Effects of Taoren-Honghua drug pair on degeneration of cervical disc cartilage endplate in rat model of dynamic and static forces imbalance / 中国药理学通报

Aim To investigate the effect of TaorenHonghua drug pair on intervertebral disc degeneration (IVDD) in rats.Methods Fifty healthy Wistar rats were randomly divided into control group,model group,sham group,meloxicam group and Taoren-Honghua drug pair group,with 10 rats in each group.We established dynamic and static forces imbalance of cervical disc degeneration model or sham surgery in rats.12 weeks later,rats were intragastrically administered with meloxicam,Taoren-Honghua drug pair or saline for 30 days.C4/5 and C6/7 discs were harvested from rats.ABOG staining was used for observation of intervertebral disc morphology,real time PCR for mRNA expressions of type Ⅱ collagen (Col Ⅱ) and type Ⅹ collagen (Col Ⅹ),and immunohistochemical staining for Col Ⅱ and Col Ⅹ.Results Compared with model group,Col Ⅱ expression increased,while Col X expression decreased in chondrocyte of intervertebral disc in Taoren-Honghua-treated group(P ＜ 0.01).Conclusion Taoren-Honghua drug pair could delay the degeneration of cartilage endplate in rat intervertebral disc.

A time series analysis on under 5 mortality rate in Zhejiang Province / 预防医学

Objective To study the temporal distribution regular pattern of under 5 mortality rate(U5MR)from 1 998 to 201 4 in Zhejiang Province,and to predict the under 5 mortality rate in 201 5.Methods A time series ARIMA (p,d,q) forecasting model for U5MR was conducted using IBM SPSS Statistics 20.0 statistical analysis software.Results The UMAR showed downward trend.The ARIMA(2,1 ,2)model of U5MR from 1 998 to 201 4 in Zhejiang Province is yt =-0.696 +0.636yt -1 +0.024yt -2 +0.340yt -3 +αt -0.003αt -1 +0.997αt -2 ,and the model fitting was good.Each of the actual mortality was consistent with the trend of model prediction,and was within the 95% confidence interval.The predicted value of U5MR was 4.08‰ (95% CI:1 .52‰ -6.64‰)in 201 5.Conclusion Time series analysis is an effective way to analyze the temporal distribution regular pattern of U5MR,which could be used for short -term prediction.

Peri-articular lymphatic system and "Bi" theory of Chinese medicine in the pathogenesis and treatment of arthritis / 中国结合医学杂志

Rheumatoid arthritis (RA) and osteoarthritis (OA) are the two most common joint diseases, and they have characterization of synovial inflammation and cartilage destruction, associated with the accumulation of numerous catabolic mediators and inflammatory cells in the synovial space and surrounding soft tissues. How these factors are cleared and if the "clearance" process contributes to pathogenesis of arthritis are not known. Recently, we found the existence of the peri-articular lymphatic system in mouse joints. The blockade of lymphangiogenesis and lymphatic draining function accelerates while stimulation of lymphatic function attenuates the severity of joint tissue lesions in mouse models of RA and OA. More importantly, we noticed the similarity between the dysfunction of lymphatic drainage in arthritic joints and "Bi" theory of Chinese medicine (CM), and demonstrated that several Bi disease-treated herbal drugs directly affect the function of lymphatic endothelial cells. Here we review the advances about the interactions between joint inflammation and changes in the peri-articular lymphatic system and discuss our view of linking "Bi" theory of CM to lymphatic dysfunction in arthritis.

Mechanism of resveratrol on the promotion of induced pluripotent stem cells / 中西医结合学报

<p><b>OBJECTIVE</b>To investigate the effects of resveratrol (RV) in reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPSCs) and the related mechanism.</p><p><b>METHODS</b>Primary MEFs were isolated from E13.5 embryos and used within three passages. Retroviruses expressing Sox2 and Oct4 were produced by transfecting GP2-293t cells with recombinant plasmids (MSCV)-Sox2 and MSCV-Oct4. Supernatants containing retroviruses were obtained after 48-hour transfection and MEFs were then infected. Different concentrations (0, 5, 10 and 20 μmol/L) of RV were added to embryonic stem cell (ESC) medium to culture MEFs 48 h post-infection. iPSC clones emerged and were further cultured. Expression of pluripotent markers of iPSCs was identified by cell immunofluorescence and reverse transcription-polymerase chain reaction. Both cytotoxicity and cell proliferation were assayed by Western blot analysis after RV was added into ESC medium. The ultrastructure change of mitochondria was observed by electron microscopy.</p><p><b>RESULTS</b>More than 2.9-fold and 1.3-fold increases in colony number were observed by treatment with RV at 5 and 10 μmol/L, respectively. The reprogramming efficiency was significantly decreased by treatment with 20 μmol/L RV. The proliferation effect on MEFs or MEFs infected by two factors Sox2/Oct4 (2 factors-MEFs, 2F-MEFs) was investigated after RV treatment. At 20 μmol/L RV, induced cell apoptosis and proliferation inhibition were more obvious than those of 5 and 10 μmol/L treatments. Clones were selected from the 10 μmol/L RV-treated group and cultured. Green fluorescent protein expression from one typical clone was silenced one month later which expressed ESC-associated marker genes Gdf3, Nanog, Ecat1, Fgf4 and Foxd3. Electron transmission microscope showed obvious cavitations in mitochondria. The expression of hypoxia-inducible factor-1α was up-regulated when 2F-MEFs were treated with RV compared to the control group.</p><p><b>CONCLUSION</b>RV improved the efficiency of reprogramming 2F-MEFs into iPSCs at low and moderate concentrations (5 and 10 μmol/L). The effect of 10 μmol/L RV on reprogramming was much greater than that of 5 μmol/L RV. However, high concentration of RV (20 μmol/L) led to more severe cavitations in mitochondria and caused cytotoxic effects. Taken together, these findings suggest that RV mimics hypoxia in cells and promotes reprogramming at a low concentration.</p>

