The tumor necrosis factor(TNF) secreted by gene modified tumor cells can lead to very effective tumor rejection. This effect of TNF on tumor growth is mediated mainly by the induction of an antitumor immune response. It requires a local and continuous presence of TNF at the tumor site. Tumor suppression induced by TNF is close dependent and a complete tumor eradication must not be accompanied by systemic toxic side effects. A complex pattern of tumor infiltrating cells has been observed in TNF producing tumors, consisting of macrophages, CD4+ and CD8~(+)T cells. For efficient tumor inhibition maccophages and CD8~(+)T cells are needed,whereas CD4~(+)T cells seem to be innocent bystander cells. TNF is also effective in T eell deficient mice, but in most cases for complete tumor elimination T ceils have to be present. Depending on the cell lines used or the levels of TNF secreted by transduced tumor cells, systemic toxicity of TNF has also been observed including cachexia or wasting of the experimental animals. In some cases, TNF gene transfected tumors did not show growth inhibition in vivo, but rather, their metastases were enhanced. Using TNF producing tumor cells as vaccine, no systemic protective immunity against a parental tumor cell challenge has been observed.

Objective To explore the expression and function of myosin light streptokinase (MLCK) in small intestine mucosa of acute necrotizing pancreatitis (ANP) rats.Methods Fifty-six male SD rats were randomly assigned to control group and ANP group.A rat model of ANP was reproduced by retrograde injection of 4％ sodium taurocholate into the biliopancreatic duct,while the control group underwent a sham operation.The rats were sacrificed at 6th,12th,24th,48th hour after ANP induction.Serum amylase、TNF α,IL 1β,diamine oxidase (DAO) were measured.The pathological scores in the pancreas and small intestine were observed.The ultrastructure and tight junction (TJ) changes in the small intestine mucosa were observed with an electron microscope.The localization and expression of MLCK in small intestine mucosa was determined by immunohistochemistry method.Results Compared to the control group,the serum amylase,TNF-α,IL-1 β,DAO level,in the ANP group were all significantly increased;[(4 978 ± 1 574) U/L vs (1 176 ± 124))U/L,(47.88 ± 15.85) μg/L vs (17.24 ± 1.99) μg/L,(132.48 ± 68.54) μg/L vs (23.51 ± 6.44) μg/L,(95.96 ± 30.84)μg/L vs (38.06 ± 17.73)U/L at 12 h],and the pathology scores of pancreas and small intestine were both significantly elevated [12 h:(12.2 ± 1.80) vs (4.68 ± 0.35),(2.58 ± 0.52) vs (0.58 ±0.26)] (P ＜0.05);the MLCK protein expression in small intestine mucosa was significantly increased in ANP group (12 h:0.1863 ± 0.0230 vs 0.1636 ± 0.0049),and the difference was statistically significant (P ＜0.05).The small intestine ultrastructure was seriously damaged and TJ was widened significantly in ANP Group.Conclusions The increased serum TNF alpha and IL-1β concentration and DAO activity and up-regulated MLCK protein expression in small intestine mucosa may damage the integrity of tight junction of intestinal epithelial cell and cause intestine mucosa barrier dysfunction.

Objective To investigate the expression of pancreatic thioredoxin-1 (TRX-1) in rats with acute necrotizing pancreatitis (ANP) and the effect of pretreatment of melatonin on its expression. Methods Male Spraque-Dawley rats (n = 12) were randomly divided to ANP group, melatonin group, control group with 24 rats in each group. The rats in ANP group received three intraperitoneal injections of 25 ml/kg body weight 6% L-arginine at an interval of 1 h to induce ANP. The rats in melatonin group received intraperitoneal injections of 25 ml/kg body weight 6% melatonin 30 min before ANP induction; rats in ANP group and control group received intraperitoneal injections of same amount of saline. Rats were sacrificed at 6 h, 12 h and 24 h after ANP induction. The serum level of amylase was measured and the pathological evaluation of pancreatic tissues was performed. The concentrations of malondialdehyde (MDA) and myeloperoxidase (MPO) in pancreatic tissues were measured. The expressions of TRX-1 protein were detected by immunohistochemistry and the expressions of TRX-1 mRNA in pancreatic tissues were determined by RT-PCR.Results In ANP group, serum level of amylase, MDA, MPO, TRX-1 mRNA and TRX-1 protein in pancreatic tissues were (3 012 ±1 425) U/L, (4.13 ± 1. 85)nmol/mg prot,(7.45 ± 1.26)nmol/mg prot, 0.68 ±0. 18, 66.8 ±8. 1, while they were (1 835±499)U/L, (3.03 ±2.12) nmol/mg prot, (5. 32 ± 1.06) nmol/mg prot, 0.50±0.09, 80. 29 ±8. 14, respectively in melatonin group, the values in melatonin group were significantly lower thanthose in ANP group (P < 0.05). The peak value of TRX-1 mRNA and TRX-1 protwein expressions shifted from 12 h after ANP induction in ANP group to 6 h after ANP induction in melatonin group. Conclusions The expression of pancreatic TRX-1 protein and TRX-1 mRNA in rats with ANP was significantly increased. Melatonin pretreatment could promote pancreatic tissues to express TRX-1 protein and TRX-1 mRNA, and may be protective for pancreatic tissues damages.

