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 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 analyze the pregnancy outcome of laparoscopic cervical cerclage before pregnancy, discuss the clinical value of the treatment of cervical incompetence.Methods 40 cases with cervical insufficiency during 2012.1 -2016.3 before pregnancy underwent laparoscopic ligation.All patients had a history of abortion or premature birth,and the average gestational age was 23.7 weeks.Preoperative cervical length was less than 2.5cm. The operation condition and the pregnancy outcome after operation were observed.Results The average time of oper-ation was 35.6(40.2 ±10.4)min and the amount of bleeding was 10.3(15.7 ±3.2)mL.Postoperative pregnancy cervical average length was 3.61 (4.35 ±0.52)cm.Pregnancy outcome:37 weeks of gestation and above delivery 20 cases (50.0%).34 -37 weeks 16 cases (40.0%),28 -34 weeks in 3 cases,received cesarean section,the aver-age gestational was 35.6 weeks.Compared with the preoperative abortion,the gestational week of delivery was pro-longed by 12.1 weeks.Neonatal survival in 38 cases (95.0%),1 case of gestational 28 +3 weeks premature death. Neonatal birth weight was 2 800(3 079 ±500)g;1 case (2.5%)was difficult to avoid abortion at 23 weeks of gesta-tion.Conclusion Laparoscopic pre -pregnancy cervical cerclage suture in cervix mouth,postoperative can maintain effective cervical length,prolong gestational age,improve the pregnancy outcome,can be used as effective treatment for the pregnant cerclage failure or cervical length less than 2.5cm of cervical insufficiency patients.

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.

BACKGROUND:Drug therapy can partly reduce and delay the progress of Alzheimer’s disease, but only 30%with the single drug treatment obtain clinical cure. OBJECTIVE:To study the therapeutic effect of bone marrow mesenchymal stem cel s transplantation for rats with Alzheimer’s disease. METHODS:Amyloidβ-protein was injected into the hippocampus of Sprague-Dawley rats to construct the model of Alzheimer’s disease. And bone marrow stromal stem cel s were transplanted into the hippocampus of the rat models. RESULTS AND CONCLUSION:At 2 weeks after modeling, compared with the control group, the escape latency in the model and experimental groups was significantly longer (P<0.05), which indicating that Alzheimer’s disease models were successful y established. At 4 weeks after cel transplantation, compared with the model group, the average escape latency in the experimental group was significantly decreased, but retention time on the platform quadrant was significantly prolonged (P<0.05). Besides, at 4 weeks after cel transplantation, expression of choline acetyltransferase in the experimental group was significantly higher than that in the model group (P<0.05). In conclusion, bone marrow mesenchymal stem cel s cannot only differentiate and survive in the hippocampus of rats with Alzheimer’s disease, but also improve the learning and memory ability.

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.

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.