Interleukin-1 Beta (IL-1<i>β</i>) in the Peripheral Blood of Dogs as a Possible Marker for the Detection of Early Stages of Inflammation

Background: Cytokines are mediators of disease. Expression levels in the blood could be of clinical relevance. Objective: Aim of this study was to show if serum levels of IL-1β could be of any clinical relevance concerning dogs. IL-1β was measured in serum samples of healthy dogs to find a reference range for healthy individuals. Measurements of IL-1β should show if this substance was a possible marker for early stages of inflammation. Therefore, a possible relation between serum levels and grades of leukocytosis was analyzed. Methods: IL-1β concentrations in the blood were assessed by the use of a human enzyme linked immunosorbent assay (ELISA). 39 dogs with different inflammatory diseases were analyzed to figure out if there was a correlation between IL-1β serum levels and the number of leukocytes in peripheral blood. The control group consisted of 16 healthy dogs. Results: about half of the samples IL-1β were detected. Most of the patients showed no detectable amounts of IL-1β. The IL-1β levels measured in the serum were stable for at least nine weeks when stored at ?20?C. The patients tested positively on IL-1β had mostly lower-grade leukocytosis compared to those who had no IL-1β in serum. All the dogs which were suffering from disease but still had no traceable IL-1β, showed a leukocytosis as a common symptom. Conclusion: This study showed that IL-1β could become an interesting marker for the detection of early stages of inflammation when leukocytosis does not yet appear in peripheral blood. Nonetheless, the possible use in diagnosis is restricted. This is due to the fact that there are only low amounts of IL-1β to be detected in the serum, even concerning patients are suffering from disease.

Interleukin-1β gene polymorphism associated with hepatocellular carcinoma in hepatitis B virus infection

AIM： To examine the effect of interleukin-l-beta （IL-113） promoter region C-511T and IL-1 receptor antagonist （IL-1RN） polymorphism among the patients with chronic hepatitis B virus （HBV） infection （HCC and non-HCC）. METHODS： Genomic DNA from 136 Thai patients with chronic HBV infection （HCC =46 and non-HCC= 90） and 152 healthy individuals was genotyped for IL-113 gene polymorphism （-511） using polymerase chain reaction with sequence specific primers （PCR-SSP）. The variable number of tandem repeats （VNTR） of IL-1RN gene was assessed by a PCR-based assay. The association between these genes and status of the disease was evaluated by X^2 test. RESULTS： IL-1B-511 genotype c/c was found to be significantly different in patients with HCC when compared with healthy individuals （P = 0.036, OR = 2.29, 95%CI = 1.05-4.97） and patients without HCC （P=0.036, OR= 2.52, 95%CI=1.05-6.04）. Analysis of allele frequencies of IL-1B-511 showed that IL-1B-511 C allele was also significantly increased in patients with HCC, compared to that in healthy control （P=0.033, OR= 1.72, 95%CI=1.04-2.84）. However, no significant association in IL-1RN gene was found between the two groups. CONCLUSION： IL-1B-511C allele, which may be associated with high IL-1B production in the liver, is a genetic marker for the development of HCC in chronic hepatitis B patients in Thai population.

妊娠合并精神分裂症患者血清中IL-1β、IL-6的表达水平及其与妊娠结局的相关性

【目的】探讨妊娠合并精神分裂症患者血清白细胞介素-1β(IL-1β)和白细胞介素-6(IL-6)的表达水平及其与妊娠结局的相关性。【方法】选取2020年1月至12月在延安大学附属医院产科分娩的妊娠合并精神分裂症患者37例(观察组)和正常孕妇28例(对照组)。采用酶联免疫吸附法(ELISA)检测所有受试者血清IL-1β和IL-6表达水平;采用Pearson分析IL-1β和IL-6表达水平的相关性及IL-1β、IL-6表达水平与不良妊娠结局的相关性;采用受试者工作特征曲线(ROC曲线)分析IL-1β、IL-6表达水平对不良妊娠结局的预测价值。【结果】与对照组比较,观察组血清IL-1β和IL-6表达水平升高(P<0.05);血清IL-1β与IL-6表达呈正相关(r=0.54,P<0.05);观察组早产、胎膜早破、胎盘早剥、产后出血发生率显著高于对照组(P<0.05);胎儿低体重、胎儿畸形、胎儿窘迫、围生儿死亡和重度窒息发生率显著高于对照组(P<0.05);血清IL-1β、IL-6表达与不良妊娠结局呈正相关(P<0.05);ROC曲线分析显示:血清IL-1β、IL-6预测不良妊娠结局的曲线下面积(AUC)分别为0.883、0.864,二者联合预测不良妊娠结局的AUC为0.891。【结论】妊娠合并精神分裂症可增加妊娠不良结局发生率,血清IL-1β和IL-6在妊娠合并精神分裂症患者血清中呈高表达,其与妊娠合并精神分裂症患者不良妊娠结局密切相关,可为妊娠合并精神分裂症病情监控提供依据。