AIM: To investigate the significance and function of IFN-γ on the changes of peripheral blood platelet count during tumor-rejection induced by a low dose of melphalan in C57BL/6 mice. METHODS: Mouse tumor rejection model induced by a single dose of melphalan was used in this experiment. Different gene-type tumor-bearing mice (IFN-γ~(+/-) and IFN-γ~(-/-)), which had the same genetic background of C57BL/6, were treated intraperitoneally with melphalan (7.5 mg/kg). Tumor size was observed and recorded every one to three days in these different gene-type mice subsequently. Blood samples were obtained from orbital venous sinus on different days before and after melphalan treatment, and then complete blood counts were performed. The function of IFN-γ on the efficacy of chemotherapy and the changes of blood platelet count in IFN-γ~(+/-) and IFN-γ~(-/-) mice after melphalan treatment was analyzed. RESULTS: There was no significant difference in tumor sizes and blood platelet count between IFN-γ~(-/-) and IFN-γ~(+/-) mice (P>0.05). On the first day after melphalan (7.5 mg/kg) treatment, there were no significant changes in tumor sizes between mice in these two groups (P>0.05). Tumors shrank a little in IFN-γ~(-/-) mice and then grew gradually. Tumors relapsed in 2 w after melphalan injection in all IFN-γ~(-/-) mice, while tumor volumes decreased progressively and tumor cured at last in IFN-γ~(+/-) mice. The number of blood PLT in IFN-γ~(+/-) mice increased to (1935±378)×10~9/L 6 h after melphalan treatment, significantly higher than before (P<0.01); While in IFN-γ~(-/-) mice it was (1183±186)×10~9/L 6 h after melphalan treatment, no obvious increase than before. There was significant difference in blood PLT 6 h after melphalan treatment between IFN-γ~(+/-) and IFN-γ~(-/-) mice (P<0.01). Later, the numbers of blood PLT in IFN-γ~(+/-) mice decreased gradually and it dropped to normal (1158±270)×10~9/L on 11th day after melphalan treatment (P>0.05); While it sustained in normal range in IFN-γ~(-/-) mice. There was no significant difference in blood platelet count between IFN-γ~(-/-) and IFN-γ~(+/-) mice. CONCLUSION: Peripheral blood platelet count increased on the first day after melphalan treatment and tumors cured in IFN-γ~(+/-) mice; While tumors relapsed and there is no increase in blood platelet count on the first day after melphalan treatment in IFN-γ~(-/-)mice. These data indicated that the increase of blood PLT count was related to the function of IFN-γ in tumor-bearing mice in vivo during tumor rejection induced by a low dose of melphalan.

