The effect of triptolide (TP) on the expression of receptor activator of nuclear factor-kappaB ligand (RANKL) and osteoprotegerin (OPG) was explored in rat adjuvant induced arthritis (AA). AA was induced in Wistar rats. Arthritis rats were treated with TP and methotrexate (MTX) at the onset (day 9) of arthritis. On the peak of arthritis (day 24), the expression of RANKL and OPG protein in the joints and RANKL mRNA in peripheral blood mononuclear cells (PBMC) was detected. TNF-alpha and IL-1beta levels in peripheral blood were determined. Bone erosion scores were also evaluated. The results showed that bone erosion scores in TP and MTX groups were lower than in AA group (P < 0.01); The expression levels of RANKL in the synovium (P < 0.01) and bone (P < 0.05), and OPG level in synovium (P < 0.05) were lower in TP group than in AA group (P < 0.05). In TP group, the expression levels of RANKL mRNA and TNF-alpha, IL-1beta in PBMC were lower than in AA group (all P < 0.01). It was concluded that TP could inhibit rat adjuvant arthritis bone erosion by suppressing the expression of RANKL.

The relationship between tumour necrosis factor-alpha (TNF-alpha) gene polymorphism and inhibitory effects of triptolide on TNF-alpha production from peripheral blood mononuclear cells (PBMC) of healthy humans was investigated. Genomic DNA from 41 healthy people was typed for TNF-alpha--308 polymorphism by allele-specific polymorphism chain reaction (AS-PCR). The TNF-alpha concentration in the supernatant was measured by ELISA. The results showed that the production of TNF-alpha from TNF-alpha--308 non-G/G genotype PBMC was higher than that from TNF-alpha--308 G/G genotype PBMC after stimulated by LPS. Triptolide could lower the production of TNF-alpha from G/ G genotype PBMC, but had no effect on the level of TNF-alpha from non-G/G genotype PBMC. It was concluded that TNF-alpha gene polymorphism was related to the TNF-alpha production from triptolide-inhibited PBMC culture in healthy humans.

Objective To observe the effect of osteopontin (OPN) and integrin αtvβ3 in collageninduced arthritis (CIA) and the possible mechanism of Tripterygium wilfordii polyglycoside (TWP) in the treatment of rheumatoid arthritis (RA). Methods CIA rats model were developed and were randomly divided into the experimental group and the TWP group.And tissue samples were obtained 4 weeks later.Then the expressions of OPN and integrin αvβ3 in the synovium,synovium fluid and serum of each group were determined by immunohistochemical stain and ELISA.Variance analysis was used for data analysis.Results The concentrations of OPN of the normal controls,experimental group and the TWP group in the serum were (5.7±2.9), (7.8±6.2), (5.0±1.9) ng/ml respectively and there were significant differences between these 3 groups (F=6.74,P=0.016).The concentration of OPN (measured by mean grey value) in the synovium and cartilage of the three groups were 229±15,81±15,93±13 and 211±17,91±19,100±15 and there were significant differences between the three groups (F=52.48,P＜0.01; F=18.98,P=0.01).The concentrations of protein αvβ3 (measured by mean grey value) in the synovium and cartilage were 235±16,91±16,131±14 and 198±10,99±15,113±14,respectively and there were significant differences between the three groups (F=23.03,P=0.002; F=12.04,P=0.008).The expressions of OPN and integrin αvβ3 in the synovium,synovium fluid and serum of the experimental group were markedly higher than that of the controls.The expressions of OPN and integrin αvβ3 in the synovium,synovium fluid and serum of the treatment group were obviously lower than the experimental group.Conclusion OPN and integrin αtvβ3 are involved in the hyperplasia of the synovium,cartilage and bone destruction in CIA rats.The underlying molecular mechanism that TWP is effective in treating synovitis and bone destruction of RA is possibly related to down-regulation of the expression of OPN protein and integrin αvβ3.

