identified have been discovered in recent years. It is obvious that oGPCRs might be the most important targets for innovating drugs.

In order to decrease the mortality of pre-hospital sudden death events(SDEs),their features especially those of cardiac sudden death were analyzed. 1030 pre-hospital emergent cases from 1973～1999 were analysed systematically. The total SDEs occurred in 106 cases, among the 68 cases of cardiovascular diseases, 89.7% had coronary heart diseases(CHD), AMI or OMI with re-infarction. There were two peak age stages of SDEs: the first was 46-55 years old stage(non-retaired age stage,22.6%), and the second stage was 66-75 years old(27.3%). The pre-hospital SDEs are the most emergent cases. Great attention must be paid to those with the history of cardiovascular diseases, especially those with the history of coronary cardiac events

Objective To investigate the role of phosphatidylinositol 3 kinase (PI3K)/protein kinase B (Akt) signaling pathway in Dexamethasone (DEX) inhibiting podocytes apoptosis which was induced by Puromycin (PAN).Methods Mouse glomerular podocytes were cultured in vitro,and were divided into control group,dimethyl sulfoxide (DMSO) group,PAN group,DEX group,and LY294002 (inhibitor of PI3 K) group.The mRNA expression of CD2-associated protein (CD2AP) was measured by using real time fluorescent quantitative polymerase chain reaction,and intracellular distribution was detected by using indirect immunofluorescence staining.Co localization of CD2AP and p85 was detected by using confocal fluorescence microscopy.The expressions of Akt,phosphorylated (p)-Akt,glycogen synthase kinase-3β (GSK3 β) and phosphorylated (p)-GSK3β were evaluated by using Western blot.Results The expressions of CD2AP mRNA in PAN group at each time point (8 h,24 h,48 h) (1.11 ± 0.16,0.78 ±0.09,0.56 ± 0.43) were significantly lower than those in the control group (1.90 ± 0.26,2.09 ± 0.12,2.28 ±0.95),and the differences were statistically significant (all P ＜ 0.05);CD2AP distributed in foot process with uniform filament and discontinuous coarse particle around perinuclear;CD2AP and p85 distributed in cell membrane and cytoplasm evenly in control group,but accumulated in nuclei in the PAN group.The expressions of CD2AP mRNA in DEX group at each time point (8 h,24 h,48 h) (1.53 ± 0.14,2.15 ± 0.27,2.13 ± 0.15) were significantly higher than those in the PAN group,and the differences were also statistically significant (all P ＜ 0.05);the distribution density and range of CD2AP were greater than those in the PAN group,and the accumulation with p85 in nuclei decreased obviously.The expressions of p-Akt and p-GSK3β were inhibited by PAN in a dose-dependent manner (P ＜0.05).The expressions of p-Akt and p-GSK3 β were lowest after PAN stimulated at 15 min and 30 min respectively.However,the expressions of p-Akt and p-GSK3 β increased depending on the concentration of DEX (P ＜ 0.05).In addition,the expressions of p-Akt and p-GSK3 β could be blocked by LY294002 (P ＜ 0.01).Conclusion DEX can protect podocytes and inhibit podocytes apoptosis through stabilizing the expression and distribution of CD2AP.The stale expression of PI3K/Akt signaling pathway is the key factor in DEX protecting podocytes.

Objective To investigate the application value of combined detection of antikeratin (AKA) antibody,anti-cyclic citrullinated peptide(anti-CCP) antibody,anti-RA33 antibody,RF and ESR in the diagnosis of rheumatoid arthritis(RA).Methods One hundred cases of RA and 60 cases of suspected RA in our hospital from June 2014 to May 2015 were collected.One hundred and twenty cases of non-RA other autoimmune diseases served as the control group.The indirect immunofluorescence assay (IFA) was used to detect anti-KA antibody,anti-CCP antibody and anti-RA33 antibody were detected by using ELISA.The rate scatter turbid assay was used to detect RF.The Westergren method was used to detect ESR.The diagnostic performance of each diagnostic indicator was evaluated.Results The detection rates of anti-KA antibody,anti-CCP antibody,anti-RA33 antibody,RF and ESR in the RA group were in turn 64%,75%,44%,84% and 51% respectively,the detection rate(sensitivity) of 5-indicator parallel detection were 97%, the specificity of 5-indicator series detection was 89.2%;in the suspected RA group,the detection rates of 5-indicator were in turn 16.7%, 31.7%,13.3%,20%,15% respectively,which in the control group were in turn 0.8%,2.5%,1.7%,0.8% and 5.8% respectively.The detection rates of 5-indicator in the RA group was significantly higher than that in the suspected RA group,showing extremely significant difference (P<0.001),the detection rate of 5-indicator had extremely significant difference between the suspected RA group and control group (P<0.001),the sensitivity of five-indicator parallel detection was 97%,which was significantly higher than that of single indicator detection (P<0.05),the detection specificity of 5-indicator series detection was up to 100%,which was also significantly higher than that of single indicator detection (P<0.05),the missed diagnosis rate of parallel detection was minimal,while the negative predictive value was highest,the misdiagnosis rate of series detection was lowest,the positive predictive value was highest,the Youden index of parallel detection was largest.Conclusion The single detection of anti-CCP antibody and RF have good sensitivity and specificity, but 5-indicator combined detection has higher sensitivity and specificity,which can better reduce the missed diagnosis rate and misdiagnosis rate,can greatly improve the efficiency of diagnosis,and has an important clinical significance for the early diagnosis of RA.