Objective To investigate the relationship between surgical timing and outcomes of aneurysmal subarachnoid hemorrhage (aSAH).Methods The patients with aSAH retrospectively received clipping or endovascular embolization.Their demographic and clinical data were collected.The modified Rankin Scale (mRS) was used to evaluate the outcomes at 6 month after procedure.Univariate and multivariate logistic regression analyses were used to determine the risk factors that influencing clinical outcomes of patients.Results A total of 198 patients with aSAH were enrolled,118 had good outcome (mRS score0-2),80 had poor outcome (mRS score ＞2; 20 of them died); 32 were early operation (operated within 2 d after onset) and 166 were late operation.Univariate analysis showed that the proportions of hypertension (29.66％ vs.52.50％ ; x2 =10.464,P =0.001),cerebral infarction (11.86％ vs.35.00％ ;x2 =15.269,P ＜0.001),cerebral hemorrhage (9.32％ vs.31.25％ ;x2 =15.410,P ＜0.001),Fisher grade 3-4 (22.88％ vs.47.50％ ; x2 =13.104,P ＜ 0.001),Hunt-Hess grade Ⅳ to Ⅴ (19.49％ vs.52.50％ ;x2 =23.557,P ＜0.001),cerebral vasospasm (5.93％ vs.25.0％ ;x2 =14.719,P ＜0.001),hydrocephalus (5.08％ vs.17.50％ ;x2 =8.093,P =0.004),and late operation (78.81％ vs.91.25％ ; x2 =5.442,P =0.020) in patients of the good outcome group were significantly higher than those of the poor outcome group.Multivariatelogistic regression analysis showed that Fisher grade 3 to 4 (odds ratio [OR] 9.13,95％confidence interval [CI] 2.98-13.45; P ＜0.001),Hunt-Hess grade Ⅳ to Ⅴ (OR 6.86,95％ CI 1.57-12.34; P＜0.001),accompanied with hydrocephalus (OR 2.59,95％ CI 1.17-4.31; P=0.024),and late operation (OR 2.17,95％ CI 1.12-3.95; P=0.029) were the independent risk factors for patients with poor clinical outcomes.The univariate analysis for both early operation and late operation groups showed that only the good outcome rate of the early operation group was significantly higher than that of the late operation group (78.13％ vs.56.2％ ;x2 =5.442,P =0.020),and there were no significant differences in the incidences of rebleeding (6.25％ vs.13.25％; x2=1.235,P=0.266),cerebral vasospasm (12.50％ vs.19.28％;x2 =0.042,P=0.834),and hydrocephalus (12.50％ vs.9.64％;x2 =0.242,P=0.623).Conclusion Early operation may significantly improve the outcomes in patients with aSAH.

Mounting evidence has demonstrated that CD4(+) T cells play an important role in anti-tumor immune responses. Thus, adoptive transfer of these cells may have great potential for anti-cancer therapy. However, due to the difficulty to generate sufficient tumor-specific CD4(+) T cells, the use of CD4(+) T cells in tumor therapy is limited. It has been found that IL-15 transfection enhances the proliferation and anti-tumor activity of tumor-specific CD8(+) T cells, but the effect of IL-15 transfection on CD4(+) T cells remains unknown. Here, the effects of retrovirus-mediated IL-15 expression in Ova-specific CD4(+) T cells from Do11.10 mice were evaluated and it was discovered that IL-15 transfected CD4(+) T cells expressed both soluble and membrane-bound IL-15. Retrovirus-mediated IL-15 expression led to a selective expansion of antigen-specific CD4(+) T cells by inhibiting their apoptosis. In vivo IL-15 transfected CD4(+) T cells were more effective in suppressing tumor growth than control retroviral vector transfected ones. To ensure the safety of the method, the employment of thymidine kinase gene made it possible to eliminate these transgenic CD4(+) T cells following ganciclovir treatment. Together, we show that IL-15 transfection induced a selective expansion of antigen-specific CD4(+) T cells ex vivo and enhanced their tumor-suppression effects in vivo. This has an important significance for improving the efficacy of adoptive T cell therapy.
Animals ; Antineoplastic Agents ; pharmacology ; CD4-Positive T-Lymphocytes ; cytology ; drug effects ; immunology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Gene Expression Regulation ; Genetic Vectors ; genetics ; Humans ; Interleukin-15 ; metabolism ; Mice ; Mice, Transgenic ; Neoplasms ; drug therapy ; Recombinant Fusion Proteins ; genetics ; Retroviridae ; genetics

