Upregulation of stromal cell-derived factor-1 alpha/CXCR4 axis-induced migration of human neural progenitors by tumor necrosis factor-alpha and interleukin-8

BACKGROUND： Studies of several animal models of central nervous system diseases have shown that neural progenitor cells （NPCs） can migrate to injured tissues. Stromal cell-derived factor 1 alpha （SDF-la）, and its primary physiological receptor CXCR4, have been shown to contribute to this process. OBJECTIVE： To investigate migration efficacy of human NPCs toward a SDF-1α gradient, and the regulatory roles of tumor necrosis factor-α （TNF-α） and interleukin-8 （IL-8） in SDF-1α/CXCR4 axis-induced migration of NPCs. DESIGN, TIME AND SETTING： An in vitro, randomized, controlled, cellular and molecular biology study was performed at the Laboratory of Department of Cell Biology, Medical College of Soochow University between October 2005 and November 2007. MATERIALS： SDF-1α and mouse anti-human CXCR4 fusion antibody were purchased from R＆D Systems, USA. TNF-αwas purchased from Biomyx Technology, USA and IL-8 was kindly provided by the Biotechnology Research Institute of Soochow University. METHODS： NPCs isolated from forebrain tissue of 9 to 10-week-old human fetuses were cultured in vitro. The cells were incubated with 0, 20, and 40 ng/mL TNF-α, or 0, 20, and 40 ng/mL IL-8, for 48 hours prior to migration assay. For antibody-blocking experiments, cells were further pretreated with 0, 20, and 40 μg/mL mouse anti-human CXCR4 fusion antibody for 2 hours. Subsequently, the transwell assay and CXCR4 blockade experiments were performed to evaluate migration of human NPCs toward a SDF-1α gradient. Serum-free culture medium without SDF-1α served as the negative control. MAIN OUTCOME MEASURES： The transwell assay was performed to evaluate migration of human NPCs toward a SDF-1α gradient, which was blocked by fusion antibody against CXCR4. In addition, CXCR4 expression in human NPCs stimulated by TNF-α and IL-8 was measured by flow cytometry. RESULTS： Results from the transwell assay demonstrated that SDF-1α was a strong chemoattractant for human NPCs （P 〈 0.01）, and 20 ng/mL produced the highest levels of migration. Anti-human CXCR4 fusion antibody significantly blocked the chemotactic effect （P 〈 0.05）. Flow cytometry results showed that treatment with TNF-α and IL-8 resulted in increased CXCR4 expression and greater chemotaxis efficiency of NPCs towards SDF-1α（P 〈 0.01）. CONCLUSION： These results demonstrated that SDF-la significantly attracted NPCs in vitro, and neutralizing anti-CXCR4 antibody could block part of this chemotactic function. TNF-α and IL-8 increased chemotaxis efficiency of NPCs towards the SDF-1αgradient by upregulating CXCR4 expression in NPCs.

THE CHANGE OF SERUM TUMOR NECROSIS FACTOR-α LEVEL AND ITS CLINICAL SIGNIFICANCE DURING THE TREATMENT OF CHRONIC HEPATITIS B WITH LAMIVUDINE

Objective To evaluate the effect of lamivudine on immunity of chronic hepatitis B by observing the sequential changes of serum TNF-α and HBV-DNA level. Methods 31 CHB patients with elevated serum ALT/AST level and HBVDNA level higher than 106 copies/mL were treated with lamivudine (100mg/day) for one year. The sequential serum samples, which were taken at the 0, 3 rd, 6 th, 12 th month after initiation of therapy, were used to detect serum level of TNF-α and quantity of HBV-DNA respectively. Results ① The serum TNF-α levels were higher than normal value before treatment in all patients; ② At In the 3 rd month of treatment, The the serum HBV-DNA level began to decline and became negative in the 54.9% of all patients. At the end of treatment, HBV-DNA was negative in 48.4% of all patients; ③ The decrease of TNF-α level was later than HBVDNA level drop. TNF-α level began to decline after 6 months treatment. At the end of treatment, TNF-α level was lower than that at in 6 th month, TNF-α level returned to normal in the 38.7% of all patients; ④ The TNF-α level decreased significantly after 6 months treatment in the patients with ALT>80IU/L at the beginning of treatment. But in the patients with ALT≤80IU/L, the TNF-α level decreased just after 12 months treatment; ⑤ TNF-α level fell obviously and early in patients whose HBVDNA became negative at in the 3 rd month. Conclusion Lamivudine can suppress the replication of HBVDNA quickly, and decrease TNF-α level in the serum TNF-α level. It suggests that lamivudine can modulate immune response directly or indirectly. The changes of serum TNF-α level may be used to evaluate the clinical efficacy of lamivudine.

