Regulatory Effect of Guilu Bugu Prescription in Treating Th17/ Treg Cell Factors of Patients with Postmenopausal Osteoporosis Based on Syndrome Differentiation / 中国实验方剂学杂志

Objective:To observe the clinical efficacy Guilu Bugu prescription in treating postmenopausal osteoporosis （PMO） with deficiency of liver and kidney Yin based on syndrome differentiation and the effect on Th17/Treg cell factors. Method:One hundred and forty patients were randomly divided into observation group （70 cases） and control group （70 cases） by random number table. Both groups' patients got basic treatment of western medicine. Patients in control group got Jintiange capsules， 3 grains/time， 3 times/day. Patients in observation group got Guilu Bugu prescription， 1 dose/day. The treatment lasted for 6 months. And the 6-month follow-up was recorded. Before treatment， at the 6th month after treatment and at the 6th month of follow-up， bone density of lumbar vertebra L2-4 were detected by DXA， and Lumbar BMD were detected by QCT. Before treatment， at the 3rd and 6th month after treatment， deficiency of liver and kidney Yin and Chinese Osteoporosis-targeted quality of life questionnaire （COQOL） were scored. Before and after treatment， Estradiol （E2）， procollagen I amino terminal pro peptide （PINP）， osteoprotegerin （OPG）， collagen I cross linked C-terminal peptide （S-CTX）， tartrate resistant acid phosphatase （TRACP）， interleukin-17 （IL-17）， IL-22， IL-10， transforming growth factor-β1 （TGF-β1） were detected， and CD4+ cells， Th17 cells and Treg cells were calculated. And the safety was evaluated. Result:At the 6th month after treatment and the 6th month of follow-up， DXA （bone mineral density and T-value of lumbar L2-4） and QCT bone mineral density increased （P<0.01）， and the figures in observation group were all higher than those in control group （P<0.01）. At the 3rd and 6th month after treatment， scores of deficiency of liver and kidney Yin and quality of life were all lower than those in control group （P<0.01）. Levels of PINP， S-CTX， TRACP， Th17 cells， ratio of Th17 and Treg， IL-17 and IL-22 were all lower than those in control group （P<0.01）， and levels of OPG， E2， Treg， IL-10 and TGF-β1 were all higher than those in control group （P<0.01）.There was no adverse reaction related to Guilu Bugu prescription. Conclusion:Based on the supplementation of calcium and vitamin D， Guilu Bugu prescription can further improve the bone mineral density， raise the estrogen level， regulate the expressions of bone metabolism markers， Th17， Treg and relevant factors， reverse the imbalance of Th17/Treg， relieve the clinical symptoms and improve the quality of life， with a better efficacy than that of Jintange capsule and a high safety.

Efficacy of Wenyangbushen Formula on mRNA Expression of Osteoprotegerin, Receptor Activator of Nuclear Factor-kappa B and Its Ligand in Rabbits with Steroid-induced Avascular Necrosis of Femoral Head / 中国康复理论与实践

Objective:To observe the effect of Wenyangbushen formula on mRNA expression of osteoprotegerin (OPG), receptor activator of nuclear factor- kappa B (RANK) and receptor activator of nuclear factor- kappa B ligand (RANKL) in rabbits with steroid-induced avascular necrosis of femoral head (SANFH). Methods:A total of 46 healthy conventional New Zealand white rabbits were randomly divided into normal group (n = 10) and model building group (n = 36). The modified method of horse serum plus methylprednisolone was used to establish the SANFH model. Two rabbits from the normal group and four from the model building group were used for HE staining. Then the other models were randomly divided into model group, and low-dose, medium-dose and high-dose treatment groups, with eight rabbits in each group. The normal group was given normal saline 10 ml/d, and the treatment groups were given Wenyangbushen formula 6.44 g/(kg·d), 9.66 g/(kg·d) and 12.88 g/(kg·d), respectively, for eight weeks. The mRNA expression of OPG, RANK and RANKL was detected by reverse transcription- polymerase chain reaction. Results:The empty lacuna rate was significantly higher in the model group than in the normal group (t = 17.085, P < 0.001). Compared with the model group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the treatment groups. Compared with the low-dose treatment group, the mRNA expression of OPG increased (P < 0.01), and the mRNA expression of RANK and RANKL decreased (P < 0.01), in the medium-dose and high-dose treatment groups. There was no significant difference in the mRNA expression of OPG, RANK and RANKL between the low-dose treatment group and the high-dose treatment group (P > 0.05). Conclusion:Wenyangbushen formula could increase the mRNA expression of OPG and inhibit the mRNA expression of RANK and RANKL in the femoral head tissue of the rabbits with SANFH.

