Objective:To analyze the causal relationship between lifestyle-based factors and the occurrence and development of hepatobiliary malignancies by Mendelian randomization study method,and to provide the potential clinical evidence for the prevention and treatment of hepatobiliary malignancies.Methods:The data from large-scale,independent genome-wide association studies(GWAS)were selected,and seven-step inclusion criteria for the instrumental variable screening were set up.The exposure lifestyles included the percentage of carbohydrate intake,percentage of fat intake,percentage of protein intake in the diet,coffee intake,weekly alcohol consumption times,leisure electronic screen exposure time,moderate to vigorous intensity physical activity(MVPA)during leisure time,sedentary behavior at work,age at first smoking,daily smoking quantity,current smoking status,and past smoking status,totaling 12 phenotypes.The primary analysis method used was the random effect model of the inverse variance weighted(IVW)method,and the heterogeneity was detected by Cochrane's Q test and the horizontal pleiotropy was detected by MR-Egger intercept method.Results:The current smoking status was significantly positively correlated with the increasing risk of extrahepatic cholangiocarcinoma(OR=1.607,95%CI:1.113-2.322,P=0.011).Higher coffee intake was causally linked to a higher risk of liver cancer and intrahepatic cholangiocarcinoma(OR=1.000,95%CI:0.999-1.000,P=0.012).In the physical activity,more MVPA was associated with the lower risk of liver cancer and intrahepatic cholangiocarcinoma(OR=0.998,95%CI:0.996-0.999,P=0.002).The Cochrane's Q test results showed that there was mild heterogeneity between MVPA and extrahepatic cholangiocarcinoma(Q=18.354,P=0.049)as well as the percentage of protein intake and intraphepatic cholangiocarainoma(Q=12.715,P=0.026),and the MR-Egger intercept method results showed there was no horizontal pleiotropy.Conclusion:There is a causal relationship between current smoking status and extrahepatic cholangiocarcinoma,and there is a causal relationship between more MVPA and the lower risk of liver cancer and intrahepatic cholangiocarcinoma.Education on smoking and physical activity for the patients may offer potential benefits for the prevention of hepatobiliary malignancies.

Objective To Establish a loop-mediated isothermal amplification(LAMP)assay for detection of Salmonella in fecal samples of tree shrews, and report the result of preliminary application of this method. Methods LAMP primers were designed and synthesized according to the conserved sequence of Salmonella specific gene invA (invasive protein gene A). To optimize the reaction time and temperature by setting 10 reaction times(24 to 42 min)and temperature(57℃ to 66℃)and tested its specificity and sensitivity. At the same time, a conventional PCR test was performed to verify and compare with the LAMP assay. 91 fecal samples of wild-derived tree shrews were detected by the LAMP assay. Results The experimental condition was confirmed as 62℃ and 34 min. The sensitivity of Salmonella was 3.36×101CFU/mL, which was 10 to 100 times higher than that of conventional PCR assay. In 10 kinds of intestinal bacteria for LAMP amplification,Salmonella enteritidis and Salmonella paratyphi B were positive,the others were negative. Among the 91 samples of tree shrew fecal samples detected by the LAMP assay, the positive detection rate was 20.88%. The LAMP assay can be completed within 40 min, the result can be observed and judged visually by color changes. Conclusions The LAMP assay established in this study is convenient,rapid,sensitive and specific. It can be used as a rapid measure for large-scale detection of Salmonella in feces of tree shrews.

Objective To establish a quick and accurate method for detection of tree shrew adenovirus(TAV) using TaqMan real-time fluorescence quantitative PCR. Methods Based on the published TAV genome sequence, a 3' conserved sequence was used to design specific probe primers. A standard curve was prepared using a recombinant plasmid containing the target gene fragment. A real-time fluorescence quantitative PCR method was established for detecting TAV based on TaqMan probe. Results The detection method was specific and was not cross-reactive with other common pathogens. The detection limit of the method was 3.7 copies/μL,showing a high sensitivity. The correlation coefficient was 0.998, and the efficiency was 95.7%. The amplification result showed a fine linear relationship,and the repeatability test effect was good. Conclusions The TAV real-time quantitative PCR detection method based on TaqMan probe has been successfully established. It has high sensitivity and reproducibility and can be applied to early detection of TAV infection.

