Objective Explore the conditions of the cloning,expression and purification of FMRP.Methods The plasmid pET22b(+)-FMR1,constructed by molecular cloning,was transformed into E.coli BL21(DE3) competent cells and induced to express FMRP by IPTG.Recombinant FMRP was purified by affinity chromatography,verified by Western-blot,and tested for its RNA binding ability.Results FMR1 cDNA was successfully cloned into pET22b(+) vector and expressed in E.coli BL21(DE3).A protein with Mr 79 000 was purified and confirmed to be FMRP.This protein retained the RNA binding ability of FMRP.Conclusion We successfully expressed recombinant hFMRP with high purity and activity in E.coli,which provided a reliable material to study the function of FMRP.

Objectives: The survivors of first-ever stroke are at a high risk of recurrent stroke. The time interval between first-ever stroke and first recurrence of stroke, however, have not been well studied. The aim of the present study was to evaluate the time interval between first-ever and first recurrence of stroke and the risk factors of stroke that were related to the length of time interval. Methods: Patients admitted in our hospitals during 2014 with first recurrence of stroke were included in the study. A total of 377 patients were enrolled and a standardized questionnaire was used to collect data in this retrospective study. Results: The mean time interval among all the subjects was 58.42 months (median, 36.01 months; range 0.16months to 455.98months). The mean time interval was shorter in hemorrhagic stroke group (56.78 months) than in ischemic stroke group (58.75 months), but the difference was not significant (p=0.819). The median of time interval was similar in the two groups. The associated risk factors to the length of time interval were age over 60 years, subtype of first-ever stroke, the length of history of hypertension. Age was the only associated risk factor to the time interval among patients with first recurrence of hemorrhagic stroke. Conclusion: Factors associated with the time interval are different among different types of recurrent stroke. This provides the basis for preventive treatment for recurrent stroke after their first-ever stroke
Stroke

Objective To sunmarize the clinical experience of emergency laparoscopic choledocholithotomy plus T-tube drainage for acute calculous cholangitis.Methods We retrospectively analyzed the clinical data of 98 cases with acute calculous cholangitis undergoing emergency laparoscopic choledocholithotomy between Mar 2007 to Jul 2011 at our hospital.Results In this study,93 cases with definite diagnosis of acute calculous cholangitis underwent at their acute stage totally laparoscopic choledocholithotomy and T-tube drainage,2 cases received laparoscopic transeystic common bile duct exploration and 3 cases were treated with primary closure of the common bile duct following laparoscopic common bile duct exploration and intra-bile duct stent.All procedures were successful without conversion to open surgery.The average of the operative time and the hospital stay was 96 minutes and 12 days respectively.The major complications included residual stones in 8 cases,bile leakage in 4 cases and intraperitoneal bleeding in I case.All were cured by conservative therapy.Conclusions For most patients of acute calculous cholangitis,emergency T-tube drainage after laparoscopic chuledocholithotomy is safe and effective.

Objective To evaluate the clinical effect of IPSe. max press ceramic endocrowns being used in preserving endodontically treated molars. Methods Forty-five endodontically treated molars were selected from 45 patients in this study. All the molars had been endodontically treated and performed with ultrasonic scaling. After 1-2 weeks, the crowns and pulp chambers were prepared. The impressions and master casts were gained and IPSe. max press ceramic endocrowns were fabricated in the lab. The endocrowns were cemented with resin cement. The further consultations were offered to these molars after 6 months and 1 year. The difference of sulcus bleeding index among these endodontically treated molars and the same teeth in the opposite sides was compared. Results In these cases, 100 % of endocrowns were

OBJECTIVETo investigate the main factors that influence the results of sperm alkaline single-cell gel electrophoresis (SCGE), optimize the conditions, and standardize its procedures.

METHODSUsing alkaline SCGE, we detected the DNA fragments of sperm treated with different concentrations of H2O2 and determined the influences of the number of agarose gel layers, pH during DNA unwinding and electrophoresis, the time of DNA unwinding and electrophoresis, and cumulative sperm number on the results of sperm alkaline SCGE. Then we optimized the procedures, analyzed the repeatability of the optimized method, and examined 40 semen samples using the method.