To explore the model of the standardized training for health management talents by referring to the standardized training for resident-doctors.This article analyzed the necessities of the model of the standardized training for health management talents from two aspects.After that,it elaborated on the significance of standardized training.At last,it proposed some ideas about constructing the standardized training for health management talents.

BACKGROUND:The rabbits were widely used as experimental animal models in the research on etiology and pathological mechanism of steroid-induced osteonecrosis of the femoral head. It is stil a valuable and realistic research topic to improve and to innovate the modeling technology nowadays. OBJECTIVE:To improve the modeling technology on osteonecrosis of the femoral head in rabbits induced by glucocorticoid combined with lipopolysaccharide, with the focus on its reduced mortality and the guaranteed successful rate of modeling. METHODS:A total of 28 New Zealand white rabbits were randomly divided into the control group (n=10) and improvement group (n=18). Models of steroid-induced osteonecrosis of the femoral head were established according to different methods. In the improvement group, rabbits were injected with sodium penicilin (5.0 mg/kg) and amikacin sulfate (1.63×104 U/kg) in the left gluteus muscle. Twenty-four hours later, al rabbits were injected with prednisolone acetate (20 mg/kg) in the right gluteus muscle. Forty-eighthours later, 5.0 μg/kg of lipopolysaccharide was intravenously injectedvia the ear. From then on, two injections of prednisolone acetate (20 mg/kg) were respectively performed alternately in the left and right gluteal muscle at an interval of each 24 hours. Sodium penicilin (5.0 mg/kg) and amikacin sulfate (1.63×104 U/kg) were intraperitonealy injected for 2 consecutiveweeks. In the control group, 10 μg/kg lipopolysaccharide was injectedvia the ear vein of rabbit. From then on, prednisolone acetate (20 mg/kg) was intramuscularly injected at an interval of each 24 hours, totaly three times. Benzylpenicilin sodium 20×104 U/rabbit was intramuscularly injected once a week. RESULTS AND CONCLUSION:Rabbit models of steroid-induced osteonecrosis of the femoral head were successfuly established in both groups. Compared with the control group, the mortality was significantly reduced after model establishment in the improvement group, and the bone lacuna and osteonecrosis of the femoral head were apparent. These findings indicated that the improved technology of model establishment of steroid-induced osteonecrosis of the femoral head could be used to aleviate the damage degree on the gluteal muscles, to guarantee the successful rate of modeling, and to noticeably reduce the mortality of rabbits.

Objective:To evaluate the influence of prophylactic intra-coronary injection nitroglycerin with diltiazem in patients of acute ST segment elevation myocardial infarction(STEMI) Methods:This study was conducted in 212 patients with acute STEMI who underwent primary percutaneous coronary inter- vention(PCI)in our center from January 2001 to December 2006.The patients were randomly assigned to receive intra-coronary nitroglycerin with dihiazem(n = 87)or none of them(125)before intervention.The clinical data,coronary angiography data after intervention,ST-segment resolution,area under the curve of CK-MB,ejection fraction(EF)and left ventricular end-diastolic di- mension several days alter operation and the major adverse cardiac events(MACE)were analyzed systemically. Results:Ten(11.5%)patients in study group and 28(22.4%)in control group had MACE(P = 0.042).The difference of re-hospitalization was also significant(19.5% vs 37.6%,P = 0.008).No/slow reflow occurred less in study group than in con- trol group,but the difference was not significant.Corrected TIMI frame count was lower(34.4?11.3 vs 38.9?17.2,P = 0.034) and myocardial blush grade was higher(2.65?0.6 vs 2.41?0.7,P =0.018)in study group.ST-segment resolution,area under the curve of CK-MB in 5 days after operation,EF and left ventricular end-diastolic dimension in study group were more significant than those in control group(P