Parkinson's disease(PD)is a progressive neurodegenerative disorder, with an etiology that is now considered to be due to interaction between genetic and environmental factors. Typical PD features include loss of dopaminergic neurons in the nigrostriatal region, with typical motor traits of PD associated with dopamine deficiency. Animal models have contributed to determining PD etiology and pathogenesis,as well as testing new therapeutic schedules and novel drug research. Rodents, tree shrews, primates, and other animal models of PD have been established by different method. These models each have their own advantages and limitations, showing different clinical features and pathological mechanisms to those in humans. Therefore, the appropriate model for scientific research must be carefully considered. This article reviews the main neurotoxic and transgenic models of PD.

Objective To evaluate the gene amplification and protein expression of HER-2 in gastric carcinoma with or without neuroendocrine differentiation (NED), and to explore the difference in HER-2 gene profile between these two neo?plasms. Methods Cases of gastric carcinoma with NED (n=70) and cases of gastric carcinoma without NED (n=150) were retrospectively reviewed. Gene amplification and protein expression of HER-2 genes in gastric carcinoma with or without NED were detected by combination of immunohistochemical method (IHC) and double color silver dye for in situ hybridiza?tion (DISH). Prognosis of gastric cancer patients with NED were predicted using Kaplan-Meiers survival analysis. Results Expression rates of HER-2 in gastric cancer with and without NED are 20.00%and 21.33%respectively. Amplification of HER-2 in gastric cancer with and without NED are 8.57%and 14.67%respectively. Gene amplification and protein expres?sion of HER-2 between gastric cancer with NED and without NED showed no statistical difference. Chromosome 17 multi-body positively correlated with gene HER-2 amplification in Gastric carcinoma with NED. Postoperative survival period in patients of gastric carcinoma with NED and HER-2 amplification was shorter than that in patients of gastric carcinoma with?out NED but with HER-2 amplification. Gastric carcinoma with or without NED, HER-2 gene amplification, lymph node me?tastasis and operation method obviously correlate with prognosis of gastric carcinoma patients (P<0.05). Conclusion The gastric cancer with NED is a special type of gastric cancer, there was no difference of gene amplification and protein expres?sion of HER-2 gene between gastric carcinoma with NED and without NED. Poor prognosis would be expected in gastric can?cer patients with NED and HER-2 amplification.

OBJECTIVETo investigate risk factors associated with lymph node metastasis and prognosis of rectal neuroendocrine tumor (NET).

METHODSClinicopathological data of 69 patients with rectal NET in our department from April 2003 to October 2011 were retrospectively analyzed. Associations of clinicopathological factors with lymph node metastasis and prognosis were examined using univariate and multivariate analysis.

RESULTSOf the 69 patients, 9 cases had lymph node metastasis. The lymph node metastasis was significantly associated with tumor size, T stage and G grade by univariate analysis. Multivariate analysis showed that T stage was the only risk factor associated with lymph node metastasis. The overall 5-year survival rate was 90.3%. Prognosis of rectal NET was significantly associated with tumor size, T stage, N stage, M stage, TNM stage and G grade by univariate analysis. Multivariate analysis showed that M stage was significantly associated with long-term survival in rectal NET patients (P=0.000, HR=2.285, 95%CI:1.484~3.518). There was no significant difference in patients with stage I between local and radical resection, while there were significant differences in those with stage II or higher between the two operations (P=0.046).

CONCLUSIONT stage is associated with lymph node metastasis and both TNM stage and M stage can affect the prognosis of patients with NET, which may be used as potential predictive factors for rectal NET. Local resection should be recommended for patients with stage I and radical resection should be recommended for patients with stage II or higher.

