AIM: To observe the change of nitric oxide (NO) generation system in the vascular adventitia, media and intima in septic shock rats. METHODS: The septic shock model was made in rats by caecal ligation and puncture. The intima, media and adventitia of the rat aorta were separated. NO production (NO - 2) , nitric oxide synthase(NOS) activity and L-arginine (L-Arg) transport were measured, separately. Inducible NOS (iNOS) distribution was detected by immunohistochemistry. RESULTS: Both in early and late stage of septic shock, NO - 2 from the intima was decreased by 66.1% and 78.9%( P

Tissue engineering composites for bone defect treatment is currently a hot spot of the research field.Mechanical loading on tissue engineering composites is important to bone formation and related research has attracted more and more attention.In the field of tissue engineering research,the establishment of a standardized animal model is the basis of experimental research for exploring the effects of different mechanical loading on in situ repair of bone defect and its action mechanism.This paper reviews the approaches used to establish experimental model of bone defect under mechanical loading such as animal selection,defect preparation,fixation and loading mode in order to provide a reference for related research.

OBJECTIVE To investigate CD25 expression in peripheral blood T lymphocytes from patients with chronic hepatitis B,and explore its significance in the pathogenesis of chronic hepatitis B.METHODS To detect CD25 expression in peripheral blood T lymphocytes of patients with chronic hepatitis B by means of flow cytometry.CD25 expression was observed in chronic hepatitis B patients.In the meantime,CD25 expression in T cells from severe chronic hepatitis or acute hepatitis B patients and asymptomatic carriers of HBV was also observed.RESULTS Varying degrees of CD25 were expressed in T cells from hepatitis B patients.The expression in CD3+ T and CD8+T cells was higher than that in CD4+T cells.CD25 expression in CD4+T was lower.The average of CD25 expression in CD3+T cells from patients with chronic hepatitis B,acute hepatitis B,and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.92?0.13)%,(0.51?0.36)%,(1.60?0.07)%,and(0.95?0.23)%,respectively.The average of CD25 expression in CD4+T cells from patients with chronic hepatitis B,acute hepatitis B and chronic severe hepatitis B and asymptomatic carriers of HBV was(2.58?0.50)%,(0.34?0.07)%,(1.45?0.02)%,and(0.83?0.13)%,respectively.CD3+T and CD4+T CD25 expression in patients with chronic hepatitis and,severe chronic hepatitis B was increased compared with that of acute hepatitis B patients and asymptomatic carriers of HBV.Compared with chronic severe hepatitis B,the expression of chronic hepatitis B was higher.CONCLUSIONS CD4+ CD25+T cells in chronic hepatitis B virus infection are increased compared with acute hepatitis,CD4+ CD25+T cells may be related to immune tolerance.

0bjective To observe the cell membrane penetration of protein transduction domain (PTD)-HBeAg fusion protein in vitro.Methods The sequence of trans-activator of transcription (Tat)-PTD was synthesized and the whole HBcAg gene was amplified by polymerase chain reaction (PCR).Overlap extension PCR was employed to fuse Tat-PTD and whole HBcAg gene.Then the fusion gene was cloned into prokaryotic expression vector pMAL-c2X.The correct vector was transformed into E.coli Rosetta-gamiTM 2(DE3),and the protein was induced by isopropyl β-D-1-thiogalactopyranoside(IPTG).Western blot was used tO identify the protein. Furthermore,the fusion protein PTD-HBcAg was purified by affinity chromatography.HBcAg protein expressed using the same methods was employed as eontr0l.The purified protein was added tO HuH-7 cell culture,then the transduction of PTD-HBcAg and HBcAg in cells were detected by indirect immunofluorescence assay (IFA).Results The fusion protein was effectively expressed in E. Coli and purified by affinity chromatography.Both purified PTD-HBcAg and HBcAg could be recognized by HBeAg monoclonal antibody in Western blot analysis.IFA visualization showed that PTD-HBeAg could be introduced into HUH-7 ceils while HBcAg only could not be detected in cells.Conclusions PTD-HBcAg fusion protein can be expressed effectively and purified in prokaryotic expression system.PTD could mediate HBcAg penetrating eell membrane into the cells.

