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OBJECTIVE:To study the stability of baicalin magnesium salt. METHODS:The stability of baicalin magnesium salt at high temperature(60 ℃),high humidity(90%),strong illumination(4000 lx),different temperatures(20,37,50,60 ℃) and pHs(6.80,5.70,4.60,4.30,3.90,3.60,3.20) was investigated,and HPLC was used to detect the drug contents. RESULTS:High humidity test indicated that the quality of baicalin magnesium salt was increased by (6.17 ± 0.12)% in the 5th day and in-creased by(6.92±0.05)% in the 10th day. Drug contents in the 10th day were respectively(94.78±0.12)%,(94.79±0.20)%, (94.66±0.15)% in the high temperature,high humidity,strong illumination tests(n=3). In phosphate buffer solution(pH 6.80), baicalin magnesium salt was stable only when the temperature was below 20 ℃;and it was yet stable in pure water(pH 6.76)at 37 ℃. pH stability test showed that the most stable pH was 4.30. CONCLUSIONS:Baicalin magnesium salt has hygroscopicity to some extent. Strong illumination affects the stability more seriously than high temperature and high humidity. The stability of ba-icalin magnesium salt in pure water is superior to in phosphate buffer solution,and the salt is stable in weak acid solution.
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Objective:To observe chicken typeⅡcollagen-induced arthritis( CIA) in serum of mice with the dynamic changes of IL-27,IL-17,IL-10. Methods: 54 DBA1/J mice were randomly divided into control group(n=6) and model group(n=12), according to the progress of CIA mouse course early,middle and late(7,14,and 35 days after booster immunization) ,taking the eyeball for blood and separating the serum under sterile condition. The dynamic levels of cytokines IL-27,IL-17,IL-10 were detected by flow cy-tometry. Results:The level of IL-27 in the model group was significantly declined in the the progress of CIA mouse course early and middle(P 0. 05);the level of IL-17 in the disease early and late was no significant difference(P> 0. 05) compared with the control group,while in the mid course significantly higher than control group(P 0. 05). Conclusion: IL-27,IL-17,IL-10 paticipate in the pathogenesis of CIA and their alterations at different stages of the disease have a relation to the development of arthritis.
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Objective To study the effects of medicated serum with total saponins from Rhizoma Dioscreae Nipponicae (RDN) on VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α. To investigate the mechanism about total saponin from RDN inhibition of angiogenesis. Methods Medicated serum of total saponins from RDN and tripterygium (positive control) were prepared. Rat synovial cells RSC-364 were divided into four groups: the blank control,IL-17+TNF-α model,tripterygium medicated serum,and total saponins medicated serum groups. After one hour of incubation,all groups except for the blank control were incubated with both IL-17(10 μg·L-1 ) and TNF-α(10 μg·L-1 ) for 24 hours. VEGF mRNA expression in RSC-364 was detected by PrimeScriptTM real-time quantitative PCR (RT-PCR) detection kit,and the AP-1 DNA-binding activity was detected by electrophoretic mobility shift assay (EMSA). Results Compared with the control blank group,both of the VEGF mRNA expression and AP-1 activity in rat synovial cell strain RSC-364 induced by IL-17 and TNF-α increased remarkably (P<0. 05,P<0.01). The VEGF mRNA expression and AP-1 activity in tripterygium medicated serum group and total saponins medicated serum group were remarkably lower than those of the model control group (P<0.05). There was no significant difference between the two medicated serum groups. Conclusion Serum medicated with total saponins from RDN can remarkably decrease VEGF mRNA expression and AP-1 activity,indicating that the total saponins from RDN could influence VEGF secretion by inhibiting the AP-1 signal transduction pathway,VEGF is the key factor of angiogenesis,thereby to restrain angiogenesis.
