1.Effects of salvia miltiorrhiza bone-setting capsule on fracture healing in a rat model of closed femoral fractures
Xiqiang WANG ; Renguang SUN ; Shirun SUN ; Jun SUN ; Yongmei WANG ; Taijian LIU ; Huijun DU ; Feng PAN
Chinese Journal of Tissue Engineering Research 2016;20(5):622-627
BACKGROUND:Salvia miltiorrhiza bone-setting capsule is a traditional Chinese medicine for the treatment of fractures due to activating blood circulation to dissipate blood stasis, reducing sweling and pain. OBJECTIVE: To observe the effects of Salvia miltiorrhiza bone-setting capsule on the fracture healing in a rat model of closed femoral fractures. METHODS: Rats were randomly divided into salvia miltiorrhiza bone-setting capsule group, physiological saline group and normal group. In the salvia miltiorrhiza bone-setting capsule group and physiological saline group, rat models of closed femoral fractures were prepared, and then given physiological saline and salvia miltiorrhiza bone-setting capsule 2 pils by intragastric administration. In the normal group, rats were housed normaly. At 7, 14 and 28 days after fractures, hematoxylin-eosin staining conditions, serum osteocalcin, the expression of colagen type I, and the expression of protein and mRNA calus transforming growth factor-beta 1 were observed in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. RESULTS AND CONCLUSION: (1) Hematoxylin-eosin staining demonstrated that at 7 days after fractures, no significant difference was found in pathological changes of femoral fracture in salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 14 and 28 days after fractures, pathological repair was more obvious in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (2) At 3 and 7 days after fractures, serum osteocalcin and the expression of type I colagen were significantly increased in the salvia miltiorrhiza bone-setting capsule group and physiological saline group (P < 0.05), and the expression trend was consistent in both groups. The expression was always higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group, and significant differences were found at 14 and 28 days after fractures (P < 0.01). (3) Transforming growth factor beta 1 expression reached a peak at 3 days after fractures, gradualy reduced, increased at 14 days (the second peak), and diminished at 28 days in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. The expression trend of transforming growth factor beta 1 was consistent in the salvia miltiorrhiza bone-setting capsule group and physiological saline group. At 7, 14 and 28 days, the transforming growth factor beta 1 expression was higher in the salvia miltiorrhiza bone-setting capsule group than in the physiological saline group. (4) Results showed that salvia miltiorrhiza bone-setting capsule could promote fracture healing, and its mechanism was probably associated with serum osteocalcin, the expression of colagen type I and transforming growth factor-β1.
2.Association of the TGF-β1 gene promoter polymorphisms with hepatocellular carcinoma
Zhongheng WEI ; Jianxun LU ; Jian PU ; Zixi WANG ; Hannian YA ; Renguang TANG ; Xianke LONG
Cancer Research and Clinic 2012;24(7):447-450
ObjectivesTo study the relationship between the allele frequencies and genotype distribution of transforming growth factor-beta 1(TGF-β1) gene promoter polymorphisms in Chinese patients with heptocellular carcinoma(HCC),and to analyze the association of the serum levels and genotype of TGF-β1 with HCC.MethodsThe polymorphisms of TGF-β gene,including polymorphisms of TGF-[β1 gene -800G/A、-509C/T,were analyzed by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP)methods in 102 patients with HCC and 110 healthy controls,and the serum level of TGF-β1 was determined by enzyme-linked immunosorbent assay (ELISA). ResultsThe HCC group showed significantly higher serum levels of TGF-β1 than control group [(51.06 ± 9.74)μg/L,(22.12 ± 8.67 )μg/L,t=22.884, P<0.01], The distributions of TGF-β gene -800G/A polymorphisms were not different significantly between HCC group and control group, but TGF-β1 -509C/T gene polymorphism was significantly different. The relative risk suffered from HCC of C allele was 1.822 times of the T allele (OR=1.822,95 %CI:1.238-2.682,t=22.884,P<0.01), the serum level of TGF-β1 T allele carriers was significantly higher than that of no carriers [(53.52:±:10.07)μg/L,(43.57±9.89)μ.g/L,t=3.898, P<0.01]. ConclusionTGF-β1 gene -509C/T polymorphism is associated with HCC, and T allele may be a risk factor for HCC, in which the TGF-β1 T allele carriers may have increased risk by enhancing the TGF-β1 expression in the pathogenesis of HCC.
