Ecological charactcristics and biological specific feature of Helleborus thibetanus Fran ch. were briefly described and cultivation technique to transform the wild plant into itscultivated variety was studied to provide a basis for the protection and expliotation ofthe wild resource of H. thibetanus.

BACKGROUND:Basic fibroblast growth factor (bFGF) plays an important role in the ligament tissue healing process, and bone marrow mesenchymal stem cel s transfected with growth factors can be used as seed cel s in ligament tissue engineering.
OBJECTIVE:To observe biological effect of bone marrow mesenchymal stem cel s transfected with recombinant adenovirus vectors carrying bFGF in three-dimensional co-culture with ligament fibroblasts.
METHODS:Passage 3 bone marrow mesenchymal stem cel s were divided into three groups:control group, Ad-EGFP group and Ad-bFGF group. Under a phase contrast microscope, the changes in cel morphology were observed and the rate of transfection was analyzed by flow cytometry. Proliferation of bone marrow mesenchymal stem cel s and ligament fibroblasts was analyzed by MTS, the expression of bFGF protein in bone marrow mesenchymal stem cel s was determined by ELISA. Scleraxis, col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein levels were detected in BMSCs and ligament fibroblasts using real-time fluorescent quantitative PCR.
RESULTS AND CONCLUSION:Recombinant adenovirus-mediated bFGF gene could transfect bone marrow mesenchymal stem cel s efficiently. After co-culture for 3, 6 days, compared with the control group and Ad-EGFP group, in the Ad-bFGF group, the proliferation ability of bone marrow mesenchymal stem cel s and ligament fibroblasts was enhanced (P<0.01), the expression of bFGF protein in supernatant was obviously higher (P<0.01), the col agen type I, col agen type III, decorin and cartilage oligomeric matrix protein mRNA expression decreased in the ligament fibroblasts (P<0.01), but the mRNA expression of Scleraxis, col agen type I, col agen type III in bone marrow mesenchymal stem cel s increased (P<0.01). The results suggest that the co-culture of Ad-bFGF-transfected bone marrow mesenchymal stem cel s with ligament fibroblasts promotes the proliferation of ligament fibroblasts while decreases the col agen synthesis at the same time, and stimulates the proliferation and differentiation of bone marrow mesenchymal stem cel s into ligament fibrolasts.

Objective To investigate the feasibility of Ad-hTGF-β1 transfected bone marrow mesenchymal stem cell(BMSCs) into chondrocytes differentiation and the change of TAZ mRNA. Methods Rats BMSCs were obtained and cultured by whole bone marrow method, and then the third-generation cells were seeded into cell culture plate, and divided into three groups:Ad-hTGF-β1 transfected group,Ad-EGFP transfected group and the control group. The control group was added in common medium without any treatment while the other two groups were respectively added in serum-free medium containing Ad-hTGF-β1 or that containing Ad-EGFP. Seven days later, real-time fluorescent quantitation PCR and Western blot were employed for detecting the expression of TGF-β1 ,while immunohistochemical and Western blot for the expression of collagen Ⅱ , and real-time fluorescent quantitation PCR for the expression of TAZ mRNA. Results Seven days after the transfection, real-time fluorescent quantitation PCR revealed that the average relative expression of TGF-β1 was:Ad-hTGF-β1 group 0.863, Ad-EGFP group 0.183, and the control group 0.180; The average relative expression of TAZ was:Ad-hTGF-β1 group 0.810, Ad-EGFP group 0.416, and the control group 0.366.The expression difference of TGF-β1 and TAZ were statistically significant (P ＜ 0.05). Western blot and immunohistochemical proved strong collagen Ⅱ expression in Ad-hTGF-β1 group while it was detected a little in the other two groups. Conclusion BMSCs could be successfully and stably induced into chondrocytes differentiation by Ad-hTGF-β1. Meanwhile, the mRNA of TAZ is up regulate during the differentiation,so it is suppose that TGF-β1 improve BMSCs into chondrocytes differentiation by TAZ.

