Objective:To investigate the alteration of the expression of PTENin rats with aging.Methods:Wistar rats were divided into 3 groups:3-month-old group,12-month-old group and 24-month-old group(n=6).Immunohistochemistry of CD34 was employed to detecte angiogenesis in the kidneys with aging.The protein expression of PTEN was examined by Western Blot.Immunohistochemistry was also employed to detect the expression of PTEN in the kidneys with aging.Results:(1)Compared with 3-month-old rats and 12-month-old rats,the level of PTEN protein expression in kidneys of 24-month-old rats was higher than that of 3-month-old rats and 12month-old rats,as examined by Western Blot(P

Objective To investigate the possibility of measuring ABO blood group antibody levels using renal cortical tubular epithelial cells (RCTECs) of cynomolgus monkey.Method The primary RCTECs were isolated from cynomolgus monkey kidneys and identified by Western blotting and flow cytometry (FACS).FACS was applied to detect the expression of ABO histo-blood group antigens on RCTECs,the binding of blood group antibodies in human serum to RCTECs,and to compare the difference of measuring ABO blood group antibody levels between using human erythrocytes and RCTECs as target cells.Result The majority of cells derived from the kidney cortex were renal tubular epithelial cells.39.90％-73.80％ of RCTECs were found to express the ABO histoblood group antigens with the capability to bind the human blood group antibodies specifically.The expression level of ABO histo-blood group antigen on RCTECs was not influenced by long-term cryopreservation,and the percentage of B antigen positive cells was 39.90％ before frozen storage and 39.65％ after a 10-month cryopreservation respectively.The normal serum and allergic serum presensitized to synthetic A antigen-keyhole limpet hemocyanin (KLH-A) of cynomolgus monkey were used to compare the abilities of RCTECs and human erythrocytes to detect antibody titers.No agglutination was observed in RCTECs test group.The mean fluorescence intensity (MFI) values of antibody were highest when the dilution fold was lowest (1 ∶ 16) and gradually decreased with increased serum dilution in both serum groups.MFI fell towards baseline value at 1 ∶ 128 dilution in normal serum group while at 1∶8 192 dilution in allergic serum group.Between 1 ∶ 16 and 1∶8 192 dilutions,MFIs of normal serum group were all lower than those of allergic serum.In human red cells test group,obvious agglutination appeared at high concentrations of antibodies,and MFIs reached the peak at 1∶64 dilution in normal serum group and at 1∶32 dilution in allergic serum group,but fluctuated irregularly thereafter.Between 1∶64 and 1∶512 dilutions,MFIs of normal serum group were all higher than those of allergic serum.Data above showed that MFI values in RCTECs group could reflect the levels of blood group antibodies more exactly.Conclusion The RCTECs of cynomolgus monkey express ABO tissue-blood group antigens and would be more suitable to be used to measure ABO blood group antibody levels by FACS.

Clinical practice, the necessary way which must be passed by medical students, is a key period of medical education and a vital part to realize the goal of higher medical education. Our hospital has taken a series of effective measures to strengthen clinical practice teaching for many years, and we are ex-ploring the concentrated training model to medical students all the time. Every year before clinical practice, the Inspection Section develops theoretical and basic clinical skills training courses, selects experienced teachers to teach, check and evaluate the training result according to students' performance in medical and teaching activities in the ward, the clinical practice syllabus and Medical Qualification Examination aims to improve medical students' theoretical knowledge, skills, medical ethics, humanity, and clinical thinking, etc. Effect evaluation shows that our training model has played a positive role in improving overall qualities of medical students and got good feedback from them.

Objective:To investigate the effects of acupoint massage on cognitive function and sleep quality in elderly patients with hypertension. Methods:A total of 68 elderly patients with hypertension were enrolled, and their ages were all over 60 years old. They were divided into a control group and a treatment group by simple random method, with 34 cases in each group. Patients in the control group were treated by routine psychological health guidance and sleep knowledge education, while patients in the treatment group were treated by acupoint massage on the base of the interventions given to the control group. Both of their treatment courses were 3 months. Pittsburgh sleep quality index (PSQI) scores and mini-mental state examination (MMSE) scores before and after treatment were recorded and compared. Results:Two groups’ PSQI scores before treatment had no significant difference (P>0.05). After 3 months of treatment, their PSQI scores were statistically different (P<0.05). There was also no between-group statistical difference in MMSE score before treatment (P>0.05), but there was a significant difference after treatment (P<0.05). Conclusion:Acupoint massage for elderly patients with primary hypertension can improve their sleep quality and enhance their cognitive function, thus it is worthy of being applied in clinical nursing work.