Caspase-1 inhibition alleviates acute renal injury in rats with severe acute pancreatitis

AIM: To assess the effect of inhibition of caspase-1 on acute renal injury in rats with severe acute pancreatitis (SAP).

High SIOOB levels in cerebrospinal fluid and peripheral blood of patients with acute basal ganglial hemorrhage are associated with poor outcome

S100B is involved in brain injury. This study aimed to determine plasma and cerebral spinal fluid （CSF） levels of S100B in patients with spontaneous intracerebral hemorrhage （ICH）, and to correlate S100B levels with Glasgow Coma Scale （GCS） scores, ICH volumes, presence of intraventricular hemorrhage （IVH）, and survival rate, and to correlate CSF S100B levels with plasma 100B levels as well as CSF interleukin-1 beta （IL-1β）levels. Ten patients with suspicion of subarachnoid hemorrhage and 38 patients with spontaneous basal ganglia hemorrhage were included in the study. Their plasma and CSF samples were collected. The concentrations of IL-1β in CSF and S100B in plasma and CSF were analyzed by enzyme-linked immunosorbent assay. Plasma or CSF S100B levels in the ICH group were significantly higher than those in the control group （178.7±74.2 versus 63.2±23.0 pg/ml; P〈0.001 or 158.1±70.9 versus 1.8±0.7 ng/ml; P〈0.001）. S100B levels were highly associated with GCS scores, ICH volumes, presence of IVH, and survival rate （all P〈0.05）. CSF S100B levels were highly associated with plasma S100B levels as well as CSF IL-113 levels （both P〈0.01） in patients with ICH. A receiver operating characteristic curve identified CSF and plasma S100B cutoff levels that predicted 1-week mortality of patients with a high sensitivity and specificity. The areas under curves （AUCs） of GCS scores and ICH volumes were larger than those of CSF and plasma $100B levels, but the differences were not statistically significant （P〉0.05）. High levels of S100B are present in the cerebrospinal fluid and peripheral blood of patients with ICH and may contribute to the inflammatory processes of ICH. The levels of CSF and plasma S100B after spontaneous onset of ICH seem to correlate with clinical outcome in these patients. Increases in peripheral S100B properly reflect brain injury, and plasma S100B level may serve as a useful clinical marker for evaluating the prognosis of ICH.

I_κB kinase-beta inhibitor attenuates hepatic fibrosis in mice

AIM: To investigate the anti-fibrosis effect of IκB kinase-beta inhibitor (IKK2 inhibitor IMD0354) in liver fibrosis. METHODS: Twenty male C57BL6 mice were divided into four groups. Five high-fat fed mice were injected with lipopolysaccharide (LPS, 10 mg/kg) intraperitoneally and five high-fat fed mice were without LPS injection to build models of liver injury, and the intervention group (five mice) was injected intraperitoneally with IKK2 inhibitor (IMD 30 mg/kg for 14 d), while the remaining five mice received a normal diet as controls. Hepatic function, pathological evaluation and liver interleukin-6 (IL-6) expression were examined. Western blotting and real-time polymerase chain reaction were used to detect the expressions of nuclear factor-κB (NF-κB), alpha-smooth muscle actin (α-SMA), tumor growth factor-beta1 (TGF-β1), tumor necrosis factor-alpha (TNF-α), typeⅠand type Ⅲ collagen proteins and mRNA. RESULTS: A mouse model of liver injury was successfully established, and IMD decreased nuclear transloca-tion of NF-κB p65 in liver cells. In the IMD-treated group, the levels of alanine aminotransferase (103 ± 9.77 μ/L vs 62.4 ± 7.90 μ/L, P < 0.05) and aminotransferase (295.8 ± 38.56 μ/L vs 212 ± 25.10 μ/L, P < 0.05) were significantly decreased when compared with the model groups. The histological changes were significantly ameliorated. After treatment, the expressions of IL-6 (681 ± 45.96 vs 77 ± 7.79, P < 0.05), TGF-β1 (Western blotting 5.65% ± 0.017% vs 2.73% ± 0.005%, P < 0.05), TNF-α (11.58% ± 0.0063% vs 8.86% ± 0.0050%, P < 0.05), typeⅠcollagen (4.49% ± 0.014% vs 1.90% ± 0.0006%, P < 0.05) and type Ⅲ collagen (3.46% ± 0.008% vs 2.29% ± 0.0035%, P < 0.05) as well as α-SMA (6.19 ± 0.0036 μ/L vs 2.16 ± 0.0023 μ/L, P < 0.05) protein and mRNA were downregulated in the IMD group compared to the fibrosis control groups (P < 0.05). CONCLUSION: IKK2 inhibitor IMD markedly improved non-alcoholic fatty liver disease in mice by lowering NF-κB activation, which could become a remedial target for liver fibrosis.