Experimental study on lumbar intervetebral disc degeneration model with kidney deficiency by ovariectomizing / 中国骨伤

<p><b>OBJECTIVE</b>To observe effects of removing arms and ovarian on lumbar intervertebral disc and vertebral bone mineral density (BMD) by establishing rat model of lumbar intervetebral disc degeneration (IDD) with kidney deficiency, and to explore internal mechanism of disc degeneration, relationship between disc degeneration and osteoporosis.</p><p><b>METHODS</b>Thirty Sprague-Dawley female rats aged one month were randomly divided into control group, lumbar IDD group and lumbar IDD with kidney deficiency group (combined group), 10 rats in each group. Lumbar IDD group removed double arms, lumbar IDD with kidney deficiency group removed double arms after 3 months, both ovaries were removed. Vertebral bone mineral density were observed by Micro-CT scan; morphological changes were tested by safranine O-fast green staining; II, X collagen protein expression in the intervertebral disc were obsevered by immunohistochemistry; extracellular matrix gene expression were obsevered by real-time polymerase chain reaction (RT-PCR), in order to evaluate the effects of removed of forelimbs and double ovarian on degeneration and vertebral bone mineral density of intervertebral disc.</p><p><b>RESULTS</b>Micro-CT scan showed osteoporosis in kidney deficiency group was obviously worse than other two groups; safranine O-fast green staining showed that intervertebral space became narrowed, intervertebral disc tissue degenerated obviously, chondral palte was underdeveloped in kidney deficiency group; immunohistochemistry showed that X collagen expression increased, type II collagen expression decreased in kidney deficiency group; RT-PCR showed that type II collagen expression in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P=0.000); Age 1 in lumbar IDD group and kidney deficiency group was lower than control group, and had statistical meaning among three groups (P=0.000, P= 0.000); while type X collagen expression was higher than control group, but no significant meaning; MMP-13 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P= 0.000, P=0.000); aggrecanase-2 in lumbar IDD group and kidney deficiency group was higher than control group, with significant meaning compared among three groups (P=0.006, P=0.008).</p><p><b>CONCLUSION</b>Rats model of lumbar disc degeneration established by removed forelimbs and ovariectomized can occure "bone like"--osteoporosis, which is similar with clinical kidney lumbar disc degeneration in tissue morphology, molecular cell biology expression.</p>

Effect of Galla chinensis on the remineralization of two bovine root lesions morphous in vitro / 国际口腔科学杂志·英文版

The present study aims to evaluate the effect of Galla chinensis compounds on the remineralization of two artificial root lesions morphous in vitro. Sixty bovine dentine blocks were divided into two groups and individually treated with two levels of demineralization solutions to form erosive and subsurface artificial carious lesions in vitro. Each group was then divided into three subgroups, each of which were treated with a remineralization solution (positive control), deionized water (negative control), or 4 000 mg⋅L(-1) aqueous solutions of Galla chinensis extract. The dentine blocks were then subjected to a pH-cycling regime for 7 days. During the first 4 days, the daily cycle included 21-h deal and 3-h demineralization applications. The dentine blocks were dealt with the entire day during the remaining 3 days. Two specimens from each of the treatment groups were selected and observed under a polarized light microscope. Data collected using a laser scanning confocal microscope were computerized and analyzed. Galla chinensis extract clearly enhanced the remineralization of both erosive lesion and subsurface lesion patterns in the specimens (P<0.05). The level of remineralization of the erosive lesion by Galla chinensis extract was lower than that of the subsurface lesion (P<0.05). In addition, the remineralization of the subsurface lesion by Galla chinensis extract was higher than that of the remineralization solution (P<0.05). No significant difference between the remineralization of erosive lesions by Galla chinensis extract and the remineralization solution was observed (P>0.05). So Galla chinensis extract has the potential to improve the remineralization of artificial root lesions under dynamic pH-cyclic conditions, indicating its potential use as a natural remineralization medicine.

Effects of centrifugating pressure on the function of preosteoblast OCT-1 / 医用生物力学

Objective To investigate the feasibility of simulating gravity pressure and observe its effect on preosteoblast OCT 1 by centrifugation. Method OCT-1 cells were cultured in 1% agarose gel with a final concentration of 2×107 cells per millilitre. The experiment was divided into two groups according to the duration: one day (1 d) and five days (5 d). Each group was further divided into three subgroups: 0 r/min (control), 200 r/min, and 500 r/min. The 200 r/min group and 500 r/min group were centrifuged for three hours once a day for one day or five days respectively. The control group was kept in the same environment without centrifugation. Results The ColⅠpositive staining was slightly strengthened in the 1d 200 r/min group while it was even more strengthened in the 1d 500 r/min group. Conversely, the positive staining was stronger in the 5d 200 r/min group than that in the 5d 500 r/min group. The markers related to osteoblast differentiation such as Alkaline phosphatise (ALP), Type I collagen α2 (Col1α2), Osteocalcin (OC) and runt-related transcription factor-2 (Runx2) mRNA expression were all up regulated after centrifugation. ConclusionsCentrifugation is a practical method for cell pressure and plays a significant role in promoting the differentiation of preosteoblast OCT-1, which can be used as a new method to simulate the gravity.