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OBJECTIVE To investigate the improvement effect mechanism of Xibining prescription （XBN） on knee osteoarthritis （KOA） model rats based on AMP-activated protein kinase（AMPK）/mammalian target of rapamycin （mTOR） signaling pathway. METHODS Totally 36 rats were randomly divided into blank group， model group， XBN group （12.56 g/kg）， XBN+metformin （AMPK agonist） group （12.56 g/kg XBN+100 mg/kg metformin）， with 9 rats in each group. Except for blank group， KOA model was induced by anterior cruciate ligament transection in other groups. After modeling， each group was given relevant medicine/normal saline， XBN and normal saline intragastrically， once a day， and metformin intraperitoneally， every other day， for 4 consecutive weeks. The pathomorphological changes of cartilage tissue in rats were observed and Mankin scoring was conducted. The expression level of Aggrecan in rat cartilage， mRNA and protein expressions of platelet reactive protein disintegrin and metalloproteinase with thrombospondin motifs 4 （ADAMTS-4）， ADAMTS-5， matrix metalloproteinase 3 （MMP-3） and MMP- 13， and the phosphorylation level of AMPK and mTOR proteins were detected. RESULTS Compared with blank group， the structure of cartilage tissue in the model group was disordered， the matrix of cartilage layer was lightly stained，the tide line was distorted or interrupted， and Mankin score was significantly increased （P＜0.05）. The protein expression of Aggrecan in cartilage tissue and the phosphorylation level of AMPK protein were all decreased significantly （P＜0.05）； mRNA and protein expressions of ADAMTS-4， ADAMTS-5， MMP-3 and MMP-13 and the phosphorylation levels of mTOR protein were significantly increased in cartilage tissues （P＜0.05）. Compared with model group， the pathological morphology of cartilage was improved significantly in each administration group， and above score or indexes were reversed significantly （P＜0.05）. Compared with XBN group， the degree of cartilage lesions in rats was further alleviated in XBN+ metformin group， and the levels of above score or indicators were further improved （P＜0.05）. CONCLUSIONS XBN can ameliorate cartilage injury in KOA model rats， promote cartilage synthesis and reduce cartilage degradation， the mechanism of which may be associated with activating AMPK/mTOR signaling pathway.
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OBJECTIVE To optimize the extraction technology for the raw drugs of Sanse powder gel paste. METHODS SD rats were divided into blank group， model group， traditional technology group， water extraction group and ethanol extraction group， with 5 rats in each group. Anterior cruciate ligament transection was used to construct knee osteoarthritis model， and the pharmacodynamic effects of different extraction methods on arthritic rats were investigated. Analgesic experiments were conducted using cold and hot pain thresholds and pain mediators calcitonin gene-related peptide （CGRP）， cyclooxygenase-2 （COX-2）， substance P （SP）， and prostaglandin E2 （PGE2） contents as indicators. HE staining was performed on the synovial membrane of rats to observe the degree of synovial cell proliferation， inflammatory infiltration and vascular invasion， and anti-inflammatory experiments were conducted using protein and mRNA expressions of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α） and IL-6 as indicators. The analgesic and anti-inflammatory effects were compared among those groups. In the orthogonal test， ethanol dosage， extraction time and extraction times were used as evaluation factors， and the contents of casticin， strychnine and toxiferine were taken as evaluation indicators； comprehensive score was calculated. The validation experiments were carried out after optimizing the extraction technology of the raw drugs of Sanse powder gel paste. RESULTS Compared with the model group， the cold and heat pain thresholds of drug administration groups （except for the traditional technology group） were all increased significantly （P＜0.05）， while the contents of pain （No.Y2021rc02） mediators CGRP， COX-2， SP and PGE2 were all decreased significantly （P＜0.05）. HE staining showed that inflammatory cell infiltration， fibrosis and collagen deposition were 炎。E-mail：liuzixiu3221@126.com decreased in the administration groups； a small amount of capillary proliferation could be found； the protein and mRNA expressions of inflammatory factors such as IL-1β， IL-6 and TNF-α were decreased significantly in synovial tissue of rats in administration groups （P＜0.05）. Compared with the traditional technology group， most indicators of the ethanol extraction group were significantly reduced （P＜0.05）， and only heat pain threshold and mRNA expression of IL-6 in rats were decreased significantly in the water extraction group （P＜0.05）. The optimal extraction technology of the raw drugs of Sanse powder gel paste included suitable dose of Sanse powder， 8-fold 55% ethanol， heating reflux extraction for 90 minutes， extracting twice. The results of 3 times of verification experiments showed that the average contents of casticin， strychnine and toxiferine were 0.007%， 0.092%， and 0.214%， respectively； RSD were all less than 5%. CONCLUSIONS The optimized extraction technology for the raw drugs of Sanse powder gel paste is stable and feasible， which can improve the efficacy of the preparation.