Effect of strophanthidin (Str) on intracellular calcium concentration ([Ca2+]i) was investigated on isolated ventricular myocytes of guinea pig. Single ventricular myocytes were obtained by enzymatic dissociation technique. Fluorescent signal of [Ca2+]i was detected with confocal microscopy after incubation of cardiomycytes in Tyrode's solution with Fluo3-AM. The result showed that Str increased [Ca2+]i in a concentration-dependent manner. The ventricular myocytes began to round-up into a contracture state once the peak level of [Ca2+]i was achieved in the presence of Str (10 μmol·L-1), but remained no change in the presence of Str (1 and 100 nmol·L-1). Tetrodotoxin (TTX), nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str (1 and 100 nmol·L-1), but had no obvious effects on the action of Str (10 μmol·L-1). The elevation of [Ca2+]i caused by Str at all of the detected concentrations was partially antagonized by rynodine (10 μmol·L-1) or the removal of Ca2+ from Tyrode's solution. In Na+, K+-free Tyrode's solution, the response of cardiomycytes in [Ca2+]i elevation to Str (10 μmol·L-1) was attenuated, while remained no change to Str (1 and 100 nmol·L-1). TTX, nisodipine, and high concentration of extracellular Ca2+ changed the response of cardiomycytes to Str at all of the detected concentrations in Na+, K+-free Tyrode's solution. The study suggests that the elevation of [Ca2+]i by Str at the low (nomomolar) concentrations is partially mediated by the extracellular calcium influx through Ca2+ channel or a "slip mode conductance" of TTX sensitive Na+ channel. While the effect of Str at high (micromolar) concentrations was mainly due to the inhibition of Na+, K+-ATPase. Directly triggering the release of intracellular Ca2+ from sarcoplasmic reticulum (SR) by Str may be also involved in the mechanism of [Ca2+]i elevation.

Rheumatoid arthritis (RA) is a chronic autoimmune disorder primarily occurred in small joints.Our previous studies suggested that sufficient lymphatic drainage was favorable for the recovery of RA.This study aimed at exploring the effect of Er Chen combined with Tao Hong Si Wu Tang (ECSWT) on RA in TNF transgenic (TNF-Tg) mice.Ten-week old TNF-Tg mice and WT littermates were detected with indocyanine green-nearinfrared (ICG-NIR) lymphatic imaging system before and after accepting the 12-week intragastric administration of ECSWT.All ankle joints were assessed by micro-CT scanning.According to three dimensional images of Micro-CT,it was found that the ankle joints in the TNF-Tg group were much eroded compared with the control group.The bone mass and structure were protected after the treatment of ECSWT.ICG-NIR results showed that lymphatic clearance rate of the TNF-Tg group decreased compared with those of the control group.In comparison with the TNF-Tg group,ECSWT promoted the repair of lymphatic function.Compared with the control group,the pulse value of the TNF-Tg group declined;while this condition could be rescued by ERSWT.In conclusion,ECSWT mitigated bone erosion of astragalus bone area in TNF-Tg mice in contrast to the saline mice,while promoted the pulse value and clearance of lymphatic vessels afferent from footpad to popliteal lymph node,implying that ECSWT was a promising agent for treating RA through its promoting lymphatic drainage function effects.

To observe the effects of Qishe Pill, a compound traditional Chinese herbal medicine, on lumbar vertebral bone formation induced by long-time upright posture in rats and to investigate the potential mechanism.

To investigate the effects of serum containing Yiqi Huayu Bushen Recipe, a compound Chinese herbal medicine, on expressions of aggrecan and type X collagen mRNAs of the degenerated intervertebral disc cells in patients with cervical spondylotic myelopathy.

In order to explore the role of activator protein-1 (AP-1) in the transcription of interleukin-5 (IL-5) gene regulated by protein kinase C (PKC) signal in peripheral blood T lymphocytes from asthmatic patient, T lymphocytes were isolated and purified from peripheral blood of each asthmatic patient. The T lymphocytes were randomly divided int9 4 groups: group A (blank control), group B (treated with PKC agonist phorbol 12-myristate 13-acetate (PMA)), Group C (treated with PMA and AP-1 cis-element decoy oligodeoxynucleotides (decoy ODNs)), and group D (treated with PMA and AP-1 mutant decoy ODNs). The ODNs were transfected into the T cells of group C and D by cation liposome respectively. Reverse transcription-polymerase chain reaction (RT-PCR) was employed to assess IL-5 mRNA expression, and electrophoretic mobility shift assays (EMSA) for the activation of AP-1. The results showed that the activation of AP-1 (88 003.58±1 626.57) and the expression of IL5 mRNA (0. 8300±0. 0294) in T lymphocytes stimulated with PMA were significantly higher than these in blank control (20 888.47±1103.56 and 0. 3050±0. 0208, respectively, P＜ 0.01), while the indexes (23 219.83±1 024.86 and 0. 3425±0. 0171 respectively) of T lymphocytes stimulated with PMA and AP-1 decoy ODNs were significantly inhibited, as compared with group B (P＜0.01). The indexes (87 107. 41±1 342.92 and 0. 8225±0. 0222, respectively) in T lymphocytes stimulated with PMA and AP-1 mutant decoy ODNs did not exhibit significant changes, as compared with group B (P＞0.05). The significant positive correlation was found between the activation of AP-1 and the expression of IL-5 mRNA (P＜ 0.01). It was concluded that AP-1 might participate in the signal transduction of PKC-triggered transcription of IL-5 gene in asthmatic T lymphocytes. This suggests the activation of PKC/AP-1 signal transduction cascade of T lymphocytes may play an important role in the pathogenesis of asthma.