红细胞压积及胎盘功能变化与胎儿预后关系的探讨

本文对202例正常和异常妊娠孕妇测定尿E/C比值和血SP_1值,其中测定红细胞压积者71例。结果表明胎儿预后不良组E/C比值多数为下降型和水平型;联合测定表明两项指标测定可提高预测胎儿预后的准确性。红细胞压积≥0.35时胎儿预后不良的发生率明显高于<0.35组(P<0.01)。本文认为测定红细胞压积可间接了解胎盘功能情况及预测胎儿预后。

Study of the mechanisms of acupuncture and moxibustion treatmentfor ulcerative colitis ratsinview of the gene expression of cytokines

AIM:To observe the effect of acupuncture and moxibustion on the expression of IL-1beta and IL-6 mRNA in ulcerative colitis rats.METHODS:The SD rat ulcerative colitis model was created by immunological method associated with local stimulation. Colonic mucosa was prepared from human fresh surgical colonic specimens, homogenized by adding appropriate amount of normal saline and centrifuged at 3000r/min. The supernatant was collected for measurement of protein conentration and then mixed with Freund adjuvant. This antigen fluid was first injected into the plantae of the model group rats, and then into their plantae, dorsa, inguina and abdominal cavities (noFreund adjuvant for the last injection) again on the 10th, 17th, 24th and 31st day. When a certain titer of serum anti colonic antibody was reached, 2% formalin and antigen fluid (no Freund adjuvant) were administered separately by enema. The ulcerative colitis rat model was thus set up. The animals were randomly divided into four groups: model control group (MC, n = 8), electro acupuncture group (EA, n = 8), herbs partition moxibustion group (HPM 8), normal control group (NC,n = 8). HPM: Moxa cones made of refined mugwort floss were placed on the medicinal pad (medicinal pad dispensing: Radix Aconiti praeparata, cortex Cinnamomi, etc) for Qihai (RN 6) and Tianshu (ST 25, bilateral) and ignited. Two moxa cones were used for each acupoint once a day and 14 times in all. EA: Tianshu (bilateral) and Qihai were stimulated by the intermittent pulse with 2Hz frequency, 4mA intensity for 20 minutes once a day and 14 times in all. After treatment, rats of all four groups were killed simultaneously. The spleen was separated and the distal colon was dissected. Total tissue RNA was isolated by the guanidinium thiocyanate phenol chloroform extraction method. RT-PCR technique was used to study the expression of IL-1 beta and IL-6 mRNA.RESULTS:IL-1 beta and IL-6 mRNAs were not detected in the spleen and colonic mucosa of the NC rats, whereas they were significantly expressed in that of the MC rats.IL-1 beta and IL-6 mRNAs were markedly lower in the EA and HPM rats than that in MC rats. There was no significant difference between the levels of IL-1 beta and IL-6 mRNAs in the EA and HPM rats. The expressions of IL-1 beta and IL-6 mRNAs were nearly the same in the spleen and colon of all groups.CONCLUSION:Acupuncture and moxibustion greatly inhibited the expression of IL-1 beta and IL-6 mRNA in the experimental ulcerative colitis rats.

