Ghrelin is a small active peptide secreted by gastric fundus cell. As an endogenous ligand of growth hormone receptor,it helps to promote growth hormone secretion,increase food intake and participate in energy metabolism. Recently,studies have indicated that Ghrelin,together with other peripheral signals,participate in the regulation of glucose and lipid metabolism. Insulin resistance (IR) is an important pathophysiological basis of the development of type 2 diabetes mellitus (T2DM),while obesity is the common risk factor of both IR and T2DM. It has been confirmed that Ghrelin has a close and complex connection with obesity and T2DM. Further study about Ghrelin may open up a new way to the diagnosis and treatment of obesity and T2DM.

Objective To establish the sperm specific Sleeping Beauty ( SB ) transposase-expression transgenic mouse for the study of the genetic modification mediated by transposon system in mouse .Methods Prm1 promoter was cloned from mouse genomic DNA to drive the expression of SB transposase .The transgenic mice were generated by microinjection .The gene type of transgenic line was identified by PCR .The expressing level in testis was determined by western blot and immunohistochemistry (IHC) staining.Results Five lines of transposase transgenic mice were obtained by microinjection and three can be germline .One mouse line with higher expression level of transposase in the testis was obtained.Conclusion One transgenic mouse model with Sleeping Beauty transposase - expression was successfully established .This model will greatly contribute to the research of genetic modification mediated by transposon in mouse.

Objective To evaluate the value of circular anastomotic stapler for sigmoid colostomy in laparoscopy abdominoperi-neal resection .Methods 62 patients of low colorectal cancer treated in our hospital from October 2010 to February 2013 was retro-spectively analyzed .All the patients were treated by laparoscopy abdominoperineal resection ,34 patients received sigmoid colostomy with circular anastomotic stapler(group A) ,and the other 28 patients received sigmoid colostomy with conventional suturing tech-niques(group B) .The medical records including operation time ,the time of return of bowl function ,postoperative hospital stay and postoperative complication rate were analysed statistically .Results All patients received the operations successfully .No conversion to open procedure and no operative death occured in two groups .The result of statistical analysis showed that in group A ,the opera-tion time ,the time of return of bowl function ,postoperative hospital stay time were shorter than group B ,and the rate of edema of sigmoid in group A were lower than group B(P<0 .05) .Conclusion The application of circular anastomotic stapler for sigmoid co-lostomy in LAPR is a safe ,effective and minimally invasive technique ,which can shorten operation time ,postoperative hospital stay and reduce the related complications of colostomy .

Objective To investigate whether the administration of the ultra-filtration extract mixture from Angelica sinensis and Hedysarum polybotrys is able to protect cardiomyocytes from oxidative injury of rats induced by H2O2 and its potential mechanism. Methods Myocardial cells from 2—3 d neonatal rats were cultured in DF medium and the cellular injury was induced by H2O2. The ultra-filtration extract mixture from A. sinensis and H. polybotrys was given in three doses of 3.75,7.5,and 15 mg/mL. Morphological changes of cardiomyocytes were observed by microscope. Survival rate of myocardial cells was assessed using MTT. The cardiomyocyte damages were estimated by detecting lactate dehydrogenase (LDH) and creatine kinase (CK) releases in the medium,superoxide dismutase (SOD) activities and intracellular malondialdehyde (MDA),and myeloperoxidase (MPO) contents. The levels of caspase-3 and hsp70 expression in cardiomyocytes were measured by RT-PCR. Results The ultra-filtration extract mixture could protect the cardiomyocytes from H2O2 injury in a dose-dependent manner (3.75,7.5,and 15 mg/mL). The ultra-filtration extract mixture could significantly decrease LDH and CK leakages and intracellular MDA and MPO contents,increase SOD activity,upregulate hsp70 expression,and downregulate caspase-3 expression. Conclusion The ultra-filtration extract mixture has protection on cardiomyocytes injured by H2O2 through improving cell antioxidant ability,upregulating hsp70 expression,and inhibiting caspase-3 activity.

AIM:To investigate the pooled association between estrogen receptor α( ESRα) rs2234693 poly-morphism and prostate cancer risk .METHODS:A systematic literature search was performed to identify the related stud-ies (up to April 2014) in several online databases including PubMed , the CNKI and Wanfang online libraries .Odds ratios with 95%confidence intervals were used to calculate the strength of association in the random effect model .RESULTS:A total of 20 studies including 4 623 cases and 9 850 controls were enrolled in the final meta-analysis.The results indicated that ESRαrs2234693 polymorphism was significantly associated with prostate cancer risk (P<0.05) in a dominant genetic model.In the subgroup analysis by ethnicity , there were significant associations between ESRαrs2234693 polymorphism and prostate cancer risk in Caucasians ( P<0.05 ) , but not in Asians and Africans ( both P>0.05 ) .CONCLUSION:This meta-analysis suggests that ESRαrs2234693 polymorphism is significantly associated with prostate cancer risk , espe-cially in Caucasians .

Objective To study the relationship of insulin receptor substrate-1 (Irs1) and metabolic disease, we generated Irs1 gene knockout rat by CRISPR/Cas9 system.Methods Two sgRNA targeting sites were designed for Irs1 targeting.The Cas9 and sgRNAs were transcribed by T7 RNA polymerase in vitro.Cas9 mRNA and sgRNA mixtures were pooled and microinjected into one-cell fertilized eggs of SD rats to generate rats with targeted mutation .Results Five rats with the mutations were detected with the efficiency of 83%.Conclusion The Irs1 gene knockout rats generated in this study can be transmitted by germline .