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This paper was aimed to study effects of diosgenin on expressions of activator protein-1 (AP-1) and vascular endothelial growth factor (VEGF) in synovial tissues of collagen-induced arthritis (CIA) rats induced by bovine type II collagen, in order to investigate the possible mechanism of herbal medicine diosgenin in the treatment of rheumatoid arthritis (RA). After the CIA rats models were successfully established, rats were randomly divided into the blank control group, CIA model group, diosgenin group, and positive medicine control (tripterygium) group. The in situ hybridization was used to detect the expressions of AP-1 (c-fos and c-jun) in synovial tissues of the knee joint. The real-time PCR was used to detect the VEGF mRNA expression in synovial tissues of rats’ knee joints. The results showed that c-jun and c-fos, VEGF mRNA expressions in synovial tissues of rats’ knee joints were obviously higher than that of the blank control group (P<0.01). After treatment of diosgenin and tripterygium, the expressions of c-jun and c-fos, VEGF mRNA were significantly reduced (P<0.01). It was concluded that diosgenin may regulate the expression of VEGF in synovial tissues through c-jun and c-fos of AP-1 in order to inhibit synovial angiogenesis for the treatment of RA.
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This study was aimed to evaluate the subchronic toxicity of diosgenin in mice. A total of 80 mice were divided into 4 groups, which were 0 (control), 100, 200, and 400 mg·kg-1 by the random number table. Intragastric administration was given once a day for 90 days in the assessment of subchronic toxicity of diosgenin among mice. The observed indexes contained body weight, fur color, diet, feces, and etc. The detected indexes contained blood routine analysis, blood biochemistry and pathological examination. The results showed that compared with the control group, the body weights of mice in the male medication group were slight reduced. There were no significant hematologic and pathologic abnormalities. It was concluded that the subchronic toxicity of diosgenin with no observed adverse effect dose level was more than 400 mg·kg-1. The oral administration was relatively safe.
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This study was aimed to observe the influence of Discornin Tablets on activation nuclear transcription factor NF-κBp65 of rheumatoid arthritis (RA) cell model as well as the expression of MMP-9, VEGF and tumor necrosis factor-α (TNF-α). Interleukin-17 (IL-17) and TNF-α were used for stimulating RSC-364 cells. Discornin Tablets at different concentrations were used for intervention. The influence of Discornin Tablets in different concentrations on cell viability was detected by MTT method. Expressions of NF-κBp65 and its inhibitory protein (IκB-α) in each group were detected by western blot method. Changes in VEGF, MMP-9 and TNF-α protein levels in cell broth supernatant were checked by ELISA. The results showed that Discornin Tablets can promote the expression of κB inhibitory pro-tein, reduce the high expression of NF-κB protein level, and inhibit the cellular secretion of VEGF, MMP-9 and TNF-α. It was concluded that Discornin Tablets had negative regulation effect on nuclear transcription factor κB of RSC-364 cells. It can increase the expression of IκB-α, as well as reduce the secretion of inflammation factors and blood vessel newborn factors. It suggested that Discornin Tablets may have the potential regulation effect on RA.
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This study was aimed to observe the influence of water-solubility nipponica saponin on activation of TNF-α+IL-17-induced rat fibroblast-like synovial cell line RSC-364 cellular model nuclear transcription factor NF-κB pathway as well as TNF-α, IL-1, ICAM-1, MMP-2, MMP-3 secretion. IL-17+ TNF-α were used for stimulating RSC-364 to establish rheumatoid arthritis (RA) cellular model. Water-solubility nipponica saponin in different con-centrations was used for intervention. The influence of water-solubility nipponica saponin in different concentrations on cell viability was detected by semi-quantitative RT-PCR method. Changes in the level of TNF-α, IL-1, ICAM-1, MMP-2, and MMP-3 of culture supernatant were detected by ELISA. The results showed that the activation of NF-κB p65 in RSC-364 stimulated by TNF-α+ IL-17 can be inhibited by water-solubility nipponica saponin ac-cording to its concentration. It improved IκB-α expression, and inhibited TNF-α, IL-1, ICAM-1, MMP-2 and MMP-3 secretion. It was concluded that water-solubility nipponica saponin can inhibit the activation of NF-κB pathway, hinder the secretion and activation of multiple downstream genes, which may be its effect in inhibiting syn-ovial inflammation in RA.
