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 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.

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.

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 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 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 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 expression of MUC-1 and CD44v6 in breast carcinoma and its diagnostic value.Methods 93 cases of breast carcinoma were examined by immunohistochemical method for MUC-1 and CD44v6.Results The positive rates of MUC-1 and CD44v6 were 71.0 ％ (66/93) and 63.4 ％ (59/93),respectively.The expression of MUC-1 in breast carcinoma was correlated with histologic differentiation (14.3 ％,69.4 ％,100.0 ％) and lymph node metastasis (22.9 ％,100.0 ％) (x2 =36.147,63.047,both P ＜ 0.0001),but had no relationship with the type of breast carcinoma,tumor clinical stages,and tumor size (all P ＞ 0.05).The expression of CD44v6 in breast carcinoma was correlated with tumor clinical stages (6.3 ％,64.6 ％,93.1％) and lymphnode metastasis (20.0 ％,89.7 ％) (x2 =9.507,45.662,both P＜0.05),but had no relationship with tumor type,tumor clinical stages and tumor size (all P ＞ 0.05).Conclusion MUC-1 and CD44v6 may play important roles in the development of breast carcinoma and the biologic behavior of breast carcinoma may be closely related with over expression of MUC-1 and CD44v6 protein.MUC-1 and CD44v6 can be used to predict the prognosis and direct the treatment of breast carcinoma.

Objective To investigate the methods of reducing radiation dose in CT coronary angiography through optimizing individualized scan dosage protocol.Methods Two hundred patients (group A)underwent coronary CTA examination which was performed with fixed 120 kV and variable mA according to their BMI.The mA was set as 150-300 mA(BMI ＜ 18.5 kg/m2),300-500 mA (18.5 kg/m2 ≤ BMI ＜ 25.0 kg/m2),and 500-800 mA(BMI ≥ 25.0 kg/m2).When all examinations were finished,a linear regression was employed to analyze the correlation between mA and BMI,body surface(Suf),image noise(SD)respectively.The results of the analysis were used to formulate a regression equation,which was further used to establish a table list for quick search on how much mA that individualized coronary CTA scan would need.Another 200 patients(group B)enrolled for the individualized scan were scanned under new protocol that previous study established.The tube voltage was 100 and 120 kV.The tube current was variable according to the data in the table list.One-way ANOVA and Kruskal-wallis H test were used for statistics.Results Regression equation between mA and BMI,Suf,SD was:mA =17.984 × BMI + 169.149 × Suf-2.282 × SD-361.039.The SD(group A:32.08 ± 5.80,group B:28.60±4.47),dose index volume(CTDIvol)[group A:(41.97 ± 11.37)mGy,group B:(33.18±10.07)mGy],effective dose(ED)[group A:(10.91 ±3.07)mSy,group B:(8.83 ±2.72)mSv]had significant differences between the two groups(F =43.45,63.71,49.07 respectively,P ＜0.01 for all).The SD and ED results obtained in group B were better than those in group A.Conclusion Better performances were obtained when BMI combined Suf was used as a new individualized protocol than when BMI was used only,which means good image quality and lower radiation dosage in coronary CTA examination.

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.