AIM: To explore whether the inhibitory effect of triptolide on IL - 1β production by PBMC is asso ciated with IL - 1β gene polymorphisms. METHODS: IL - 1β gene polymorphism was analyzed in 31 healthy volunteers. From genomic DNA, the C - T polymorphism at IL - 1 β - 511 was typed by PCR - RFLP. Meanwhile the IL - 1 β was also measured in the supernatants of the cultured and stimulated peripheral blood mononuclear cells (PBMC) by ELISA. RE SULTS: After LPS stimulation in PBMC cultures of healthy subjects, the secretion levels of IL - 1 β in 9 volunteers who carried IL - 1β -511 T/T genotype were higher than in volunteers who are not T/T genotype (P ＜0.05). Triptolide sup pressed the production of IL - 1β significantly in LPS - treated human PBMC carried C/C and C/T genotype ( P ＜ 0.05 ), but this significant inhibitory effect of triptolide was not seen in T/T genotype ( P ＞ 0.05 ). CONCLUSION: The gene polymorphism at IL - 1β - 511 was related to the production of IL - 1β, and the inhibitory effect of triptolide on the produc tion of IL - 1β was different in C/C, C/T, T/T genotype of IL - 1β -511, which may be one of the reasons for the phe nomenon that people respond differently to triptolide.

BACKGROUND: Common threewingnut root has the functions of anti-inflammation and immune inhibition, etc., and it has been used at present to treat various autoimmune diseases including rheumatoid arthritis.. Common threewingnut root has complex components, and triptolide is acknowledged as one of the important effective components of common threewingnut root.OBJECTIVE: To establish rat models of type Ⅱ collagen induced arthritis, and observe the effects of triptolide on the contents of interleukin-6,interleukin-10 and tumor necrosis factor alpha (TNF-α) in peripheral serum and synovial fluid.DESIGN: A randomized control animal experiment.SETTING: Department of Integrated Traditional and Western Medicine,Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiments were carried out in the Tongji Hospital from November 2004 to July 2005. Fifty healthy male Wistar rats of clean degree were purchased from the experimental animal center of Tongji Medical College, Huazhong University of Science and Technology [qualification number of animal [scxk(E)2004-2007]. Triptolide (nobatch number because of temporary production) was bought from Fujian Institute of Medical Sciences, and the purity was above 98.5%.METHODS: ① Ten of the 50 rats were randomly selected as the normal controls, and the others were made into models. Type Ⅱ collagen emulsion was injected intradermally at five points along the back and tail of the rats,0.05 mL for each point, and injected intradermally at two points after 15 days. The rats in the normal control group were treated with saline in the same way. The effects of the model establishment were evaluated according to the scoring standards of arthritis index at 30 days after the first immunity, and the rats scored 6 points or above were taken as successful models and enrolled in the experiments. Twenty successful rat models were randomly divided into arthritis model group (n=10) and triptolide treated group (n=10). ② Triptolide (100 mg/L)was dispensed into parenteral solution with propylene glycol (0.05 in volume fraction), and then intramuscularly injected into hindlimb of rats in the triptolide treated group (0.04 mL/100 g), once every three days for 30 days. The rats in the normal control group were given isovolume saline, and those in the arthritis model group were treated with isovolume propylene glycol (0.05 in volume fraction). ③ The materials were removed at 30 days after administration. The contents of interleukin-6, interleukin-10 and TNF-α in peripheral serum and synovial fluid were detected with enzyme-linked immunosorbant assay(ELISA).MAIN OUTCOME MEASURES: The effects of triptolide on contents of TNF-α, interleukin-6 and interleukin-10 in peripheral serum and synovial fluid were observed.RESULTS: Fifty male Wistar rats of clean degree were selected, 10 were used as normal controls, and 20 of the other 40 rats were successfully made isto models and enrolled in the analysis of results. ① The TNF-α contents in peripheral serum and synovial fluid were the highest in the arthritis model group, and obviously decreased after treatment of triptolide [(35.09±8.82), (15.35±3.56) ng/L; (44.17±8.94), (22.54±4.76) ng/L; P＜ 0.01], which were similar to those in the normal control group (P ＞ 0.05).② The contents of interleukin-6 in peripheral serum and synovial fluid were the highest in the arthritis model group, and were obviously decreased after treatment of triptolide [(76.58 ±6.81), (42.45 ±5.72) rig/L;(88.69±10.56), (48.67±5.97) ng/L; P ＜ 0.01], but did not recover to the levels in the normal control group (P ＜ 0.05). ③ The contents of interleukin-10 in peripheral serum and synovial fluid were the lowest in the arthritis model group, and obviously increased after treatment of triptolide[(17.53±2.07), (21.23±2.91) ng/L; (10.59±2.96), (14.74±1.85) ng/L; P＜ 0.01], which were similar to those in the normal control group (P ＞ 0.05).CONCLUSION: Triptolide can treat arthritis by modulating the contents of cytokines.

