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 To analyze the related risk factors for Lisfranc injury resulting from low energy violence.Methods A retrospective study was performed for 61 cases (35 males,26 females) with low-energy foot injury hospitalized from June 2008 to June 2014.Mean age was 36.7 years (range,16-57 years).Fall injuries were noted in 24 cases,sports injuries in 21 cases,and twist injuries in 16 cases.The cases were divided into Lisfranc injury group(n =23) and non-Lisfranc injury group (n =38) according to the different diagnosis.Univariate analysis and multi-factor logistic regression analysis were used to identify the factors that may lead to the occurrence of Lisfranc injury including age,gender,body mass index,operation history,smoking,alcohol abuse,injury reason,medial depth of the mortise/ second metatarsal length (b/a),lateral depth of the mortise/ second metatarsal length (c/a),first metatarsal-to-talus angle,first intermetatarsal angle,second metatarsal length/foot length(a/g),calcaneal inclination angle and cuboid-navicular overlap/cuboid vertical height (e/e + f).Results Univariate analysis showed between-group differences were significant in age (x2 =7.385,P ＜0.05),injury reason (x2 =8.663,P ＜ 0.05),calcaneal inclination angle (t =3.958,P ＜ 0.05),b/a (t =5.051,P ＜ 0.05) and a/g(t =4.618,P ＜ 0.05).Logistic regression analysis identified b/a(OR =1.036,95 ％ CI 0.018-0.450,P ＜ 0.01) and a/g(OR =1.013,95％ CI 0.005-0.374,P ＜ 0.01) as independent risk factors for low-energy Lisfranc injury.Conclusion Low-energy Lisfranc injury is independently associated with b/a and a/g,and may relate to the decreased medial depth of the mortise and increased foot length.

[ ABSTRACT] AIM:To investigate the effect of artemisinin on lipopolysaccharide ( LPS)-induced intestinal epi-thelial barrier damage in IEC-6 cells and its molecular mechanism.METHODS:Cultured IEC-6 cells were divided to 5 groups:control group, LPS (100 mg/L) group and LPS +Artemisinin (30, 50 and 100μmol/L) groups.The cytotoxici-ty was detected by MTT assay.The releases of TNF-α, IL-1βand IL-6 in the IEC-6 cells were measured by ELISA.The transepithelial electrical resistance ( TER) was detected by electrical resistance tester, and the horseradish peroxidase (HRP) flux permeability were analyzed by a microplate reader.The expression of tight junction proteins, ZO-1, claudin-1 and occludin, and the expression of TLR4/MyD88/NF-κB at mRNA and protein levels were determined by RT-qPCR and Western blot.RESULTS:Artemisinin alone (up to 100 μmol/L) or in combination with LPS (100 mg/L) was not toxic to IEC-6 cells.Compared with control group, the releases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells significantly increased after treatment with LPS.The expression of TLR4/MyD88/NF-κB was activated by LPS.LPS down-regulated the protein expression of ZO-1, claudin-1 and occludin.However, artemisinin treatment decreased the re-leases of TNF-α, IL-1βand IL-6 in the culture supernatant of IEC-6 cells.The expression of TLR4/MyD88/NF-κB at mR-NA and protein levels was gradually reduced after treatment with artemisinin.In addition, artemisinin upregulated the pro-tein expression of ZO-1, claudin-1 and occludin significantly (P<0.01) in a dose-dependent manner.CONCLUSION:Artemisinin attenuates LPS-induced intestinal epithelial barrier damage by inhibiting TLR4/MyD88/NF-κB activation in the IEC-6 cells.

Objective To introduce a new modified percutaneous dilational tracheostomy and compare the application effect of percutaneous dilational tracheostomy with modified percutaneous dilational tracheostomy in the treatment of critically ill patients. Methods A total of 60 critically ill patients undergoing tracheotomy were selected , and they were randomly divided into two groups according to the methods of tracheotomy. Sex, age, weight, body mass index, acute physiology and chronic health evaluation, operation time, incision size, intraoperative blood loss, incision healing time, incidences of complications after operation were compared between the two groups. Results There were not statistically significant differences of in sex, age, weight, body mass index, and acute physiology and chronic health evaluation between percutaneous dilational tracheostomy group and modified percutaneous dilational tracheostomy group (P>0.05). Operation time, incision size and intraoperative blood loss of modified percutaneous dilational tracheostomy group was statistically significantly shorter than that of percutaneous dilational tracheostomy group [(5.80 ± 1.19) min vs. (7.65 ± 1.05) min, (8.33 ± 3.30) ml vs. (11.33 ± 4.34) ml, (1.08 ± 2.96) cm vs. (1.27 ± 2.54) cm] (P<0.05). The incision healing time and incidence of complications after operation of percutaneous dilational tracheostomy group had no statistical significance compared with modified percutaneous dilational tracheostomy group (P>0.05). Conclusions The modified percutaneous dilational tracheostomy can save operation time, and reduce intraoperative blood loss, so it can be widely used.

