Objective To investigate the effects of CpG oligodeoxynucleotide (ODN) on cellular immune responses in suckling mice infected with human rotavirus (HRV). Methods Forty ICR suckling mice were randomly divided into four groups: control group. HRV infection group, CpG ODN pretreatment group and CpG ODN treatment group. Suckling mice were sacrificed four days after rotavirus challenge. Small intestine and spleen samples were collected under sterile condition. Thedegree of small intestinal mucosal injures was evaluated with standard scoring criteria. The spleen index was calculated and spleen lymphocyte stimulation index was detected by methyl thiazolyl tetrazolium ( MTT) assays. Levels of gamma interferon (IFN-γ), interleukin-4 ( IL-4) in the supernatant of spleen lymphocyte culture were detected by enzyme linked immunosorbent assay (ELISA). T lymphocyte subsets were analyzed by flow cytometry. The data was compared by one way ANOVA. Results The scores of mucosal injures of mice in HRV infection group, CpG ODN pretreatment group and CpG ODN treatment group were 4. 00 ±1. 31, 2. 75 ±1. 28 and 2. 87 ±0. 99, respectively, and the differences among groups were statistically significant (F=ll. 32,P＜0. 01). The scores of mucosal injures in CpG ODN pretreatment group and CpG ODN treatment group were both lower than that in HRV infection group (P＜0. 05). Compared with control group, spleen lymphocyte stimulation index and the percentage of CD8+ T lymphocytes were higher, the percentage of CD4+ T lymphocyte and ratio of CD4+ /CD8+ T cells were lower, levels of T helper (Th)1 type cytokine IFN-y increased significantly in HRV infection group; while the spleen index, spleen lymphocyte stimulation index, percentages of CD4+ and CD8+ T lymphocytes, IFN-y levels were all increased in CpG ODN pretreatment group and CpG ODN treatment group (P＜0. 05). The spleen index, spleen lymphocyte stimulation index, the percentage of CD4+ T lymphocytes, CD4+/CD8+ ratios and IFN-y levels in CpG ODN pretreatment group and CpG ODN treatment group were all significantly higher than HRV infection group (P＜0. 05). Conclusion CpG ODN potently enhances cellular immune responses in ICR suckling mice infected with HRV and CpG ODN could induce dominant Th1 response.

Objective To establish an animal model of human rotavirus(HRV)-induced diarrhea. Methods Sixty ICR suckling mice were randomly divided into two groups, and each was further divided into 3 subgroups(A, B, C for group Ⅰ, and D, E, F for group Ⅱ);group B, C, E and F were assigned as experimental groups. while group A and D were controls. Mice in group B and C were inoculated with3 × 10~5 PFU and 3×10~6 PFU HRV Wa strain fluid respectively when they were 4-day old. Mice in group E and F were inoculated with 3×10~5 PFU and 3×10~6 PFU HRV Wa strain fluid respectively when they were10-day old. The symptoms, fecal viral excretion, intestinal histopathologies and ultrastructures of animals were observed. One-way ANOVA was used to compare the differences among the groups. Results There were significant differences in the duration of diarrhea and viral excretion, jejunal villous height, the weight at d4 and d7 after inoculation among three subgroups in group Ⅰ(F=204.38, 86.60, 7.18, 18.41 and10.08, P<0.01). Diarrhea was not observed in group Ⅱ, and the differences in jejunal villous height and the weight at d4 and d7 after inoculation among three subgroups were not significant(F=0.16, 0.13 and 1.03, P>0.05). Compared with the group D, the duration of viral excretion was longer in group E and F(F=8. 34, P<0.01). Conclusion Animal model of HRV diarrhea can be established in 4-day-old ICR suckling mice infected with 3×10~6PFU HRV.

Objective Because of local infiltrated growth and spread,the operative resection of hilar (cholangiocacinoma) is very difficult. Recently,combined extended hepatectomy and vascular resection had been performed for treatment of hilar cholangiocarcinoma and has greatly increased the resection rate and survival rate.However, it is associated with high operative morbidity and mortality. The aim of this study is to explore a reasonable hepatic resection strategy, that is safe and beneficial for the patient. Method Sixteen (consecutive) cases of hilar cholangiocarcinoma with involvement of hilar vessels have been treated in our hospital since 1977. En bloc resection of the hilar tumor that included hepatic segment I,IV and involved blood (vessel), as well as hepato-duodenal ligament skeletonization was performed in 15 patients. Results All 15 cases were successfully operated on with a resection rate of 93.8%, and 12 cases with R_0 resection. The operative mortality and in-hospital mortality rate were 0. Temporary bile leak and abdominal infection (respectively) developed in 1 patienteach,with an overall morbidity of 13.3%,and both were cured by non-(operative) therapy. No case of liver failure occurred. Follow up showed the median survival was 22 months and 7 are still alive. Conclusions (1)The resection rate and survival rate for hilar cholangiocarcinoma could be improved by combination of liver and hilar vascular resection.(2)Hilar cholangiocarcinoma mainly spreads to the medial segment(S4) and caudate(S1), and these segments need to be resected in the combined (operation).(3)Although resection of the middle part of liver takes little more time than hemi-hepatectomy and trisegmentectomy, it could preserve more liver parenchyma and reduce postoperative morbidity (such as liver failure) and mortality rates.(4)The resection of the hilar blood vessel involved by tumor was necessary to improve the resection rate and cure rate. Reconstruction of the blood vessel was made selectively by taking the circumstances into consideration.

Objective: To investigate the relationship between the EBV-induced liver injury and caspase-3/6 in children. Methods: Data of 249 patients seen from July 2016 to June 2017 who got infection with EBV were collected in second affiliated hospital of Wenzhou Medical University and the patients were divided into two groups; 168 patients who were diagnosed with hepatitis were selected for abnormal liver function group, laboratory tests were performed in acute phase and convalescent phase. Meanwhile the 81 patients, whose liver function were normal were selected for normal liver function group. Two ml of blood plasma was collected from each patient from both groups at the beginning and after a week’s treatment of the abnormal liver function group. The patients were aged from 1 to 14 years. The abnormal liver function group was further divided into four groups: young children (1-3 years old), preschool children (4-6 years old), school children (7-10 years old), teenagers (10-14 years old). Firstly, we recorded their alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), gamma glutamyl transpeptidase (GGT) and direct bilirubin (DBIL), then by the use of enzyme linked immunosorbent assay (ELISA) we measured the protein level of caspase-3 and caspase-6. Results: In the group of abnormal liver function group, in the acute phase of the teenagers the level of ALT, caspase-3 and caspase-6 are improved significantly(927.2±82.5 vs. 158.5±41.4, P<0.001, 169.8±52.8 vs. 91.5±10.1, P<0.001, P<0.05, 82.2± 2.6, P<0.001, 965.8±31.5, P<0.001, 184.8±14.3, P<0.001), while the AST, TBLL, GGT and DBLL had no significant difference. Conclusions In the process of EBV-induced liver injury, there was a linear correlation between the increase of caspase-3/6 and ALT in the acute stage, suggesting that EBV-induced liver injury is closely related to caspase-3/6.