Liuwei Dihuang Pill(六味地黄丸)Treats Postmenopausal Osteoporosis with Shen(Kidney) Yin Deficiency via Janus Kinase/Signal Transducer and Activator of Transcription Signal Pathway by Up-regulating Cardiotrophin-Like Cytokine Factor 1 Expression

Objectives： To investigate the mechanism of Liuwei Dihuang Pill （六味地黄丸, LDP） in treating postmenopausal osteoporosis （PMOP） with Shen （Kidney） yin deficiency. Methods： In this study, 205 cases of PMOP were divided into the PMOP Shen-yin deficiency group （Group A）, PMOP Shen-yang deficiency group （Group B）, PMOP without Shen deficiency group （Group C）, and control group （Group N）. Real-time polymerase chain reaction （RT-PCR） and Western blot techniques were used to observe the effects of LDP treatment on the cardiotrophin-like cytokine factor 1 （CLCF1）, ankyrin repeat and SOCS box containing 1 （ASB1）, and proldneticin 2 （PROK2） genes and the Janus kinase/signal transducer and activator of transcription （JAK/STAT） signaling pathway. Results： The mRNA （P〈0.05） and protein （P〈0.01） expression levels of the CLCF1 gone in Group A were significantly lower than the corresponding levels in Group N. After LDP treatment for 3 months, the mRNA expression levels of the CLCF1 gone were obviously up-regulated （P〈0.01）. After 6-month treatment, the expression levels of CLCF1 mRNA and protein were significantly up-regulated （both P〈0.01）, and the average bone density of the top femur had significantly increased （P〈0.05）. In vitro, CLCF1 overexpression resulted in a significant increase in the total protein and phosphorylated protein levels of JAK2 and STAT3. Conclusions： The CLCF1 gone is an important gone associated with PMOP Shen-yin deficiency and the therapeutic effects of LDP may be mediated by up-regulation of CLCF1 gone expression and activation of the JAK/STAT signaling pathway.

Effects of Bifidobacterium infantis on cytokine-induced neutrophil chemoattractant and insulin-like growth factor-1 in the ileum of rats with endotoxin injury

BACKGROUND The digestive tract is the maximal immunizing tissue in the body, and mucosal integrity and functional status of the gut is very important to maintain a healthy organism. Severe infection is one of the most common causes of gastrointestinal dysfunction, and the pathogenesis is closely related to endotoxemia and intestinal barrier injury. Bifidobacterium is one of the main probiotics in the human body that is involved in digestion, absorption, metabolism, nutrition, and immunity.Bifidobacterium plays an important role in maintaining the intestinal mucosal barrier integrity. This study investigated the protective mechanism of Bifidobacterium during ileal injury in rats.AIM To investigate the effects of Bifidobacterium on cytokine-induced neutrophil chemoattractant(CINC) and insulin-like growth factor 1(IGF-1) in the ileum of rats with endotoxin injury.METHODS Preweaning rats were randomly divided into three groups: Control(group C),model(group E) and treatment(group T). Group E was intraperitoneally injected with lipopolysaccharide(LPS) to create an animal model of intestinal injury.Group T was intragastrically administered Bifidobacterium suspension 7 d before LPS. Group C was intraperitoneally injected with normal saline. The rats were killed at 2, 6 or 12 h after LPS or physiological saline injection to collect ilealtissue samples. The expression of ileal CINC mRNA was evaluated by reverse transcription-polymerase chain reaction(RT-PCR), and expression of ileal IGF-1 protein and mRNA was detected by immunohistochemistry and RT-PCR,respectively.RESULTS The ileum of rats in Group C did not express CINC mRNA, ileums from Group E expressed high levels, which was then significantly decreased in Group T(F =23.947, P < 0.05). There was no significant difference in CINC mRNA expression at different times(F = 0.665, P > 0.05). There was a high level of IGF-1 brown granules in ileal crypts and epithelial cells in Group C, sparse staining in Group E, and dark, dense brown staining in Group T. There was a significant difference between Groups C and E and Groups E and T(P < 0.05). There was no significant difference in IGF-1 protein expression at different times(F = 1.269, P > 0.05). IGF-1 mRNA expression was significantly different among the three groups(P < 0.05),though not at different times(F = 0.086, P > 0.05).CONCLUSION Expression of CINC mRNA increased in the ileum of preweaning rats with endotoxin injury, and exogenous administration of Bifidobacterium reduced CINC m RNA expression. IGF-1 protein and mRNA expression decreased in the ileum of preweaning rats with endotoxin injury, and exogenous administration of Bifidobacterium prevented the decrease in IGF-1 expression. Bifidobacterium may increase IGF-1 expression and enhance intestinal immune barrier function in rats with endotoxin injury.

