Objective To investigate the effect of N-methyl-N-nitrosourea (MNU) on expressions of interphotoreceptor retinoid-binding protein (IRBP) and recoverin in rat retinal.Methods Thirty female Sprague-Dawley (SD) rats were randomly divided into five groups,including normal control group,and MNU treatment one day,three days,seven days and 10 days groups,each group had six rats.Rats in MNU-treatment groups received a single intraperitoneal injection of 40 mg /kg MNU,while rats in the normal control group received intraperitoneal injection of saline 5 ml/kg.Reverse transcription-polymerase chain reaction (RT-PCR) and immunofluorescence were used to detect the retinal expression of IRBP and recoverin.Results RT-PCR results indicated that retinal IRBP levels decreased after MNU injection compared with normal control (P＜0.01),while recoverin levels increased (one day:P＞0.05; three days:P＜0.05; seven days:P＜0.5; 10 days:P＜0.05).Immunofluorescence assays demonstrated that normally IRBP protein is mainly in the inner and outer segments of photoreceptors.After MNU treatment,IRBP protein was detected in all retinal layers but much faint.Recoverin was expressed in the inner nuclear layer,inner plexiform layer and ganglion cell layer,and its expression was increased after MNU injection.Conclusion MNU suppresses IRBP expression and promotes recoverin expression in rat retina.

Objective To study the optimal dose and reaction time of human leukocyte antigen B27(HLA‐B27)antibody in flow cytometry .Methods Take 52 cases of whole blood in patients with ankyl‐osing spondylitis(AS) .According to HLA‐B27 antibody doses ,samples were divided into two groups:5 μL group and 10 μL group .HLA‐B27‐positive rate were tested after 5 ,10 ,15 min , respectively .Results The HLA‐B27 positive rate of 5 μL group at different reaction time were (84 .16 ± 1 .21)% ,(94 .81 ± 1 .33)% ,(94 .10 ± 1 .26)% ;the positive rate of 10 μL group at different reaction time were (85 .40 ± 1 .27)% ,(96 .76 ± 1 .31)% , (95 .36 ± 1 .45)% .The positive rate of HLA‐B27 in 10 μL group was higher than 10 μL group(F=90 .08 ,P<0 .05) .The positive rate of HLA‐B27 after reacting for 10 and 15 min were higher than that after reacting for 5 min(F=60 .25 ,P<0 .05) .There was not statistically significantly different between the reaction time of 10 min and 15 min(F=1 .08 ,P>0 .05) .Conclusion The opti‐mal dose and reaction time of HLA‐B27 antibody in flow cytometry are 10μL and 10 min;There is not any interaction between anti‐body dose and the reaction time of HLA‐B27 antibody .

Aim To prepare soluble human C1 q and tumor necrosis factor related protein-6 in Escherichia coli and analyze the bioactivity. Methods Recombi-nant plasmid was transformed into E. coli expression strain, and the recombinant protein Trx-hCTRP6 was expressed induced by IPTG and then purified. Results Trx-hCTRP6 was expressed efficiently and purified using Ni-NTA affinity chromatography and Superdex G-75 column. The purified Trx-hCTRP6 was shown to be active under in vivo and in vitro assay conditions. Con-clusion Active Trx-hCTRP6 is efficiently prepared from E. coli protein expression system.

Objective To investigate the expression and clinical significance of ROC1 in ovarian cancer tissues ,to provide new ideas for the gene therapy of ovarian cancer .Methods 261 women with ovarian cancer underwent resection from December 2005 to December 2007 in the Affiliated Hospital of Zunyi Medical College and with 5 years follow-up were enrolled .ROC1 mRNA transcription and protein expression level of ovarian cancer tissue and surrounding normal tissue were tested ,and later the correla-tion between the results and patient′s prognosis was statistically analyzed .Results ROC1 in ovarian cancer tissue was over-ex-pressed ,but almost no expression in surrounding normal ovarian carcinoma ;ROC1 expression levels had a negative correlation with the survival rate of the patients .Conclusion ROC1 in ovarian cancer tissue was over-expressed ,but almost no expression in sur-rounding normal ovarian carcinoma ;ROC1 expression levels had a negative correlation with the survival rate of the patients .