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BACKGROUND: The invasion of highly metastatic carcinoma cells on reconstituted basement membrane component is considered as a common model for in vitro experimental studies of invasion and metastasis of malignant tumors. It has been reported that synthetic peptide influences some tumor cells. However, the effect of synthetic peptide on highly metastatic human lung giant-cell carcinoma cell(PG) has been rarely studied.OBJECTIVE: To study the effects of synthetic peptide RGD, YIGSR and their derivatives on invasive ability of PG in vitro.DESIGN: A completely randomized controlled trial was conducted.SETTING and MATERIALS: The human lung highly metastatic tumor and NIH3T3 cell line were both supplied by the Department of Pathology of Beijing Medical University. The synthetic peptides were provided by the Synthetic Laboratory of the Institute of Materia Medica of the Chinese Academy of Medical Sciences and Chengde Medical College. Transwell invasion chamber was purchased from Costor Company. The whole experiment was conducted at the Department of Biochemistry of Chengde Medical College.INTERVENTIONS: NIH3T3 cell line was used to prepare chemotactic factor. PG cells were seeded in Transwell invasion chamber. Under the effect of chemotactic factor, the effects of synthetic peptides on PG invasive ability were examined.MAIN OUTCOME MEASURES: PG cell number of passing through the reconstituted basement membrane was used as an index to evaluate the invasive ability of tumor cells.RESULTS: An appropriate number of PG cells were cultured for 10 hours with chemotactic factor, and the number ofinvasive cells reached a number range that can be applied to statistical analysis(50 to 100 invasive cells). After 100 mg/L of synthetic peptides RGD, YIGSR were incubated for 48 hours with PG cells,the rate of inhibition on invasive ability was 53.63% and 36.86% respectively, and 0% and 6.48% respectively after incubation for 10 hours.CONCLUSION: The PG cells were the proper cells for in vitro experimental studies of invasive ability of cancer cells for the purpose of screening of anti-tumor medicine. RGD and Juncha/Zacha-peptide YIGSR showed a strong activity in inhibiting tumor invasion and a potential value in prevention and treatment of malignant tumor development.
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Objective To analysis the relationship between the multipathogens infection and unhealthy pregnancy.Methods All patients was selectively divided into three groups:group A including 106 patients with unhealthy pregnancy,group B:55 gravidas with normal induced abortion,group C:55 normal parturient with term labor.The ten kinds of pathogens from abortion tissues and placentas were tested by PCR method respectively.Anti-pathogen IgM antibodies were detected by ELISA in the blood specimens.Results The distribution status of the pathogens were as followed:Human parvovirus B19(B19V 23.6%)＞Cytomegalovirus(CMV,16.0%)＞Toxoplasma (TOX，15.1%)＞Rubella virus(RV，10.4%)＞Ureaplasma urealyticum(9.4%)＞Coxsackie(7.5%)＞Herpes simplex virus(HSV,6.6%)＞Adenovirus,Eprstan Barr virus(3.8%)＞Chlamydia trachomatic(CT 2.8%) in the group A.The positive rates of the pathogens infection in both group C and group B were lower.The positive rates of B19V,CMV,RV,HSV,TOX were significant difference(χ2 test,P＝0.000～P＝0.031) among the three groups,hovever others were not(χ2 test,P＝0.121～P＝0.724).Conclusions The pathogen infection is an important factor to occur the unhealthy pregnancy,the major pathogens of unhealthy pregnancy are B19V，CMV，TOX，RV，HSV.These pathogens infection should be detected timely in gestational period.
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Chlamydia trachomatic(CT 2 8%) in the group A.The positive rates of the pathogens infection in both group C and group B were lower.The positive rates of B 19 V,CMV,RV,HSV,TOX were significant difference(? 2 test,P=0 000～P=0 031) among the three groups,hovever others were not(? 2 test,P=0 121～P=0 724).Conclusions The pathogen infection is an important factor to occur the unhealthy pregnancy,the major pathogens of unhealthy pregnancy are B 19 V,CMV,TOX,RV,HSV.These pathogens infection should be detected timely in gestational period.