Objective To investigate the effect of butylphthalide on Nrf2 signaling pathway after cerebral ischemia-reperfusion in rats and neuroprotective effect.Methods A model of middle cerebral artery ischemia-reperfusion injury in rats was induced by suture method.The rats were randomly divided into 4 groups:a sham operation,an ischemia-reperfusion,a low-dose (100 mg/kg) butylphthalide,and a high-dose (400 mg/kg) butylphthalide.Neurological deficit score was performed at 24 h after referfusion.Western blotting was used to detect the Nrf2 expression,the superoxide dismutase activity and malondialdehydecontent in the ischemic brain tissue.TUNEL assay was used to detect the nerve cell apoptosis.Immunohistochemical staining was used to detect the expression of cleaved caspase-3.Results Butylphthalide significantly upregulated the Nrf2 protein expression with a dose-dependent manner (P ＜ 0.05),increased superoxide dismutase activity (P ＜ 0.05),decreased malondialdehyde content (P ＜ 0.05),decreased numbers of cleaved caspase-3 positive cells and apoptotic cells (P ＜ 0.05).Conclusions Butylphthalide may play a significant neuroprotective effect after cerebral ischemia-reperfusion injury in rats.Its role may be associated with the upreglation of Nrf2 signaling pathway and enhancing antioxidant activity

OBJECTIVE:To optimize the extraction craft of gentiopicrin from gentian.METHODS:The extraction craft was optimized by orthogonal test with gentiopicrin as an index,and the content of gentiopicroside was determined by TLC scanning method.RESTLUTS:The quantity of menstruum and the extraction times of gentiopicrin had significant influence in the gentiopicrin extraction(P

Acute Disease ; Humans ; Paraquat ; poisoning

[Abstract ] Objective Bone marrow mesenchymal stem cells(BMSCs) can be induced to the differentiation into vascular smooth muscle cells in many induction conditions.We sought to explore the possibility of the differentiation of mesenchymal stem cells into vascular smooth muscle cells by continuous cell culture in vitro. Methods Rat BMSCs were isolated from the bilateral tibial and femoral bones by the method of whole bone marrow adherence, followed by ex vivo expansion.BMSCs were identified by flow cytometry and three-lineage differentiation.After continuous five days'cell culture of BMSCs, the specific surface antigens of VSMCs were detec-ted by immunofluorescence, western blot and real-time PCR. Results BMSCs expressed CD29、90, in contrast, they did not express CD45、34、49d.After induction of osteogenesis, adipogenesis and chondrogenesis, alizarin red、oil red and alcian blue staining pro-duced a strong reaction in cells.The expressions ofα-SMA、Calponin1、SM-MHC and SM22 in the cells of experimental group were no-tably increased, which indicated that BMSCs were differentiating towards VSMCs. Conclusion In the absence of exogenous stimula-tion, BMSCs can be successfully induced to differentiate into VSMCs by continuous cell culture.

BACKGROUND: Chinese medicine Gukang prescription has a clear effect on clinical treatment of osteoporosis, but the therapeutic pathway is stil unclear.
OBJECTIVE:To investigate the effects of Chinese medicine Gukang on the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin by regulating core binding factor alpha 1 expression to control the growth and development of osteoblasts.
METHODS:Sprague-Dawley neonatal rats within 24 hours after delivery were used for the separation and culture of osteoblasts. Adult Sprague-Dawley rats were used to prepare drug-containing serum, and then divided into two groups randomly:normal control group and Gukang group. Rats in the normal control and Gukang groups were intragastrical y administrated with extract of Gukang prescription and normal saline based on rat’s body surface area, for 1 consecutive week. Two hours after the last administration, blood samples were taken from the heart. Then the serum was col ected. Osteoblasts at passage 3 were confirmed with alkaline phosphatase assay and digested. After counting and planting, al osteoblasts were divided into two groups and treated with col ected
serum for 72 hours. Proliferative rate of osteoblasts was detected by using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide test. Secretion of alkaline phosphatase was detected by using enzyme linked immunosorbent assay and corrected with the corresponding absorbance value. mRNA expression of core binding factor alpha 1, receptor activator of nuclear factor kappa B ligand and osteoprotegerin were detected by using reverse
transcription-PCR in al groups.
RESULTS AND CONCLUSION:mRNA expression of osteoprotegerin and core binding factor alpha 1 in the Gukang group was significantly higher than that in the normal control group, but protein and mRNA expression of receptor
activator of nuclear factor kappa B ligand were dramatical y lower in the Gukang group compared with the normal control group (P<0.01). These findings indicate that Chinese medicine Gukang prescription can modulate the expression of
core binding factor alpha 1, thereby adjusting the expression of receptor activator of nuclear factor kappa B ligand and osteoprotegerin, which may be one of the mechanisms underlying Gukang treatment for osteoporosis.

