Objective To explore the repetitive expressions of autophagy marker protein-rnicrotubule-associ-ated protein 1 light chain 3 (LC3) in hippocampus in newborn rats with recurrent seizure and the influence of 3-methyladeine (3-MA) on LC2 expressions. Method Seventy-two 6-day-old SD rats were randomly (random nam-ber) divided into the recurrent neonatal seizure group (RS group, n = 24), the 3-MA-treated seizure group (3-MA group, n = 24) and control group (n = 24). Rats in RS group were subjected to 55 attacks of seizure induced by flurothyl in 9 successive days from the 6th postnatal day (P6). In 3-MA group, 2 μL of 3-MA was injected every day till seizure induced. Western blot analysis was used to determine LC3 protein level in hippocampus at different intervals of 1.5 h,3 h,6 h and 24 h after the last convulsion. The LC3 protein level was analyzed with Dunnett test after ANOVA. Results LC3 protein levels in RS group at the different intervals were significantly higher than those in the control group and in 3-MA group (F =4.70,5.28,8.51 and 5.89, respectively, P <0.05), and there were no significant differences in LC3 protein level between 3-MA group and control group at those intervals (P > 0.05). Conclusions The autophagy/lysosomal pathway is immediately activated after recurrent seizure evidenced by the elevated expressions of LC3 in hippocampus. The 3-MA is involved in the regulation of autophagy/ lysosomal pathway by down-regulating the expressions of LC3.

10.3969/j.issn.1007-3969.2013.05.011

0.05). Conclusions Autop-hagy/lysosomal pathway was activated immediately after recurrent seizures as indicated by the elevated expression of Beclin1 in cerebral cortex. 3-MA was involved in the regulation of autophagy/lysosomal pathway by down-regulating the expression of Beclin1.

Objective To study the role of cutaneous nerve and protease activated receptor 2 (PAR2)in the development of pruritus in atopic dermatitis(AD).Methods Dermal sheets were prepared from chronically pruritic skin lesions of 7 patients with AD,as well as from the normal skin of 7 healthy human controls.Double labeled immunofluorescence was performed using mouse anti-protein gene product 9.5(PGP9.5)monoclonal antibody and rabbit anti-substance P(SP)polyclonal antibody to observe the morphological changes in cutaneous nerve fibers,and Image-Pro Plus 6 software was used to semiquantitively assess the length,diameter of nerve fibers,integral optimal density of PAR2 and SP in dermal sheets.Results Immunofluoresence double staining showed that PAR2 co-expressed with PGP9.5 or SP in cutaneous nerve fibers.Compared with the normal control skin,both the total length and average diameter of PGP 9.5-expressing nerve fibers were increased(11051.8±1900.9 μm vs 7264.0±2659.9 μm,4.23±0.15 μm vs 3.95±0.15 μm,both P＜0.01)in pruritic lesions,while only the average diameter of SP-expressing nerve fibers was up-regulated(3.99±0.20 μm vs 3.80±0.07 μm,P＜0.05),and the total length of them remained unchanged(4304.7±1455.0 μm vs 3380.0±1735.4 μm,P＞0.05).Also,increased integral optimal density was observed for SP and PAR in pruritic lesions in comparison with the normal control skin (27.71±16.52 vs 12.63±4.31.35.99±8.63 vs 22.69±9.56.both P＜0.05).Conclusion Our results indicate a hyper-plasia of cutaneous nerve fibers in chronic itchv skin lesions of AD and an increase in the expression of PAR2 and SP in the cutaneous nerve fibers,suggesting that the signal enhancement in PAR2 pathway may be related to the mechanism of pruritus in patients with AD.

