OBJECTIVE:To investigate the paracrine effects of bone marrow mesenchymal stem cells on osteoblast biological function. METHODS:Bone marrow mesenchymal stem cells were isolated using standard Ficol-Paquedensity gradient centrifugation. Mesenchymal stem cellconditioned medium was prepared to cultivate osteoblasts, MG63. Proliferation of MG63 cells was analyzed by cellcounting kit-8. Migration of MG63 cells was analyzed by cellscratch method. Alkaline phosphatase activity of MG63 cells was analyzed by microplate test kit. Real-time PCR was performed to evaluate osteoblast differentiation markers, alkaline phosphatase, col agen type I and osteocalcin. Alizarin red staining was performed to evaluate osteoblast mineralization. RESULTS AND CONCLUSION:The cells were strongly positive for CD44, CD73 and CD90, but negative for CD34. MG63 cells cultured in the conditioned medium showed better proliferation and migration than those cultured in the Dulbecco’s modified Eagle’s medium. The activity and mRNA expression of alkaline phosphatase were much higher after induction of 4, 7 days (P<0.01). There was no significant difference in expression of col agen type I and osteocalcin after induction of 4 days, but they were significantly higher than those in the control group after induction of 7 days (P<0.05). Alizarin red staining showed that the number of calcium nodules was increased and the mineral apposition was enhanced after induction of 21 days with the conditioned medium. These findings suggest that the paracrine substance of bone marrow mesenchymal stem cells can significantly promote osteoblast proliferation, migration, differentiation and mineralization.

Objective To isolate and purify high-purity neohesperidin from Fructus Aurantii extract.Methods Fructus Aurantii extract was treated by macroporous resin and extracted by n-butanol, then concentrated and some solid was precipitated.After crystallizing the precipitate three times, neohesperidin was obtained.The concentration of n-butanol extract was optimized, which was of great importance in the whole preparation.Results The yield of neohesperidin was 48.6% and purity of neohesperidin was up to 98.7% in Fructus Aurantii extract.Conclusion High-purity neohesperidin could be prepared feasibly and economically by proposed method.

Objective To investigate the efficiency,decision of intra-operative puncture route,treatment of perioperative complications and discuss the other relative problems of the treament for lumbar disc herniation with percutaneous transforaminal endoscopic discectomy.Methods To excise the nucleus pulpesus under percutaneous transforaminal endoscopic discectomy,use the Macnab standard,visual analogue scale and infrared thermal imaging to estimate the efficiency.Results Among followed-up of 208 patients,182 patients were excellent and good outcome,23 patients favorable,2 patients fair,0 patient poor.The leg and back VAS was a significant improvement 1 week post-operation compared with pre-operation (P ＜ 0.05),but no statistical difference among 3 months,6 months,12 months and 1 week post-operation (P ＞ 0.05).The infrared thermal imaging point out that the legs skin temperature of D-value was a significant improvement post-operation compared with pre-operation (P ＜ 0.05).Conclusions The method excised the nucleus pulpesus,and provided the spine maximum protection about the stability and flexibility.Intra-operative puncture route of individuation design can reduce the complications of intervertebral foramen perioperative,and the key to improve the effectiveness.

Objective To select the optimum conditions of the vacuum belt drying process of Radix Salvia Miltiorrhiza(RSM) extract.Methods The process was studied by using orthogonal test design and grading method for multi-index on the parameters of the water content of dried product and drying rate of RSM extract,the average quantity of vapour during unit time span,as the index.Results The optimum process determined by the grading method was listed as follows: water content of the extract before drying was 40%,the feeding speed was 1.5 mL/s,the belt speed was 5 cm/min.Conclusion This technology can increase the average quantity of vapour during unit time span and the drying product has high quality with lower water content and desirable drying rate.

Objective Near-infrared (NIR) spectroscopy was used as a fast analytical technique in the ethanol reflux-extraction process of red ginseng. Methods The NIR spectra of the extracting solution of red ginseng were obtained and the reference measurements of the active constituent in the extracting solution were performed by the colorimetric method. Firstly, the interference information in the spectra was detected by orthogonal signal correction (OSC) method. Then a calibration model between NIR spectra and reference measurements was established by partial least square regression. Results The results showed that the predictive accuracy of NIR calibration model used for the determination of ginsenoside in ethanol extracting process of red ginseng was good. Conclusion NIR Spectroscopy could be applied to the fast analysis for ethanol extracting processes of red ginseng.

Objective Near-infrared (NIR) reflectance spectroscopy was used to develop a fast quality assessment method by simultaneous material identification and moisture quantification of Radix Ginseng Rubra (RGR). Methods The sample was identified by the comparison of its spectrum with a standard NIR spectral library. Similarity measurement was used as the discriminating parameter. The moisture content of sample was quantified by a partial least square (PLS) calibration model, correlative spectrum calculation was used for wavelength selection and multiplicative signal correction (MSC) was applied for pretreatment in the calibration model. Primary reference data were obtained using the traditional loss on drying (LOD) method. Results The NIR library can distinguish RGR from the counterfeit successfully. The optimized eight-factor PLS calibration model of NIR spectra has a high correlation coefficient (R=(0.999 7).) Conclusion The proposed method is rapid, accurate and can be used routinely in the traditional Chinese materia medica manufacturers for quality control of raw materials.