Promising Effects of Zerumbone on the Regulation of Tumor-promoting Cytokines Induced by TNF-α-activated Fibroblasts

Inflammation plays an important role in the development of several cancers.Inflammatory cytokines,including tumor necrosis factor-α(TNF-α),are associated with the induction of inflammation.Chronic inflammation contributes to the progression of cancer through several mechanisms,including increased cytokine production and activation of transcription factors,such as nuclear factor-κB(NF-κB).Zerumbone(ZER),a component of subtropical ginger(Zingiber zerumbet Smith),seems to have anti-inflammatory,anti-cancer,and antioxidant activities.In this study,we aimed to explore the protective function and mechanisms of ZER against TNF-α-induced cancer-promoting cytokines.We found that the viability of stimulated human fibroblast cell lines was reduced after treatment with ZER(IC50=18µmol/L),compared to un-stimulated fibroblasts(IC50=40µmol/L).Besides,ZER inhibited mRNA expression and protein secretion of transforming growth factor-β(TGF-β),interleukin-33(IL-33),monocyte chemoattractant protein-1(MCP-1),and stromal cell-derived factor 1(SDF-1),which were produced by TNF-α-induced fibroblasts,as measured by quantitative real time-PCR(qRT-PCR)and ELISA assays.The mRNA expression levels of TGF-β,IL-33,SDF-1,and MCP-1 showed 8,5,2.5,and 4-fold reductions,respectively.Moreover,secretion of TGF-β,IL-33,SDF-1,and MCP-1 was reduced to 3.65±0.34 ng/mL,6.3±0.26,1703.6±295.2,and 5.02±0.18 pg/mL,respectively,compared to the untreated group.In addition,the conditioned media(CM)of TNF-α-stimulated fibroblasts increased the NF-κB expression in colorectal cancer cell lines(HCT-116 and Sw48),while in the vicinity of ZER,the expression of NF-κB was reversed.Considering the significant effects of ZER,this component can be used as an appropriate alternative herbal treatment for cancer-related chronic inflammation.

酸脂清胶囊对尿酸性肾病大鼠肾功能及TNF-α的影响

目的:观察酸脂清胶囊对尿酸性肾病大鼠肾功能及TNF-α的影响,探讨其作用的机理。方法:将6周龄SD雄性大鼠分为5组,分别为正常组、模型组、生理盐水组、酸脂清治疗组、别嘌醇治疗组。后4组用腺嘌呤、乙胺丁醇造模,成功后分别给予生理盐水、酸脂清、别嘌醇灌胃给药,3周后检测各组大鼠血中尿素氮(BUN)、肌酐(Cr)、尿酸(UA),同时用酶联反应法(Elisa法)测量各组肾脏组织中肿瘤坏死因子α(TNF-α)的含量。结果:1)造模2周后,与正常组比较,模型组大鼠血中BUN、Cr、UA均明显升高(P<0.05),证实造模成功。2)治疗3周后,与生理盐水组相比,酸脂清治疗组、别嘌醇治疗组大鼠血中BUN、UA均明显下降(P<0.05),酸脂清治疗组和别嘌醇治疗组Cr均有下降,但只有酸脂清治疗组有统计学意义(P<0.05)。3)酸脂清治疗组和别嘌醇治疗组血中BUN、Cr、UA相比,差异无统计学意义(P>0.05)。4)Elisa法测定各组TNF-α的浓度,与正常组相比,生理盐水组TNF-α明显升高(P<0.05);与生理盐水组相比,酸脂清治疗组、别嘌醇治疗组TNF-α含量明显减少(P<0.05);5)肾组织病理切片显示:与模型组相比,酸脂清治疗组、别嘌醇治疗组尿酸盐结晶沉积、炎性细胞浸润均明显减少,肾小管扩张改善。结论:酸脂清胶囊能够降低尿酸性肾病大鼠血中BUN、Cr、UA的含量,减少尿酸盐结晶在肾小管中沉积及炎性细胞浸润,降低TNF-α在肾组织中的含量,从而达到治疗的作用。