Objective:To study the relation between triptolide inhibit peripheral blood mononuclear cell to secret TNF-? and tumour necrosis factor-? gene polymorphism.Methods:Genomic DNA from 41 healthy people was typed for TNF-? -308 polymorphism by allele-specific polymorphism chain reaction(AS-PCR); the TNF-? concentration in the supernatant was measured by ELISA.Results:The TNF-? production of TNF-? -308 non-G/G genotype in LPS-inhibited peripheral blood mononuclear cell(PBMC) culture was more than that of G/G genotype; Compared with TNF-? -308 non-G/G genotype peripheral blood mononuclear cell(PBMC), triptolide can lower the production of TNF-? in G/G genotype peripheral blood mononuclear cell(PBMC).Conclusion:Tumour necrosis factor ?(TNF-?) gene polymorphism might influence the TNF-? secretion of peripheral blood mononuclear cell(PBMC) in healthy humans. We speculate that it may be relative to the different curative effect of Tripterygium Wilfordii Hook.F.(TWHF) to RA patients.

AIM:To explore whether the inhibitory effect of triptolide on IL-1? production by PBMC is associated with IL-1? gene polymorphisms.METHODS:IL-1? gene polymorphism was analyzed in 31 healthy volunteers.From genomic DNA,the C-T polymorphism at IL-1?-511 was typed by PCR-RFLP.Meanwhile the IL-1? was also measured in the supernatants of the cultured and stimulated peripheral blood mononuclear cells(PBMC)by ELISA.RESULTS:After LPS stimulation in PBMC cultures of healthy subjects,the secretion levels of IL-1? in 9 volunteers who carried IL-1?-511 T/T genotype were higher than in volunteers who are not T/T genotype(P0.05).CONCLUSION:The gene polymorphism at IL-1?-511 was related to the production of IL-1?,and the inhibitory effect of triptolide on the production of IL-1? was different in C/C,C/T,T/T genotype of IL-1?-511,which may be one of the reasons for the phenomenon that people respond differently to triptolide.