Objective To evaluate the accuracy and feasibility of 64-slice spiral CT (64SCT) in assessing coronary artery disease.Methods A total of 30 suspected patients (male 21 cases, female 9 cases, and mean age of 54.6 years) were undergone both 64SCT and selective coronary angiography (SCA). Volume redering (VR) ,multiplanar reconstruction (MPR), maximum intensity projection (MIP) and transverse section were reconstructed. The results of coronary reconstructions were compared with SCA to analyze the accuracy of the 64SCT in detecting coronary artery stenoses.Results In the 396 segments of coronary artery(diameter≥2 mm)of 30 patients, 385 were judged to be evaluable by 64SCT. The evaluable rate was 97.2%. The sensitivity, specificity, positive predictive value and negative predictive value of the 64SCT in detecting coronary artery stenoses(≥50% of stenosis) were 96.22%, 94.56%, 89.44% and 96.88%. The accuracy rate of 64SCT in detecting ≥50% stenosis of coronary artery was 95.90%.Conclusion 64SCT has high accuracy in detecting coronary artery stenoses, as a noninvasive method,it can be used for screening patients with known or coronary artery diseases.

Objective To investigate the clinical significance of serum lactate dehydrogenase (LDH) and its isoenzymes,M or H subunits in the retreated patients with multiple myeloma(MM).Methods 141 medical records of retreated patients with multiple myeloma were collected,and the relationship of LDH and its isoenzymes,subunits with DS staging system,ISS staging system,creatinine,bone marrow plasma cell percentage,immunoglobulin and microglobin was analyzed.Results Serum total LDH activity was not significantly related to DS staging system and ISS staging system.LDH1 percentages was negatively correlated with ISS staging system (P＜0.01),and LDH4 percentages positively associated with ISS staging system (P＜0.05).LDH activity,absolute value of LDH1 ,LDH2,LDH3,LDH4,and H subunits were positively linked to serum creatinine in myeloma kidney disease patients with renal inadequacy (P＜0.05).Both percentage and absolute value of LDH5 were negatively correlated with bone marrow plasma cell percentage (P＜0.01 and P＜0.05 respectively).Conclusion The detection of serum LDH isoenzyme is more valuable than total LDH activity in retreated patients with multiple myeloma.

This study aimed at evaluating the effect of different processing methods on the contents of antiosteoporotic compositions in P.corylifolia.HPLC method adopted to analyze the contents of antiosteoporotic compositions,including coumarin (psoralen and isopsoralen),flavonoids (neobavaisoflavone,corylifolin,corylifolinin and corylifol A) and meroterpenes (bakuchiol) in P.corylifolia.The contents were analyzed before and after the processing methods.Results:The results showed that the content of coumarin,flavonoids and meroterpenes obviously changed in different processed P.Corylifolia in comparison with the crude drugs.In conclusion,it was demonstrated that the therapeutic efficacy of osteoporosis was enhanced by CP method which increased all of the three compositions.The testimony tallied with the Chinese medical theory:stir-frying with salt-water could enhance the effect of the tonification of spleen and kidney.

Objective To explore the roles and possible mechanism of calcium-sensing receptor(CaSR) in cell cardiac hypertrophy model using angiotensin Ⅱ (Ang Ⅱ ).Methods The cultured neonatal rat ventricular myocytes were treated with Ang Ⅱ as cell cardiac hypertrophy model.Hypertrophic neonatal rat cardiomyocytes were treated with GdCl3(a specific agonist of CaSR) and/or with Ro318220(a specific inhibitor of PKC pathway).To evaluate the status of cardiac hypertrophy,cell diameter was observed by HE dyeing,and protein content was determined through coomassie brilliant blue protein kit.The intracellular calcium concentration( [ Ca2+]i) was determined by laser scanning confocal microscope.The protein expression of CaSR and PKC pathway were analyzed using Western blotting.Results ①Compared to the control group(0.1263 ± 0.0443),the protein expression of CaSR was increased in Ang Ⅱ group and in GdCl3 group(0.1963 ± 0.0375,0.2778 ± 0.0564,all P＜ 0.05).Moreover,compared with Ang Ⅱ alone,the increase was significant in GdCl3 group(P ＜ 0.05).②Compared to control group(222.70 ± 22.09),AngⅡ group(392.16 ± 36.85) remarkably increased [Ca2+]i(P＜ 0.05),and this increase of [Ca2+]i was further enhanced in GdCl3 group (502.60 ± 44.21) versus Ang Ⅱ group (P ＜ 0.05).③Compared to control group,Ang Ⅱ could induce cardiomyocyte hypertrophy,and GdCl3 enhanced the effect.Moreover,this enhancement was attenuated by Ro318220.④Compared to control group(0.27 ± 0.07,0.69 ± 0.06,0.87 ± 0.04),the protein expression of PKCα,PKCε and PKCδ was increased in Ang Ⅱ group(0.60 ± 0.16,1.02 ± 0.13,1.20 ± 0.18,all P＜ 0.05) and the protein expression of PKCα,PKCε was increased in GdCl3 group(0.82 ± 0.16,1.34 ± 0.12,all P ＜ 0.05).Moreover,compared with Ang Ⅱ group,the protein expression of PKCα,PKCε was obviously increased in GdCl3 group (all P ＜ 0.05);compared with GdCl3 group,the protein expression of PKCα,PKCε(0.41 ± 0.10,0.85 ± 0.14) was obviously decreased in Ro318220 group(all P ＜ 0.05).Conclusions CaSR is involved in cardiac hypertrophy induced by Ang Ⅱ through PKC pathway in cultured neonatal rat cardiomyocytes.