Screening of strains with high content of secondary production in endophytic fungi and fermentation liquid from wild soybean / 中草药

Objective: Using 42 endophytes separated and purified from wild soybean as materials, the accumulation of secondary metabolites in hypha and fermentation liquid of endophytic fungi was analyzed so as to screen the strains with high content of secondary production. Methods: Liquid fermentation cultivation with shaking flask and ultraviolet spectrophotometer was utilized to detect the contents of total triterpenoids, flavonoids, polysaccharides, polyphenols, and total phenolic in different endophyte mycelia and their ferment liquids. Results: For 42 endophytic fungi from wild soybean, strains Y2S9, WDL3, and WDL2 had high content of polysaccharide up to 85.51 mg/g, then Y1S9 and Y6S8 had higher content of the flavonoid with 2.99 and 2.24 mg/g, respectively; In addition Y6S8, Y2R1, and Y6R2 would produce more triterpenoid with 25.55, 17.89, and 16.39 mg/g; Meanwhile WDS7 had the highest content of total phenol with 22.65 mg/g. For the fermentation liquid of endophytic fungi, strains Y1S8 and SYS4 had higher contents of flavonoid with 3.111 and 4.809 mg/L; But strains YSYL2, Y1S8, and WDL3 had the higher levels of total triterpenoids up to 17.46, 35.00, and 42.98 mg/L, respectively. Conclusion: Different habitats might be the main factor causing difference in the contents of secondary production. Eleven strains with high yield secondary metabolites selected from wild soybean would lay the foundation for mining endophytic fungi function and screening new drugs.

Effects of huogu I formula (I) on correlated factors of bone regeneration in chickens with steroid-induced necrosis of femoral head / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the mechanism of Huogu I formula (I) in treating osteonecrosis of femoral head.</p><p><b>METHODS</b>Forty-eight healthy female Leghorn chickens were randomly divided into control group, model group and Huogu I group, and each group consisted of 16 chickens. At the meantime of model establishment, chickens of the Huogu I group were administrated with decoction, while the model and control group with distilled water by gavage. At the 8th and 16th week after medication, blood samples were obtained for blood lipid detection while both sides of femoral head were harvested for the rest of examinations. Specifically, expressions of bone morphogenetic protein-2 (BMP2), transforming growth factor beta1 (TGFβ(1)), Smad4 and Smad7 were evaluated by immunohistochemistry, while expression of osteoprotegerin/receptor activator of nuclear factor kappaB ligand (OPG/RANKL) mRNA was detected by in situ hybridization.</p><p><b>RESULTS</b>Compared with the control group, serum levels of total cholesterol (TC), triglyceride (TG) and low-density lipoprotein cholesterol (LDL-C) in the model group rose significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the model group dropped prominently. Positive cell counting of Smad7 and RANKL increased dramatically. In contrast with the model group, levels of TC, TG and LDL-C in Huogu I group reduced significantly. Positive cell counting of BMP2, TGFβ(1), Smad4 and OPG in femoral head of the Huogu I group increased prominently. Indices of Smad7 and RANKL both decreased significantly. Especially at the 8th week, these variations were more significant.</p><p><b>CONCLUSION</b>Huogu I formula is effective in promoting repair of necrotic femoral head by regulating the expressions of BMP2, TGFβ(1), Smads and OPG/RANKL of osteoclast in femoral head.</p>