Objective To study the effects of NRG2 on cardiac structure and function , we established the cardiac-specific human NRG2 transgenic mice and investigate the effect of NRG2 on cardiac structure and function under pressure overload situation .Methods The transgenic vector was constructed by insertion of the human NRG2 gene under the α-MHC promoter.The transgenic mice were generated by microinjection and were all maintained on a C57BL/6J genetic background .The genotype of transgenic mice was identified by PCR and the expression level of target gene was determined by western blot .Transverse aortic constriction ( TAC) was applied to prepare the pressure overload induced cardiomyopathy mice model .The cardiac structure and function of the transgenic mice were compared and analysized by echocardiographic and pathological observation .Results Transgenic mice with high level of NRG2 in heart tissues were established.The left ventricular wall thickness (LVPWD) was increased, and to 15.6% at 3 months old compared with that of the non transgenic ( NTG) mice.The hypertrophy of left ventricular wall caused by pressure overload was removed due to the expression of NRG2 .Meanwhile, cardiac disarray and fibrosis were increased obviously compared with that of the NTG mice.Conclusion The transgenic expression of NRG2 in heart tissues could shorten the pathological process of hypertrophy, but accelerated the process of heart failure (HF).

OBJECTIVE:To establish a method for the content determination of saccharide in polysaccharides containing galact-uronic acid based on phenol-sulfuric acid method combined with correction factor method. METHODS:The determination condi-tion of phenol-sulfuric acid was optimized. A series of standard curves were drawn with glacturonic aid-glucose mixed control with different mass ratio. The content of saccharide in Codonopsis pilosula polysaccharides CPP1b samples containing galacturonic acid was determined. According to regression equation of galacturonic acid-glucose ratio of 0-100%,the correction factor was calculated by using C. pilosula polysaccharides CPP1b as the reference polysaccharide,and the results of content determination of saccharide were corrected. The rationality of this method was verified by using mixed monosaccharide control with same composition as C. pi-losula polysaccharides CPP1b. RESULTS:The correction factor of C. pilosula polysaccharide CPP1b to glucose was 3.33;in vali-dation test,the content of saccharide in mixed monosaccharide control with same composition as C. pilosula polysaccharides CPP1b was 103.47%. RSDs of precision and stability tests was <1%. The recoveries ranged 93.52%-107.35%(RSD=5.09%,n=6). CONCLUSIONS:The established method can accurately determine the content of saccharide in C. pilosula polysaccharides CPP1b containing galacturonic acid.

Objective To study the effects of different drying processes on the effective constituents in Wen Codonopsis pilosula (WPP) Decoction Pieces; To develop the optimized producing and preparing integration process based on fresh WPP.MethodsFresh WPP in harvest period was prepared respectively as follows:① Fresh WPP was dried to different percentage (30%–100%) of original moisture contents of crude drugs at 80℃ in oven, then sliced and dried at 50℃ to obtain eight decoction pieces of WPP (XⅠYP1–8).② The fresh WPP was baked to 50% of water content of crude drug under different temperatures (50–120℃), respectively, then sliced and dried at 50℃ to obtain eight decoction pieces of WPP (XⅡYP1–8).③ Dried WPP was moistened, sliced and naturally dried, then were renamed as traditional decoction pieces. The contents of lobetyolin and atractylenolideⅢ was determined by HPLC. The phenol-sulfuric acid and colorimetric method were applied respectively to detect contents of polysaccharide and total flavonoids. The contents of aqueous and alcoholic extracts were determined simultaneously. Results The contents of alcoholic extracts (55.36%), aqueous extracts (54.91%) and atractylenolideⅢ (10.95 μg/g) in XⅠYP3 pieces were higher than other decoction pieces.Conclusion The optimized process was that fresh WPP was baked to water content of 50% at 80℃, then sliced and dried. Compared with conventional preparing methods,the integration process was time-saving and effort-saving. Meanwhile, the prepared pieces have higher content of active ingredients.

Objective To explore the feasibility and efficacy of laparoscopic total mesorectal excision(TME) with preservation of anal sphincter for rectal cancer.Methods From October 2001 to March 2004,54(patients) with rectal cancer underwent laparoscopic total mesorectal excision(TME) with preservation of anal sphincter.In 51 of the 54 cases,the operation was completed laparoscopically,including 14 cases of(laparoscopic) anterior resection(AR) with the anastomosis level above the peritoneal reflection;20 cases of laparoscopic low anterior resection(LAR) with the anastomosis level more than 2cm above the dentate line;16 cases of laparoscopic ultralow anterior resection(ULAR) with the level of anastomosis within 2cm of the dentate line;and 2 cases of laparoscopic coloanal anastomosis(CAA) with the level of the anastomosis at or below the dentate line.whereas conversion to an open approach was required in three cases.Results The average operating time was 145 minutes(range 110~210min),and mean operative blood loss was 50 mL(range 30~80mL).Bowel function was restored and diet was resumed at 48 to 36 hours after operation.The average hospital stay was 9 days(range 7~14d).TME was completed successfully in 51 patients.Postoperative analgesics were used in 20 patients.No intraoperative or postoperative complications were(observed).Follow-up time was from 6~36 months in 51 patiebts,and there was no port-site or local tumor recurrence.Conclusions Laparoscopic TME is feasible and safe.It is a perspective technique with the(benefits) of minimally invasive technique and lower blood loss during operation,and rapid recovery.