[ ABSTRACT] AIM:To study the influence of insulin resistance on fatty liver in the mice fed with high-fat diet (HFD).METHODS:Male 8-week-old C57BL/6J mice were randomly divided into HFD group (with 60% calories by high saturated fatty acid) and control group (with chow diet).The mice in both groups were fed for 12 weeks.The body weight, liver weight, serum triglyceride (TG) and total cholesterol (TC), and blood glucose and insulin levels were meas-ured.Hyperinsulinemic euglycemic clamp experiment was applied to reflect insulin sensitivity .The lipid deposition in the liver was analyzed by HE staining , Sudan IV staining and measurement of liver fat content .The phosphorylation levels of IRS1 and Akt, and the protein levels of SREBP-1 and FAS were determined by Western blot to reflect the activities of insu-lin signaling and lipid synthesis .RESULTS:Compared with control group , the body weight and liver weight were signifi-cantly increased in HFD group .TG and TC contents in serum and liver tissues were remarkably increased in HFD group . High-fat diet induced insulin resistance , as evidenced by increased serum insulin levels , reduced glucose infusion rate and decreases in IRS1 and Akt phosphorylation levels .In livers of HFD group, HE staining showed that the cytoplasm of hepa-tocytes was filled with vacuoles .Sudan IV staining also displayed that many different sizes of red lipid drops existed in the hepatocytes , and the protein levels of SREBP-1 and FAS were significantly increased .In primary normal hepatocytes with exogenous oleic acid intervention for 48 h, the phosphorylation levels of IRS 1 and Akt were reduced , and the protein ex-pression of SREBP-1 and FAS was significantly increased in a dose-dependent manner .CONCLUSION: Feeding with HFD leads to insulin resistance , resulting in activation of lipid synthesis and accumulation of lipid deposition in the liver , thus inducing fatty liver .

Objective To investigate the difference of leucorrhea routine tests results among 3 periods of pregnancy in pregnant women.Methods Vaginal cleanliness,the presence of clue cells,trichomonas and fungi were tested for 3 103 obstetric outpatients who were pregnant and treated in the hospital.The test results were analyzed and compared.Results Among pregnant women in different periods of pregnancy,the difference in the proportion of vaginal cleanliness degree Ⅲ and fungal infection rates were statis-tically significant(P <0.05).Among pregnant women of different pregnant periods with vaginal cleanliness degree Ⅱ,difference of fungi infection rate was statistically significant(P <0.05).Among pregnant women of different pregnant periods with vaginal clean-liness degree Ⅳ,the difference of positive rates of fungi,trichomonas and clue cells were statistically significant(P <0.05).In addi-tion,there are a number of mixed infections.Conclusion Vaginitis during pregnancy should be early detected,diagnosed and trea-ted.

Objective Detect the infection rate of herpes simplex virus (HSV ) by jointly using chemiluminescence assay and PCR ,and provides reference for clinical diagnosis .Methods The serum samples were collected from the pregnant women who had routine examination records in the hospital .Chemiluminescence assay was used to detect HSV IgM and IgG in those samples .Cervi‐cal secretions were collected from pregnant women with positive results and qualitatively tested for HSV DNA .Results The posi‐tive rate of HSV1 DNA was 0 .5% (7/1 422) ,the positive rate of HSV2 DNA was 1 .1% (16/1 422) .For pregnant women whose HSV IgM and IgG were both positive ,positive rate of HSV1 DNA was 0 .4% (4/1 008) and that of HSV2 DNA was 0 .6%(6/1 008);for those who only had HSV IgM positive ,the positive rate of HSV1 DNA was 0 .8% (1/130) ,and that of HSV2 DNA was 3 .1% (4/130);for those who only had HSV IgG positive ,the positive rate of HSV1 DNA was 0 .7% (2/284) ,that of HSV2 DNA was 2 .1% (6/284) .Among those three HSV antibody positive cases ,the difference in HSV1 DNA positive rate was not sta‐tistically significant(P>0 .05) ,while the difference in HSV2 DNA positive rate was statistically significant(P<0 .05) .Conclusion The test of HSV antibodies during pregnancy can be used as a routine test ,and HSV DNA test can be used as further test for those with HSV antibody positive ,which could improve the accuracy of diagnosis .Early screening ,detection ,and treatment are im‐portant for pregnant women with HSV infection .