Effects of different concentrations of gubishu containing serum on the proliferation of rabbit articular chondrocytes in vitro culture / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the effects of different concentrations of Gubishu containing serum on the proliferation of rabbit articular chondrocytes cultured in vitro.</p><p><b>METHODS</b>Articular chondrocytes were obtained from the cartilage of 1-month rabbit and cultured in vitro. They were randomly divided into 8 groups,blank and Gubishu groups in different concentrations (5%, 10%,15%, 20%), MTT assay method was adopted to observe the influence of Gubishu containing serum with different concentrations to the proliferation of chondrocytes after incubated 1, 3, 5, 7 and 9 days.</p><p><b>RESULTS</b>The proliferation of chondrocytes was dependent on the concentration in Gubishu groups. At same time point,there was significant value between every groups, 20% concentration was greatest (P<0.05); There was significant differences between 5%, 10% and 20% concentration of the blank groups at same time point (P<0.05), and was not between 15% and 20% concentration at the 1, 3, 5 and 7 days (P>0.05), 20% concentration of the blank group was greatest. 20% concentrations of Gubishu containing serum was significantly greater than 20% concentrations of blank group at the 1, 3, 5 and 7 days (P<0.05).</p><p><b>CONCLUSION</b>20% concentrations of Gubishu containing serum can significantly increase the proliferation of chondrocytes, and bring the logarithmic growth period forward to the 3 day.</p>

Experimental research progress of warming yang and reinforcing kidney of Chinese medicine to promote the differentiation of bone marrow stromal cells / 中国骨伤

Bone marrow stromal cells (BMSCs), a kind of stem cells residing in bone marrow, have self-renewal, high proliferative capacity and the potential of multilineage differentiation. It has a good prospect in application of the cell replacement therapy, the gene therapy and the tissue engineering and so on. As the content of BMSCs is extremely low in bone marrow, BM-SCs must be amplified in vitro and induced to differentiation to meet the clinical needs. Researches of the recent years suggest there is a very promising way that Chinese medicine could induce BMSCs proliferation, differentiation. Based on the Chinese medicine theory, "the kidney generating marrow and dominating bone" and "kidney storing essence, essence and marrow", the TCM scholars have done some researches to explore the function of warming yang and reinforcing kidney of Chinese medicine to promote bone marrow stromal cells and found that these drugs can promote the BMSCs to proliferate and to differentiate into osteogenic, cartilage and nerve cells. This article elaborates and presents the researches on this aspect.

Progress on integrated Chinese and Western medicine in the treatment of osteoarthritis / 中国结合医学杂志

Osteoarthritis (OA) is a chronic degenerative disease of the joints caused by wide variety of factors. factors. This paper provides a review of the clinical and experimental research on integrated Chinese and Western medicine in the treatment of osteoarthritis. Western medicine in the treatment of osteoarthritis. (1) Clinical research: integrated Chinese and Western medicine therapies were used including physiotherapy, medications, acupuncture, functional training, intra-articular injection of sodium hyaluronate therapy, and arthroscopic debridement with Chinese medicine articular iontophoresis therapy. articular iontophoresis therapy. (2) Experimental research: modern methods were used in studying the mechanism of Chinese medicine in slowing down cartilage degeneration, promoting articular cartilage repair, inhibiting synovial inflammation, and blocking cartilage destruction. inflammation, and blocking cartilage destruction. In addition, this article also reviews the advantages, prospects, and problems of the therapies. and problems of the therapies.

Protection of apoptosis of osteoblast cultured in vitro by Morinda Root Polysaccharide / 中国骨伤