TRAF2-MLK3 interaction is essential for TNF-α-induced MLK3 activation

Mixed lineage kinase 3 （MLK3） is a mitogen-activated protein kinase kinase kinase that is activated by tumor necrosis factor-α （TNF-α） and specifically activates c-Jun N-terminal kinase （JNK） on TNF-a stimulation. The mecha- nism by which TNF-α activates MLK3 is still not known. TNF receptor-associated factors （TRAFs） are adapter molecules that are recruited to cytoplasmic end of TNF receptor and mediate the downstream signaling, including activation of JNK. Here, we report that MLK3 associates with TRAF2, TRAF5 and TRAF6; however only TRAF2 can significantly induce the kinase activity of MLK3. The interaction domain of TRAF2 maps to the TRAF domain and for MLK3 to its C-terminal half （amino acids 511-847）. Endogenous TRAF2 and MLK3 associate with each other in response to TNF-α treatment in a time-dependent manner. The association between MLK3 and TRAF2 mediates MLK3 activation and competition with the TRAF2 deletion mutant that binds to MLK3 attenuates MLK3 kinase activity in a dose-dependent manner, on TNF-α treatment. Furthermore the downstream target of MLK3, JNK was activated by TNF-α in a TRAF2-dependent manner. Hence, our data show that the direct interaction between TRAF2 and MLK3 is required for TNF-α-induced activation of MLK3 and its downstream target, JNK.

In silico prediction of phytoconstituents from Ehretia laevis targeting TNF-αin arthritis

Objective Rheumatoid arthritis(RA)is an autoimmune disease involving the synovial lining of the major joints.Current therapies have noteworthy side effects.Our study involved in silico evaluation of Ehretia laevis(E.laevis)phytoconstituents targeting tumor necrosis factor-α(TNF-α).Methods Molecular docking studies performed to investigate the binding pattern of the plant E.laevis phytoconstituents along with the crystal structure of TNF-α(PDB ID:2 AZ5)using AutoDock Vina followed by a study of interacting amino acid residues and their influence on the inhibitory potentials of the active constituents.Further the pharmacokinetic profile and toxicity screening carried out using Swiss ADME and pk CSM.Results The docked results suggest that lupeol(-9.4 kcal/mol)andα-amyrin(-9.4 kcal/mol)has best affinity towards TNF-αcompared to standard drug thalidomide(-7.4 kcal/mol).The active chemical constituents represents better interaction with the conserved catalytic residues,leading to the inhibition/blockade of the TNF-α-associated signaling pathway in RA.Furthermore,pharmacokinetics and toxicity parameters of these phytochemicals were within acceptable limits according to ADMET studies.Conclusion The binding potential of phytoconstituents targeting TNF-αshowed promising results.Nonetheless,it encourages the traditional use of E.laevis and provides vital information on drug development and clinical treatment.

慢性阻塞性肺疾病急性加重期血清瘦素与肺功能关系的临床研究

目的：探讨血清瘦素（ leptin ）与慢性阻塞性肺疾病（ chronic obstructive pulmonary disease， COPD）急性加重期患者肺功能的关系。方法选择COPD急性加重期患者35例、COPD稳定期患者38例及健康志愿者30例，抽取空腹血。用放免法测定血清瘦素、用酶联免疫吸附法（ELISA）测定肿瘤坏死因子-α（TNF-α）及用全自动生化仪检测C-反应蛋白（CRP）；检测肺功能，记录1 s用力呼气量（ FEVl ）、最大通气量（ MVV）、肺一氧化碳弥散量（ DLCO ）等指标、分析COPD急性加重期患者血清瘦素水平与TNF-α、CRP、肺功能等的相关性。结果①COPD急性加重期患者血清瘦素、TNF-α、CRP均明显高于COPD稳定期组和对照组，而FEVl、MVV、DLCO则明显低于后者；②COPD急性加重期患者血清瘦素与TNF-α、CRP呈正相关，与FEVl、MVV、DLCO呈负相关。结论血清瘦素可判断COPD急性加重期预后，可以评估肺功能损害风险。