AIM:To investigate the influence of calorie restriction ( CR) on contractility and mitochondrial bi-osynthesis in different types of rat skeletal muscles .METHODS:CR rat model was set up by feeding 60%normal food in-take of control rat every day for 8 weeks.Soleus (SOL) and extensor digitorum longus (EDL) were isolated under anesthe-sia.The twitch tension, titanic tension and fatigue resistance of SOL and EDL in response to electrical stimulation were measured to reflect the contractile function of the muscles .The copy number ratio of mitochondrial gene cytochrome C oxi-dase subunit I ( COX I) to nuclear gene β-actin was determined to evaluate the mitochondrial biosynthesis .ATP content was measured to mirror mitochondrial function .RESULTS:Compared with control group , CR for 8 weeks significantly in-creased twitch tension and titanic tension of both SOL and EDL , but only improved fatigue resistance in SOL .Markedly in-crease in ATP content in both skeletal muscles by CR intervention was observed , especially in SOL .Although CR activated AMP-activated protein kinase (AMPK) in both 2 muscles, up-regulation of mitochondrial biosynthesis and transcription of mitochondrial regulatory genes peroxisome proliferator-activated receptor γcoactivator 1α( PGC-1α) and nuclear respirato-ry factor ( NRF) was only observed in SOL .CONCLUSION:CR for 8 weeks enhanced the contractility of both rat SOL and EDL in response to electrical stimulation , especially in SOL composed of slow-twitch fibers.The mechanisms may be related to the activation of AMPK and the promotion of mitochondrial biosynthesis in SOL .

Objective:To observe the expression of Ki-67,P53 and LAT1 in tissues of esophageal squamous cell carcinoma and precancerous lesions and to investigate its clinical significance.Methods: Immunohistochemical method was used to detect the expressions of Ki-67,P53 and LAT1 in tissues from 20 cases of normal esophageal mucosa,68 cases of precancerous lesions including 21 cases of mild atypical hyperplasia,22 cases of atypical hyperplasia,25 cases of severe atypical hyperplasia and cancer tissues from 34 esophageal cancer patients.The relationship between the expression of the three in esophageal carcinoma was analysed.Results: Re-spectively,the positive expression rate of Ki-67 in normal esophageal mucosa, mild atypical hyperplasia, moderate dysplasia, severe dysplasia and carcinoma was 0%( 0/20 ) , 23.8%( 5/21 ) , 40.9%( 9/22 ) , 76.0%( 19/25 ) , 82.4%( 28/34 ) and the positive expression rate of P53 was 0%(0/20),14.3%(3/21),31.8%(7/22),48.0%(12/25),67.6%(23/34) and the positive expression rate of LAT1 was 0%( 0/20 ) , 19.0%( 4/21 ) , 36.4%( 8/22 ) , 52.0%( 13/25 ) , 76.5%( 26/34 ).The rank correlation analysis showed that the positive expression of Ki-67,P53 and LAT1 were significantly correlated with histological grade(r=0.626,0.427, 0.586,P<0.01) and the expression of Ki-67,P53 was positively correlated with LAT1 in esophageal carcinoma tissues(r=0.428, 0.596,P<0.01).Conclusion:Abnormal expression of Ki-67,P53,LAT1 protein was significant related to carcinogenesis of esophageal cancer and the combined detection of the three has important clinical significance.

Objective To discover new method studying renal deficiency and syndrome of blood stasis by gemellary research method.Methods Through clinical investigation of 300 instances of twins,we choice three twins,include mistress Yang from Qionglai city who is being later stage of older;mistress Yu from Chengdu city who is being presenium;mistress Lu from Beijing who is being Middle-age using five species of ovi-type appraisement methods,and syndrome-penthemeron investigation by blood stasis,stagnation of blood measuring scale,renal deficiency measuring scale,and eight principles measuring scale,in this text.Results(1)Get three family constellation graphs,which provide inspiration for syndrome-penthemeron studing.(2)Though investigating to three twins,we discover that this relative value of eight principles measuring scale has augmented tendency with time-lapse.3.There haven’t regularity from the stagnation of blood measuring scale and renal deficiency measuring scale.Conclusion Using gemellary research method to study Diabetes Mellitus is perfect.In spite of there haven’t ideal result,but it provid prospective thinking and suggestion for us.

To cultivate high-quality medical talents,Shanghai Jiao Tong University School of Medicine established a multidisciplinary and comprehensive basic medical practical integration curriculum Medical Functional Experiment.Over several years' exploration and renovation,insisting on student-centered,this course has established a modular experiment teaching system with emphasis on foundation,combina-tion with clinical and focusing on innovation.Integration curriculum adopts four-stage teaching pattern which is suitable for medical students,and implements diversified teaching method and evaluation system.And meanwhile,the paper explores the ideas and approaches to further deepen the construction of teaching staff,expand experimental teaching techniques,strengthen the construction of curriculum materials,and strengthen the cultivation of teachers' innovative ability.