MELD score,insulin-like growth factor 1 and cytokines on bone density in end-stage liver disease

AIM:To determine the contributions of insulin-like growth factor 1 (IGF-1),cytokines and liver disease severity to bone mineral density in patients pre-transplantation.METHODS:Serum IGF-1,tumor necrosis factor-α (TNFα) and interleukin 6 (IL-6) were measured and the Model for End-Stage Liver Disease (MELD) score calculated in 121 adult patients referred to a single centre for liver transplantation.Bone mineral density (BMD) of the lumbar spine and femoral neck were assessed via dual energy X-ray absorptiometry.Demographics,liver disease etiology,medication use and relevant biochemistry were recorded.RESULTS:A total of 117 subjects were included,with low BMD seen in 68.6%,irrespective of disease etiol-ogy.In multivariable analysis,low body mass index (BMI),increased bone turnover and low IGF-1 were independent predictors of low spinal bone density.At the hip,BMI,IGF-1 and vitamin D status were predictive.Despite prevalent elevations of TNFα and IL-6,levels did not correlate with degree of bone loss.The MELD score failed to predict low BMD in this pre-transplant population.CONCLUSION:Osteopenia/osteoporosis is common in advanced liver disease.Low serum IGF-1 is weakly predictive but serum cytokine and MELD score fail to predict the severity of bone disease.

cytokine 的角色像受体的因素 1 在肝的星形的房间和纤维变性

AIM:To elucidate the role of cytokine receptor-like factor 1(CRLF1) in hepatic stellate cells and liver fibrosis.METHODS:Rat hepatic stellate cells(HSCs) were isolated by Nykodenz gradient centrifugation and activated by culturing in vitro.Differentially expressed genes in quiescent and culture activated HSCs were identified using microarrays.Injections of carbon tetrachloride(CCl 4) for 4 wk were employed to induce liver fibrosis.The degree of fibrosis was assessed by Sirius red staining.Adenovirus expressing CRLF1 was injected through tail vein into mice to achieve overexpression of CRLF1 in the liver.The same adenovirus was used to overexpress CRLF1 in quiescent HSCs cultured in vitro.Expression of CRLF1,CLCF1 and ciliary neurotrophic factor receptor(CNTFR) in hepatic stellate cells and fibrotic livers was analyzed by semi-quantitative reverse transcription-polymerase chain reaction and Western blotting.Expression of profibrotic cytokines and collagens was analyzed by the same method.RESULTS:CRLF1 is a secreted cytokine with unknown function.Human mutations suggested a role in development of autonomous nervous system and a role of CRLF1 in immune response was implied by its similarity to interleukin(IL)-6.Here we show that expression of CRLF1 was undetectable in quiescent HSCs and was highly upregulated in activated HSCs.Likewise,expression of CRLF1 was very low in normal livers,but was highly upregulated in fibrotic livers,where its expression correlated with the degree of fibrosis.A cofactor of CLRF1,cardiotrophin-like cytokine factor 1(CLCF1),and the receptor which binds CRLF1/CLCF1 dimer,the CNTFR,were expressed to similar levels in quiescent and activated HSCs and in normal and fibrotic livers,indicating a constitutive expression.Overexpression of CLRF1 alone in the normal liver did not stimulate expression of profibrotic cytokines,suggesting that the factor itself is not pro-inflammatory.Ectopic expression in quiescent HSCs,however,retarded their activation into myofibroblasts and specifically decreased expression of type Ⅲ collagen.Inhibition of type Ⅲ collagen expression by CRLF1 was also seen in the whole liver.Our results suggest that CLRF1 is the only component of the CRLF1/CLCF1/CNTFR signaling system that is inducible by a profibrotic stimulus and that activation of this system by CLRF1 may regulate expression of type Ⅲ collagen in fibrosis.CONCLUSION:By regulating activation of HSCs and expression of type Ⅲ collagen,CRLF1 may have an ability to change the composition of extracellular matrix in fibrosis.