Objective To investigate the techniques of laparoscopic retroperitoneal lymph node dissection(LRPLND)through extraperitoneal approach. Methods Seven non-seminomatous germ cell testicular tumor(NSGCT)patients of clinical stage Ⅰ underwent LRPLND through extraperitoneal approach.The average age was 31(27-39 years old),the average weight was 62 kg(weight 58-72 kg).Pathological examination revealed 2 testis mixed carcinoma(major of embryonal carclnoma and seminoma),2 yolk sac tumor,1 ehoriocarcinoma,2 teratoma with seminoma.Two of them were in right side,and 5 in left.All the chest X-ray,abdominal CT and bone scanning of them were normal before operation.All patients were general anesthetized.Three or 4 trocars were placed,from 2 of them two gasbags were used to expand the retroperitoneal space at volume of 800-900 ml.The retroperitoneal fat was cleared off from the surface of Gerota's fascia to iliac fossa and the plane spance betwwwn anterior rena fascia and posterior peritoneum was separated In the same way the Plane between dorsal renal and the surface of psoas major and quadratus lumborum unto iliae lossa was exposed.Abdominal aorta or vena cava was exposed,then divided and dissected free from surfaee of psoas maior.The conflux of renal vein and testicular vein at the deep face of renal artere(left)was exposed,then testicular vein was ligated and divided it at its end.Fat and lymph tissue between ureter and vessels was dissected to the crotch of abdominal aorta or inferior vena cava.The primary inguinal incision of testectomy was then excided.Normal antegrade ejaculation recovered in 1 month postoperatively. Results The initial 2 operations were converted to open way as the peritoneum were penetrated largely.The other 5 operations were successful.The average operating time was 285 min(245-350 min),intraoperative blood loss was 100-250 ml.Four patients'pathologic results accorded with clinical staging,and 2 positive lymph nodes were found in the other one.The average number of lymph node resected was 25.6 counts(22-31).Follow-up for 3-20 months,chest X-ray and abdominal CT revealed no evidence of recurrence or distant metastasis,and serum tumor markers were in normal range.Normal antegrade ejaculation recovered in 1 month postoperatively. Conclusions The technique through extraperitoneal approach could be applied for LRPLND.It might be an approach for diagnosis and treatment of stage Ⅰ NSGCT.

Objective To evaluate the long?term effect of the comprehensive control strategy for schistosomiasis with empha?sis on infectious source control in marshland?type endemic areas of Poyang Lake. Methods Three heavy endemic villages with schistosomiasis in the lake areas were selected as pilots for the comprehensive control strategy which included replacing bovine with tractors and forbidding grazing on marshlands. The data of schistosome infections of residents and Oncomelania hupensis snails were collected and analyzed statistically from 2009 to 2014. Results There were no patients with schistosomiasis through feces examinations in the 3 pilot villages. The average density of live snails in Niuzhou marshland showed a decline trend and the density was stable in Liulingwei marshland. There were no infected snails in the 2 marshlands and no schistosome adult worms were found in the sentinel rats Conclusion The implantation of the comprehensive control strategy for schistosomiasis with em?phasis on infectious source control can control the prevalence of schistosomiasis during a long period.

Over one half the patients with rheumatoid arthritis (RA) are being treated with methotrexate (MTX). Although well proven, the efficacy of MTX varies in individual patients. This study examined the metabolic biomarkers that can be used to predict the therapeutic effect of MTX by using metabolomic analysis. Rats were immunized with collagen to rapidly cause collagen-induced arthritis (CIA) and then treated with 0.1 mg/kg MTX for 4 weeks. The clinical signs and the histopathological features of CIA were observed to evaluate the therapeutic effects. Urine samples of CIA rats were collected, and analyzed by using 600 M (1)H-nuclear magnetic resonance ((1)H-NMR) for spectral binning after the therapy. The urine spectra were divided into spectral bins, and 20 endogenous metabolites were assigned by Chenomx Suite. Multivariate analyses were performed to identify the spectral pattern of endogenous metabolites related to MTX therapy. The results showed that the clustering of the spectra of the urine samples from the responsive rats (n=20) was different from that from the non-responsive rats (n=11). Multivariate analysis showed difference in metabolic profiles between the responsive and non-responsive rats by using partial least squares-discrimination analysis (PLS-DA) (R(2)=0.812, Q(2)=0.604). In targeted profiling, 13 endogenous metabolites (uric acid, taurine, histidine, methionine, glycine, etc.) were selected as putative biomarkers for predicting therapeutic response to MTX. It was suggested that (1)H-NMR-based metabolomic analysis can be used to predict the therapeutic effect of MTX, and several metabolites were found to be related to the therapeutic effects of MTX.