目的探讨选择性脊神经后根切断术(SPR)治疗小儿脑性瘫痪(CP)的疗效。方法对我院获取1年完整随访资料的62例SPR患儿进行回顾性分析。结果按AshWorth 5级肌力标准判断,术后下肢肌张力平均下降2.4级,上肢肌张力平均下降0.4级。术后1年患肢肌张力缓解62例(缓解率100%),行走功能和步态改善48例(77.4%)。伴随症状部分改善。结论SPR能有效缓解CP患儿肢体肌张力,改善康复条件。

Objective To explore renin expression in cytomegalovirus(CMV) infected juxtaglomerular cells(JG) and its biological significance. Methods JG model cell line As4.1 cells derived from kidney tissue were respectively incubated with murine CMV at multiplicities of infection(MOI) of 10, 0.1 and 0 for 5 d, and the control was mock infection with the same amount of ultraviolet inactive CMV as MOI 10, then the cells were harvested. CMV immediate early gene(IE1) mRNA in the cells was tested by RT-PCR. The renin positive cells and the renin fluorescence granules in the cells were examined by immunofluorescence stain. Whether or not re-nin antigen and CMV antigen were showed in the same cells by FITC and TRITC immunofluorescence. The renin gene expression in the cells was individually detected by real-time RT-PCR and Western blot. Results The cells infected by CMV showed typical cytopathic effect(CPE) and viral plaques in the cell monolayer. CMV IE1 mRNA was found in the viral infected cells by RT-PCR. The mass or ring granules of renin positive fluorescence appeared in the cytoplasm of the CPE cells. The renin positive cells congregated around the viral plaques. Renin positive granules and CMV positive granules showed in the same cells. Renin expression in the CMV infected cells exhibited in a dependent manner of ratio of infectious virus particles to cells. Conclusion CMV infection of the cells derived from kidney tissue induces renin expression related to a new pathogenesis of cardiovascular diseases.

BACKGROUND:Estrogen signaling pathway for interaction between aromatase and estrogen-related receptor may exist in bone marrow mesenchymal stem cel s, which is used for regulating biological activity of bone marrow mesenchymal stem cel s. OBJECTIVE:To observe the expression of aromatase and estrogen-related receptors in adult bone marrow mesenchymal stem cel s during osteogenic differentiation. METHODS:Bone marrow mesenchymal stem cel s were respectively cultured in low-glucose DMEM medium (control group) and osteogenic induction medium (induction group). Cel proliferation and calcium deposition were determined by MTT assay and alizarin red staining, respectively. The expression of aromatase, estrogen receptorα, estrogen receptorβ, and estrogen-related receptorαduring osteogenic differentiation were determined by real-time PCR and western blot analysis. Estradiol levels in supernatants and lysates were detected by ELISA method. RESULTS AND CONCLUSION:In the induction group, the proliferation ability of bone marrow mesenchymal stem cel s was the strongest at 72 hours of culture;while there were a great amount of calcium nodules formed at 21 days of culture. Results from PCR and western blot assay showed that the expression of aromatase and estrogen receptorαwas improved in the induction group, but the expression of estrogen-related receptorαwas inhibited. There was no difference in the expression of estrogen receptorβbetween the two groups. ELISA results indicated that the level of estradiol in the supernatant of induction group was the highest. These findings indicate that aromatase, estrogen receptorα, estrogen receptorβand estrogen-related receptorαare al involved in osteogenesis of bone marrow mesenchymal stem cel s. Moreover, estradiol can be synthesized and secreted in bone marrow mesenchymal stem cel s, and most likely, promote the osteogenic differentiation of bone marrow mesenchymal stem cel s by related receptor pathway.

BACKGROUND:Jumonji domain-containing histone demethylase (JMJD) can promote osteoblast differentiation, and estrogen-related receptor alpha (ERRα) can promote osteoblast differentiation and increase bone formation. However, little is reported on the association between postmenopausal osteoporosis andJMJD and ERRα. OBJECTIVE: To study the changes in the JMJD2 family expression in patients with postmenopausal osteoporosis. METHODS: Postmenopausal patients with osteoarthritis of the hip scheduled for total hip arthroplasty, aged 50-70 years, were enroled, including 10 postmenopausal osteoporosis patients (experimental group) and 10 patients with no postmenopausal osteoporosis (control group). During the arthroplasty, the cancelous bone specimens from the femoral head were colected. Then, immunohistochemistry and western blot assay were used to detect expression of histone demethylase (JMJD2A, JMJD2B), histone methylation (H3K9me3, H3K36me3) and ERRα. RESULTS AND CONCLUSION:In the experimental group, the expressions of JMJD2A, JMJD2B and ERRαwere from weakly positive to positive; these expressions were significantly lower in the experimental group than the control group (P < 0.05). The expressions of H3K9me3 and H3K36me3 were significantly higher in the experimental group than the control group (P < 0.05). These findings indicate that the expression of JMJD2A and JMJD2B is consistent with the expression of ERRα in the patients with postmenopausal osteoporosis, and JMJD is likely to serve as an antagonistic enzyme of osteoporosis.