中西医结合治疗脓毒症的临床疗效观察

目的：探讨益气活血解毒中药汤剂治疗脓毒症的临床疗效。方法将62例脓毒症患者采用信封随机法分为对照组（31例）、治疗组（31例）。对照组给予脓毒症西医常规治疗，治疗组在脓毒症西医常规治疗基础上加用益气活血解毒法中药汤剂中西医结合治疗。分析比较两组炎性因子、病情危重程度评分、住ICU时间及28 d病死率。结果治疗组治疗后IL-6、TNF -α水平较对照组明显下降（P＜0．05），住ICU时间较对照组明显缩短（P＜0．05），APACHEⅡ评分明显低于对照组（P＜0．05），28 d病死率低于对照组（26．7％vs 37．9％，P＝0．048）。结论益气活血解毒法可明显降低炎性因子IL-6及TNF-α水平，缩短住ICU时间，提高28 d生存率。

复方柴芩退热颗粒对发热大鼠模型退热作用的研究

目的:研究复方柴芩退热颗粒对2,4二硝基苯酚及细菌内毒素所致大鼠发热模型的退热作用,进一步明确该颗粒的退热效果,初步探讨其退热机制。方法:将SPF级SD大鼠随机分为正常对照组、模型组、酚麻美敏片组[给药剂量0.148 g/(kg·d)]、小柴胡冲剂组[给药剂量2.160 g/(kg·d)]、复方柴芩退热颗粒高、中、低剂量组[给药剂量分别为6.480、3.240、1.620 g/(kg·d)]。各给药组按相应剂量灌胃给药,正常对照组、模型组同法给予等体积的蒸馏水,每天1次,连续给药5天,第4天各组动物均开始禁食不禁水24 h,末次给药前开始造模。造模实验一:除正常对照组外,余各组大鼠均皮下注射2,4-二硝基苯酚17 mg/kg,正常对照组皮下注射同体积生理盐水;造模实验二:除正常对照组外,余各组大鼠均腹腔注射细菌内毒素80μg/kg。测大鼠各时间点的体温和血清中白细胞介素-1β(IL-1β)、前列腺素E2(PGE2)、肿瘤坏死因子-α(TNF-α)的含量。结果:实验一:与正常对照组比较,模型组大鼠造模后1 h、2 h、3 h体温上升值均显著升高,大鼠血清IL-1β、PGE2含量均显著升高(P<0.01)。与模型组比较,酚麻美敏片组、小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组大鼠血清IL-1β、PGE2含量均明显降低(P<0.05,P<0.01);酚麻美敏片组大鼠在造模后1 h、2 h、3 h体温上升值均显著降低(P<0.05,P<0.01);小柴胡冲剂组大鼠在造模后2 h、3 h体温上升值均显著降低(P<0.05,P<0.01);复方柴芩退热颗粒高、低剂量组在造模后1 h、2 h、3 h体温上升值均明显降低(P<0.05,P<0.01);复方柴芩退热颗粒中剂量组在造模后2 h、3 h体温上升值均明显降低(P<0.05)。实验二:与正常对照组比较,模型组大鼠造模后2 h、4 h、6 h、8 h体温上升值均显著升高,大鼠血清PGE2、TNF-α含量均显著升高(P<0.01)。与模型组比较,酚麻美敏片组、小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组大鼠血清PGE2、TNF-α含量均明显降低(P<0.05,P<0.01);酚麻美敏片组大鼠在造模后2 h、4 h、6 h体温上升值均显著降低(P<0.05,P<0.01);小柴胡冲剂组、复方柴芩退热颗粒高、中、低剂量组在造模后2 h、4 h体温上升值均明显降低(P<0.05,P<0.01)。结论:复方柴芩退热颗粒对2,4二硝基苯酚及细菌内毒素所致大鼠发热均有降温作用,能有效降低大鼠血清PGE2、IL-1β和血清TNF-α含量,对大鼠实验性高热模型有较好的解热作用。

Lipopolysaccharide “Two-hit” Induced Refractory Hypoxemia Acute Respiratory Distress Model in Rats