RESULTSThree agarose gel layers could reduce the background. The optimal pH during DNA unwinding and electrophoresis was 10, and the best times for DNA unwinding and electrophoresis were 40 min and 30 min, respectively. Fifty sperm were adequate to ensure the reliability of the results. Based on the percentage of tail DNA, the intra- and inter-assay repeatabilities of the optimized sperm alkaline SCGE were 3.12% and 7.13%, and by the DNA damage score, they were 2.38% and 6.09%, respectively. Sperm DNA fragments were significantly increased in the infertile patients with oligoasthenoteratozoospermia as compared with healthy fertile males (P <0.05).

CONCLUSIONThe optimized sperm alkaline SCGE, highly repeatable and easy to be standardized, can be applied to the clinical detection of sperm DNA fragmentation in infertile men.

Asthenozoospermia ; genetics ; Comet Assay ; standards ; DNA Damage ; DNA Fragmentation ; Humans ; Hydrogen Peroxide ; toxicity ; Male ; Oligospermia ; genetics ; Oxidants ; toxicity ; Reproducibility of Results ; Sperm Count ; Spermatozoa ; drug effects ; enzymology ; Time Factors

Objective To study the effect of annexin A1 on cardiac function,tumor necrosis factorα(TNF?α),and interleukin 1β(IL?1β)in diabetic rats. Methods Twenty?four SD rats were randomly divided into normal control and diabetic groups. The type 2 diabetes model was in?duced with a high?glucose and high?fat diet and administration of low?dose streptozotocin.Left ventricular end?diastolic volume(LVEDV),left ven?tricular end?systolic volume(LVESV),peak velocity of early diastolic mitral?to?late diastolic peak velocity(e/a)ratio,left ventricular ejection frac?tion(LVEF),and stroke volume(SV)were measured by using color Doppler ultrasonography at the end of week 8. The expression levels of TNF?αand IL?1βin blood were measured by using enzyme?linked immunosorbent assay,and the expression level of annexin A1 in blood was measured at weeks 0,4,and 8 by using real?time polymerase chain reaction. Results Compared with the normal control group,the diabetic group had de?creased LVEDV,e/a,and SV(P<0.05).The annexin A1 expression level in the diabetic group decreased significantly after 8 weeks(P<0.01). The TNF?αand IL?1βlevels in the diabetic group were significantly higher than those in the normal control group(P<0.05)and increased signifi?cantly after 8 weeks(P<0.01). Annexin A1 level correlated with the TNF?αand IL?1βlevels in the diabetic group(P<0.01). Conclusion Annexin A1 expression shows an anti?inflammatory effect that improved the cardiac function of diabetic rats.

Objective To evaluate the clinical application of selective renal artery embolization (SRAE) for the treatment of kidney diseases. Methods Seventy-four cases of renal carcinomas, 11 cases of renal angiomyolipomas (RAML) and 72 cases of traumatic renal haemorrhages were first demonstrated by renal arteriography under Seldinger technique to ensure a site, range and neighbouring relation of lesions and then followed by percutaneous catheterized selective renal arterial embolization with embolic agents. Results The edema around the carcinomas and abscesses became obvious, and bleeding were reduced, which were convenient for operation after SRAE for the preoperative adjuvant treatment of renal carcinomas. The clinical symptoms were improved obviously in 9 eases with advanced renal carcinoma after palliative treatment. The tumour volumes of 11 RAML were decreased evidently by 15 %-65 % with the average of 42 %. The bleeding in 68 cases of traumatic renal haemorrhage were ceased completely in 1-4 days after embolism, 2 cases with serious renal fragmented injury and huge perirenal hematoma combined with shock received successful operation after SRAE, 2 cases bleeding again after SRAE were cured by the second SRAE. No severe complications occurred after embolization in all the patients. Conclusions As a minimal invasive technique,selective renal artery embolization is a safe, effective method with less complications and an adjuvant pre-op-erative therapy for renal carcinoma or a palliative treatment for advanced carcinoma and an effective treatment for RAML and traumatic renal haemorrhage.

