Aim To study the chemical constituents of Cypripedium tibeticum. Methods Compounds were isolated by repeated silica gel chromatography and purified on Sephadex LH-20 and structures were determined by spectral analysis. Results Cypritibetquinones A and B were isolated from the ethyl acetate residue and their structures were determined as 7-hydroxy-2-methoxy-1, 4-phenanthraquinone ( 1 ) and 7-hydroxy-2,10-dimethoxy-1,4-phenanthraquinone ( 2 ), respectively, by extensive spectral analyses. Conclusion Cypritibetquinones A and B are two new phenanthraquinones.

Objective To investigate the influence of c-myc on the growth, proliferation, apoptosis,invasion and cell cycle of the gastric line HFE145. Methods The cDNA of c-myc was subcloned into a constitutive vector pcDNA3.1 followed by transfection in HFE145 by using liposome. Then stable expression clones (HFE-myc) were selected. The apoptosis and cell cycles were detected using flow cytometry. The growth and proliferation were analyzed by making cell growth curves and colony formation assay respectively. The ability of invasion were tested using cell migration assay. Results HFE-myc group grew faster than HFE145and HFE-pc. The cell counts of HFE-myc in five of seven days were more than those of others significantly (P＜0.05). There was no difference between the two control group. Cell cycle analysis showed that HFE-myc group proliferated faster, mean proportion of cells in G2-M was about 25 % and higher significantly (P ＜0.05)than those of the two control groups. Results of colony formation assay showed that the mean colony formation rate of HFE-myc was 0.27 and higher than those of the control groups(P ＜0.05). The results of cell migration assay suggested that the cell migration rate of HFE-myc was not higher significantly than those of the control groups (P ＞0.05). Conclusion c-myc can promote the growth, proliferation. It can increase the proportion of cells in division stage, so promote the division. But it have little influence on the invasion of cells.

Objective To study the effect of hyperbaric oxygen(HBO)therapy on abilities of learning and memory and hippocampal pyramidal cell in vascular dementia rats.MethodsThe vascular dementia model was reproduced and divided into control,sham and treatment groups.After 30 d of HBO therapy,the rats were assessed with the Morris water maze,and their hippocampal pyramidal cells were observed after HE staining.ResultsCompared with the control group,the ability of learning and memory of rats in treatment group increased,as well as the hippocampal pyramidal cells did.ConclusionHyperbaric oxygen therapy can be used to treat vascular dementia.

ABSTRACT:Objective To explore the role of PTEN in the suppression of neural stem cells so as to clarify whether neural stem cell proliferation can be promoted by regulating the PI3K-Akt/PTEN expression level. Methods We removed the hippocampus from neonatal 24 h Kunming mice,isolated and cultured the generation of neural stem cells,which were then identified by immunofluorescence test.We randomly grouped the cultured neural stem cells into normal group,ischemia model group,hypoxia group (hypoxia + ischemia model group),Lip2000 group (hypoxia+ ischemia model group + Lip2000 null transfection group,PTEN transfection group (hypoxia +ischemia model group+Ad5-PTEN transfection group),and PTEN interference group (hypoxia+ ischemia model group+Lip2000+PTENsiRNA interference group).We detected the proliferation of neural stem cells in the groups at different time points,and PTEN protein expression of neural stem cells in each group after PTEN transfection,
and the effect on iconic protein on Akt-PI3K signaling pathway.Results (1 )Nestin identification of the neural stem cells was tested by immunofluorescence.We observed green bright and typical cell spheroids under fluorescence microscope;the clear structure could be seen within spheroids.(2)We determined the proliferation of neural stem cells at different time points by MTT.After 36 h of culture,the neural stem cells had obvious proliferation in the hypoxia group and Lip2000 group compared with the model group, the PTEN transfection group and PTEN interference group (P <0.05),including the cell proliferation of PTEN interference group compared to that of the model group,and PTEN transfection group obviously,the differences were also significant (P < 0.05 );(3 ) Compared with the cells in the normal group,PTEN expression increased in the model group and PTEN gene transfection group (P <0.05);it was all lower in other groups than in the normal group (P <0.05).The trend of decrease was similar in hypoxia group,Lip2000 group,and PTEN interference group (P >0.05);(4)After PTEN plasmid transfection,there were no apparent changes in the total Akt and bad in the groups,but compared with the normal group,PI3K,p-Akt,and p-bad expressions in model group and PTEN gene transfection group reduced (P <0.05),with the more obvious changes in the PTEN transfection group.PI3K,p-Akt,and p-bad expressions in the hypoxia group, Lip2000 group and PTEN interference group increased (P < 0.05 ). Conclusion Hypoxia contributes to promoting the proliferation of neural stem cells,and the inhibition of PTEN on PI3K/Akt might be the key factor for blocked proliferation of adult neural stem cells.