Objective To observe the pregnancy by transvaginal double slit cervical pregnancy outcome after ligation,and evaluate its clinical application value.Methods The clinical data of 80 cases with cervical incompe-tence were analyzed.All patients had pregnant metaphase inevitable abortion history,the average abortion pregnant was 21.7 week.According to different treatment methods,80 cases were divided into two groups,treatment group (40 cases)in non pregnant women with silk thread woven tape by double vagina and cervix cerclage,another group of 40 cases in 15 -20 weeks of pregnancy by using polypropylene McDonald method with cerclage.The operation,preg-nancy cervical tube length and pregnancy outcome were observed.Results The average operation time and bleeding volume between the two groups had no statistically significant differences.The pregnancy cervical average length of the treatment group was 4.6(4.8 ±0.5)cm,which was significantly longer than 3.9(3.1 ±0.8)cm of the control group (t =2.732,P <0.05).In the treatment group,the full -term delivery rate,preterm delivery rate,mean gestational age at delivery were 55.0%(22 /40),40.0%(16 /40),35.8 weeks,the average abortion was extended 14.1 weeks com-pared with preoperation,fetal survival rate was 90.0%(36 /40),neonatal birth weight was (3 100.0 ±200.0)g;28 gestational weeks following the inevitable abortion rate was 2.5%(1 /40).In the control group,the term delivery rate was 35.0%(14 /40),the rate of premature birth was 50.0%(20 /40),and the rate of abortion was 15.0%(6 /40)in 28 weeks.Mean gestational age at delivery was 33.2 weeks,which was extended 11.5 weeks compared with preoperative average abortion,fetal survival rate was 85.0% (34 /40),neonatal body mass was (2 900.0 ±300.0)g. The term delivery rate,fetal survival rate and neonatal weight in the treatment group were higher than those in the con-trol group.Conclusion Pregnant by double vagina and cervix cerclage treatment cervical function can maintain preg-nancy,cervical length,prolong the time of pregnancy and improve the perinatal survival rate,simple operation,take out stitches convenient,it is worthy of clinical application.

Objective To observe the effect of pregnancy double loop cerclage for cervical incompetence. Methods 80 cases of cervical function incompetence according to the type of surgery were divided into two groups, 40 cases in each group,15 -20 weeks of surgery in pregnancy,the treatment group underwent double loop cerclage, the control group underwent McDonald single cerclage.The operation,postoperative cervical length and pregnancy outcome of the two groups were observed.Results There was no difference in operation time and blood loss between the two groups.The length of cervix in the treatment group was (4.63 ±0.52)cm,which was longer than (3.71 ± 0.82)cm of the control group(t =2.106,P <0.05).Pregnancy outcome:20 cases of the treatment group in term birth (50%),preterm birth in 18 cases (45%)(34 -37 weeks in 12 cases,28 -34 weeks in 6 cases),fetal survival in 36 cases (90%),less than 28 weeks pregnant inevitable abortion in 2 cases (5%),abortion did not survive.Average delivery gestational age was 35.2 weeks,and preoperative average abortion pregnant weeks contrast,treatment group delivery gestational weeks extended 13.5 weeks,the average weight of newborns was (3 200 ±200)g.The control group of full -term delivery was 14 cases (35%),premature in 20 cases (50%)(34 -37 weeks in 7 cases and 13 cases of 28 -34 weeks),fetal survival in 33 cases (82.5%),less than 28 weeks pregnant inevitable abortion in 4 cases (10%),abortion did not survive.Average delivery gestational age was 33.2 weeks,compared with the average abortion weeks before operation,the gestational age of the control group was extended by 11.5 weeks.The weight of newborn was (2 900 ±300)g.In the treatment group,the term labor rate,fetal survival rate,neonatal weight were higher than those of the control group,the difference was statistically significant (χ2 =7.83,P <0.05).Conclusion The second trimester cervical double cerclage can maintain effective cervical length,prolong gestational age,improve perinatal survival rate.