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 study the difference and clinical significance of serum Angiopoietin-2 (Ang-2) level in gastric cancer, precancerous disease and the changes in post operation follow-up patients.MethodsSerum Ang-2 and CEA levels were detected with ELISA method in 158 gastric cancer patients, 31 chronic astrophic gastritis patients, 38 gastric adenomatous polyp patients, 28 chronic gastritis patients with benign gastric ulcer and 30 healthy controls.The changes of serum Ang2 levels in 131 patients with radical gastrectomy were measured in 2-year post-operational followed-up and the prognosis was also evaluated.The correlation between serum Ang-2 level and the pathologic character of gastric cancer was analyzed.ResultsThe serum Ang-2 and CEA levels of gastric cancer group were (331.8± 64.3) μg/L and (42.6 ± 37.3) μg/L, respectively, which were significantly higher than those of healthy control group [(187.4±32.7) μg/L and (4.2±3.1) μg/L]and benign gastric ulcer group [(197.3±35.4) μg/L and (4.5±3.2) μg/L](all P ＜0.01).The serum Ang-2 level in chronic astrophic gastritis group was also markedly increased (P＜0.05), however there was no significant change in CEA level.There was no obvious change of Ang-2 and CEA level in gastric adenomatous polyp group (P＞0.05).Pre-operation serum Ang-2 levels of the relapse group and the metastasis group were significantly higher than that of no relapse group (P＜0.05).Post-operation serum Ang-2 level of no relapse group decreased to normal range in 1 month and without obvious increase in 2 years follow-up.The serum Ang-2 level of the relapse group also decreased to normal range in 1 month post-operation, while which was significantly higher than that of no relapse group and normal control group from 6 months to 2 years after operation (P＜0.01).Serum Ang-2 level of the metastasis group was markedly higher than that of no relapse group and normal control group during 2 years after operation (P＜0.01).The 2-year survival rate of no relapse group was significant higher than that of relapse group and metastasis group.The change of serum Ang-2 level was correlated with in the gastric cancer differentiation degree, TNM pathologic classification, lymphatic metastasis, distant metastasis, the depth of invasion and the tumor size (P＜0.01), however not correlated with histopathology type and the location of tumor.ConclusionSerum Ang-2 level may be a new and valuable gastric cancer marker, which will help to monitor tumor relapse, migration and prognosis assessment.

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.

Objective To study the methylation status of the promoter region of several tumor suppressor genes in p53-Bax mitochondrial apoptosis pathway and its role in cholangiocarcinoma. Methods The hypermethylation of the promoter region of tumor suppressors death-associated protein kinase (DAPK), p14, and target of methylation-associoted silencing-1 (TMS1/ASC) were detected by methylation-specific PCR. P53 gene status (exon 5-8 ) were examined by automated sequencing. The relationship between gene mutations and the biological behaviors of cholangiocarcinoma was analyzed. Results Methylation existed in at least one promoter region of tumor suppressor gene in the tumor tissues of 24 patients (66. 7% ). The frequencies of tumor suppressor gene methylation in cholangiocarcinoma were: p14 24%, DAPK 30. 6%, and TMS1/ASC 36. 1%. The frequencies of tumor suppressor gene methylation in the adjacent tissues were: TMS1/ASC 8.3% and DAPK 5.6%. DNA sequencing showed p53 gene mutation was found in 22 of 36 patients (61.1% ), and p53 gene mutation combined with the methylation of tumor suppressor was found in 14 (38.9%) patients, which was significantly correlated with pathologic biology, invasion, and differentiation ( P ＜ 0.05 ). The 1-year, 2-year, and 3-year survival rates were significantly higher in tumor-suppressing genes methylation group ( n = 4) (70%, 43 %, and 28%, respectively)than those in p53 gene mutation group (n = 14) (28%, 5%, and 0%, respectively) (χ2 =9. 060, P =0.03).Conclusions Promoter hypermethylation of p53-Bax mitochondrial apoptosis pathway is a common epigenetic event in cholangiocarcinoma. Although the methylations of TMS1/ASC and DAPK genes in the adjacent tissues are relatively low, they may be informative for the early detection of cholangiocarcinoma. P53 gene mutation combined with the methylation of tumor suppressor may be related with the pathologic biology of cholangiocarcinoma, making the latter trend to be with high malignancy and poor prognosis.