Objective: To investigate chronic kidney disease (CKD) and its risk factors in people receiving physical examination. Methods: hTis retrospective study included people over 20 years old who had physical examination in the Health Management Center of Third Xiangya Hospital from Janurary 2008 to June 2011. CKD and its risk factors as well as questionnaire were recorded. hTe risk factors were analyzed by multivariate logistic analysis. CKD was deifned by kidney damage (microalbuminuria≥30 mg/L) and/or hematuria and/or reduced kidney function [evaluate glomerular ifltration rate (eGFR)<60mL/(min.1.73 m2)]. We counted eGFR according to the modiifcation of diet in renal disease (MDRD). Results: A total of 5 708 physical examination reports were included. The detection rate of albuminuria, reduced renal function and hematuria was 25.0%, 1.7% and 1.1%. hTe detection rate of CKD was 25.6%, and detection rate of CKD stage 1-5 was 17.8%, 6.7%, 1.1%, 0 and 0, respectively. Multivariate logistic analysis indicated that diabetes mellitus, hypertension, hypercholesterolemia, male, age, and smoking were the risk factors for CKD. Increasing physical activity was the protective factor against CKD. Conclusion: High prevalence of CKD in people receiving physical examination is found in Changsha, especially stage 1 and 2 CKD. Physical examination is important to screen CKD. Stopping smoking, control of blood glucose, blood pressure, blood lipids and increasing physical activity may help reduce the prevalence of CKD.

Objective To evaluatethe effect of mTORC1 inhibitor on the proliferation in human pancreatic neuroendocrine tumors(pNET)cell line BON,to explore the function of glutamine metabolism in it.Methods In vitro cultured human pancreatic neuroendocrine tumors(pNET)cell line BON,BON cells were treated with different concentrations of rapamycin(1,5,10,25,50, 100 nM)for 12,24 h.Then CCK-8 assay are used to calculate the growth inhibitory rate.Rapamycin treated with BON 12 h,test the glutamine uptake level compared with control.Then deprive of glucose and/or glutamine,CCK-8 assay were used in observation of cell proliferation,cell cycle distribution was analyzed by flow cytomety.Results Rapamycin significantly inhibited the growth of BON cells in a time-and dose-dependent manner(P <0.05).Meanwhile,rapamycin can reduce the glutamine uptake level compared with control.BON obviously depends on glutamine for growth,without glucose and glutamine group have obvious difference in growth rate(P <0.05).Conclusion mTORC1 inhibitor can inhibit BON cells proliferation and influence the glutamine uptake lev-el.suggesting that mTORC1 inhibitor might inhibit BON cells proliferation by influenced the glutamine metabolic pathway.

ObjectiveTo explore the molecular epidemiological characteristics of group A rotavirus diarrhea in children in Shanghai and to provide the background data for the implementation of rotavirus vaccination.MethodsA total of 910 stool samples were collected from the outpatient children with acute diarrhea from August 2008 to July 2009.Group A rotavirus was detected by usingcommercial colloidal gold device.Rotavirus strains were characterized for G and P genotypes using the nested reverse transcription polymerase chain reaction (RT-PCR).ResultsGroup A rotavirus was detected in 268(29.4％) out of 910 stool samples.Rotavirus infection was found year-round and the peak season was from October 2008 to January 2009,with the detection rates ranging from 38.3 ％ to 70.5％.Ninety-one percent of children (244 cases) with rotavirus-associated diarrhea occurred in children ＜3 years of age.The detection rate of rotavirus was highest (36.6％) in children aged 12-23 months.Among the 268 group A rotavirus-positive strains,G1 was the most common G genotype (65 strains),accounting for 24.3％,followed by G3 (40 strains,14.9％),G mixed genotypes (37strains,13.8 ％),G2 (27 strains,10.1％),G9 (14 strains,5.2％),G4(5 strains,1.9％),other G types (5 strains,1.9％),and unclassified G type (75 strains,28.0％).P[8] and P[4] were the most common P genotypes,accounting for 54.9％ (147 strains) and 11.9％ (32 strains),respectively,followed by P mixed genotypes (6 strains,2.2％) and other P genotypes (4 strains,1.5％),unclassified P type (79 strains,29.5％).The G/P genotype combinations were found as follows:G1P [8] (13.4％),G3P[8] (13.4％),GmixP[8] (10.1％),G1P[4] (8.2％),G9P[8] (2.2％),G2P [4] (1.9％),G1Pmix (1.9％).ConclusionsGroup A rotavirus is a major causative agent of diarrhea in infants and young children in Shanghai.The peak season of rotavirus infection appears in fall and winter.The currently licensed rotavirus vaccines cover the majority of genotypes of rotavirus strains prevailing in Shanghai.