Objective To compare the effect of kidney-tonifying blood-activating recipe (KBR) and Aescuven Forte Tablets ( AFT) in improving the sperm quality of varicocele-induced male sterility, thus to optimize the therapeutic therapy for varicocele-induced male sterility. Methods A total of 102 varicocele-induced male sterility with abnormal sperm parameters after conservative treatment were randomized into KBR group (N=53) and AFT group ( N=49) . KBR group was given KBR plus natural vitamin E and AFT group was given AFT plus natural vitamin E, and the treatment lasted for 8 continuous weeks. Before and after treatment, the quality of seminal fluid was analyzed, sperm quantization parameters such as total number of sperm (TNS) , total number of progressive motility sperm ( TNPS) , total number of normal form sperm ( TNNS) and total number of nor mal form and progressive motility sperm ( TNNPS) were observed, and the improvement rate of sperm quantization parameter was compared. Results (1) Before treatment, the differences of TNS, TNPS, TNNS and TNNPS were insignificant between the two groups ( P>0.05) . After treatment, TNNS was not improved in AFT group ( P>0.05) , but TNS, TNPS, TNNPS were much improved in both groups ( P<0.01 compared with those before treatment) . The improvement of KBR group was superior to that of AFT group ( P<0.05) . ( 2) The improvement rate for TNS, TNPS, TNNS, TNNPS was 90.57%, 79.25%, 67.92%, 77.36%in KBR group, and was 75.51%, 73.47%, 28.57%, 61.22% in AFT group respectively. The improvement rate for TNS and TNNS in KBR group was superior to that in AFT group ( P<0.05 or P<0.01) . Conclusion Varicocele-induced male sterility patients usually have the syndrome of kidney deficiency and blood stasis, so KBR, which has the function of tonifying kidney and activating blood, has synergistic action on the effect of AFT in improving sperm quality of varicocele-induced male sterility patients.

Aim This study was aimed to explore the influence and mechanism of the long-term exercise on skeletal muscle contraction and mitochondrial biosyn-thesis in different muscle fibers.Methods Soleus (SOL)and extensor digitorum longus (EDL)were i-solated from SD male rats with platform running train-ing for eight weeks.The changes of contractility under different electrical stimulation were observed, mito-chondrial biosynthesis,including ATP content,mito-chondrial DNA,the gene expression of PGC-1αand NRF were also detected.Results Long-term endur-ance exercise can improve twitch tension and titanic tension of SOL and EDL ,but only enhanced the fa-tigue resistance in SOL.ATP contents were significant-ly increased in the two types of muscles,but mtDNA content,PGC-1αexpression and NRF translation were only obviously enhanced in SOL,in accompanied with an increase in p-AMPK/AMPK protein ratio.Conclu-sion Long-term endurance exercise increased skeletal muscle contractility and improved the anti-fatigue abili-ty in SOL,which may be associated with increase in mitochondrial biosynthesis via activated AMPK-PGC-1αaxis.

AIM:To clone the SPA gene promoter and construct its luciferase report vector of SPA gene and to study its transcriptional targeting activity.METHODS:① The SPA gene sequence was acquired from GenBank,of which the upstream was analyzed according to bioinformatics.The results showed that the upstream region of SPA gene sequence about 163bp has the function of promoter.② The SPA gene promoter fragment was generated by polymerase chain reaction and then subcloned into the multiple clone site(MCS)of luciferase report gene vector pGL3-basic to generate the recombined plasmid pGL3-SPA.This fragment was also subcloned into pGL3-control to generate recombined plasmid pGL3-SPA-enhancer by replacing its primary SV40 promoter.③ pGL3-SPA,pGL3-SPA-enhancer,pGL3-control,pGL3-basic were cotransfected with pRL-TK into A549 cells and H441 cells.The luciferase activities were measured by dual luciferase reportor(DLR)system.RESULTS:Sequencing and restricted digestive results showed that SPA gene promoter was successfully cloned and identified,and also correctly subcloned into plasmid pGL3-basic and pGL3-control to construct its luciferase report plasmid pGL3-SPA and pGL3-SPA-enhancer,respectively.The transcriptional activity was high in H441 cells.CONCLUSION:The luciferase report system of SPA gene promoter is successfully constructed with high transcriptional activity.