Objective To explore the regulation mechanism of autophagy-related protein, microtubule-associated protein 1 light chain 3 (LC3), via c-Jun in methotrexate resistant human choriocarcinoma JEG-3 cell lines. Methods Human choriocarcinoma JEG-3 cell lines, and methotrexate resistant choriocarcinoma JEG-3 (JEG-3/MTXR) cell lines were used in our present study. Phosphorylation c-Jun (p-c-Jun) was evaluated after exposure to 0.02 ng/ml methotrexate for 72 hours in both cells by western blot. c-Jun gene was knockdown by small interference RNA (siRNA) in JEG-3/MTXR cells, and LC3 was evaluated by western blot and reverse transcription-PCR. The binding of LC3 promoter with c-Jun protein was detected via chromatin immunoprecipitation assay (ChIP) with or without 0.02 ng/ml methotrexate exposure. Results The results showed that p-c-Jun was up-regulated after methotrexate treatment for 72 hours (1.99±0.20, versus 0.20±0.06 at 0 hour;P<0.05) by western blot analysis in JEG-3/MTXR cell lines. Further investigation demonstrated that c-Jun-siRNA could inhibit the up-regulation of LC3 formation and after methotrexate exposure (LC3 mRNA:1.24±0.17 versus 3.03±0.43;LC3 protein:0.52±0.07 verus 1.20± 0.15; all P<0.05). The binding of LC3 promoter by c-Jun protein was up-regulated after methotrexate treatment by the method of ChIP in methotrexate resistant JEG-3/MTXR cells [(2.95 ± 0.35) times]. Conclusion Autophagy-related gene LC3 expression regulated by c-Jun protein may be involved in the effect mechanism of the development of methotrexate resistance in choriocarcinoma JEG-3 cells.

Objective The purpose of this study was to investigate the reproductive characteristics of indoor-housed Meriones meridianus.Methods According to the data of Meriones meridianus from 1996 to 2002 in the Center for Laboratory Animal research of Xinjiang, the sexual maturity period of male and female animals, the gestation period of female animals, the litter size, weaning rate, survival rate, sex ratio per month and different fetal times were statistically analyzed.Results Sexual maturity period of male and female animals was 109.3 ±21.0 days and 106.3 ±21.7 days, and gestation period of female animals was 21.3 ±1.4 days.The litter size, weaning rate of different fetal times showed no significant difference compared between those of the first to seventh litter ( P>0.05) , and the survival rate of fourth and sixth litter were lower than that of the average offspring per litter, but the difference was no significant (P>0.05).The sex ratio of from first to sixth litter indicated no significant difference ( P>0.05 ) , and the average proportion of male and female was 1.4:1.0.During a year, the animals almost stopped to reproduce from September to November, however, the differences of litter size between the other months were statistically not significant ( P >0.05 ) , the weaning rate and survival rate per month suggested a significant difference ( P <0.05 ) between some of the months.Conclusions Compared with the background data of wild Meriones meridianus, the laboratory reproduction of Meriones meridianus show some differences, mainly, the season of breeding is shifted to December, and the number of reproduction increased by one or two litters.Our results provide useful reference for laboratory animalization of Meriones meridianus.

Objective To study the relationship between p16,p27 proteins and clinicopathological characteristics of breast cancer and their prognostic value.Methods p16 protein and p27 protein were detected by two step immunohistochemical method in 93 tissues of breast cancer.Results The positive rates of p16 protein and p27 protein were 46.2 ％ (43/93) and 54.8 ％ (51/93).The positive rate of p16 protein in different tumor location (left/right),the maximal diameter and histological type had no statistically significant difference (all P ＞ 0.05),but in the different age groups and axillary lymph node metastasis,the differences were statistically significant (both P ＜ 0.05).The positive rate of p27 protein in different age groups,tumor location (left/right) and the maximal diameter had no significant difference (all P ＞ 0.05),but was correlated with the infiltration and axillary lymph node metastasis (both P ＜ 0.05).Conclusions The negative expression of p16 and p27 is significantly related with the biological behaviors of breast cancer.Detection of both p16 protein and p27 protein may play an important role in predicting the prognosis of breast cancer.