Ligustrazine alleviates gastric mucosal injury in a rat model of acute necrotizing pancreatitis

BACKGROUND: Acute necrotizing pancreatitis (ANP) leads to a systemic inflammatory response characterized by widespread leukocyte activation and, as a consequence, distant organ injury. The aim of this study was to explore the relationship between gastric microcirculatory impairment and inflammatory mediators released in rats and to evaluate the therapeutic effect of ligustrazine extracted from Rhizoma ligusticum wallichii on gastric mucosa injury in a rat model of ANP. METHODS: Ninety-six Sprague-Dawley rats were randomly divided into three groups: normal control (group Q; ANP without treatment (group P); and ANP treated with ligustrazine (group T). The ANP model was induced by injection of 50 g/L sodium taurocholate under the pancreatic membrane (4 ml/kg). Group C was given isovolumetric injection of 9 g/L physiological saline by the same route. Group T was injected with ligustrazine (10 ml/kg) via the portal vein. The radioactive biomicrosphere technique was used to measure the blood flow 2 and 12 hours after the induction of ANP. Samples of the pancreas and stomach were taken to assess pathological changes by a validated histology score; meanwhile, the levels of serum interleukin-1 beta (IL-1 beta) were determined. Gastric tissues were also used to measure the level of myeloperoxidase (MPO), which is expressed intracellularly in the azurophilic granules of neutrophils. RESULTS: Blood flow in group P was significantly lower than that in group C (P < 0.01). Pathological changes were significantly aggravated in group P. The gastric MPO activity in group P was significantly higher than that in group C (P < 0.01). The level of serum IL-1 beta in group P increased more significantly than that in group C (P < 0.01). Blood flow of the stomach in group T was significantly higher than that in group P after 2 hours (P < 0.01). The pathological changes were significantly alleviated in group T. The MPO activity of group T was significantly lower than that of group P (P < 0.01). Although serum IL-1 beta level of group T, was higher than of group C (P < 0.01), it was lower than that of group P (P < 0.01). There was a negative correlation between gastric blood flow and MPO activity (r=-0.983, P < 0.01), and between gastric blood flow and pathological score (r=-0.917, P < 0.05). CONCLUSIONS: Decreased gastric blood flow and increased inflammatory mediators can be seen early in ANP, and both are important factors for gastric and mucosal injury. Ligustrazine can ameliorate microcirculatory disorder and alleviate the damage to the pancreas and stomach.

Efficacy of suspended moxibustion in stroke rats is associated with a change in tail temperature

Suspended moxibustion-produced heat can transfer from the acupoint to other sites of the body. The suspended moxibustion should be terminated when clinical propagated sensation disappears, because this implies that the quantity of moxibustion is sufficient. We wanted to investigate if this phenomenon also occurs in experimental animals. In the present study, a rat model of stroke was established and treated with suspended moxibustion at Dazhui （DU14）for 60 minutes. Results showed that the increase in tail temperature began at 15 minutes after suspended moxibustion and decreased gradually at 40 minutes. In addition, neurological function was significantly improved in stroke rats with tail temperature increase following suspended moxibustion, and this effect was associated with significantly reduced tumor necrosis factor a and interleukin 113 mRNA. However, there was no significant difference between 40- and 60-minute suspended moxibustion. The findings indicate that elevated tail temperature began to decrease at 40 minutes after suspended moxibustion, and further suspended moxibustion was not useful in the recovery of stroke rats.

Fermented Chinese formula Shuan-Tong-Ling attenuates ischemic stroke by inhibiting inflammation and apoptosis