AIM To explore the effects of glycyrrhetinic acid on the gastric ulcer rats infected by Helicobacter pylori (Hp) and its action mechanism.METHODS Gastric ulcer rat models were induced by acetic acid stress and then followed by Hp infection.After treatment with low and high doses of glycyrrhetinic acid,the ulcer index,gastric acid and proteinase activities in gastric ulcer rats were analyzed.The effects of glycyrrhetinic acid on the expressions of BCL2 and Caspase-3,the GSK3β activity in gastric mucosa and gastric epithelial cells,and the cell apoptosis level were then detected.RESULTS Glycyrrhetinic acid reduced the ulcer index,gastric acid and proteinase activities in rats.Besides,the expression of BCL2 was significantly up-regulated by glycyrrhetinic acid in gastric mucosa and gastric epithelial cells,whereas the expression of Caspase-3,level of cell apoptosis,and GSK3β activity were significantly reduced.After the treatment with GSK3 β activator LY294002,the level of BCL2 was down-regulated,Caspase-3 expression was increased,and the level of cell apoptosis was enhanced.CONCLUSION Glycyrrhetinic acid promotes the healing of gastric ulcer infected by Hp via regulating GSK3β activity and inhibiting apoptosis of gastric epithelial cells.

Near infrared photoimmunotherapy (NIR-PIT) is a highly selective molecularly targeted phototherapy for cancer which is based on injecting a conjugate of IRDye700DX,a water-soluble near-infrared silicon-phthalocyanine dye,and a monoclonal antibody that targets an antigen on the cancer cell surface. Subsequent local irradiation of NIR light causes the rapid and specific tumor cell death. Due to the good clinical translation prospects of NIR-PIT,this paper summarizes the influencing factors,antitumor mechanism,main challenges and recent strategies,which may benefit for its research and clinical application.

Objective: To compare the anti-inflammatory effect of Fraxini Cortex pieces between integrated production process and traditional processing method. Methods: The model of rat paw swelling induced by carrageenan was used to study the anti-inflammatory and swelling reliving effects of water extracts of Cortex Fraxini with different methods. The main chemical components of the 2 kinds of Cortex Fraxini herbal pieces were determined by high performance liquid phase. Results: Compared with the blank group, the water extracts of the 2 kinds of Cortex Fraxini could reduce the swelling of the rats and improve the various indicators of inflammation. However, the anti-inflammatory and swelling reliving effects of the integrated processing of Cortex Fraxini were more significant. The contents of 4 main active ingredients of esculine, fraxin, aesculetin and fraxetin in the integrated processing of Cortex Fraxini were higher than that of the traditionally processed Cortex Fraxini. The total amount of 4 kinds of coumarins in the integrated Cortex Fraxini was about 1.5 times that of the traditionally processed water extract of Cortex Fraxini. Conclusion: The integrated processing and traditional processing of Cortex Fraxini have similar effects on anti-inflammatory effects, and have the superiority of reducing the loss of active ingredients, which is worthy of popularization and application.

Objective To investigate the prokaryotic expression and immunoreactivity of BspE,a type Ⅳ secretion protein of Brucella,and the effect of recombinant protein BspE on cytokines.Methods According to the BspE gene of Brucella M5-90 published in GenBank,the gene fragments were synthesized by a company and then ligated into PUC57 vector for sequencing.The sequenced gene was cloned into a prokaryotic expression vector pET-28α and transformed.Induced expression was performed in E.coli DE3 competent cells.The obtained target protein was purified by a Ni-NTA affinity column,and its reactogenicity was analyzed by Western blotting.Mouse RAW264.7 cells were treated with 25 g/L BspE recombinant protein for 12,24,48 h,and the control group was treated with the same amount of BSA instead of BspE,and enzyme-linked immunosorbent assay (ELISA) was used to detect interleukin (IL)-1β level.Results The recombinant expresed plasmid of pET-28α-BspE was successfully obtained.The results of Western blotting showed a single band with a relative molecular mass of about 30.1 × 103,and the recombinant protein BspE had good reactogenicity,and IL-1β levels (ng/L)were significantly elevated by the recombinant protein BspE (12 h:43.27 ± 2.13 vs 30.24 ± 1.66,24 h:57.78 ± 3.44 vs 41.22 ± 1.22,48 h:72.52 ± 3.04 vs 46.77 ± 2.75,t =8.38,7.86,10.89,P ＜ 0.05).Conclusions BspE recombinant protein has better immunoreactivity and can increase the expression level of IL-1β in mouse macrophages.This study provides a scientific basis for the role of effector proteins in the pathogenesis of Brucella.

Cardiovascular disease is a leading cause of death among the elderly and the incidence of coronary artery disease progressively increases with advancing age.Traditional risk factors are incompletely predictive of cardiovascular disease development.With the advent of high-throughput next-generation genome sequencing technologies in recent years, some studies have indicated that aging is associated with an increased frequency of somatic mutations of hematological neoplasm-related genes in the hematopoietic system, providing a competitive growth advantage for mutant hematopoietic cells and thus allowing for their clonal expansion, a phenomenon known as clonal hematopoiesis of indeterminate potential(CHIP). CHIP is common in middle-aged and elderly populations and is associated with increased risks of hematological cancer and all-cause death.There is growing evidence that CHIP is involved in the development and progression of multiple cardiovascular disorders through the activation of inflammatory responses.In this review, we will give an overview of current advances in the understanding of clonal hematopoiesis in cardiovascular disease.