Objective To explore the expression levels of YBX1 and FOXA1 in gastric cancer tissues and determine their relationship with prognosis. Methods A total of 131 patients with gastric cancer were studied, and the corresponding adjacent normal tissues of each patient were selected as the control. qRT-PCR and immunohistochemistry were used to detect the expression levels of YBX1 and FOXA1 in cancer tissues and adjacent tissues. The correlation between YBX1 and FOXA1 protein expression in gastric cancer tissues was expressed by Crammer's V coefficient, and the correlation between YBX1 mRNA and FOXA1 mRNA was analyzed by Pearson method. Kaplan-Meier method was used to analyze the relationship between YBX1, FOXA1 protein expression in gastric cancer tissues and the 5-year overall survival rate of patients. Univariate and multivariate Cox regression analyses were used to analyze the factors affecting the prognosis of patients with gastric cancer. Results Compared with paracancerous tissue, the levels of FOXA1 and YBX1 in cancer tissues were lower and higher, respectively (P < 0.05). A negative correlation was observed between YBX1 mRNA and FOXA1 mRNA in gastric cancer (r=-0.675, P < 0.05). The expression of YBX1 and FOXA1 proteins in gastric cancer tissues was negatively correlated (Crammer's V=-0.497, P < 0.001). The expression of YBX1 and FOXA1 proteins in gastric cancer tissue was related to the degree of differentiation, lymph node metastasis, and TNM staging (P < 0.05). The 5-year survival rate of the YBX1 negative expression group and the FOXA1 positive expression group were higher than those of the YBX1 positive expression group, and the FOXA1 negative expression group both P < 0.05). TNM staging and YBX1 were independent risk factors for death in patients with gastric cancer (P < 0.05), and FOXA1 was a protective factor (P < 0.05). Conclusion YBX1 is highly expressed and FOXA1 is lowly expressed in gastric cancer tissues; they are closely related to the disease progression and prognosis of patients with gastric cancer.

Endoplasmic reticulum (ER) stress is involved in the development and progression of tumors.In recent years, great attention has been paid to the study of the interplay of ER stress and T cell differentiation and functionality.Intense ER stress in the tumor-infiltrating T cells exacerbates T cell exhaustion and impairs T cell anti-tumor immunity.Therefore, a variety of ER stress inhibitors have been developed and utilized to alleviate T cell exhaustion, which improves T cell function in tumor microenvironment.Furthermore, the downregulation of several circadian clock genes like Per1 and Per2 also aggravates T cell exhaustion, and the key downstream effector molecules in ER stress regulate the transcription of Per family, thus enhancing the T cell function.In the present manuscript, we particularly summarize how ER stress impacts the anti-tumor immunity of T cells, and further discuss potential strategies for improving tumor immunotherapy via targeting ER stress.

Background and Objectives: Tracking of the tumor progression by MSCs-based therapy is being increasingly important in evaluating relative therapy effectively. Herein, Bioluminescence imaging (BLI) technology was used to dynamically and quantitatively track the hepatocellular carcinoma suppressive effects by human umbilical cord mesenchymal stem cells (UC-MSCs). Methods: and Results: The stem cells present typical phenotypic characteristics and differentiation ability by morphology and flow cytometry analysis of marker expression. Then, the growth inhibition effect of conditioned medium and UC-MSC on H7402 cells was studied. It is found both the conditioned medium and UC-MSC can effectively decrease the proliferation of H7402 cells compared with the control group. Meanwhile, the relative migration of UC-MSC to H7402 is also increased through the transwell migration assay. In addition, a mice hepatoma tumor model was built by H7402 cells which can express a pLenti-6.3/DEST-CMV-luciferase 2-mKate2 gene. The effect of stem cells on growth inhibition of tumor in a mice transplantation model was dynamically monitored by bioluminescence imaging within 5 weeks. It has shown the bioluminescence signal intensity of the tumor model was significantly higher than that of the UC-MSC co-acting tumor model, indicating that the inhibition of UC-MSC on liver cancer resulted in low expression of bioluminescent signals. Conclusions The microenvironment of UC-MSCs can effectively inhibit the growth of liver cancer cells, and this therapeutic effect can be dynamically and quantitatively monitored in vivo by BLI. This is of great significance for the imaging research and application of stem cells in anticancer therapy.