Heat shock factor 1 promotes neurite outgrowth and suppresses inflammation in the severed spinal cord of geckos

The low intrinsic growth capacity of neurons and an injury-induced inhibitory milieu are major contributo rs to the failure of sensory and motor functional recovery following spinal cord injury.Heat shock transcription factor 1(HSF1),a master regulator of the heat shock response,plays neurogenetic and neuroprotective roles in the damaged or diseased central nervous system.However,the underlying mechanism has not been fully elucidated.In the present study,we used a gecko model of spontaneous nerve regeneration to investigate the potential roles of gecko HSF1(gHSF1) in the regulation of neurite outgrowth and inflammatory inhibition of macrophages following spinal cord injury.gHSF1 expression in neurons and microglia at the lesion site increased dramatically immediately after tail amputation.gHSF1 ove rexpression in gecko primary neuro ns significantly promoted axonal growth by suppressing the expression of suppressor of cytokine signaling-3,and fa cilitated neuro nal survival via activation of the mitogen-activated extracellular signal-regulated kinase/extracellular regulated protein kinases and phosphatidylinositol 3-kinase/protein kinase B pathways.Furthermore,gHSF1 efficiently inhibited the macrophagemediated inflammatory response by inactivating 1kappa B-alpha/NF-kappaB signaling.Our findings show that HSF1 plays dual roles in promoting axonal regrowth and inhibiting leukocyte inflammation,and provide new avenues of investigation for promoting spinal co rd injury repair in mammals.

枢经推拿对脊髓神经结扎大鼠模型炎症反应及SOCS1、TLR4蛋白表达的影响

目的本研究旨在探讨枢经推拿对神经病理性疼痛(NP)的抑炎镇痛机制。方法30只SPF级SD大鼠随机分为正常组、模型组、推拿组、假推拿组、假手术组,每组6只。除了正常组不进行处理,假手术组暴露L5神经外,其余3组大鼠采用脊神经结扎(SNL)法建立NP大鼠模型。造模后第1天开始,推拿组给予枢经推拿治疗,假推拿组给予抚摸,持续干预7 d,在造模前1 d及造模后1 d、3 d、7 d检测各组大鼠热痛缩足反应潜伏期(PWL)和机械性刺激缩足反应阈值(PWT)。干预7 d后,采用Western blot检测大鼠脊髓背角中细胞因子信号转导抑制因子1(SOCS1)、Toll样受体4(TLR4)、核因子κB(NF-κB)表达水平,采用ELISA检测脊髓背角组织白细胞介素6(IL-6)水平。结果造模前1 d,各组大鼠的PWT、PWL没有显著差异(P>0.05);造模后1 d、3 d、7 d,与正常组、假手术组相比,假推拿组、推拿组、模型组大鼠的PWT、PWL均降低(P<0.05),且造模后3 d和7 d,与模型组、假推拿组相比,推拿组大鼠的PWT、PWL均升高(P<0.05)。与正常组、假手术组相比,假推拿组、推拿组、模型组大鼠的SOCS1蛋白水平均明显升高(P<0.05);推拿组大鼠与模型组、假推拿组相比,SOCS1水平升高(P<0.05)。与正常组、假手术组相比,假推拿组、推拿组、模型组大鼠的脊髓背角TLR4、NF-κB、IL-6水平明显增加(P<0.05);与模型组相比,推拿组大鼠的TLR4、NF-κB、IL-6水平明显降低(P<0.05)。结论枢经推拿可以改善由SNL诱导的NP,其机制可能与通过上调脊髓背角中SOCS1蛋白表达,负反馈下调TLR4及NF-κB蛋白表达,降低促炎因子IL-6产生有关。

Interleukin-1α, 6 regulate the secretion of vascular endothelial growth factor A, C in pancreatic cancer