Objective To explore the lymph nodes (LN) metastasis characters of the endometrial carcinoma and its relation with the patients' prognosis. Methods A retrospective study was carried out on 227 cases of endometrial carcinoma who admitted to our department and underwent LN excision from Jul. 2000 to Feb. 2008. Results Among 227 cases who underwent pelvic LN excision, there were 22 cases (9.7%) presented LN metastasis. There were 12 cases with positive external iliac LN from 20 cases of patients with data in LN grouping. Para-aortic LN excision was carried out on 138 patients. There were 6 cases with positive para-aortic LN, 5 cases of them together with pelvic LN metastasis. Those patients with cervix involvement, annex metastasis, deep myometrium infiltration, grade 2-3 and negative estrogen receptor occurred pelvic LN metastasis more frequently than the others ( P ＜ 0. 05 ). Among the 6 cases with positive para-aortic LN, there were 3 cases ( 3/6) with deep myometrium infiltration. For those whose paraaortic LN was negative, it was only 16. 7% (22 cases). But there were no difference statistically between them ( P＞ 0. 05 ). There were significant difference in 3 years disease-free survival rate between patients with positive pelvic LN or negative pelvic LN [(81. 8 ± 8. 2)% vs ( 97. 4 ± 1. 2 ) % , P = 0. 004]. While there were not significant difference in 3 years disease-free survival rate between patients with positive para-aortic LN or negative para-aortic LN [100% vs ( 96. 7 ± 1. 6) % , P ＞ 0. 05]. Single factor analysis showed that the age more than 50 years, annex metastasis and pelvic LN metastasis related with the recurrence (P ＜0. 01). But cervix involvement, deep myometrium infiltration, para-aortic LN metastasis, pathology type, tumor grade and estrogen receptor did not relate with the recurrence ( P ＞ 0. 05 ). Cox regression analysis showed that annex metastasis and the age of patients were independent risk factors affecting the recurrence ( P = 0. 011, P = 0. 025 ). Conclusions The most common site of pelvic LN metastasis is the external iliac LN for endometrial carcinoma patients. The patients with positive para-aortic LN always accompanied pelvic LN metastasis. Those patients with cervical involvement, annex metastasis, deep myometrium infiltration, poor differentiation and negative estrogen receptor be more likely exist pelvic LN metastasis. Pelvic LN metastasis may affect the prognosis of endometrial carcinoma patients.

Objective To investigate the relationship between apolipoprotein E(Apo E) gene poly-morphism and mild cognitive impairment (MCI) in patients with type 2 diabetes mellitus (T2DM), and e-valuate the correlative risk factors. Method 40 cases of type 2 diabetes with MCI and 80 cases of type 2 diabetes without MCI were enrolled in this study. The polymorphism of the Apo E gene was detected by PCR-restriction fragment length polymorphism(PCR-RFLP). According to the clinical data such as course of disease, plasma glucose, plasma fat and body mass index (BMI), the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression. Results The frequency of Apo E ε_4 allele in the group of type 2 diabetes with MCI was higher than that without MCI ( 25.0% vs 10. 0% ), and the difference had statistical significance( P < 0. 01 ). The indexes of the statistical significant difference be-twcen the two groups were age, course of disease, postprandial blood glucose ( P2BG), HBA1C, BMI,family history of T2DM, hypertension, diabetic retinopathy, diabetic peripheral neuropathy, Apo E gene. The independent risk factors included diabetic retinopathy ( OR = 3. 452, P < 0. 05 ), diabetic peripheral neuropathy( OR = 3. 252, P <0. 05), Ape E gene( OR = 2. 441, P < 0.01 ), HBA1C ( OR = 1. 372, P <0.05), P2BG(OR = 1. 194, P <0.05), age(OR = 1. 194, P <0.01) and course of disease(OR =1. 142, P <0. 05). Conclusion Apo E ε_4 allele has significant relationship with T2DM and MCI. The age, course of disease, control of plasma glucose, and microvascular complication of diabetes have relation-ship with the cognitive function.

Objective To investigate the changes of serum apolipoprotein A-I and its clinical significance to acute respiratory infection.Method Totally 44 patients with acute respiratory infection were divided into three groups according to various concentration of Serum apolipoprotein A-I.They were procaleitonin(PCT)＜0.5 ng/ml group,0.5 ng/ml≤PCT＜2 ng/ml group and PCT≥2 ng/ml group.We measured apolipoprotein A-I,C-reactive protein,procalcitonin and albumin within 24 hours after admission.Results With the increase of serumPCT,the production of ApoA-I and albumin were down-regulated,while CRP up-regulated.Conclusions Apolipoprotein A-I has a sound relationship with the acute respiratory infection.It can be used as one of the diagnostic criteria in severe infection patients who have disorders of lipometabolism.

s:The partical coding sequence of E2 gene of 13 Hog Cholera Virus(HCV) field isolates, Shimen strain, Chinese vaccine strain(C strain) and Thiverval strain attenuated by low temperature in France,were obtained by reverse trancriptase -polymerse chain reation (RT-PCR) and sequenced.All size were 251bp.The obtained 224bp sequences were analysed by DNA star and compared with the previously published sequences of Alfort strain ,Brescia strain and other references strains.The results showed that those sequencing fragments of 13 HCV field strains were the sequence of E2 gene of HCV.Compared with Shimen strain,the base substitute of all stains were randomly distributed in the entire sequence,and had not base insert and base gap.The variation most occurred at 3' end. The identity of nucleotide sequence and amino acid sequenceof 22 HCV strains were 78.1%～100%?78.4%～100%. The identity of nucleotide sequence and amino acid sequence of 13 HCV field strains were 78.1%～100%?78.4%～100%.The identity of nucleotide sequence and amino acid sequence of 4 HCV field strains isolated in the 1970s～1980s were 79.0%～88.3%?81.1%～87.8%.The identity of nucleotide sequence and amino acid sequence of 9 HCV field strains isolated in the 1990s were 80.8%～100%?83.8%～100% respectively. This paper showed that the genetic variation of HCV was diversity.