Objective To study the effect of γ-rays irradiation on the differentiation potential of the human peripheral blood monocytes (PBMCs) into osteoclast-like cells (OCLs) in vitro.Methods PBMCs were isolated by density gradient centrifugation,treated by receptor activator of nuclear factor-κ B ligand (RANKL) and macrophage-colony stimulating factor (M-CSF) and exposed to 137Cs γ-rays with different radiation doses (0,0.75,2 Gy).After seven days of incubation,the cells were stained for tartrate resistant acid phosphatase (TRAP) and bone slices were stained by toluidine blue on the tenth day.Meanwhile,the characteristic osteoclast markers including Cathepsin K and integrin β3 were analyzed by real-time PCR.Tartrate resistant acid phosphatase 5b (TRAcP-5b) in the culture supernatant wasdetermined by ELISA.Results PBMCs were differentiated into OCLs by the treatments of RANKL and MCSF.The number of TRAP positive multinucleated OCLs was significantly higher in the dose of 0.75 Gy group than in control (0 Gy) group (t =3.451,P ＜ 0.05).Compared with the control group,the expression levels of Cathepsin K and integrin β3 and the concentration of TRAcP-5b were significantly elevated (t =2.343,2.728,3.631,P ＜ 0.05).However,in the 2 Gy group,there was a decrease in the number of osteoclasts,mRNA expression level of osteoclast characteristic markers and TRAcP-Sb,but no statistically significant differences compared with the control group.Conclusions Ionizing radiation may influence the osteoclastogenesis during the PBMCs differentiation to OCLs.At low dosage,ionizing radiation promotes osteoclastogenesis and enhances the resorptive activity of osteoclasts,but a decline of differentiation potential was observed at high dosage of radiation.

Objective To detect the expression of microRNA(miRNAs)in the late stage terminal hindgut de-velopment in fetal rats.Methods Twenty -four female rats were randomly divided into 2 groups.They were matched in proportion of male and female 21 .The experimental rats(n =1 2)received 1 0 g/L ethylenethiourea (1 25 mg/kg)by gavage on gestational day 1 1 ,and the control rats (n =1 2)received same amount of distilled water.The fetal rats were obtained by caesarean section on the gestational day 1 6 in each group.One centimeter rectum was taken out from 2 similar weight fetal rats for extracting total RNA by Trizol method.There were 24 fetal rats in each group.Chip hybri-dization was conducted after Poly(A)and biotin added to the RNA.Then,the chip was washed and dyed,and scanned thereafter.According to the differences in the expression profile of miRNAs and target gene analysis results,the miRNAs probably regulating the key gene of hindgut development was selected for target genes expression analysis.Results Compared with control group,expressions of 1 1 1 miRNAs in terminal hindgut of experimental group were up -regulated on the gestational day 1 6,and 1 1 7 miRNAs were down -regulated.Ten miRNAs of biggest differential expression be-tween them were selected for target genes prediction,pathway analysis,and Gene Ontology analysis.The results showed that some genes were closely related to rat fetus terminal hindgut growth and development,such as Shh,Hoxd13,and so on.According to the differential expression of miRNAs and target genes analysis,miR -1 93 might have an important role in the Hoxd13 gene for anorectal development.Real -Time PCR and Western blot showed that the expression of Hoxd13 and its protein level were significantly decreased when miR -1 93 highly expressed in rat intestinal epithelial cells,and the difference was statistically significant compared with the control group(1 .00 ±0.1 2 vs 0.71 ±0.1 0) (0.88 ±0.06 vs 0.75 ±0.08)(t =3.329,3.1 30;P =0.029,0.01 1 ).Conclusions miRNAs probably have an im-portant regulatory role in their target gene expression in terminal hindgut development of fetal rats,and miR -1 93 can significantly inhibit the expression of Hoxd13 gene in rat intestinal epithelial cells.

Objective:To explore the role of indoleamine-pyrrole 2,3-dioxygenase (IDO),an immunomodulatory enzyme,in renal cell carcinoma (RCC).Methods:A total of 40 patients diagnosed as RCC in the Second Xiangya Hospital were included in this study.All patients received nephrectomy.The histopathological features of samples were assessed semi-quantitatively.IDO mRNA level in RCC and non-RCC renal tissues was determined by real-time quantitative PCR (RT-qPCR).And the expression of IDO protein in endothelial cells was examined by immunohistochemistry; a Kaplan-Meier survival curves was calculated on the basis of IDO mRNA level.Results:Level of IDO mRNA in RCC samples was significantly higher than that in tumor-free samples with P＜0.001.Patients with high IDO expression had an significantly longer survival time than those with low IDO expression (P=0.01).There was a statistically significant inverse correlation between IDO and proliferation marker Ki67.Patients with high IDO level were of low Ki67 level,and vice versa (P＜0.01).Conclusion:IDO might be a prognostic biomarker for patients with RCC.