Vascular endothelial growth factor (VEGF, namely VEGF-A) is an angiogenic polypeptide and VEGF-C is a lymphangiogenic polypeptide that has been implicated in cancer growth, invasion and metastasis. Several cytokines and growth factors play an important part in cancer progression. These cytokines and growth factors are the principal mediators of cancer cells-stromal cell interaction , which is critical for invasion of cancer cells to the surrounding tissues and metastatic dissemination to distant organs. In this study, we studied VEGF-A, C expression in cultured human pancreatic cancer cell lines and whether the presence of VEGF-A, C in the cell lines is regulated by cytokines interleukin-lct (EL-1α), and interleukin-6 (IL-6). METHODS: We used Northern blot and Western blot methods to analyze expression of the gene and protein of VEGF-A, C in all 6 tested cell lines (ASPC-1, CAPAN-1, MIA-PaCa-2, PANC-1, COLO-357 and T3M4) respectively. To analyze what is the regulator for this VEGF-A, C expression in pancreatic cancer,we used the reverse transcription -polymerase chain reaction (RT-PCR) method to analyze VEGF-A, C expression in cultured human pancreatic cancer cell lines (CAPAN-1 and COLO-357) under the stimulation with IL-1α (10μg/L) or IL-6 (100 μg/L). RESULTS:Northern blot analysis revealed the presence of the 4.1-kb VEGF-A mRNA transcript and 2.4-kb VEGF-C mRNA transcript in all 6 tested cell lines. Immunoblotting with highly specific anti-VEGF-A, anti-VEGF-C antibody revealed the presence of a molecular weight of 43-kDa VEGF-A protein and 55-kDa VEGF-C protein in all the cell lines. RT-PCR analysis revealed the levels of the VEGF-A and VEGF-C gene were 1-2 fold and a 1-fold increase in the COLO-357 cell line by stimulation with IL-la, however, no effect was found in the CAPAN-1 cell line. The levels of the VEGF-A and VEGF-C gene were 2-5 fold and a 1-fold increase in the CAPAN-1 cell line by stimulation with IL-6, but, no effect was found in the COLO-357 cell line. CONCLUSION:These findings suggested that the expression of VEGF-A, C and their regulation by IL-1α, IL-6 in pancreatic cancer contributes to the lymphatic and distant metastasis and the disease progression.

Adipokines and proinflammatory cytokines, the key mediators in the pathogenesis of nonalcoholic fatty liver disease

Nonalcoholic fatty liver disease(NAFLD)is a condition in which excess fat accumulates in the liver of a patient with no history of alcohol abuse or other causes for secondary hepatic steatosis.The pathogenesis of NAFLD and nonalcoholic steatohepatitis(NASH)has not been fully elucidated.The"two-hit"hypothesis is probably a too simplified model to elaborate complex pathogenetic events occurring in patients with NASH.It should be better regarded as a multiple step process,with accumulation of liver fat being the first step,followed by the development of necroinflammation and fibrosis.Adipose tissue,which has emerged as anendocrine organ with a key role in energy homeostasis,is responsive to both central and peripheral metabolic signals and is itself capable of secreting a number of proteins.These adipocyte-specific or enriched proteins,termed adipokines,have been shown to have a variety of local,peripheral,and central effects.In the current review,we explore the role of adipocytokines and proinflammatory cytokines in the pathogenesis of NAFLD.We particularly focus on adiponectin,leptin and ghrelin,with a brief mention of resistin,visfatin and retinol-binding protein 4 among adipokines,and tumor necrosis factor-α,interleukin(IL)-6,IL-1,and briefly IL-18 among proinflammatory cytokines.We update their role in NAFLD,as elucidated in experimental models and clinical practice.

Adhesion molecule and proinflammatory cytokine gene expression in hepatic sinusoidal endothelial cells following cecal ligation and puncture

INTRODUCTIONMultiple organ dysfunction syndrome (MODS) isthought to be a frequent consequence of sepsis[1-3].Despite substantial advances in our knowledge and understanding of the basic pathophysiologic mechanisms[4-7], in critically ill patients infections and sepsis are still associated with a high mortality[8,9].