Objective To analyze the clinical features of infantile cytomegalovirus(CMV) hepatitis with cholestasis and investigate intrahepatic cholestasis due to hepatocytic impairment caused by CMV infection.Methods Forty-seven children with CMV cholestatic he-patitis were divided into 2 groups according to the level of total bilirubin(TB):22 cases with serum TB lower than 136.8 ?mol/L(groupⅠ),and 25 cases with serum TB higher than 136.8 ?mol/L(groupⅡ).All children were treated with both gangciclovir and routine met-hods,and serum biochemistry were checked before and after treatment.SPSS 13.0 software was used to analyze the data.Results Forty-seven cases of infantile CMV cholestatic hepatitis had different degrees of jaundice,hepatosplenomegaly and abnormal liver functions.The differences of serum ALT and AST between the 2 groups had statistical significance,the levels of serum gamma glutamy transferase(GGT) and alkaline phosphatase(ALP) were lightly higher in groupⅡcompared with those in groupⅠ,but there were no statistical significance.TB,direct bilirubin(DB),ALT and AST were decreased in the 2 groups after treatment,GGT and ALP hadn′t decreased significantly after treatment.Conclusions CMV infection can injure hepatocytes and epithelials on each grade of bile duct,thus CMV hepatitis causes intrahepatic cholestasis.Cholestasis due to hepatocytic impairment deserves emphasis and intervention should be done as early as possible.Gangciclovir therapy for CMV infection manifest effective and safe in short-term.

Objective In order to lay the foundation for establishment of a clean grade Meriones meridianus popu-lation, and provide reference for the formulation of Meriones meridianus parasite detection standard.Methods A survey of parasite infection in Meriones meridianus was carried out, and three kinds of antiparasitic drugs, tetramethrin/per-methrin, praziquantel, and tinidazole were used in high-, medium-and low-doses for eradication of the target parasites. Results Three types of parasites were detected among the animals.The infection rate of surface parasite Myobia musculi was 82.1%(23/28), that of intestinal parasite Hymenolepis nana was 78.1% (25/32) and Tritrichomonas muris was 90.6%(29/32).Significant results of parasite eradication were achieved with the antiparasitic drugs.Medicated bath u-sing 1%tetramethrin/permethrin for 30 s every 4 days for 5 times and 1.5%tetramethrin/permethrin for 30 s every 4 days for 3 times completely eradicated the surface Myobia musculi infection.Praziquantel in a dose of 31 mg/kg by daily gastric gavage for consecutive 7 days was the optimal dosage to completely eradicate the intestinal Hymenolepis nana.Tinidazole in a dose of 42 mg/kg administered by gastric gavage daily for consecutive 7 days was the optimal dosage for eradication of in-testinal Tritrichomonas muris infection.Conclusions Three types of parasites are detected in Meriones meridianus in our laboratory, i.e.surface infection with Myobia musculi, and intestinal infection with Tritrichomonas muris and Hymenolepis nana.Tetramethrin/permethrin, praziquantel and tinidazole can be used to effectively eradicate these three types of para-sites, respectively.

Objective:To investigate the relationship between tumor necrosis factor-α (TNF-α)-308A/G gene polymorphism and mild cognitive impairment(MCI)in patients with type 2 diabetes mellitus(T2DM),and their correlative risk factors thereof.Methods:Forty cases of T2DM with MCI and 80 cases of T2DM without MCI were selected for this study.The polymorphism of the TNF-α-308A/G was detected by PCR-restriction fragment length polymorphism (PCR-RFLP).According to the clinical data,such as course of disease,plasma glucose,plasma fat and body mass index(BMI),the independent risk factors of T2DM and MCI were analyzed by non-conditional logistic regression.Results:The frequency of TNF-α2 allele was significantly higher in the group of T2DM with MCI than that without MCI (P＜0.01).The indexes of the statistical significant difference between the two groups were the age,course of disease,postprandial blood glucose(P2BG),glycosylated hemoglobin,body mass index,family history of T2DM,hypertension,diabetic retinopathy,diabetic peripheral neuropathy and TNF-α.The independent risk factors included TNF-α,diabetic peripheral neuropathy,diabetic retinopathy,age and P2BG.Conclusion:There is a significant relationship between TNF-α2 allele and T2DM with MCI.There is a significant relationship between the age,control of plasma glucose and microvaseular complication of T2DM with the cognitive funotion.