<p><b>OBJECTIVE</b>To explore the protection on apoptosis and the mechanism of promoting the cytoactive of osteoblast by Morinda Root Polysaccharide through the observations of the cultured osteoblast in vitro.</p><p><b>METHODS</b>Prepared blood serum with Morinda Root Polysaccharide and Morinda Root aqueous extract and cultured Osteoblast in vitro with it. The second generation osteoblasts in vitro were separated from the cranium of 24-hours newborn SD rat, which were divided into control group (adding only rat serum during cultivation), induction apoptosis group (adding trans-retinoic acid in control group), Morinda Root aqueous extract group (adding serum prepared by Morinda Root aqueous extract in induction apoptosis group) and Morinda Root Polysaccharide group (adding serum prepared by Morinda Root Polysaccharide in induction apoptosis group). Adopting fluorescence microscope, apoptosis detected by flow cytometry and gene expression of Bcl-2 and Bax detected by RT-PCR, to evaluate the effect of Morinda Root Polysaccharide on the course of osteoblast apoptosis.</p><p><b>RESULTS</b>The apoptotic rate of Morinda Root aqueous extract group and Morinda Root Polysaccharide group were significantly lower than that of induction apoptosis group (P < 0.01). The apoptosis ratio of Morinda Root Polysaccharide group was lower than that of Morinda Root aqueous extract group (P < 0.05). Expression level of Bcl-2 mRNA of apoptosis cell: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01). Expression level of Bax mRNA: induction apoptosis group > Morinda Root aqueous extract group > control group > Morinda Root Polysaccharide group (P < 0.01). Bcl-2/Bax: control group > Morinda Root Polysaccharide group > Morinda Root aqueous extract group > induction apoptosis group (P < 0.01).</p><p><b>CONCLUSION</b>Morinda Root can inhibit the apoptosis of osteoblast induced by trans-retinoic acid in some extent. The above role of Morinda Root Polysaccharide is significant better than that of Morinda Root aqueous extract. It is indicated that Morinda Root Polysaccharide is one of the essential component of inhibiting osteoblast apotosis.</p>

Experimental study on the effects of pharmaco-serum of diabetic rats fed with Chinese herbs qianggubao decoction (Chinese characters: see text) on osteoblast culture in vitro / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the optimum phase and dose of pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) on the differentiation and mineralization of osteoblast (OB). METHODS (OB) was isolated from the skull of 10 newly born SD rats aged 1 to 2 days by means of Trypsin-collagenase digestion. After the OB was identified, different kinds of pharmaco-serum of diabetic rats fed with inactive Qianggubao decoction ([Chinese characters: see text]) of different phase (rats were fed with medicine three days or five days after last fed with medicine one hour or three hours) and concentration (5%, 10%, 20%) were added to the OB and incubated. After 7 days and 18 days of culture,the effects of the differentiation and mineralization of osteoblast were detected.</p><p><b>RESULTS</b>The secretion of ALP and formation of mineralized nodules of osteoblast in the different doses of pharmaco-serum groups were almost the same as that of normal control group, but were superior to that in the model control group. And the group with concentration of 20% pharmaco-serum was the best in the secretion of ALP and formation of mineralized nodules of osteoblast. As to the phases of pharmaco-serum, the best one on the differentiation and mineralization of osteoblast was the serums from diabetic rat-model fed with Qianggubao decoction ([Chinese characters: see text]) three days or five days, after one hour of last fed with medicine.</p><p><b>CONCLUSION</b>The pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) can promote the differentiation and mineralization of osteoblast. Allow for time and the cost of experiment,we presume that pharmaco-serum of diabetic rats fed with Qianggubao decoction ([Chinese characters: see text]) three days, after one hour of last fed, with concentration of 20% and not-inactivation is the optimum on the differentiation and mineralization of osteoblast.</p>

Effect of compound duzhong jiangu granule on knee joint osteoarthritis: a report of 400 cases / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To study the clinical efficacy and adverse reaction of compound Duzhong Jiangu granule (CDJG) on knee joint osteoarthritis (KJO) of Gan-Shen deficiency with stasis in tendon and muscle syndrome.</p><p><b>METHODS</b>The randomized controlled method was adopted in this study, comparative study was conducted in 400 patients in the treated group treated with CDJG and 200 patients in the control group treated with Zhuanggu Guanjie pill (ZGP).</p><p><b>RESULTS</b>The total effective rate in the treated group was 92% and the curative-markedly effective rate was 47%, which were higher than those in the control group respectively. Moreover, CDJG showed superiority in improving symptoms and with shorter initiating time of action as compared with ZGP. However CDJG had the effect more favourable for patients of mild condition.</p><p><b>CONCLUSION</b>CDJG is a kind of effective and safe medicine for treatment of KJO.</p>