ObjectiveTo investigate the influence of cobalt chloride ( CoCl2 )-mimetic hypoxia on theproliferation,apoptosis and migration of human pancreatic cancer cell fine PANC1.MethodsPANC1 cells were treated with 0(control),100,200,400,800 μmol/L CoCl2 respectively for 24 h.Real-time RT-PCR and Western blot were used to determine hypoxia induced factor ( HIF)-1o mRNA and protein expression respectively,and cell counting kit-8(CCK-8) assays,flow cytometry and cell scratch test were used to examine the proliferation,apoptosis and migration of PANC1 cells,respectively.ResultsIn the control group and 100,200,400 and 800 μmol/L CoCl-2 groups,the expressions of HIF-1t mRNA were 1,1.08 ±0.12,1.12 ± 0.09,1.04±0.11,0.66 ±0.07,and the expressions of VEGF mRNA were 1,2.69±0.35,4.81 ±0.54,2.19 ± 0.21,0.79 ± 0.08,while the expressions of HIF-1 α protein were 0.23 ± 0.03,0.36 ± 0.04,1.15 ± 0.11,1.08 ± 0.09,0.44 ± 0.04; and the expressions of VEGF protein were 0.14 ± 0.02,0.12 ± 0.01,0.95 ±0.09,0.87 ±0.09,0.55 ±0.06; and cell viability rates were 100％,(98.43 ±2.88)％,(76.15 ± 0.70)％,(53.87 ±0.77)％,(35.23 ±0.67)％ ; while cell apoptotic rates were (5.2 ±1.12)％,(5.74 ± 1.07)％,(6.82 ± 1.85)％,(12.09 ±3.53)％,(31.88 ±6.95)％ ; the cell migration distance of PANC1 cells were (43.24 ±3.67)％,(59.46 ±5.39)％,(80.56 ±8.05)％,(63.89 ±5.96)％,(9.09 ± 1.59 ) ％.Compared with those of control group,the expressions of VEGF mRNA,VEGF and HIF-1 α protein,cell migration distance showed a two-way variation ( ascending first and descending later) (P ＜0.05 ),and the expression of HIF-1α mRNA and cell proliferation rate was decreased in a dose-dependent manner,while the cell apoptosis was increased in a dose-dependent manner.Conclusions CoCl2 significantly inhibits the proliferation and promotes apoptosis of PANC1 cells at certain level.CoCl2 has a two-way effect on the migration of PANC1 cells,and it may be related to the direct injury of high concentration of CoCl2 on cells.

Objective To investigate the expressions of RGC-32 and E-cadherin in pancreatic cancer and analyze their clinicopathological significance and the correlation with each other.Methods SP immunohistochemistry was used to detect the expressions of RGC-32 and E-cadherin in 42 cases of pancreatic cancer tissues,12 cases of chronic pancreatitis tissues and 8 cases of normal pancreatic tissues.Results The positive staining for RGC-32 was predominantly observed in the cytoplasm of pancreatic acinar cells.The positive staining for E-cadherin was mainly observed in the cytomembrane of normal pancreatic and chronic pancreatitis acinar cells,but aberrant expression ( cytoplasm expression and ( or ) weaker expression) could be found in pancreatic cancer cells.The positive expression rate of RGC-32 and aberrant expression rate of E-cadherin were 78.6％ (33/42) and 54.8％ (23/42),respectively,in pancreatic cancer tissues,which were significantly higher than those in normal pancreatic tissues [37.5％ (3/8) and 0] and chronic pancreatitis [41.7％ (5/12)and 8.3％ (1/12) with statisticai significance,P ＜0.05].The expression of RG C-32 in pancreatic cancer was associated with lymph node metastasis and TNM staging (P =0.016,0.025,respectively),but not with age,gender and differentiation degree ( P =0.831,1.000,0.629,respectively).The aberrant expression of E-cadherin was associated with differentiation degree,lymph node metastasis and TNM staging ( P =0.024,0.004,0.004,respectively),but not with age and gender ( P =0.970,1.000,respectively).A significantly positive correlation was found between positive expression rate of RGC-32 and aberrant expression rate of E-cadherin (r =0.458,P ＜0.01 ).Conclusions Both positive expression rate of RGC-32 and aberrant expression rate of E-cadherin are up-regulated significantly in pancreatic cancer tissues and RGC-32 may be involved in the invasion and metastasis of pancreatic cancer by regulating epithelial mesenchymal transition.