To establish a stable and reliable model of refractory hypoxemia acute respiratory distress syndrome （ARDS） and examine its pathological mechanisms, a total of 144 healthy male Wistar rats were randomized into 4 groups： group Ⅰ （saline control group）, group Ⅱ （LPS intravenous ＂single-hit＂ group）, group Ⅲ （LPS intratracheal ＂single-hit＂ group） and Group IV （LPS ＂two-hit＂ group）. Rats were intravenously injected or intratracheally instilled with a large dose of LPS （10 mg/kg in 0.5 mL） to simulate a single attack of ARDS, or intraperitoneally injected with a small dose of LPS （1 mg/kg） followed by tracheal instillation with median dose of LPS （5 mg/kg） to establish a ＂two-hit＂ model. Rats in each group were monitored by arterial blood gas analysis and visual inspection for three consecutive days. Arterial blood gas values, lung wet/dry weight ratio and pathological pulmonary changes were analyzed to determine the effects of each ALI/ARDS model. Concentrations of TNF-α, IL-1 and IL-10 in the bronchoalveolar lavage fluid （BALF） and blood plasma were meastired by using enzyme-linked immunosorbent assays （ELISA）. Our resulsts showed that single LPS-stimulation, whether through intravenous injection or tracheal instillation, could only induce ALl and temporary hypoxemia in rats. A two-hit LPS stimulation induces prolonged hypoxemia and specific pulmonary injury in rats, and is therefore a more ideal approximation of ARDS in the animal model. The pathogenesis of LPS two-hit-induced ARDS is associated with an uncontrolled systemic inflammatory response and inflammatory injury. It is concluded that the rat ARDS model produced by our LPS two-hit method is more stable and reliable than previous models, and closer to the diagnostic criteria of ARDS, and better mimics the pathological process of ARDS.

Protective Effect of Diallyl Trisulfide on Liver in Rats with Sepsis and the Mechanism

The protective effects of diallyl trisulfide on liver were examined in rats with sepsis. Sepsis was reproduced in rats by cecum ligation and puncture (CLP). Fifty-six male Wistar rats were randomly divided into sham-operated group (group S, n=8), sepsis model group (group C, n=24), diallyl trisulfide (DATS)-treated group (group D, n=24). Animals in groups C and D were further divided into three subgroups according to different observation time points, with 8 rats in each sub-group.Rats in group D and C were intravenously injected with normal saline or DATS respectively at a dose of 20 mg/kg after the establishment of sepsis model. Eight rats in groups C and D were sacrificed at 3, 6 and 24 h post-CLP and their livers were harvested for detection of interleukin (IL)-1 receptor associated kinase-4 (IRAK-4), nuclear factor-κB (NF-κB), c-fos, c-jun, malondialdehydethhe (MDA) and superoxide dismutase (SOD), tumor necrosis factor alpha (TNF-α) and for pathological examination. The results showed that the levels of serum IRAK-4, NF-κB and TNF-α in hepatic tissues were higher in group C than group S (control group) (P<0.05). After DATS treatment, the levels of IRAK-4 and NF-κB in the hepatic tissues and serum TNF-α in group D were lower than those in group C (P<0.05). The levels of c-fos and c-jun and MDA in the hepatic tissues were higher in group C than in group S (P<0.05). After DATS treatment, the levels of c-fos and c-jun and MDA in the hepatic tissues were significantly lower in group D than in group C (P<0.05). When compared with group S group, concentration of SOD in the hepatic tissues in group C was significantly lower (P<0.05). After DATS treatment, the concentration of SOD in the hepatic tissues was higher in group D than in group C (P<0.05). These findings suggested that treatment with DATS could ameliorate sepsis-induced liver injury in rats. The protective effect might be related to its ability to inhibit the signal pathway of IRAK-4 and NF-κB, thereby decreasing the production of oxygen free radicals and down-regulating the expression of c-fos and c-jun.

Inflammatory Mechanism of Total Flavonoids of Chrysanthemum and Medicated Serum on Castrated Dry Eye Animal and Cell Models