3.Optimization of the Honey-stir-fired Technology of Chelidonium majus
Renguang WANG ; Xintong SUI ; Yanqun WANG ; Xinrong WANG ; Xiujun ZHONG ; Tianyi ZHAO ; Shumin WANG
China Pharmacy 2021;32(12):1473-1479
OBJECTIVE:To optimize the honey-stir-fried technology of Chelidonium majus . METHODS :Taking the mass ratio of water to honey ,the ratio of honey water to C. majus ,stir-fired temperature ,stir-fired time as the factors ,the total contents of chelidonine ,coptisine hydrochloride ,sanguinarine,berberine,chelerythrine as response values ,Box-Behnken response surface method was used to optimize the processing technology ,and valifation test was conducted. RESULTS :The optimum process conditions were as follows the ratio of water to refined honey 1∶1.9(g/g),the ratio of honey water to C. majus 21∶100(g/g), stir-fried temperature 122 ℃,stir-fried time 10.40 min. After 3 times of validation ,average total contents of 5 components was 10.37 mg/g(RSD=0.23%),relative error of which with predicted value (10.39 mg/g)was 0.19%. CONCLUSIONS :The optimized honey-stir-fried technology of C. majus is stable and feasible.
4.Optimization of Water Extraction Technology of Saikosaponin a and Saikosaponin d from Bupleurum chinense by Electroma- gnetic Cracking
Renguang WANG ; Jingyao YANG ; Xinshu ZHANG ; Zhidong QIU ; Xin CHEN ; Ailing JIA
China Pharmacy 2019;30(18):2546-2551
OBJECTIVE: To establish the method for simultaneous determination of saikosaponin a and saikosaponin d in Bupleurum chinense water extract, and to optimize its water extraction technology for electromagnetic cracking. METHODS: HPLC method was used. The determination was performed on SB-C18 column with mobile phase consisted of acetonitrile-water (gradient elution) at the flow rate of 1.0 mL/min. The column temperature was 40 ℃. The detection wavelength was set at 210 nm, and the sample size was 10 μL. Based on single factor experiment, using extraction time, particle size, solide-liquid ratio as factors, total extraction rate of saikosaponin a to saikosaponin d as indexes, the extraction technology was optimized by using Box-Behnken response surface methdology, and compared with the results of ultrasound method and decoction method. RESULTS: The linear range of saikosaponin a and saikosaponin d were 50.70-202.80 μg/mL (r=0.999 9) and 50.50-202.00 μg/mL (r=0.999 9), respectively. The quantitation limits were 0.16 and 0.13 μg/mL, respectively. The detection limits were 0.05 and 0.04 μg/mL,respectively. RSDs of precision, stability and reproducibility tests were all lower than 2%. The average recoveries were 98.23-102.47% (RSD=1.80%, n=6) and 98.84%-102.06% (RSD=1.60%, n=6). The optimal extraction technology was as follows: the extraction time of 2.50 min; the particle size of 80 mesh, solid-liquid ratio of 1 ∶ 28 (g/mL). Results of 3 times of validation tests showed that the optimal technology included the average total extraction rates of saikosaponin a and saikosaponin d were 8.42 mg/g, which was higher than that of ultrasonic method (8.34 mg/g) and decoction method (8.06 mg/g), and the extration time was shorter. CONCLUSIONS: Established method is simple and accurate, and can be used for simultaneous determination of saikosaponin a and saikosaponin d in B. chinense water extract. The optimized water extraction technology for electromagnetic cracking is stable and feasible.