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OBJECTIVE：To discuss the mechanism of synovitis of knee osteoarthritis （KOA）relieved by “Sanse powder ” volatile oil based on the activation of Nod-like receptor family 3（NLRP3）inflammasome. METHODS ：“Sanse powder ”volatile oil was extracted by distillation- condensation process and the components of it were analyzed by GC-MS. Fibroblast-like synovial cells（FLS）were extracted from the knee joint of male SD rats. The effects of 10，25，50，100，250，500，1 000 μg/mL“Sanse powder”volatile oil on the viability of FLS were examined by CCK-8 assay. KOA inflammatory cell model was induced by lipopolysaccharide for 12 h. The effects of 10，250 μg/mL“Sanse powder ”volatile oil on the protein and mRNA expression of NLRP3，caspase-1 and apoptosis-associated speck-like protein （ASC）were detected. The levels of IL- 1β and IL-18 in supernatants of FLS were determined . RESULTS ：Totally 0.51-0.61 g volatile oil with yellow clear and unique aromatic odor were extracted from“Sanse powder ”，and the extraction rate was 0.33％-0.41％（n＝9）. A total of 41 chemical components were isolated ，and 30 of them were identified and their peak area accounted for 90.073 6％ of the total peak area. The components with high relative content were gingerol flavonoids （17.573 9％），δ-junionene（15.434 5％），gingerol（11.509 5％），etc. When the concentration of volatile oil was 10-250 μg/mL，there was no significant effect on survival rate of FLS （P＞0.05）. Compared with blank group ，the relative protein and mRNA expression of NLRP 3，caspase-1 and ASC and the levels of IL- 1β and IL-18 in supernatant were significantly increased in model group （P＜0.05）. Compared with model group ，relative expression or levels of above indexes were all decreased significantly in FLS and supernatant of “Sanse powder ”volatile oil 10 and 250 μg/mL groups（P＜0.05）. CONCLUSIONS：“Sanse powder ”volatile oil can inhibit NLRP 3 inflammasome activation in FLS and reduce the downstream inflammatory cascade response ，thus exerting its efficacy in ameliorating synovial inflammation of KOA.
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Objective To evaluate the efficacy and safety of Qianggu capsule and calcium drugs for osteoporosis and to provide reference to choose drug for patients with osteoporosis in clinic. Methods Databases including PubMed, Embase, The Cochrane Library, CNKI, WanFang Data, VIP and SinoMed were searched to collect randomized controlled trials (RCTs) of Qianggu capsule and calcium supplements for osteoporosis from inception to the 31th of January 2017. Two reviewers independently screened literature, extracted data and evaluated the risk of bias of included studies. Meta-analysis was performed using RevMan 5.2 software. Results A total of 8 RCTs were included, which involved 594 patients. The results of meta-analysis showed that the effective rate of the Qianggu capsule group was significantly superior to that of the calcium group [RR (95％ CI) was 1.14 (1.06-1.24)]. The bone mineral density (BMD) at lumbar vertebra and femoral intertrochanter of the Qianggu capsule group were statistically higher than those of the calcium group after three months'treatment [MD (95％ CI):0.10 (0.08-0.12), 0.09 (0.06-0.12)]. The bone mineral density (BMD) at lumbar vertebra, femoral neck, ward triangle and femoral intertrochanter of the Qianggu capsule group were statistically higher than those of the calcium group after six months' treatment [SMD (95％ CI): 0.26 (0.03-0.49), 0.56 (0.32-0.80), 0.64 (0.38-0.91); MD (95％ CI): 0.03 (0.02-0.04)]. The incidence of adverse events of the Qianggu capsule group was statistically higher than that of the calcium group [OR (95％ CI) was 2.03 (1.08-3.82)]. Conclusions Comparing to the calcium related drugs, the Qianggu capsule showed a better effect, and could effectively increase the bone mineral density, but the adverse events should be surveillanced.