Objective To investigate the incidence of osteoporosis (OP) in patients with ankylosing spondylitis and the relationship between OP and the clinical data. Methods Serum levels of bone alkaline phosphatase (BALP) and tartrate resistant acid phosphatase 5b (TRACP5b) were detected by enzyme linked immunosorbent assay ( ELISA ) in 60 cases with ankylosing spondylitis, and it was compared with normal controls. Bone mineral density (BMD) was measured through dual-energy X-ray absorptiometry ( DXA), including lumbar ( L2 - L4), bilateral femoral neck and greater trochanter. Some clinical data was collected and analyzed at the same time. Results The incidence of OP in AS patients was 35%, and the incidence of OP in the femoral proximal end was higher than that in lumbar. Compared with normal controls[ ( 1.06 ±0. 18 )U/L ], the levels of serum TRACP5b in AS[ (1.31 ± 0. 82 )U/L] patients was significantly higher ( P <0. 05 ). The levels of serum BLAP in OP combined AS group[ ( 21.65 ± 5.41 ) U/L]were significantly lower than non-OP group[ (32. 37 ± 16. 5 ) U/L] ( P <0. 05 ). The disease duration was negatively correlated with the BMD of femoral neck ( P < 0. 01 ). Conclusions There was higher incidence of OP in AS patients, which were related with the abnormality of bone metabolism and the disease duration.Multiple factors participated in the regulation of bone metabolism of AS.

Objective The goal of this study was to investigate the correlation between the level of serum vitamin D and bone metabolism and disease activity in ankylosing spondylitis(AS) patients,and thus to explore the role of vitamin D in bone metabolism in AS patients.Methods The serum levels of BALP,TRACP-5b,25-(OH)D3 and blood lymphocytes VDR of 80 AS patients were measured by enzyme-linked immunosorbent assay (ELISA) and compared with those of the control group.Bone mineral density (BMD)was measured by dual-energy X-ray absorptiometry (DEXA).The ESR and CRP level of AS patients were also measured.The correlation between those parameters was analyzed and evaluated.Patients were divided into normal,insufficient and deficient subgroups according to the serum 25-(OH)D3 levels for further comparison.Indepondent saimple t test,t'test andx2 test were used for statistical analysis.Results The 25-(OH)D3 of AS patients [(11.9±2.7) μg/L] was significantly lower than that of the control groups [(22.3±7.9) μg/L] (P＜0.05),while the serum levels of BALP [(3.9±2.7) μg/L] and TRACP-5b [(46±25) ng/L] of AS patients were significantly higher than those of the control group [(2.4±1.0) μg/L] (P＜0.05).According to linear correlation analysis,25-(OH)D3 was negatively correlated with CRP (r=0.324,P=0.003).The ESR,BALP,TRACP-5b in the deficient subgroup were higher than those in the normal and insufficient subgroups(P＜0.05).Conclusion The plasma 25-(OH)D3 may decrease in AS patients,and this may activate bone metabolism,results in increased morbidity of osteoporo-sis,and negatively affect disease activity.

Objective To study the effects of EP4 and EP2 antagonists on the differentiation of Treg/ Th17 cells and disease progression in mice of collagen-induced arthritis (CIA) model.Methods DBA/1 mice wereimmunized subcutaneously twice at the root of the tail with type Ⅱ collagen emulsified in Freund's complete adjuvant.EP2 and EP4 antagonist therapies were intraperitoneally administrated for 14 consecutive days after the second immunization.Clinical signs,histological manifestation,serum interleukin (IL)-17 and quantity of CD4+CD25+Foxp3+ Treg cells were determined.ANOVA and t-test were used for statistical analysis.Results Clinical signs of the disease appeared on day 27 and peaked on day 35 after the first immunization.The quantity of CD4+CD25+Foxp3+ Treg cells in spleens [(1.67±0.15)％] and draining inguinal lymph nodes [(3.30±0.36)％] isolated from CIA mice were significantly lower than those of normal DBA/1 mice [(2.77±0.45)％ and (4.73 ±0.45)％ respectively,P＜0.05].Serum IL-17 level of CIA mice [(27±7) pg/ml] was significantly higher than that of normal DBA/1 mice [(14±4) pg/ml,P＜0.05].Intra-peritoneal injection of EP4 but not EP2 antagonist to CIA mice decreased paw edema and swelling,and alleviated the histological manifestations (1.8±1.0 vs 3.5±0.6,P＜0.05) on day 35 after the first immunization.The percentages of CD4+CD25+Foxp3+ Treg cells in both inguinal lymph nodes [(4.20±0.32)％] and spleens [(2.63±0.40)％] were significantly higher in EP4 antagonist-treated but not EP2 antagonist-treated CIA mice compared with CIA mice group [(3.30±0.36)％ and (1.67±0.15)％ respectively,P＜0.05].The level of serum IL-17 was significantly lower in EP4 antagonist-treated [(15±7) pg/ml] but not EP2 antagonist-treated CIA mice compared with CIA mice group [(27±7) pg/ml,P＜0.05].Conclusion EP4 antagonist therapy alleviates clinical symptoms of CIA,improves the histological manifestations,decreases the serum IL-17 level and increases the percentages of CD4+CD25+Foxp3+ Treg cells in both spleens and draining inguinal lymph nodes,so targeting EP4 receptor may be a new possible therapeutic possibility in the prevention and treatment of rheumatoid arthritis.