Effect of cholecystokinin on cytokines during endotoxic shock in rats

AIM To study the effect of cholecystokinin-octapeptide (CCK-8) on systemic hypotension and cytokine production in lipopolysaccharide (LPS)-induced endotoxic shock (ES) rats.``METHODS The changes of blood pressure were observed using physiological record instrument in four groups of rats: LPS (8 mg. kg-1, iv) induced ES; CCK-8 (40 μg.kg- 1 iv) pretreatment 10 min before LPS (8 mg. kg- 1);CCK-8 (40 μg.kg-1, iv) or normal saline (control) groups.Differences in tissue and circulating specificity of the proinflammatory cytokines (TNF-a, IL-l3 and IL-6) were assayed with ELISA kits.``RESULTS CCK-8 reversed LPS-induced decrease of mean artery blood pressure (MABP) in rats. Compared with control, LPS elevated the serum level of IL-6 significantly (3567_-687 ng.L-1 vs 128_+22 ng.L-1, P＜0.01), while contents of TNF-a and IL- lβ elevated significantly (277 _± 86ng.L-1 vs not detectable and 43 ± 9 ng.L-1 vs notdetectable, P<0.01) but less extent than IL-6, CCK-8significantly inhibited the LPS-induced increase in serum TNF-a, IL-lβ and IL-6. LPS elevated spleen and lung content of IL-Iβ significantly (5184 ± 85 ng.L-1 vs 1047 ±21 ng.L-1 and 4050 ± 614 ng.L-1 vs not detectable,P＜0,01). while levels of TNF-a and IL-6 also rosesignificantly but in less extent than IL-lβ. CCK-8 inhibited the LPS-induced increase of the cytokines in spleen and lung. in the heart, CCK-8 significantly inhibited LPS.induced increase of TNF-a (864 ± 123 ng. L-1 in CCK-8 +LPS group vs 1599_-227 ng-L-1 in LPS group, P＜0.01),and IL-lβ (282 ± 93 ng-L-1 in CCK-8 + LPS group vs 621 ±145 ng.L-1 in LPS group, P＜0.01).``CONCLUSION CCK-8 reverses ES, which may be relatedto its inhibitory effect on the overproduction of cytokines.

Cytokine levels in patients with chikungunya virus infection

Objective: To investigate cytokine profile in patients with chikungunya virus (CHIKV) infection. Methods: Twenty eight pairs of serum samples collected from CHIKV infected patients during the outbreak of chikungunya fever in South Thailand in 2008 were obtained. A multiple cytokine assay for detection of 17 cytokines was performed. Results: In the acute stage of CHIKV infection, the patients had significantly higher levels of interleukin-6, granulocyte colony-stimulating factor, granulocyte-macrophage colony-stimulating factor, monocyte chemotactic protein 1 and tumor necrosis factor alpha than the control ( P<0.001, P=0.023, P=0.015, P <0.001 and P=0.024, respectively). When the disease developed to the recovery stage, the patients had significantly lower levels of interleukin-6, granulocyte-macrophage colony-stimulating factor, monocyte chemotactic protein 1 and macrophage inflammatory protein beta than in the acute stage ( P<0.001). Conclusions: This study provides additional information that these cytokines could play roles in pathogenesis of CHIKV infection and could be used as disease biomarkers or drug targets.

HCV-specific cytokine induction in monocytes of patients with different outcomes of hepatitis C

AIM：Cytokine release by macrophages critically determines the type of immune response to an antigen Therefore.we studied hepatitis C virus (HCV0-Specific induction of interleukins-1β,-10,-12(IL-1β,il-10,IL-12),and tumor necrosis factor-α(TNF-α) in monocytes.METHODS:Intracellular cytokine expression was studied by flow cytometry in 23 patients with chronic hepatitis C,14 anti-HCV seropositives without viremia and 11 controls after stimulation of peripheral blood mononuclear cells with recombinant core,NS3,NS4 NS5a and NS5b proteins .RESULTS:Patients with HCV viremia revealed greater spontaneous exprssion of IL-1β,TNF-α,and IL-10,Furthermore,greater than twofold higher IL-10 epression was induced by the HCV antigens in chronic hepatitis C than in the other two groups (P<0.05) In contrast,neither IL-12 noir TNF-α was induced preferentially.CONCLUSION:In chonic hepatitis C antigen-specific cytokine induction in monocytes is apparently shifted towards predominant IL-10 induction-not counterbalanced by antiviral type 1 cytokines,This may contribute to persistent viral replication.

Tumor-related cytokine release syndrome in a treatment-naïve patient with lung adenocarcinoma:A case report

BACKGROUND Cytokine release syndrome(CRS)is defined as systemic inflammation that usually occurs following chimeric antigen receptor T-cell therapy administration;however,it has not been reported in patients with untreated non-small cell lung cancer to date.CASE SUMMARY A 44-year-old nonsmoking woman presented to the hospital due to fever,palpitation,nausea,and cough for 1 mo and was diagnosed with stage cT3N3M0(IIIc)adenocarcinoma of the lung.Auxiliary examinations revealed elevated cytokine[tumor necrosis factor-α,interleukin(IL)-1β,and IL-6]and inflammatory factor levels,which decreased after treatment with corticosteroids and immunoglobulin and when tumor growth was controlled following chemotherapy,radiotherapy,and antiangiogenesis therapy.However,tumor recurrence was observed.After administration of nivolumab as third-line treatment,the patient’s condition was transiently controlled;however,CRS-like symptoms suddenly emerged,which led to a resurgence of cytokines and inflammatory factors and rapid death.CONCLUSION CRS can develop in treatment-naïve lung cancer patients.Patients with tumorrelated CRS may be at risk of CRS recurrence,aggravation,and onset of immune checkpoint inhibitor-related adverse events.