Objective: To study the effects of Ruanmailing Oral Liquid, a compound traditional Chinese herbal medicine, on spatial learning and memory ability and expression of apurinic/apyrimidinic endonuclease/redox factor-1 (APE/Ref-1) in hippocampal CA1 region in rats with experimental vascular dementia (VaD). Methods: VaD was induced in rats by permanent occlusion of bilateral common carotid arteries. Forty-five VaD rats were randomly divided into untreated group, nimodipine group, low-dose Ruanmailing group and high-dose Ruanmailing group. Another 15 rats underwent a sham operation consisting of similar skin incision and manipulation but without occlusion of carotid arteries. From the next day after occlusion, the rats were intragastrically administered with normal saline, nimodipine suspension or Ruanmailing Oral Liquid respectively for 30 days. Morris water maze experiment was adopted to test learning and memory of rats in each group. Expression of APE/Ref-1 protein in the hippocampal CA1 region was measured by immunohistochemical method. Results: Escape latency was significantly shortened and number of entries in the target area of rats was significantly increased in the high-dose Ruanmailing group as compared with those in the untreated group (P<0.01). Compared with the untreated group, count of APE/Ref-1 positive cells was significantly increased in the hippocampal CA1 region in the high- and low-dose Ruanmailing groups (P<0.01). Compared with the low-dose group and the nimodipine group, the count of APE/Ref-1 positive cells was remarkably increased in the hippocampal CA1 region in rats of the high-dose Ruanmailing group (P<0.01). There was no statistical difference between the low-dose Ruanmailing group and the nimodipine group. Conclusion: Ruanmailing Oral Liquid can improve the learning and memory ability and enhance the lowered expression level of APE/Ref-1 in the hippocampal CA1 region of rats with VaD.

BACKGROUND:As a lipid-lowering drug, simvastatin has been proved to be effective in promoting bone formation. Previous studies have demonstrated that local y applied simvastation accelerated fracture healing at middle phase in osteoporotic rats, while no study focuses on the influence of local y applied simvastatin on fracture healing at late period in an osteoporotic rat.
OBJECTIVE:To investigate the effect of simvastatin local y applied from a bioactive polymer coating of implants on osteoporotic fracture healing at late period.
METHODS:Female Sprague-Dawley rats were divided into sham group, osteoporotic fracture group and simvastatin group. In the sham group, the abdominal cavity was exposed without ovariectomy. Six weeks later, femur fracture models were established in normal or osteotoporotic Sprague-Dawley rats, and intramedul ary stabilization was achieved with uncoated titanium Kirschner wires in normal rats (sham group),with polylactic acid coated titanium Kirschner wires (osteoporotic fracture group) and with simvastatin/polylactic acid coated titanium Kirschner wires (simvastatin group). Femurs were harvested after 12 weeks, bone mineral density was determined with dual X-ray absorptiometry, and then radiographic and histological analysis was performed for analysis of fracture healing. Immunohistochemical evaluation was employed for bone morphogenetic protein 2 expression.
RESULTS AND CONCLUSION:The bone mineral densities of both the total fractured femur and fractured site 12 weeks after fracture in the osteoporotic fracture group and simvastatin group were markedly decreased compared to normal fractured rats. The bone mineral density of the fractured site was significantly higher in the simvastatin group than the osteoporotic fracture group. Radiographic results demonstrated completely finished cal us remodeling in the sham group;poor healing, pale cal us density and blurred fracture line were seen in the osteoporotic fracture group;disappearance of fracture line, bone defects fil ed with cal us, and deep periosteal reaction were found in the simvastatin group. X-ray scores in the sham and simvastatin groups were significantly higher than that in the osteoporotic group (P<0.05). Hematoxylin-eosin staining showed a delayed healing process in the osteoporotic group, and revealed a significantly processed cal us with regular-shaped newly formed bone trabeculae in the simvastatin group. Immunohistochemical evaluation showed no significant difference in the bone morphogenetic protein 2 expression between any two groups. These findings suggest an improved fracture healing under local application of simvastatin in osteoporotic rats.