Objective To evaluate the effect of penehyclidine hydrochloride on Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway in the myocardium of pediatric patients undergoing radical correction of tetralogy of Fallot with cardiopulmonary bypass (CPB).Methods One hundred pediatric patients of both sexes,aged 5 months-3 yr,with body mass index of 13.9-16.0 kg/m2,of American Society of Anesthesiologists physical status Ⅱ or Ⅲ (New York Heart Association classification Ⅱ or Ⅲ),with the left ventricular ejection fraction＞50％,scheduled for elective radical correction of tetralogy of Fallot with CPB,were divided into 2 groups (n =50 each) using a random number table:penehyclidine hydrochloride group (group P) and control group (group C).Penehyclidine hydrochloride was intravenously injected at a dose of O.04 mg/kg immediately after successful internal jugular vein puncture in group P,and the equal volume of normal saline was given instead at the same time in group C.Before anesthesia induction,at 10 min after induction,after re warming to 36 ℃,at 1 h after termination of CPB,at the end of surgery and at 24 h after surgery,venous blood samples were collected to detect the concentrations of tumor necrosis factor-alpha,interleukin-6 (IL-6),IL-8 and cardiac troponin T in plasma (by enzyme-linked immunosorbent assay).Myocardial specimens were obtained from the right auricular appendage after opening of pericardium and at 1 h after aortic unclamping for microscopic examination and for determination of activated NF-κB and TLR4 protein and mRNA expression (by Western blot and real-time polymerase chain reaction,respectively).Results Compared to group C,the concentrations of plasma tumor necrosis factor-alpha,IL-6,IL-8 and cardiac tropnin T were significantly decreased after re-warming to 36 ℃C,at 1 h after termination of CPB,at the end of surgery and at 24 h after surgery,the expression of activated NF-κB and TLR4 protein and mRNA was down-regulated at 1 h after aortic unclamping (P＜0.05),and the pathological changes of myocardium were significantly attenuated in group P.Conclusion The mechanism by which penehyclidine hydrochloride reduces inflammatory responses is related to inhibition of the activation of NF-κB/TLR4 signaling pathway in the myocardium of pediatric patients undergoing radical correction of tetralogy of Fallot with CPB.

Thirteen novel oleanolic acid (OA) derivatives were designed and synthesized with modification at positions of C-3, C-12 and C-28 of OA. Their structures were confirmed by MS, 1H NMR and elemental analysis. Their in vitro cytotoxicities against various cancer cell lines (SGC7901, MCF-7 and A549) were evaluated by MTT assay. The results indicated that the tested derivatives were found to have stronger cell growth inhibitory activity than OA. Among them, compounds II2 and II3 showed more potent cytotoxicity on MCF-7 and A549 tumor cells than gefitinib (positive control). They are worthy to be studied further.
Antineoplastic Agents, Phytogenic ; chemical synthesis ; pharmacology ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; Drug Design ; Humans ; Oleanolic Acid ; chemical synthesis ; pharmacology

Objective To investigate the prevalence of obesity and metabolic syndrome(MS)of children and adolescents in Longquan mountainous area in Zhejiang province.Methods A representative sample involving 2 135 children and adolescence aged 10 to 15 years were randomly surveyed and a total of 2 125 had available data(male/female ratios as:1 109/1 016).Using the standard methods,we measured the weight,height,waist circumference,hip circumference,blood pressure,detected fasting plasma glucose (FPG),high density lipoprotein cholesterol (HDL -C),triglyceride(TG),total cholesterol(TC),and calculated non-high density lipoprotein cholesterol(non-HDL). The prevalence of obesity and MS among the 10 to 15 years old children and adolescence in Longquan was compared with that in six cities in China(Beijing,Tianjin,Shanghai,Zhejiang,Chongqing and Guangxi).Results The preva-lence of obesity was 4.7% in 10 -15 -year -old teenage,in which the male obesity prevalence was 6.3%(70/1 109),female obesity prevalence was 3.0%(30 /1 016).The prevalence of overweight was 9.4% and the prevalence of boys was 11.0%,while the girl was 7.6%.The prevalence of obesity and overweight was 16.4%(299/2 125).The prevalence of MS was 2%(42/2 125)in the survey,but 42.0%in the obesity.The prevalence of obesity in Longquan(4.7%)was lower than that in the national six cities(8.1%)among the children and adoles-cents from 10 to 15 years old(χ2 =31.09,P=0.000).But in the obesity students,the prevalence of MS(42.0%)in Longquan was higher than that in six cities(28.8%)(χ2 =5.43,P=0.02).Conclusion The prevalence of obesity in Longquan mountainous area was lower than that in the national six cities among the children and adolescents from 10 to 15 years old.However,the prevalence of MS among the obesity in Longquan was higher than that in six cities.