Effects of aminoguanidine on nitric oxide production induced by inflammatory cytokines and endotoxin in cultured rat hepatocytes

AIM To study the effects of aminoguanidine(AG) and two L-arginine analogues Nω-nitro-L-arginine methyl ester (L-NAME) and Nω-nitro-L-arginine (L-NNA) on nitric oxide (NO) productioninduced by cytokines (TNF-α, IL-11β, and IFN-γ)and bacterial lipopolysaccharide (LPS) mixture(CM) in the cultured rat hepatocytes, andexamine their mechanisms action.METHODS Rat hepatocytes were incubatedwith AG, L-NAME, L-NNA, Actinomycin D (ActD)and dexamethasene in a medium containing CM(LPS plus TNF-α, IL-1β, and IFN-γ) for 24 h. NOproduction in the cultured supernatant wasmeasured with the Griese reaction. IntracellularcGMP level was detected with radioimmunoasey.RESULTS NO production was markedlyblocked by AG and L-NAME in a dose-dependentmanner under inflammatory stimuli conditiontriggered by CM in vitro. The rate of themaximum inhibitory effects of L-NAME (38.9%)was less potent than that obtained with AG(53.7%, P<0.05). There was no significantdifference between the inhibitory effects of AGand two L-arginine analogues on intracellularcGMP accumulation in rat cultured hepatocytes.Non-specific NOS expression inhibitordexamethasone ( DEX ) and iNOS mRNAtranscriptional inhibitor ActD also significantlyinhibited CM-induced NO production. AG(0.1mmol.L-1) and ActD (0.2ng@Lt) wereequipotent in decreasing NO production inducedby inflammatory stimuli in vitro, and botheffects were more potent than that induced bynon-selectivity NOS activity inhibitor L-NAME(0. 1 mmol@ L- 1) under similar stimuli conditions(P＜O.O1).CONCLUSION AG is a potent selectiveinhibitor of inducible isoform of NOS, and themechanism of action may be not onlycompetitive inhibition in the substrate level, butalso the gene expression level in rathepatocytes .

The Effect of Chloroquine on Pro-Inflammatory Cytokines Levels in Graves’ Disease: Historical Cohort from a Pilot Randomized Controlled Trial

Objectives: Analyzing the trend in the serum inflammatory cytokines levels in a historical cohort of patients treated with combination of chloroquine and methimazole. Material and methods: We analyzed the pro-inflammatory serum cytokines level [Interleukin-6(IL-6), Tumor Necrosis Factor alpha (TNF-α), Interleukin 1 alpha (IL-1 α) and Interferon gamma (INF-γ)] in the stored blood samples of 22 patients with Graves’ disease who previously randomized to receive either chloroquine and methimazole combination therapy or methimazole monotherapy. Total T3, T4 and TSH levels were measured by an enzyme linked immunosorbent assay (ELISA) method (DRG, New York, USA) and the result was published previously. In this study we used an ELISA method (Bender Medsystem Vienna Austria) to measure serum pro-inflammatory cytokines in the first 6 months of trial. Results: No significant differences in serum cytokines concentration were observed between the two treatment groups (p > 0.05). Although it was not statistically significant, serum INF-gamma concentration tended to be lower in the chloroquine group after four months of therapy (p = 0.12). Conclusion: In this study we found changes in the serum thyroid hormones level did not accompany concomitant changes in the serum cytokines levels in two treatment groups. Therefore it is possible that chloroquine reduce serum thyroid hormones levels independent of its immunomodulatory effect.