The fermented Chinese formula Shuan-Tong-Ling is composed of radix puerariae（Gegen）,salvia miltiorrhiza（Danshen）,radix curcuma（Jianghuang）,hawthorn（Shanzha）,salvia chinensis（Shijianchuan）,sinapis alba（Baijiezi）,astragalus（Huangqi）,panax japonicas（Zhujieshen）,atractylodes macrocephala koidz（Baizhu）,radix paeoniae alba（Baishao）,bupleurum（Chaihu）,chrysanthemum（Juhua）,rhizoma cyperi（Xiangfu） and gastrodin（Tianma）,whose aqueous extract was fermented with lactobacillus,bacillus aceticus and saccharomycetes.ShuanTong-Ling is a formula used to treat brain diseases including ischemic stroke,migraine,and vascular dementia.Shuan-Tong-Ling attenuated H_2O_2-induced oxidative stress in rat microvascular endothelial cells.However,the potential mechanism involved in these effects is poorly understood.Rats were intragastrically treated with 5.7 or 17.2 m L/kg Shuan-Tong-Ling for 7 days before middle cerebral artery occlusion was induced.The results indicated Shuan-Tong-Ling had a cerebral protective effect by reducing infarct volume and increasing neurological scores.Shuan-Tong-Ling also decreased tumor necrosis factor-α and interleukin-1β levels in the hippocampus on the ischemic side.In addition,Shuan-Tong-Ling upregulated the expression of SIRT1 and Bcl-2 and downregulated the expression of acetylated-protein 53 and Bax.Injection of 5 mg/kg silent information regulator 1（SIRT1） inhibitor EX527 into the subarachnoid space once every 2 days,four times,reversed the above changes.These results demonstrate that Shuan-Tong-Ling might benefit cerebral ischemia/reperfusion injury by reducing inflammation and apoptosis through activation of the SIRT1 signaling pathway.

Role of Toll-like receptor 4 in inflammatory reactions of hippocampal neurons

Lipopolysaccharide stimulates Toll-like receptor 4 on immune cells to produce immune mediators. Toll-like receptor 4 is also expressed by non-immune cells, which can be stimulated by lipopolysaccharide. However, whether Toll-like receptor 4 is expressed by primary cultured hippocampal neurons and its specific role in lipopolysaccharide-induced neuroinflammation is currently undefined, in this study, Toll-like receptor 4 antibody blocking was used to analyze the Toll-like receptor 4 signaling pathway and changes in inflammation of lipopolysaccharide stimulated hippocampal neurons. Immunofluorescence showed that Toll-like receptor 4 protein was mainly located in the membrane of hippocampal neurons. Quantitative reverse transcription-PCR and western blot assay showed that after stimulation of lipopolysaccharide, the mRNA and protein levels of Toll-like receptor 4 and the mRNA levels of interleukin-ll3 and tumor necrosis factor-（] were significantly increased. In addition, there was increased phosphorylation and degradation of kappa B a inhibitor in the cytosol and increased nuclear factor-KB p65 expression in the nuclei. Pretreatment with Toll-like receptor 4 antibody could almost completely block this increase. These experimental findings indicate that lipopolysaccharide participates in neuroinflammation by stimulating Toll-like receptor 4/nuclear factor-KB pathway in hippocampal neurons, which may be both ＂passive victims＂ and ＂activators＂ of neuroinflammation.

Elevated levels of inflammatory cytokines and high-sensitivity C-reactive protein in periodontitis patients in Kosovo: A pilot study

The following article has been retracted due to the investigation of complaints received against it. The Editorial Board found that the same contents have been published in another journal at the same time. The scientific community takes a very strong view on this matter, and the Open Journal of Stomatology treats all unethical behavior such as plagiarism seriously. This paper published in Vol.3 No.1 32-38, 2013 has been removed from this site. Title: Elevated levels of inflammatory cytokines and high-sensitivity C-reactive protein in periodontitis patients in Kosovo: A pilot study Authors: Zana Sllamniku-Dalipi, Hasan Mehmeti, Fatmir Dragidella, Ferit Kocani, Metush Disha, Kastriot Meqa, Luljeta Begolli, Gramos

Amentoflavone protects hippocampal neurons: anti-inflammatory, antioxidative, and antiapoptotic effects

Amentoflavone is a natural biflavone compound with many biological properties, including anti-inflammatory, antioxidative, and neuroprotective effects. We presumed that amentoflavone exerts a neuroprotective effect in epilepsy models. Prior to model establishment, mice were intragastrically administered 25 mg/kg amentoflavone for 3 consecutive days. Amentoflavone effectively prevented pilocarpine-induced epilepsy in a mouse kindling model, suppressed nuclear factor-κB activation and expression, inhibited excessive discharge of hippocampal neurons resulting in a reduction in epileptic seizures, shortened attack time, and diminished loss and apoptosis of hippocampal neurons. Results suggested that amentoflavone protected hippocampal neurons in epilepsy mice via anti-inflammation, antioxidation, and antiapoptosis, and then effectively prevented the occurrence of seizures.