Objective To observe the effects of total flavonoids of chrysanthemum and medicated serum on the expression of related proteins in the lacrimal tissue and dry-eye cell models of male rabbits with dry eye caused by castration.Methods(1)150 male Japanese rabbits were randomly divided into five groups,with 30 rabbits in each group:normal control group(group A),sham group(group B),model group(group C),androgen control group(group D)and total flavonoids of chrysanthemum treatment group(group E).The androgen deficiency dry-eye model was established by bilateral castration in groups C,D and E.Normal saline was administered to groups A,B and C by gavage;androgen(testosterone propionate)was injected into muscle in group D;and group E was given total flavonoids of chrysanthemum by gavage.All white rabbits were tested the Schirmer I test(SIT)and tear break-up time(BUT).After euthanasia,tear gland tissue was harvested so that we could observe pathological changes in the expression of related inflammatory factors in the lacrimal gland tissue.The expression of interleukin-1β(IL-1β),tumor necrosis factor-α(TNF-α)and transforming growth factor-β1(TGF-β1)was detected in the lacrimal gland tissue by immunohistochemistry.Reverse transcription PCR was used to quantitatively detect expression of TGF-β1 mRNA.(2)Male Wistar rat lacrimal epithelial cells were used to establish a model of eye stem cell apoptosis caused by androgen levels.The blank control group was set up without androgen culture,the control group with androgen culture,and the total flavonoids of chrysanthemum group without androgen.The MTT method was used to determine the optimal intervention dosage of drug-containing plasma.Western blot and QPCR were used to detect the expression of AR mRNA,NF-κB phosphorylated protein and TGF-β1 in lacrimal epithelial cells,and the androgen-like effect of total flavonoids of chrysanthemum was observed.Results(1)Immunohistochemistry showed that groups A,B,D and E had significantly lower expression of IL-1βand TNF-αthan group C(P<0.05);among these,group E had slightly higher expression than group D(P>0.05).RT-PCR results showed that the relative expression of TGF-β1 mRNA in groups A,B,D and E was significantly higher than in group C(P<0.05),and the relative expression of TGF-β1 mRNA in groups D and E was higher than that in groups A and B(P<0.05).(2)Using the MTT method,the final concentration of interfering cells was calculated to be 13.2%.The expression of AR protein,NF-κB and TGF-β1 in the chrysanthemum flavonoid plasma intervention and testosterone propionate intervention groups was enhanced,and there were significant differences relative to the blank group(P<0.01).The expression level of NF-κB in the total flavonoid containing plasma intervention group was lower than that in the testosterone propionate intervention group(P<0.01).Conclusions The total flavonoids of chrysanthemum can inhibit IL-1βand TNF-αexpression in the lacrimal gland tissue of castrated male rabbits with dry eye to increase synthesis of TGF-β1 mRNA and TGF-β1,thereby inhibiting the inflammatory response.The medicated plasma with total flavonoids of chrysanthemum promotes expression of AR mRNA,upregulating expression of NF-κB,further promoting upregulation of TGF-β1 protein expression in lacrimal epithelial cells,inhibiting inflammation by regulating related proteins,and ultimately alleviating the symptoms of dry eye.

Dual therapy of rosiglitazone/pioglitazone with glimepiride on diabetic nephropathy in experimentally induced type 2 diabetes rats

Diabetic nephropathy is a major cause of end-stage renal disease （ESRD） in the general population. It is estimated that diabetic nephropathy will eventually develop in about 40% of all patients with diabetes; therefore, prevention is critical for delaying the development and progression of diabetic kidney disease. Despite extensive efforts, medical advances are still not successful enough to prevent the progression of the disease. In the present study, we focused on the comparison of combination therapies and whether they offered additional renoprotection. Type 2 diabetes mellitus was induced by intraperitoneally administering streptozotocin （90 mg/kg） in neonatal rats and then these rats were treated with rosiglitazone （1.0 mg/kg） in combination with glimepiride （0.5 mg/kg） or with pioglitazone （2.5 mg/kg） in combination with glimepiride （0.5 mg/kg）. Diabetic nephropathy markers were evaluated by biochemical and ELISA kits and renal structural changes were examined by light microscopy and transmission electron microscopy. Results show that the combination of pioglitazone with glimepiride is more effective in amelioration of diabetic nephropathy than rosiglitazone with glimepiride drug therapy due to glycemic control, suppressing albumin excretion rate, total protein excretion rate and augmented TNF-α signaling during the development of streptozotocin induced type 2 diabetic nephropathy.