Objective: To investigate the pathological changes of human hepatocellular carcinoma (HCC) after continuous passaging in nude mice. Methods: The mice model of HCC SMMU-LTMN were continuously observed for 20 years (1987-2007) . The subcutaneously transplanted carcinoma had been passaged for 228 generations. The pathological data of abdominally transplanted HCC, orthotopically transplanted HCC in nude mice, and orthotopically transplanted HCC in NOD-SCID mice were recorded. The pathological studies were conducted by light microscope, electron microscope, image analysis, chromosomal analysis, and measurement of alpha-fetoprotein (AFP) in peripheral blood. Results: (1) The local invasion and metastasis of of tumors were present in all the above 4 models for a long time. The local invasion rate and the pulmonary metastasis rate of subcutaneously transplanted tumors were 59.70% (40/67) and 37.10% (23/62), respectively. The pulmonary metastasis rate of abdominally transplanted tumors was 59.02%(36/61). The intra-hepatic and pulmonary metastasis rate of the othotopically transplanted tumors were 18.18%(4/22) and 31.82% (7/22), respectively. The pulmonary metastasis rate of HCC in NOD-SCID mice was 53.85%. (2) The tissue structure and the differentiation of the 10th generation tumor cells was similar to those of primary HCC, with grade 2 differentiation and coarse trabecular pattern as the main characteristics. From the 11th generation to the 228th generation, the main characteristics of tumor cells were grade 3 differentiation and lump pattern. Electron microscope also showed worse differentiation. (3)The AFP level was 92 500 ?g/L in cells before the 32th generation; it decreased to 6 729?g/L from the 33th-130th generation cells; and the level of the 220th generation was 1 000-5 000 ?g/L.(4)The DNA contents had a wide distribution (from 2c to 6c) in abdominally transplanted tumors and the pulmonary metastatic tumors; the mean DNA index in the former tumors (2.60?0.20) was wider than the that in the latter (2.10?0.26) . (5)From the 55th generation to 206th generation, it was found that tumor cells had integrated into the chromosome of the nude mice. Conclusion: The subcutaneously transplanted HCC in nude mice can be stably expressed for 20 years, with no change in the local invasion and metastasis ability of HCC. The differentiation of the tumor cells worsenes and the AFP level is decreased in the blood; some chromosome of tumor cells integrate into the chromosome of nude mice, which may be related to the internal environment of nude mice and the multi-potential differentiation of the tumor cells.

Objective To detect the expression of MAGE-A9 in human hepatocellular carcinoma (HCC), and to investigate the association between expression of MAGE-A9 and the clinicopathological characteristics of HCC.Methods Reverse transcription-polymerase chain reaction (RT-PCR), one-step quantitative-PCR (qPCR) and immunohistochemistry (IHC) methods were performed to characterize the expression of MAGE-A9 in HCC cell lines and tissues.Kaplan-Meier survival analysis and Cox regression were employed to evaluate the prognosis of 100 HCC patients.Results The expression of MAGE-A9 mRNA in HCC (4.44±0.342) was significantly higher than that in non-cancerous cells and tissues (1.73±0.178) (P ＜ 0.05).The expression level of the MAGE-A9 protein in HCC was related to the pathological grade (P =0.003), portal vein invasion (P =0.001), distant metastasis (P =0.022) and TNM stage (P =0.005).Cox regression analysis revealed that MAGE-A9 expression is an independent prognostic factor for disease-free survival (P =0.006) and overall survival (P =0.022).Conclusion MAGE-A9 is a valuable prognostic biomarker for HCC patients, and its high expression suggests unfavorable survival outcomes in HCC patients.

0 05) Conclusion N CTCP had a better visualization of the whole biliary tree than PTC or ERCP in patients with obstructive biliary diseases, and had similar results in identifying obstructive location and nature with that of the direct cholangiopancreatography As one of the noninvasive techniques, N CTCP provided another way to diagnose extrahepatic bile obstructive diseases and was an effective alternative to unsuccessful direct cholangiopancreatography or for those patients who were unsuitable for the direct cholangiopancreatography