Objective To evaluate the predictive values of different systems for clinical staging of esophageal carcinoma in one group of patients and improve the criteria for T staging,and to provide a basis for accurate clinical staging. Methods A retrospective study was performed in 701 patients with esophageal carcinoma who received radical radiotherapy in our hospital. The prognosis was performed according to American Joint Committee on Cancer ( AJCC) tumor-node-metastasis staging system,Chinese 2004 staging system,the draft of Chinese 2009 staging system,and gross tumor volume of the primary tumor (GTV-T). Results In terms of T stage,patients evaluated according to the AJCC staging system were in relatively early stages;23. 1% of them were in stage T1,and the survival curves of T3 and T4 patients were close to each other;the survival curves plotted according to the Chinese 2004 staging system were well separated, but relatively few patients were in stages T1 and T4 , yielding an uneven distribution;according to the draft of Chinese 2009 staging system, the survival curve of T3 patients intersected that of T4 patients, and up to 43. 2% of patients were in stage T4.The new T staging was performed based on GTV and the extent of tumor invasion into the adjacent tissue and organ, and the results showed that there was no intersection between survival curves and a relatively balanced T stage distribution. In terms of N staging,patients were divided into stages N0 ,N1 ,and N2 . The TNM staging was performed by a combination of N staging and new T staging, resulting in significant separation between survival curves ( P=0. 000) . Conclusions The combination of T staging,which is based on GTV and the extent of tumor invasion,and N staging,which is based on metastasis of lymph nodes, can accurately predict the survival of non-surgically treated patients with esophageal carcinoma.

Objective To explore the prediction value of the modified clinical staging standard of GTV volume on non-surgical treatment esophageal carcinoma by analyzing the GTV volume of esophageal carcinoma and the invasion degree of structures and surrounding organs as the T stage standard.Methods A retrospective analysis was performed for 701 esophageal cancer patients treated by definitive radiotherapy from Jan.2006 to Dec.2012.After grouping and analysis by the previous GTV volume staging standards, we put forward the idea that considering effects of invasion degree of structures and surrounding organs of tumor on the basis of GTV volume when it came to T stage, which would be re-classified by downgrading and reevaluation of survival and prognosis.Results There was no significant survival differences between T3 and T4 on previous GTV volume staging standards (P ＞ 0.05), and also had shown an inconspicuous survival difference between stage Ⅲ and stage Ⅳ when combined with three-group N stage(P ＞ 0.05).We had modified the T stage standards of GTV volume: Based on different size of GTV volume, and in consideration of the invasion of adjacent structures and organs, new T stages had shown good separation on a corresponding survival curve(x2 =59.702 ,P ＜0.05).In clinical TNM staging which combined with the new T stage and three-group N stages, the 701 patients were divided into stage Ⅰ , Ⅱ , Ⅲ and Ⅳ, with corresponding 5-year survival rates of 33.5％ , 26.3％ , 13.4％ , 9.2％ , respectively, which strongly revealing significant differences of survival rates (x2 =82.577, P ＜ 0.05).Conclusions The new T staging standard, which combined GTV volume with invasion degree of adjacent structures and organs, could accurately predict the prognosis of patients with radical radiotherapy of esophageal carcinoma.

Objective To analyze the efficacy of chemoradiotherapy in the treatment of esophageal carcinoma and its influencing factors,and to provide an optimal combination mode of chemoradiotherapy for treating esophageal carcinoma. Methods A retrospective analysis was performed on clinical data from 232 patients with esophageal carcinoma who were admitted to our hospital from January 2006 to December 2012 and received radical chemoradiotherapy. All patients received three?dimensional conformal radiotherapy or intensity?modulated radiotherapy as well as platinum?based chemotherapy. The overall survival ( OS ) and local control ( LC) rates were calculated using the Kaplan?Meier method and analyzed using the Logrank test. Univariate and multivariate prognostic analyses were made by the log?rank test and the Cox proportional hazard model,respectively. Results In all patients,the 1?,3?,and 5?year LC rates were 66?1%,42?2%, and 38?5%,respectively;the median LC time was 24?4 months;the 1?,3?,and 5?year OS rates were 73?3%, 37?2%,and 19?5%,respectively;the median OS time was 21 months. The univariate analysis revealed that T stage,N stage,clinical stage,irradiation range,and no less than 3 cycles of chemotherapy were influencing factors for OS ( P=0?000,0?000,0?000,0?030,0?001) and LC ( P=0?112,0?031,0?009,0?074,0?218) . The multivariate analysis revealed that N stage,clinical stage,and no less than 3 cycles of chemotherapy were independent prognostic factors for OS ( P=0?006,0?000,0?001) . Conclusions The LC and long?term OS rates in patients with early?stage esophageal carcinoma can be substantially improved by radical chemoradiotherapy. The irradiation range and no less than 3 cycles of chemotherapy improve the long?term survival in patients.