Relationship between peritoneal macrophages and inflammatory reaction in a rat model of severe acute pancreatitis

Objective To investigate the relationship between peritoneal macrophages(PMAs)and inflammatory reaction in a rat model of severe acute pancreatitis(SAP).Methods Sprague-Dawley rats were randomly divided into control group and SAP group.To induce SAP in rats,40 g/L sodium taurocholate(0.1 mL/100 g)was injected into the pancreatic duct through retrograde exposure of pancreatic bile duct in hepatic porta.One-third of rats were sacrificed at 3,6 or 12 h after modeling.PMAs were extracted,and incubated for 24 h in a humidified 5% carbon dioxide incubator.The expressions of tumor necrosis factor alpha(TNF-α)and interleukin-1β(IL-1β)mRNA in PMAs were measured by semi-quantitative RT-PCR.The levels of TNF-α and IL-1β in culture medium and serum were evaluated.The histological changes of pancreas were examined.Results The expressions of TNF-α mRNA and IL-1β mRNA in PMAs were significantly higher in SAP group than in control group at each time point(P<0.01).The concentrations of TNF-α and IL-1β in culture medium and serum were significantly elevated in SAP group compared with control group(P<0.01).The histological analysis of pancreas indicated that the damage was more severe in SAP group than in control group(P<0.01).Conclusion PMAs secrete cytokines into pancreatitis-associated ascitic fluid,and this study demonstrates a correlation between SAP and the activation of PMAs.

Effects of External Use of Pereskia aculeate Miller on Foot Swelling of Adjuvant Arthritis in Rats

[Objectives] To study the anti-inflammatory effects and its possible action mechanism of Pereskia aculeate Miller on rats with adjuvant arthritis( AJ). [Methods] Fifty SD rats( half male and half female) were randomly divided into 5 groups: blank group,model group,positive control group( Glucosidorum Tripterygll Totorum( GTT),12 mg/kg),and P. aculeate high and low dose group( 0. 86 and0. 43 g/m L). Except the blank group,other groups were induced with complete Freund's adjuvant( CFA) to establish the AA rat model. On the 17 th day of modeling,the drug was externally applied for soaking,and the diameter of foot hole and foot joint before and after administration was measured to observe the degree of swelling. The enzyme-linked immunosorbent assay( ELISA) was adopted to determine the inflammatory cytokines interleukin-1β( IL-1β) and tumor necrosis factor-α( TNF-α). [Results] Compared with the model group,the degree of swelling of the foot sole and foot joints was reduced in the P. aculeate high dose group and the positive control group( P < 0. 05). According to ELISA test,compared with the model control group,the serum levels of IL-1β( P < 0. 05) and TNF-α( P < 0. 05) were significantly reduced in the P. aculeate high dose group and the positive control group. Compared with the positive control group,there was no significant difference( P > 0. 05). [Conclusions] P. aculeate has significant inhibitory effects on foot swelling of adjuvant arthritis in rats,and the action mechanism is possibly related to the decrease of IL-1β and TNF-α.

The efficacy and safety of thalidomide for treating metastatic breast cancer:a systematic review

Objective This systematic review was conducted to investigate the efficacy and safety of thalidomide in metastatic breast cancer(MBC).Methods Based on pre-defined inclusion and exclusion criteria,data were independently collected from different databases by three investigators.Overall,three studies were included.Results The included studies indicated that no patient achieved a partial or complete response from different thalidomide dose levels.Thalidomide was well-tolerated at doses of 100 mg,200 mg,and 400 mg.In all three studies,common side effects included constipation,somnolence,fatigue,peripheral neuropathy,and dry mouth.Circulating angiogenic factors were not significantly correlated with disease progression.Conclusion The available evidence indicates that single-agent thalidomide has little or no activity in patients with MBC.

Changes in peripheral blood inflammatory factors(TNF-α and IL-6) and intestinal flora in AIDS and HIV-positive individuals

Objective:In this study,we investigated the changes in peripheral blood inflammatory factors and intestinal flora in acquired immune deficiency syndrome(AIDS)and human immunodeficiency virus(HIV)-positive individuals(AIDS/HIV patients),and explored the relationships among intestinal flora,peripheral blood inflammatory factors,and CD4^+T lymphocytes.Methods:Thirty blood and stool samples from an AIDS group and a control group were collected.The levels of tumor necrosis factor-α(TNF-α)and interleukin-6(IL-6)were determined by enzyme-linked immunosorbent assay(ELISA),and the number of CD4^+T lymphocytes by a FACSCount automated instrument.Quantitative real-time polymerase chain reaction(qRT-PCR)was used to determine the messenger RNA(mRNA)levels of Bifidobacterium,Lactobacillus,Escherichia coli,Enterococcus faecalis,and Enterococcus faecium.Correlations among intestinal flora,inflammatory factor levels,and CD4^+T lymphocyte values were evaluated using the Spearman correlation coefficient.Results:The levels of TNF-αand IL-6 in the AIDS group were higher than those in the control group,while the number of CD4^+T lymphocytes was lower.The amounts of Bifidobacterium and Lactobacillus in the AIDS group were significantly lower than those in control group,while the amounts of E.coli,E.faecalis,and E.faecium were much higher.The amounts of Bifidobacterium and Lactobacillus were negatively correlated with the content of TNF-αand IL-6 and the CD4^+T lymphocyte count,while those correlations were reversed for E.coli,E.faecalis,and E.faecium.Conclusions:The intestinal microbiota of AIDS/HIV patients were disordered,and there was a correlation between the amount of intestinal flora and the number of CD4^+T lymphocytes and the levels of TNF-αand IL-6.