ObjectiveTo observe the effects of extract of Ginkgo biloba, Ginaton, on cerebral oxygen utilization coefficients (O2UCc) and lactate clearance rate (LCR) in patients with delayed encephalopathy after acute carbon monoxide poisoning (DEACMP).Methods The 196 patients with DEACMP admitted to Harrison International Peace Hospital Affiliated to Hebei Medical University from January 2011 to March 2016 were enrolled, and they were randomly divided into a control group and an observation group, 98 cases in each groups. The control group received routine treatments such as hyperbaric oxygen, measures to prevent and treat cerebral edema, and promote brain cell metabolism, etc routine therapies: while in the observation group, besides the above routine treatments, additionally intravenous drip of Ginaton 70 mg (with 0.9% sodium chloride injection 250 mL added), once a day, for consecutive 2 weeks to complete one therapeutic course. At ambient air, before treatment and 6, 24 and 72 hours after treatment, the O2UCc and LCR and the changes of mini-mental state examination (MMSE) score, clinical efficacy and hospital mortality were observed and compared between two groups. Pearson correlation test was used to analyze the correlations between O2UCc, LCR and MMSE score.Results On the day of admission before treatment, there were no differences in O2UCc, lactate and MMSE score between the two groups (allP > 0.05). At 6 hours, 24 hours after treatment, the levels of O2UCc in observation group were obviously lower than those in control group [6 hours: (40.64±9.50)% vs. (45.78±7.94)%, P = 0.000; 24 hours: (30.51±6.76)% vs. (33.34±8.19)%,P = 0.009], while the levels of LCR were significantly higher than those in control group [6 hours: (14.93±2.27)% vs. (11.45±1.88)%,P = 0.000; 24 hours: (19.86±3.42)% vs. (13.73±2.35)%,P = 0.009]. There were no statistical significant differences in O2UCc and LCR at 72 hours after treatment between the two groups (P > 0.05). The MMSE scores at 6, 24 and 72 hours after treatment in observation group were higher than those in control group (6 hours: 15.52±3.61 vs. 11.60±2.49, 24 hours: 20.05±5.79 vs. 14.85±5.71,72 hours: 23.87±5.96 vs. 18.07±6.97, allP < 0.05). The total effective rate in observation group was significantly higher [77.55% (76/98) vs. 61.22% (60/98),P < 0.05], and the mortality [3.06% (3/98) vs. 10.20% (10/98),P < 0.05] was obviously lower than their own counterparts in control group. The correlation analysis showed that at different time points, the levels of O2UCc were negatively correlated to the corresponding MMSE scores (r6 h = -0.153,r24 h = -0.158, P6 h = 0.032,P24 h = 0.027), and there were positive correlations between the levels of LCR and MMSE scores (r6 h = 0.473, r24 h = 0.242,P6 h = 0.000,P24 h = 0.001) in patients with DEACMP.Conclusions The treatment of Ginaton in patients with DEACMP can effectively elevate the LCR and MMSE score, reduce O2UCc, decrease mortality and improve the prognosis, thus the clinical curative effect is distinct.

Objective To investigate the relationship between the genetic polymorphisms, which played roles in the pathogenesis of metabolic syndrome (MS), and susceptibility of non-alcoholic fatty liver disease (NAFLD) in Han people in Guangdong province. Methods The subjects were selected from an epidemiologie survey in Guangdong province. Fifty to 117 adult NAFLD patients, who met the criteria of Chinese guideline for diagnosis of NAFLD and had typically clinical, biochemical signs and abdominal ultrasonography, were recruited in the study. By using 1 : 1 matched method of nested case-control study, same numbers of people without NAFLD were included as controls. The genetic analyses was performed by using genomic DNA extracted from peripheral blood. Polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) was applied to detect the single nucleotide polymorphisms (SNPs) at 9 sites in 7 candidate genes. Results Most SNPs of the genes were related to the susceptibility of NAFLD. Some of them had positive relation (increasing the risk) such as tumor necrosis factor (TNF)-α-238, adiponectin-45, leptin-2548, peroxisome proliferator-activated receptors (PPAR) γ-161 and phosphatidylethanolamine N-methyltransferase (PEMT)-175. Some had negative relation (decreasing the risk) including adiponectin-276 and hepatic lipase-514. And some had no relation (TNF-α-380 and PPAR g coactivator-1α-482). Conclusion Most cytokines' SNPs of candidate genes discovered in MS patients are related to the susceptibility of NAFLD.