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.

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.

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 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 purify and refold the inclusion body of a human anti-HBsAg scFv with a 6?His tag, and to determine the affinity constant of the purified recombinant product.METHODS: Solubilizing in buffers containing urea or guanidine hydrochloride (GuHCl), the inclusion body was purified by IMAC, and then refolded by dialysis against urea or GuHCl, at the same time, Ni 2+ charged chelate column was utilized for in situ refolding. The affinity constant of the refolded scFv, polished by immune-affinity chromatography, was determined by non-competitive ELISA. RESULTS: The refolded scFv with highest specific bioactivity was produced by dialysis against GuHCl. Under this condition, the recovery of target protein reached (61.08?1 45)%. The affinity constant of the polished scFv was confirmed to be(2.30?0.32) ?10 7 L/mol. CONCLUSION: The inclusion body studied in this paper can be refolded efficiently under optimal dialysis condition in vitro . The antigen-binding property of this recombinant scFv is not affected by the purification tag fused to the N terminal of the protein.

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

Objective: To investigate the dose-response relationship between hemoglobin concentration and preterm birth, during pregnancy. Methods: With Zhuang ethnicity, a total of 12 780 pregnant women and their infants that admitted to Wuming、Pingguo、Jingxi、Debao、Longan and Tiandong hospitals, were recruited, in Guangxi Zhuang Autonomous Region, from January 2015 to December 2017. Non-conditional logistic regression method was used to analyze the effect of anemia on preterm birth during pregnancy. Dose-response relationship between hemoglobin concentration and preterm birth was explored, using the restrictive cubic spline model. Results: After excluding 2 053 pregnant women with hypertension or aged 35 years and over, results from the non-conditional logistic regression analysis showed that the risk of preterm birth in the anemia group was 1.29 times (OR=1.29, 95%CI: 1.04-1.59, P=0.019) of the non-anemia group in the first trimester. Data from the restricted cubic sample showed that there appeared nonlinear "L" dose-response relationship between hemoglobin concentration and preterm birth in the first trimester and "U" shape in the third trimester (non-linearity test P<0.001). Conclusion There appeared nonlinear dose-response relationship between the hemoglobin concentration and preterm birth, both in the first and third trimesters.

Objective:To investigate the relationship between health promoting behavior and frailty in elderly patients with proliferative diabetic retinopathy(PDR),and the mediating effects of self-efficacy and loneliness.Methods:In an eye hospital of Anhui Province,214 elderly patients with PDR were selected from May 2021 to Nov 2022 by convenient sampling method.The Frailty Scale,Chinese Version of Health-Promoting Lifestyle Profile-Ⅱ,Self-Efficacy Scale and Simplified Loneliness Scale were used in this survey.Bootstrap method of Process software was used to analyze the mediating effect of self-efficacy on the relationship between health promoting behaviors and frailty in elderly PDR patients and the moderating effect of loneliness on the relationship between self-efficacy and frailty.Results:A total of 220 questionnaires were distributed and 214 valid questionnaires were returned,with valid response rate of 97.27% .Moderated mediation effect analysis suggested that health-promoting behaviors negatively predicted frailty(β=-0.508,P＜0.01).Health promoting behaviors and self-efficacy had significant predictive effects on frailty(β=-0.191 and-0.433,P＜0.01),and health promoting behaviors also had a significant predictive effect on self-efficacy(β=0.063,P＜0.01).Self-efficacy played a partially mediating role between health promoting behaviors and frailty,and the mediating effect accounted for 14.76% of the total effect.The product term of loneliness and self-efficacy significantly predicted frailty(β=0.255,P＜0.01),the mediating effect of self-efficacy on frailty was moderated by loneliness.Conclusions:The health-promoting behaviors of elderly patients with PDR affect frailty through self-efficacy,and loneliness moderates the relationship between self-efficacy and frailty.The moderated mediation model is established.

Objective To investigate the influence of the interval between transcatheter arterial embolization (TAE) and radiofrequency ablation on the treatment outcome of small hepatocellular carcinoma. Methods A total of 70 patients with hepatocellular carcinoma who received treatment in Beijing YouAn Hospital, Capital Medical University, from January 2019 to June 2020 were enrolled and divided into observation group (radiofrequency ablation was performed on the day or the second day of TAE) and control group (radiofrequency ablation was performed at 1-2 weeks after TAE) using a random number table, with 35 patients in each group. The independent samples t -test was used for comparison of continuous data between two groups, and the paired t -test was used for comparison of observation indicators before and after surgery; the chi-square test was used for comparison of categorical data between two groups. Results The observation group had a significantly shorter length of hospital stay than the control group (7.80±2.76 days vs 14.31±2.19 days, t =-10.93, P < 0.001). There were no significant differences between the observation group and the control group in complete ablation rate (95.12% vs 95.00%, χ 2 =0.001, P =0.980), incidence rate of adverse reactions (25.71% vs 20.00%, χ 2 =0.324, P =0.569), and 1-year recurrence rate (11.43% vs 14.29%, χ 2 =0.128, P =0.721), and there were also no significant differences in laboratory markers between the two groups before and after treatment (all P > 0.05). Conclusion TAE combined with sequential radiofrequency ablation at a short interval is safe and effective in the treatment of small hepatocellular carcinoma and can significantly shorten the length of hospital stay and reduce hospital costs, and therefore, it holds promise for clinical application.