Adult ; Aged ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; diagnosis ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Neuroendocrine Tumors ; diagnosis ; pathology ; Prognosis ; Rectal Neoplasms ; diagnosis ; pathology

Objective To analyze the restricted usage of TCRβ V/J subfamily and CDR3 repertoire in patients with peripheral T-cell lymphoma unspecified (U-PTL).Methods The total RNA was respectively extracted from lymph node of U-PTL and reverse transcriptase,then multi-PCR was used to amplify the complete DNA sequence(CDS) of TCR β-chain.The recombinant plasmids were sequenced and sequence was analyzed by using online TCR resources.Results There were 9 TCR β chain CDS obtained from four patients.TCRβ-chain presented specific repertoire skewing in patients with U-PTL.There were restricted usage of BV2,BV4S2,BV14,BV29S1 of BV subfamily and BJ1S4,BJ2S3,BJ2S5,BJ2S7 of BJ subfanily.The clonal proliferation T cells had different CDR3 amino acid sequences.Conclusions There are restricted usage of TCR β V/J subfamily in patients with U-PTL.The sequences of CDR3 in different TCR clone proliferation are mostly different.

Objective To analyze the restricted usage of TCR BV,BJ subfamily and their sequences in patients with breast cancer.Methods The total RNA was extracted from tissues of lymph node metastasis,then reverse transcripted.The complete DNA sequence of TCR β-chain was amplified by multi-PCR.The recombinant plasmids were sequenced by using online TCR resources.Results 5 TCR β-chain CDS were obtained in two patients.TCR β-chain presented specific repertoire skewing in metastatic lymph node.There were selected usage of BV 2,BV 14,BV 29S1 of BV subfamily and BJ1S1,BJ2S2,BJ2S3,BJ2S5 of BJ subfamily.The clonal proliferation T cells had different CDR3 amino acid sequences.Conclusions There are selected usages of TCR BV,BJ subfamily in patients with breast cancer.The sequences of CDR3 are mostly different in TCR clone proliferation.

Objective To investigate the clinical pathological features and prognosis of gastric carcinoma with neu-roendocrine differentiation (NED) and mixed gonadal neuroendocrine carcinoma of the stomach (MANECs). Methods A retrospective analysis of 61 cases of gastric carcinoma with NED and 34 cases of MANECs and their histochemistry and im-munohistochemistry were also observed. Prognosis of the 2 groups were compared by the Kaplan-Meiers survival analysis. Prognostic factors associated with patients with gastric cancer were analyzed by COX proportional hazards model. Results Tumor location, distant metastasis and lymph node metastasis were statistically different between these 2 groups (P<0.05). Syn positive expression rate is higher than CgA and CD56 in the gastric carcinoma with NED group;Postoperative survive pe-riod of the gastric carcinoma with NED is shorter than that of MANECs (P<0.05). Lymph node metastasis and distant region-al transfer is obviously correlated with prognosis (P<0.05). Conclusion Immunohistochemistry is important for the diag-nosis of these two tumor. The number of neuroendocrine cells can help to assess prognosis and guide treatment.

Objective To investigate risk factors associated with lymph node metastasis and prognosis of rectal neuroendocrine tumor (NET). Methods Clinicopathological data of 69 patients with rectal NET in our department from April 2003 to October 2011 were retrospectively analyzed. Associations of clinicopothological factors with lymph node metastasis and prognosis were examined using univariate and multivariate analysis. Results Of the 69 patients, 9 cases had lymph node metastasis. The lymph node metastasis was significantly associated with tumor size, T stage and G grade by univariate analysis. Multivariate analysis showed that T stage was the only risk factor associated with lymph node metastasis. The overall 5-year survival rate was 90.3%. Prognosis of rectal NET was significantly associated with tumor size, T stage, N stage, M stage, TNM stage and G grade by univariate analysis. Multivariate analysis showed that M stage was significantly associated with long-term survival in rectal NET patients (P=0.000, HR=2.285, 95%CI:1.484~3.518). There was no significant difference in patients with stage Ⅰ between local and radical resection, while there were significant differences in those with stage Ⅱ or higher between the two operations (P=0.046). Conclusion T stage is associated with lymph node metastasis and both TNM stage and M stage can affect the prognosis of patients with NET, which may be used as potential predictive factors for rectal NET. Local resection should be recommended for patients with stage Ⅰ and radical resection should be recommended for patients with stage Ⅱor higher.