Surface charge tunable nanoparticles for TNF-α siRNA oral delivery for treating ulcerative colitis

Nanopartide （NP） drug delivery systems have been successfully designed and implemented to orally deliver small interfering RNAs （siRNAs） for inflammatory disorders. However, the influence of surface charge on orally administered siRNA nanocarriers has not been investigated. In this study, we prepared structurally related poly（ethylene glycol）-block-poly（lactic-co-glycolic acid） （PEG5K-b-PLGA10K） NPs with the assistance of a synthesized lipid featuring surface amine groups for subsequent charge tuning. NPs were prepared by a double emulsion method, and their surface charge could be tuned and controlled by a succinylation reaction to yield NPs with different surface charges, while maintaining their size and composition. The prepared NPs were termed as aminated NPs （ANPs）, plain NPs （PNPs）, or carboxylated NPs （CNPs） based on their surface charge. All NPs exhibited the desired structural stability and siRNA integrity after enzymatic degradation. In vivo studies showed that ANPs significantly accumulated in inflamed colons, and they were successful in decreasing TNF-α secretion and mRNA expression levels while maintaining colonic histology in a murine model of acute ulcerative colitis （UC）. This study described a methodology to modify the surface charge of siRNA-encapsulating polymeric NPs and highlighted the influence of surface charge on oral delivery of siRNA for localized inflammatory disorders.

MODULATION OF MACROPHAGE-MEDIATED CYTOTOXIC ACTIVITY——EFFECT OF FUSARIN C ON Mφ-CF PRODUCTION AND TNF-α mRNA EXPRESSION

Fusarium moniliforme is a predominant fungus found in fungus-contaminated food in Linxian County, China, a high risk area of esophageal. cancer. Rats fed with corn bread inoculated with Fusarium moniliforme developed papillomas and squamous-cell carcinomas

CRP, but not TNF-α or IL-6, decreases after weight loss in patients with morbid obesity exposed to intensive weight reduction and balneological treatment

Objective：The aim of this study was to evaluate the concentrations of C-reactive protein （CRP）, tumor necrosis factor-α （TNF-α）, interleukin-6 （IL-6）, and the degree of homeostasis model assessment-insulin resistance （HOMA-IR） in patients with morbid obesity exposed to a three-week low-calorie diet and balneotherapy. Methods：The study included 33 patients （25 females and 8 males; mean age 46 years） with body mass index （BMI） values of〉40 kg/m2. Evaluations of CRP, IL-6, TNF-α, lipid profile, HOMA-IR, and fasting glucose were carried out before （baseline data） and three weeks after the treatment. The control group consisted of 20 healthy volunteers （15 females and 5 males） with a mean age of 39 years and BMI values of≤24.9 kg/m2. Results：In the blood of patients with morbid obesity we found significantly elevated levels of CRP, TNF-α, triglycerides, HOMA-IR and fasting glucose, but a de-creased level of high density lipoprotein （HDL）-cholesterol, compared with the healthy individuals. The treatment resulted in about a 9.4%reduction in body weight from 122.5 to 111.0 kg and a significant decrease in the concen-tration of CRP, but no change in TNF-αor IL-6. HOMA-IR was significantly reduced. Conclusions：The decrease in CRP level without changes in TNF-α or IL-6 concentrations after the low-calorie diet and balneological treatment, suggests that an essential amount of adipose tissue must be removed before proper adipocyte function is restored. The decrease in HOMA-IR indicates an improvement in insulin sensitivity, which is beneficial in obese patients.