Cytokine receptor-like factor 1(CRLF1)promotes cardiac fibrosis via ERK1/2 signaling pathway

Cardiac fibrosis is a cause of morbidity and mortality in people with heart disease.Anti-fibrosis treatment is a significant therapy for heart disease,but there is still no thorough understanding of fibrotic mechanisms.This study was carried out to ascertain the functions of cytokine receptor-like factor 1(CRLF1)in cardiac fibrosis and clarify its regulatory mechanisms.We found that CRLF1 was expressed predominantly in cardiac fibroblasts.Its expression was up-regulated not only in a mouse heart fibrotic model induced by myocardial infarction,but also in mouse and human cardiac fibroblasts provoked by transforming growth factor-β1(TGF-β1).Gain-and loss-of-function experiments of CRLF1 were carried out in neonatal mice cardiac fibroblasts(NMCFs)with or without TGF-β1 stimulation.CRLF1 overexpression increased cell viability,collagen production,cell proliferation capacity,and myofibroblast transformation of NMCFs with or without TGF-β1 stimulation,while silencing of CRLF1 had the opposite effects.An inhibitor of the extracellular signal-regulated kinase 1/2(ERK1/2)signaling pathway and different inhibitors of TGF-β1 signaling cascades,comprising mothers against decapentaplegic homolog(SMAD)-dependent and SMAD-independent pathways,were applied to investigate the mechanisms involved.CRLF1 exerted its functions by activating the ERK1/2 signaling pathway.Furthermore,the SMAD-dependent pathway,not the SMAD-independent pathway,was responsible for CRLF1 up-regulation in NMCFs treated with TGF-β1.In summary,activation of the TGF-β1/SMAD signaling pathway in cardiac fibrosis increased CRLF1 expression.CRLF1 then aggravated cardiac fibrosis by activating the ERK1/2 signaling pathway.CRLF1 could become a novel potential target for intervention and remedy of cardiac fibrosis.

巨噬细胞抑制因子-1、胰岛素样生长因子结合蛋白3、糖类抗原19-9检测对胰腺癌患者预后的预测价值

目的探讨糖类抗原19-9(CA19-9)、胰岛素样生长因子结合蛋白3(IGFBP3)、巨噬细胞抑制因子-1(MIC-1)检测对胰腺癌患者预后的预测价值。方法选取82例胰腺癌患者作为观察组,另选取81例健康体检志愿者作为对照组。对比两组受试者MIC-1、IGFBP3、CA19-9水平,分析胰腺癌患者预后的影响因素和MIC-1、IGFBP3、CA19-9单独及联合检测对胰腺癌患者预后的预测价值。结果观察组患者MIC-1、IGFBP3、CA19-9水平均明显高于对照组,差异均有统计学意义(P﹤0.01)。82例胰腺癌患者根据预后情况分为好转组61例与预后不良组21例,好转组与预后不良组患者分化程度、临床分期、淋巴结转移情况及MIC-1、IGFBP3、CA19-9水平比较,差异均有统计学意义(P﹤0.05)。分化程度为低分化、临床分期为Ⅲ~Ⅳ期、有淋巴结转移、MIC-1≥500 pg/ml、IGFBP3≥40 U/ml、CA19-9≥40 U/ml均为胰腺癌患者预后不良的独立危险因素(P﹤0.05)。MIC-1、IGFBP3、CA19-9联合检测预测胰腺癌患者预后的灵敏度和特异度分别为87.4%、71.9%,曲线下面积为0.816(95%CI:0.715~0.917),高于MIC-1、IGFBP3、CA19-9单独检测。结论MIC-1、IGFBP3、CA19-9水平升高均与胰腺癌患者预后不良密切相关,三者在胰腺癌患者预后预测中有较高的使用价值。

Topical biological agents targeting cytokines for the treatment of dry eye disease

Because inflammation plays a key role in the pathogenesis of dry eye disease and Sjogren's syndrome, topical anti-inflammatory agents such as corticosteroids and cyclosporine A have been used to treat inflammation of the ocular surface and lacrimal gland. Systemic biological agents that target specific immune molecules or cells such as tumor necrosis factor(TNF)-α, interferone-α, interleukin(IL)-1, IL-6, or B cells have been used in an attempt to treat Sjogren's syndrome. However, the efficacy of systemic biological agents, other than B-cell targeting agents, has not yet been confirmed in Sjogren's syndrome. Several studies have recently evaluated the efficacy of topical administration of biological agents targeting cytokines in the treatment of dry eye disease. Topical blockade of IL-1 by using IL-1 receptor antagonist could ameliorate clinical signs and inflammation of experimental dry eye. Using a mouse model of desiccating stress-induced dry eye, we have demonstrated that topical application of a TNF-α blocking agent, infliximab, could improve tear production and ocular surface irregularity, decrease inflammatory cytokines and Th-1 CD4+ cells on the ocular surface, and increase gobletcell density in the conjunctiva. Although controversy still remains, the use of topical biological agents targeting inflammatory cytokines may be a promising therapy for human dry eye disease.