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This study was aimed to evaluate clinic effect of the layers adjusting external application therapy on knee osteoarthritis (OA). A total of 240 knee OA cases were randomly divided into the treatment group and the control group, with 120 cases in each group. The layers adjusting external application therapy was used in the treatment group. And analgesic plaster of compound of rhizoma arisaematis was used in the control group. The treatment was continued for 21 days. Then, 14-day follow-up was given after the treatment course. There were a total of 35 days. Observations were made on indexes of pain, swelling and joint dysfunction with quantitative scoring. The results showed that the total effective rate was 98.2%in the treatment group, and 83.9%in the control group. The indexes of pain, swelling and joint dysfunction were markedly improved in the treatment group as compared with the control group (P<0.01). It was concluded that the layers adjusting external application therapy was effective on the treat-ment of knee OA.
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Objective To observe the changes of PPARγ expression in ventilator-induced lung injury rats and explore the role of PPARγ in the pathogenesis of ventilator-induced lung injury. Methods Sixty male Sprague-Dawley rats were randomly divided into 3 groups ( n=21 each ):group N received large tidal volume with mechanical ventilation ( Vt=12 mL/kg);group C received lower tidal volume with mechanical ventilation ( Vt=6 mL/kg);group R received room air without mechanical ventilation. Rats in every group were randomly divided into 3 subgroups respectively by 1,4 and 8 h. The samples of lung were collected at 1,4 and 8 h after ventilation. Lung pathological examina-tion, total protein and white blood cells in bronchoalveolar fluid and wet-to-dry weight were detected. The exoressions of PPARγmRNA were detected by RTPCR;PPARγ protein in lung tissues was detected by western bolt. Result After 4 and 8 h ventilation in group N,total pro-tein and WBC in bronchoalvelor fluid,W/D were markedly higher than those of group C and R (P 0. 05). Conclusion PPARγmRNA and protein expressions in the rats lung tissue of ventilator-induced lung injury were decreased and as-sociated with inflammation and damage of lung tissue.
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BACKGROUND:Studies have shown that Weilingxian can maintain and promote the synthesis of proteoglycan and collagen Ⅱ of chondrocyte,and protect artlcular cartilage and postpone the development of osteoarthdtis by inhibiting the level of intedeukin-1(1L-1)possibly.OBJECTIVE:Based on the previous studies,to observe the effect of Weilingxian on the proliferation of rabbit knee articular chondrocyte and transforming growth factor-β1 mRNA expression,and then to explore the role and possible mechanism of Weilingxian in the treatment of osteoarthdtis.METHODS:Knee cartilage was shredded after harvested from New Zealand white rabbits under sterile conditions,and chondrocytes were isolated and cultured by the way Of enzymatic digestion.After identifying by toluidine blue staining,the third-passage calls in the logarithmic growth phase were cultured in vitro and randomly divided into two groups after adherence.The experimental groups were cultured in DMEM with 0.01,0.05,0.1,0.5,and 1.0 mg/mL Weilingxian,while the control group was given with normal medium alone.Chondrecytes morphology was observed under an inverted phase contrast microscope,and the phenotype was identified by toluidine blue staining;Methyl Thiazolyl Tetrazolium(MTT)assay method was adopted to observe the influenca of Weilingxian with difierent concentrations on the proliferation of chondrocytes,and anti-transcription-polymerase chain-type reaction(RT-PCR)was used to assay the expression changes of transforming growth factor-β1 mRNA.RESULTS AND CONCLUSlON:Primary cultured chondrocyte was round-shaped,and most of It adhered after 24 hours,the appearance was polygonal and irregular-shaped;after passage,cell growth was faster than before,the typical appearance was slabstone-like;long spindle-shaped chondrocytes appeared after four generations;after six generations,most cells showed long spindle-shaped fibroblast-like appearance,the rate of growth also slowed down.Extracellular matrix of chondrocytes was stained to be blue by toluidine blue staining,and the nucleus was dark blue.Different concentrations of Weilingxian could promote the proliferation of chondrocytes,effect of 0.5 mg/mL group was significantly,and the peak of proliferation was on the third day.0.05,0.1,0.5,and 1.0 mg/mL Weilingxian group could promote the expression of transforming growth factor-β1 mRNA.and there was no significant difference between four groups(P＞0.05),but the peak was at 0.5 mg/mL group.Weilingxian can promote proliferation of chondrocyte and transfonlling growth factor-β1 mRNA expression,and these may be one of the possible mechanisms that Weilingxian can work in the treatment of osteoarthritis.