Objective To compare the characterization and myocardial differentiation capacity of amniotic fluid-derived mesenchymal stromal cells (AF MSCs) and umbilical cord Wharton's Jelly-derived mesenchymal stromal cells (WJ MSCs). Methods The human AF MSCs were cultured from amniotic fluid samples obtained by amniocentesis. The umbilical cord WJ MSCs were obtained from Wharton's Jelly of umbilical cords of infants delivered full-term by normal labor. The morphology, growth curves, and analyses by flow cytometry of cell surface markers were compared between the two types of cells. Myocardial genes (GATA-4, c-TnT, α-actin, and Cx43) were detected by real-time PCR and the corresponding protein expressions were detected by Western blot analysis after myocardial induced in AF MSCs and WJ MSCs. Results Our findings revealed AF MSCs and WJ MSCs shared similar morphological characteristics of the fibroblastoid shape. The AF MSCs were easily obtained than the WJ MSCs and had a shorter time to reach adherence of 2.7 ± 1.6 days to WJ MSCs of 6.5 ± 1.8 days. The growth curves by MTT cytotoxic assay showed the AF MSCs had a similar proliferative capacity at passage 5 and passage 10. However, the proliferative capacities of WJ MSCs were decreased at 5 passage relative to 10 passage. Both AF stem cells and WJ stem cells had the characteristics of mesenchymal stromal cells with some characteristics of embryonic stem cells. They express CD29 and CD105, but not CD34. They were positive for Class I major histocompatibility (MHC I) antigens (HLA-ABC), and were negative, or mildly positive, for MHC Class II (HLA-DR) antigen. Oct-4 was positive in all the two cells types. Both AF MSCs and WJ MSCs could differentiate along myocardium. The differentiation capacities were detected by the expression of GATA-4, c-TnT, α-actin, Cx43 after myocardial induction. Conclusions Both AF MSCs and WJ MSCs have the potential clinical application for myogenesis in cardiac regenerative therapy.

AIM:To investigate the role of B cells in CD45RB antibody-induced transplantation immune toler-ance.METHODS:Single cell suspension was made from the spleen of BALB/c nude mice disposed by CD45RB antibod-y, then mixed cultured with T cells of BALB/c mice and spleen cells of C57BL/6 mice.The Th1, Th2, Treg and Tm cells were monitored by flow cytometry during the culture process .The skin graft model was set up with B 6.μMT-/-mice as re-ceptors and BALB/c mice as donors.CD45RB antibody was intraperitoneally injected into the receptors after transplantation and then CD3+CD45RBhi cells were detected by flow cytometry .In another mixed lymphocyte culture , CD45RB antibody was added, and then B cells were isolated and injected into B6.μMT-/-mice through the tail vein.The heart transplanta-tion model was established with B 6.μMT-/-mice as receptors and BALB/c mice as donors, and then the survival and the migration of B cells to the thymus were observed .RESULTS:When T lymphocytes were co-cultured with B lymphocytes treated with anti-CD45RB monoclonal antibody (mAb) in vivo, the percentages of Th2 and Treg cells were up-regulated and Th1 cells were down-regulated, but Tm cells were not altered as compared with the control .In vivo without B lympho-cytes, anti-CD45RB mAb also down-regulated the expression of CD45RB in T lymphocytes.The reduction was faster and the percentage of CD3 +CD45RBhi T cells was not altered as compared with the control .The B lymphocytes treated with an-ti-CD45RB mAb in vitro prolonged the lifetime of receptor in heart transplantation model but failed to induce complete toler -ance.After recieving B cells treated with anti-CD45RB mAb and allogeneic heart transplantation , B cells migrated to the thymus in B6.μMT-/-mice.CONCLUSION:B lymphocytes play a definite role in the transplantation immune tolerance induced by anti-CD45RB mAb through their affection on T-cell subgroups and also in the central tolerance .However, the induction of immune tolerance can not only rely on B cells .

BACKGROUNDTo study the expression of catalylic subunit of DNA-dependent protein kinase (DNA-PKcs) in non-small cell lung cancer (NSCLC) and its relationship with apoptosis.

METHODSNSCLC tissues from 113 untreated patients were analyzed immnohistochemically with antibodies to DNA-PKcs, p53 and bcl-2.

RESULTSThere were expressions of DNA-PKcs, p53 and bcl-2 in NSCLC at different levels. The positive rate of DNA-PKcs, p53 and bcl-2 was 89.38%(101/113), 61.95%(70/113) and 59.29%(67/113) respectively. The expression of DNA-PKcs was significantly related with the histological types. Its expression in squamous carcinoma was significantly lower than that in adenocarcinoma and bronchioloalveolar carcinoma. The expression of DNA-PKcs increased with the increasing differentiated degree of NSCLC ( P < 0.05), but had no relationship with lymph node metastasis. There was no significant relation between the expression of p53 and the pathological type of NSCLC. A significant difference of bcl-2 expression existed in the histological types of lung cancer ( P < 0.01). Its expression in squamous carcinoma was significantly higher than that in adenocarcinoma, bronchioloalveolar carcinoma and adenosquamous carcinoma, but had no relationship with the differentiated degree of lung cancer and lymph node metastasis. The expressions were significantly related between DNA-PKcs and p53 ( P < 0.01), p53 and bcl-2 ( P < 0.05).

CONCLUSIONSThe expression of DNA-PKcs is fairly high in NSCLC. The high expression of DNA-PKcs and overexpressions of mutated p53 and bcl-2 may be important causes of radioresistance in NSCLC.