Effect of electroacupuncture at Lower He-Sea points including Yanglingquan(GB 34)on nuclear factor-κB and interleukin-1βin guinea pigs with acute cholecystitis

Objective:To observe the effect of electroacupuncture(EA)at Lower He-Sea points on the expression levels of interleukin-1β(IL-1β)in the serum and gallbladder tissues,and nuclear factor-κB(NF-κB)in gallbladder tissues of the guinea pigs with acute cholecystitis(AC),and to explore whether Yanglingquan(GB 34),the Lower He-Sea point pertaining to Dan Fu(gallbladder),is relatively specific for the Dan Fu(gallbladder)disorders.Methods:Eighty-two healthy guinea pigs were randomly divided into 6 groups according to the random number table method,a blank group,a model group,a Yanglingquan(GB 34)group,a Zusanli(ST 36)group,a Shangjuxu(ST 37)group,and a Xiajuxu(ST 39)group,with 12 guinea pigs in the blank group while 14 in the other groups,respectively,half males and half females in each group.Except for the blank group,guinea pigs in the other groups were injected with E.coli into the gallbladder to establish AC models.Guinea pigs in the blank group were fed routinely without special treatment;those in the model group were daily tied up for 30 min without EA treatment;those in the 4 groups receiving EA treatment were acupunctured at the corresponding Lower He-Sea points after daily binding and stimulated with the SDZ-V EA instrument.After successful modeling and treatment for 5 d,blood was collected from the abdominal aorta of the guinea pigs,and the gallbladder tissues in each group were isolated for hematoxylin-eosin(HE)staining to observe the morphological changes.Enzyme-linked immunosorbent assay(ELISA)was used to detect the serum IL-1βlevel,and immunohistochemistry(IHC)was used to detect the expression levels of NF-κB and IL-1βin gallbladder.Results:On the 3rd day after modeling,the guinea pigs in the five groups with modeling were mentally depressed with decreased appetite,significantly reduced activities,slouch,lassitude,slack and matted fur,and loose stools;two guinea pigs were selected from each group(one male and one female,not included in the final statistics)to isolate the gallbladder after sacrifice;macroscopic observation showed that the gallbladder wall was differently thickened;the bile color was dark green and opaque with particles suspended or accumulated;light microscope observation showed that the submucosal blood vessels of the gallbladder were congested,along with mucosal edema,ulceration,necrosis,shedding,and a large number of inflammatory cells infiltrating in the lamina propria,indicating that the AC model was successfully prepared.Compared with the model group,the gallbladder tissue injuries of the four groups receiving EA treatment were all differently repaired,the serum IL-1βlevels were reduced(P<0.01 or P<0.05),and the IL-1βlevels in the gallbladder tissues were reduced(P<0.01 or P<0.05).Compared with the model group,the NF-κB expression level in the Yanglingquan(GB 34)group was significantly reduced(P<0.01),but was not statistical different in the Zusanli(ST 36)group,Shangjuxu(ST 37)group and Xiajuxu(ST 39)group(all P>0.05).Compared with the Yanglingquan(GB 34)group,the gallbladder tissues of the Zusanli(ST 36)group,Shangjuxu(ST 37)group and Xiajuxu(ST 39)group were more severely damaged,and the expression levels of serum IL-1β,the NF-κB and IL-1βin the gallbladder tissues were increased(P<0.01 or P<0.05).Intervention effect of Yanglingquan(GB 34)on AC guinea pigs was superior to that of Zusanli(ST 36),Shangjuxu(ST 37)and Xiajuxu(ST 39).Conclusion:EA at the Lower He-Sea points of the stomach,large intestine,small intestine and gallbladder can produce curative effects on AC guinea pigs and reduced the inflammatory symptoms.Intervention effect of Yanglingquan(GB 34)on AC guinea pigs is superior to that of Zusanli(ST 36),Shangjuxu(ST 37)and Xiajuxu(ST 39).The mechanism of EA at Yanglingquan(GB 34)in treating AC may be regulating IL-1βand NF-κB to control the inflammatory response and improve the gallbladder tissue damage.