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BACKGROUND:Astragalus or olfactory ensheathing cell(OECs)transplantation alone can promote survival of various neurons and functional improvement of injured spinal cord.The clinical effect of astragalus in combination with OECs transplantation remains unclear.OBJECTIVE:To observe the effect of astragalus on the proliferation of OECs and the secretion of neurotrophic factors.METHODS:OECs of SD rats,24 hours old,were isolated and cultured,and identified by immunohistochemistry.OECs at culture day 8 were seeded in polylysine-coated 96-well plate and incubated in 5% CO2 at 37℃ for 24 hours,followed by 2 mg/L,20 mg/L,200 mg/L,2 g/L,20 g/L astragalus injection for 24 hours.MTT and flow cytometry were used to assess the effects of astragalus in promoting OECs proliferation;enzyme linked immunoassay(ELISA)was used to measure the content of neurotrophic factors.Serum culture medium was served as negative control.RESULTS AND METHODS:Compared with the negative control group,2 and 20 mg/L astragalus injection significantly promoted OECs proliferation(P＜0.05,P＜0.01).The result of flow cytometry showed that astragalus could promote OECs in G1 stage and reduce the percentage of cells in S and G2 stages,but the difference was not significant(P＞0.05).However,astragatus injection at each mass concentration significantly reduced the number of OECs(P＜0.05).Moreover,2 mg/L astragalus injection significantly promoted in vitro neurotrephic factor levels in OECs.Results show that astragalus injection in combination with OECs can promote recovery of spinal cord injury.
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OBJECTIVE: To introduce the present research situation of treating spinal cord injury using olfactory ensheathing cells (OECs) transplantation from the aspects of biological characteristics, culture, purify of OECs and its application.DATA SOURCES: PubMed database and Chinese Journal Full-text Database were retrieved with key words of "olfactory ensheathing cells, spinal cord injury" from 2000 to 2009. DATA SELECTION: Inclusion criteria: ①The literature was closely associated with OECs and spinal cord injury. ②Literatures about the same circle published in the near future or on the authoritative journals. Exclusion criteria: ①Repetitive study. ②Meta analysis.RESULTS: OECs has similar characteristics to Schwann cells and astrocytes. At present, the research about OECs includes axonal regeneration, cellular replacement, demyelinating prevention and so on. Furthermore, some clinical experiment had been carried on. However, some difficulties exist in clinical trials, such as transplantation dose of OECs, activity of cell factor and transplant risk. We need to do further study on OECs transplantation and more follow up after cells graft. Some research had tried to transplant with gene transfer cells, and satisfactory results were obtained.CONCLUSION: With the development of genetic engineering and research of OECs transplantation, OECs transplantation must bring more hope to patients with spinal cord injury.
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OBJECTIVE:To promote rational drug use and improve medical quality.METHODS:A comprehensive analysis was performed on the contents of drug counseling problems presented by 483 patients and medical staff.RESULTS:The use of cardio-cerebral-vascular drugs,anti-bacterial drugs and digestive system drugs were the most common drugs inquired by patients,and their questions focused on the action and clinical use,adverse reactions,administration and dosage etc of drugs.However,anti-bacterial drugs were the leading drugs inquired by medical staff,and their questions focused on drugs available for choice and combined use of drugs etc.CONCLUSIONS:Drug counseling can help promote rational drug use,improve medical quality and patients' awareness toward medical care,besides,it is conducive to the publicity for hospital pharmacists.