Impaired tumor necrosis factor receptor-1 (TNFR-1) signaling has been found in some malignant tumors with poor prognosis. However, the exact role of TNFR-1 signaling in fibrosarcoma remains unclear. Here, we explored the question by comparing the growth of TNFR-1 deficient (Tnfr1 (-)) and TNFR-1 competent (Tnfr1 (+)) fibrosarcoma FB61 cells (FB61-m and FB61-R1) in mice. TNFR-1 expression on fibrosarcoma cells delayed their growth in vivo but not in vitro. Moreover, reduced FB61-R1 tumor growth was also obtained in TNFR-1 knockout mice. The mechanism relies mainly on the TNFR-1-mediated downregulation of vascular endothelial growth factor (VEGF) production by tumor cells. Importantly, treatment of FB61-m tumors with melphalan resulted in a short delay of tumor growth, followed by a quick remission. However, when FB61-R1 tumors were treated with melphalan, tumor growth was similarly delayed at first and then completely rejected. Our results reveal evidence for TNFR-1 on tumor cells as a prerequisite in chemotherapy for fibrosarcoma, and provide novel insight into the therapeutic approach against some types of tumors using TNFR-1 angonist.
Animals ; Down-Regulation ; drug effects ; Fibrosarcoma ; drug therapy ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Melphalan ; administration & dosage ; Mice ; Molecular Targeted Therapy ; Receptors, Tumor Necrosis Factor, Type I ; genetics ; Signal Transduction ; drug effects ; Vascular Endothelial Growth Factor A ; biosynthesis

Metastasis is the leading cause of mortality for cancer patients, and lymphatic metastasis is one of the main ways of tumor metastasis. The role of CCL21 and its receptor CCR7 in lymphatic metastasis has been increasingly concerned in recent years. CCR7 is mainly expressed by both dendritic cells and T cells for immune responses. CCL21, the chemokine ligand for CCR7, secreted from lymphatic endothelial cells binds CCR7 and recruits immune cells toward lymphatic vessels and lymphatic nodes. CCR7 expressed tumor cells can also metastasize to lymphatic system by the similar way as immune cells. Targeting CCL21/CCR7 axis to inhibit lymphatic metastasis but remain potent anti-tumor immune response has increasingly become a spot light of tumor immunotherapy. In this review, we summarize the role of CCL21/CCR7 axis in lymphatic metastasis, as well as preclinical trials and clinical trials in targeting CCL21/CCR7 axis for tumor metastasis therapy, hoping to accelerate the progress on tumor metastasis therapy by targeting CCL21/CCR7 axis.
Cell Line, Tumor ; Chemokine CCL21 ; Endothelial Cells ; Humans ; Lymphatic Metastasis ; Neoplasms/therapy* ; Receptors, CCR7/genetics*

The lung is one of the most common sites for cancer metastasis. Collagens in the lung provide a permissive microenvironment that supports the colonization and outgrowth of disseminated tumor cells. Therefore, down-regulating the production of collagens may contribute to the inhibition of lung metastasis. It has been suggested that miR-29 exhibits effective anti-fibrotic activity by negatively regulating the expression of collagens. Indeed, our clinical lung tumor data shows that miR-29a-3p expression negatively correlates with collagen I expression in lung tumors and positively correlates with patients' outcomes. However, suitable carriers need to be selected to deliver this therapeutic miRNA to the lungs. In this study, we found that the chemotherapy drug cisplatin facilitated miR-29a-3p accumulation in the exosomes of lung tumor cells, and this type of exosomes exhibited a specific lung-targeting effect and promising collagen down-regulation. To scale up the preparation and simplify the delivery system, we designed a lung-targeting liposomal nanovesicle (by adjusting the molar ratio of DOTAP/cholesterol-miRNAs to 4:1) to carry miR-29a-3p and mimic the exosomes. This liposomal nanovesicle delivery system significantly down-regulated collagen I secretion by lung fibroblasts in vivo, thus alleviating the establishment of a pro-metastatic environment for circulating lung tumor cells.

Yersiniosis is one of the "other infectious diarrhea" of the notifiable infectious diseases and also an important food-borne disease. However, it lacked the basis or standard for diagnosis. The Chinese Preventive Medicine Association coordinated experienced researchers from National Institute for Communicable Disease Control and Prevention, China CDC and other institutes to produce the group standard entitled "Diagnosis of Yersiniosis" (T/CPMA 005-2019). Based on the principle of "legality, scientificity, advancement, and feasibility" , the standard gives a clear definition for Yerisiniosis, stipulates diagnosis basis, principles and main differential diagnosis and provides two informative appendixes for epidemiological and clinical characteristics and a normative appendix for laboratory detection. The standard provides accurate basis and methods of Yersiniosis diagnosis for hospitals and CDCs at all levels in China. It will solve the problems that Yersiniosis cannot be clearly diagnosed for clinical cases and in the outbreaks.