Therapeutic Mechanism of Santeng Dingtong Recipe (三藤定痛方) on Monosodium Urate Crystal-Induced Rabbit Arthritis

Objective: To study the therapeutic mechanism of Santeng Dingtong recipe (STDT) on monosodium urate crystal (MSU) induced rabbit arthritis Methods: Forty-two rabbits were randomly divided into six groups, 7 in each group. Group 1 received 0.9% saline 2. 5 ml/kg per day by gastrogavage (ig) for 10 days; Group 2, 3 and 4 received STDT 0.125 g/kg, 1.0 g/kg and 8.0 g/kg per day respectively by ig for 10 days; Group 5 received colchicine 4. 5 mg/kg per day by ig for 4 days; and Group 6 was untreated. MSU crystals 10 mg /500ul containing polymyxin B 10 u/ml was injected into the knee joints of Group 1-5 to make rabbit arthritis models. Leukocytes in synovial lavage fluids was then counted and differentiated; pathological injury of synovial membranes was observed under HE staining; interleukin-1 beta (IL-1B), tumor necrosis factor alpha (TNFa) and leukotriene B4 (LTB4) content in synovial lavage fluids were determined by ELISA. Results: MSU caused a rapid leukocyte infiltration and increased production of IL-1B, TNFa and LTB4 2 hrs after intra-articular injection. STDT inhibited neutrophil infiltration in synovial fluids dose-dependently, protected the synovial membrane against pathological injury and reduced the production of IL-1B, TNFa and LTB4; while colchicine did not decrease the level of TNFa, but significantly inhibited neutrophil infiltration in synovial fluid and reduced the production of IL-11B and LTB4. Conclusion: STDT exerts an anti-inflammatory effect in rabbit model of acute MSU arthritis, its mechanism being probably due to the decrease of XL-1B, TNFa and LTB4 synthesis.

Effect of Moxibustion on IL-1β and IL-2 in Rat Models of Rheumatoid Arthritis

Objective： To observe the influence on IL-1β and IL-2 in rat models with rheumatoid arthritis after moxibustion on Shenshu （BL 23） and Zusanli （ST 36） points, and to discuss the mechanism of moxibustion. Methods： Fifty male Wistar rats were divided randomly into 5 groups, control group, model group, drug group, moxibustion group, and laser group, 10 for each. Four groups except the normal group were built on the model of rheumatoid arthritis. The changes of body weight and plantar circumference were measured and the level of IL-1β, IL-2 in sera were examined by ELISA. Results： Compared with the model group, the weight and plantar circumference of rats in the moxibustion group were improved significantly after treatment （P〈0.01）, and the improvement of plantar circumference also had significant differences compared with the drug group and the laser group （P〈0.05）. The level of IL-1β, IL-2 in sera were down regulated in the moxibustion group and the laser group, which had statistical differences compared with the model group （P〈0.05）, but no statistical differences were found when comparing with the drug group. Conclusion： Moxibustion obviously improves the toe tumefaction of the rats with rheumatoid arthritis, which is better than CO2 laser of 10.6μm. On the aspect of decreasing the amount of IL-1β, IL-2, CO2 laser of 10.6μm is similar with moxibustion.

Effect of moxibustion plus Duhuo Jisheng decoction on hs-CRP,IL-1β and TNF-α levels in middle-aged and elderly patients with knee osteoarthritis

Objective： To observe the clinical effect of moxibustion combined with Duhuo Jisheng decoction for middle-aged and elderly patients with knee osteoarthritis（KOA） and its impact on serum high sensitive C-reactive protein（hs-CRP）,interleukin-1β（IL-1β） and tumor necrosis factor-α（TNF-α） levels.Methods： A total of 90 eligible KOA patients were randomized into an observation group（n=45） and a control group（n=45）.Cases in the observation group received moxibustion plus oral administration Duhuo Jisheng decoction, while cases in the control group received Duhuo Jisheng decoction. Then the hs-CRP, IL-1β and TNF-α levels were examined and the clinical effect was evaluated.Results： Before treatment, the hs-CRP, IL-1β and TNF-α levels showed no between-group statistical differences（all P〉0.05）. After 8 weeks of treatment, the hs-CRP, IL-1β and TNF-α levels all dropped significantly in both groups（P〈0.01 or P〈0.05）, and the treatment group showed more substantial changes than the control group（all P〈0.05）. The total effective rate was 97.8% in the treatment group, versus 86.7% in the control group, showing a statistical difference（P〈0.05）.Conclusion： Moxibustion plus Duhuo Jisheng decoction has a good clinical effect for middle-aged and elderly KOA patients,and the effect may relate to the decreased hs-CRP, IL-1β and TNF-α levels.