Objective To observe the metabolic changes of subcortical structures in children with intractable epilepsy using 18 F-FDG PET/CT,and to investigate the mechanism of subcortical structure involvement in epileptic seizures and its clinical significance.Methods Features of 18F-FDG PET/CT imaging in 611 intractable epilepsy children were analyzed.The metabolic changes of cortex and subcortical structures (basal ganglia,thalamus and cerebellum) were observed.The children were divided into three groups (young,middle and older groups) according to age,also mild group and severe group according to the number of involved lobar,respectively.The incidence of metabolic abnormalities in subcortical structures of different groups were analyzed.Results Among 611 children,unilateral cortical metabolic abnormality was found in 525,and bilateral cortical metabolic abnormalities were found in 86 children.The involvement of subcortical structures was detected in 190 children,including basal ganglia (n=64),thalamus (n=113) and cerebellum (n=105).The incidence of metabolic abnormality in subcortical structures under different age groups was not statistically different (all P＞ 0.05),while the incidence of metabolic abnormality in subcortical structures of severe group was significantly higher than that of mild group (all P＜0.001).Conclusion 18 F-FDG PET/CT might be able to detect the metabolic abnormalities of subcortical structures,therefore indicating the involvement of cerebral cortex.

Objective To investigate the characteristics of metabolic foci on 18F-fluorodeoxyglucose (FDG) PET/CT scan in pediatric patients with epilepsy.Methods Twenty-three pediatric patients (15 males,8 females,age range:0.5-13.3 years) with epilepsy were retrospectively reviewed from March 2014 to December 2016.All patients underwent 18F-FDG PET/CT and metabolic foci were found.The visual method and semi-quantitative analysis were used to analyze images.Fourteen of them underwent surgery and were followed up for 3-24 months.Results Glucose hypermetabolism were observed most frequently in the frontal and parietal lobes,with or without surrounding/remote hypometabolism.On the day of PET/CT imaging,8 patients had no seizures,14 patients had seizures,and 1 patient was uncertain.The sites of resection were consistent with the regions of hypermetabolism in 9 patients,among whom the pathological results showed 8 cortical malformations and 1 Rasmussen's syndrome.Follow-up results for the above 9 patients showed that there was 7 Engel class Ⅰ patients and 2 Engel class Ⅲ patients.Conclusion The hypermetabolism may mostly appear in the frontal and parietal lobes of pediatric patients with epilepsy,and malformations of cortical development seem to be the most common pathology results.

Objective: To observe the expression levels of PD-1/PD-L1 costimulatory molecules and explore the clinical significance in patients with chronic lymphocytic leukemia (CLL) . Methods: The expression of PD-1/PD-L1 in peripheral blood CD8⁺ T cells, CD4⁺T cells, CD19⁺B, and dendrites cells (DC) was detected by flow cytometry in 57 CLL patients and 20 healthy controls. The correlations of PD-1/PD-L1 expression with disease stage, CD38 expression, ZAP-70 expression, chromosome karyotype abnormality and β₂-MG expression were analyzed. Results: ①Compared with control, CLL patients, including 39 males and 18 females with the median age of (63.7±10.7) years, had no statistically significant difference in age and gender (P>0.05) . CLL patients had the higher PD-1/PD-L1 expression than healthy controls (P<0.05) . ②In Rai staging, the later the stage, the higher expression of PD-1/PD-L1 (P<0.05) . ③PD-1 expression in CD8⁺CD38⁺ group (11 cases) was higher than that in CD8⁺CD38^- group (46 cases) (P=0.004) , and CD8⁺ poor prognosis chromosome group (14 cases) also had significant higher PD-1 expression than CD8⁺good prognosis chromosome group (28 cases) (P=0.004) . ④The expression of PD-L1 was higher in CD38⁺group, ZAP-70⁺group, and poor prognosis group, as compared to that in CD38^-group (P=0.002) , in ZAP-70^-group (P<0.001) , in good prognosis group (P=0.023) . There was no correlation between the expression of PD-1/PD-L1 and β₂-MG (P>0.05) . Conclusion This data reveals that PD-1/PD-L1 was highly expressed in CLL patients. Its expression levels were correlated with Rai stage, CD38, ZAP-70, chromosome karyotype, but not with β₂-MG. PD-1/PD-L1 may be a prognostic factor in patients with CLL.