绝经后骨质疏松症肾阴虚证关联基因CLCF1启动子区相互作用转录因子的筛选及分析

目的检测、分析与CLCF1基因上游启动子区域相互作用的转录因子。方法采用DNA pulldown技术联合质谱鉴定的方法检测与CLCF1基因启动子区域相互作用的蛋白,对鉴定出的蛋白质进行生物信息学分析,筛选出50个转录因子,使用平行反应监测(parallel reaction monitoring,PRM)技术靶向验证转录因子的表达情况。结果试验组与对照组共筛选出251个差异蛋白,GO富集结果表明差异蛋白主要涉及核糖体生物发生、正向调控DNA模板转录等生物学过程,包含转录调节复合物、SWI/SNF超家族型复合物,具有与cAMP应答元件结合、ATP水解活性等分子功能。KEGG通路显示,差异蛋白主要参与人t细胞白血病病毒1型感染、线粒体自噬、甲状腺激素等信号通路。PRM验证发现MAFK、TFE3、MITF、JUNB、FOSL2这5个转录因子表达趋势与DNA pulldown检测结果一致。结论MAFK、TFE3、MITF、JUNB、FOSL2这5个转录因子可能与CLCF1基因启动子区相结合。

血清miR-3065-3p、CRLF1与结肠癌病理特征的关系及对术后肝转移风险的评估价值分析

目的分析血清微小RNA-3065-3p(miR-3065-3p)、细胞因子受体样因子1(CRLF1)与结肠癌病理特征的关系及对结肠癌术后肝转移风险的评估价值。方法选取2018年1月至2019年1月聊城市人民医院收治的152例接受手术的结肠癌患者(结肠癌组),根据术后是否肝转移分为肝转移组(28例)和无肝转移组(124例),另选取同期65例体检健康者作为对照组,采用实时荧光定量聚合酶链反应(qRT-PCR)检测血清miR-3065-3p、CRLF1 mRNA相对表达水平。Pearson相关系数分析结肠癌患者血清miR-3065-3p与CRLF1 mRNA表达的相关性,并分析二者与临床病理特征的关系。采用多因素Logistic回归分析结肠癌患者术后肝转移的危险因素,受试者工作特征(ROC)曲线分析血清miR-3065-3p、CRLF1 mRNA表达对结肠癌患者术后肝转移风险的评估价值。结果结肠癌组血清miR-3065-3p相对表达水平高于对照组,CRLF1 mRNA相对表达水平低于对照组(P均<0.001)。Pearson相关系数分析结果显示,结肠癌患者miR-3065-3p与CRLF1 mRNA表达呈负相关(r=-0.750,P<0.001)。不同脉管侵犯、TNM分期、淋巴结转移结肠癌患者血清miR-3065-3p、CRLF1 mRNA相对表达水平比较,差异有统计学意义(P均<0.05)。多因素Logistic回归分析显示,脉管侵犯(OR=3.644,95%CI:1.544~8.601)、TNM分期Ⅲ期(OR=3.913,95%CI:1.398~10.957)、淋巴结转移(OR=3.706,95%CI:1.547~8.877)、CRLF1 mRNA(OR=3.338,95%CI:2.001~5.568)为结肠癌患者术后肝转移的独立危险因素,miR-3065-3p(OR=0.823,95%CI:0.741~0.914)为独立保护因素(P均<0.05)。ROC曲线分析显示,血清miR-3065-3p、CRLF1 mRNA表达单独与联合评估结肠癌患者术后肝转移风险的曲线下面积(AUC)分别为0.797、0.784、0.888,灵敏度分别为57.14%、57.14%、82.14%,特异度分别为89.52%、89.52%、87.10%。血清miR-3065-3p、CRLF1 mRNA表达联合评估结肠癌患者术后肝转移风险的AUC大于二者单独评估(P均<0.05)。结论结肠癌患者血清miR-3065-3p高表达,CRLF1 mRNA低表达,二者与结肠癌脉管侵犯、TNM分期、淋巴结转移和术后肝转移密切相关,二者联合检测对结肠癌术后肝转移具有良好的辅助评估价值。