Effect of Gubi prescription on caveolin-1 expression and phosphoinositide 3 kinase/protein kinase B and Fas signal pathways in rats with knee osteoarthritis

OBJECTIVE: To investigate the effects of Gubi prescription on the expression of caveolin-1, and the phosphoinositide 3 kinase/protein kinase B(PI3 K/Akt) and Fas signal pathways in rats with knee osteoarthritis(KOA).METHODS: Forty KOA model rats were established using a modification of Hulth's method. Rats were divided into five groups by the random number method: model, positive drug(Vicolli group), and high-, medium-, and low-dose Gubi prescription groups(n = 8/group). In the sham surgery group(n = 8), only anterior and posterior cruciate ligaments of rats were exposed during surgery. A normal group(n = 8) consisted of rats with no treatment. Rats were intragastrically administered corresponding drugs once every day for eight consecutive weeks. Then, rat synovial membranes were extracted and histomorphological changes were recorded. m RNA expression was measured by q-PCR.Serum superoxide dismutase(SOD), malondialdehyde(MDA), nitric oxide(NO), and interleukin 1β(IL-1β) levels were measured. Western blotting determined the effects of Gubi prescription on protein expressions of caveolin-1, Bax, Bcl-2, Fas, and caspase-3 in chondrocytes from KOA rats. The knee cartilage of rats was excised and cultured under aseptic conditions. After coincubation of chondrocytes with Gubi prescription-containing serum, IL-1β, and si RNA, Western blotting was used to determine the protein expressions of caveolin-1, Bax, Bcl-2, Fas, and caspase-3.RESULTS: The morphological score of the articular synovium in the model group was significantly higher than in the normal group(P < 0.01). The morphological score in the high-and mediumdose Gubi prescription groups was lower than in the model group(P < 0.05). Chondrocytes from the decoction-containing serum group had a lower expression of Bax(P < 0.05), and higher expressions of Bcl-2(P < 0.05) and caspase-3(P < 0.05) compared with the model group. Chondrocytes in the decoction-containing serum group had higher expressions of Bax and Bcl-2(P < 0.01) and lower expressions of caveolin-1 and Fas(P < 0.05) compared with the model group. Compared with the model group, Bax and caspase-3 expressions were reduced in the chondrocytes of all three Gubi prescription groups(P < 0.05) whereas Bcl-2 expression was increased(P < 0.05). Compared with the model group, the expressions of caveolin-1 and Fas(P < 0.05) were reduced in groups that received high-and medium-doses of Gubi prescription. Gubi prescription increased the serum level of SOD and significantly reduced those of MDA, NO and IL-1β(P < 0.05).CONCLUSION: Gubi prescription suppressed the chondrocyte-related PI3 K/Akt and Fas signal pathways and inhibited the overexpression of caveolin-1 in rat chondrocytes.

Effect of modified Qing Long Bai Wei needling on the levels of IL-1β, IL-6 and INF-α in synovial fluid of knee osteoarthritis patients

Objective:To compare the modified Qjng Long Bai Wei needling method and ordinary acupuncture method in the effects of improving the levels of interleukin (IL)-1β,IL-6 and tumor necrosis factor (TNF)-α in the treatment of knee osteoarthritis (KOA),and to determine the advantage of the modified Qing Long Bai Wei needling method for KOA.Methods:One hundred KOA patients were randomized into a treatment group and a control group by using the random number table,with 50 cases in each group.The treatment group was intervened by the modified Qing Long Bai Wei needling method,and the control group was given ordinary acupuncture.The two groups were observed before and after the treatment to determine the changes in the levels of IL-1β,IL-6 and TNF-α in synovial fluid,and the clinical efficacies were compared between the two groups.Results:The total effective rate and clinical recovery rate were 97.9％ and 52.1％ respectively in the treatment group,versus 85.1％ and 25.5％ in the control group,and the between-group differences were statistically significant (both P＜0.01).After the treatment,the levels of Ib1β,IL-6 and TNF-cα in synovial fluid changed significantly in both groups (all P＜0.01);there were significant differences in the levels of IL-1β,IL-6 and TNF-α in synovial fluid between the two groups (all P＜0.01).Conclusion:The modified Qjng Long Bai Wei needling is an effective method for KOA and it can significantly improve the levels of IL-1β,IL-6 and TNF-α in synovial fluid.