Objective To observe the effect of basic fibroblast growth factor (bFGF) on magnetic resonance spectroscopy of vascular dementia (VD) rats.Methods After the VD model was reproduced, the 12 rats were randomly divided into the therapy group and dementia group with 6 animals in each group. Another 6 rats were selected as sham operation group. The VD rats in therapy group were treated with bFGF by hypodermic injection. After 5 weeks, abilities of learning and memory of three groups' rats were tested by Morris water maze. The changes of magnetic resonance spectroscopy and N-acetyl aspartate (NAA), NAA/（Cr＋Cho） in lobus temporalis of VD rats were also observed.Results Abilities of learning and memory of rats significantly improved in the therapy group than that in the dementia group (P<0.01). The values of NAA, NAA/（Cr＋Cho） in lobus temporalis of VD rats significantly increased in the therapy group than that in the dementia group (P<0.01).Conclusion bFGF by hypodermic injection can obviously elevate abilities of learning and memory and the values of NAA, NAA/（Cr＋Cho） in lobus temporalis of VD rats.

OBJECTIVE: To investigate androgen receptor (AR) expression and the effect of epidermal growth factor (EGF) and testosterone on AR expression level.
 METHODS: EGF or different concentrations of testosterone were incubated with the primary urethral plate fibroblasts from patients with hypospadias. The levels of AR expression in the fibroblasts were detected by immunocytochemical assays and graphical analysis.
 RESULTS: There was no significant difference in AR activation under physiological concentrations (3×10(-8) mol/L) of testosterone between the control and the distal hypospadias group (P>0.05). However, there was a significant decrease in AR activation in the proximal hypospadias group compared to that in the control group (P<0.001). Under the concentration of 3×10(-6) mol/L, the effects of testosterone on AR activation were dramatically different in the three groups (control group>distal hypospadias group>proximal hypospadias group, P<0.001). AR activation level in the group of proximal hypospadias was improved most obviously when EGF and physiological concentration of testosterone were employed in the urethral plate fibroblasts from hypospadias patients (P<0.001), and it was improved more in the distal hypospadias group than that in the control group (P=0.02).
 CONCLUSION AR expression and activation in the urethral plate fibroblasts from hypospadias patients are abnormal. EGF can be used to improve AR activation in fibroblasts from different types of hypospadias, especially in the proximal type.
Cells, Cultured ; EGF Family of Proteins ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Hypospadias ; metabolism ; Male ; Receptors, Androgen ; metabolism ; Testosterone ; pharmacology

Objective To evaluate the impact of water pollutants, water levels and meteorological factors on the Oncomelania hupensis density in Eastern Dongting Lake regions, so as to provide insights into schistosomiasis control. Methods O. hupensis snails were surveyed using a systematic sampling method in snail-infested marshlands in Eastern Dongting Lake regions from 2007 to 2014, and data pertaining to water pollutants, water levels and meteorological factors were collected. The duration of submergence and the date of the start of submergence were calculated. The snail density and its influencing factors were descriptively analyzed, and a linear mixed model was generated to examine the impacts of variables on the snail density. In addition, smooth curves were fitted to investigate the relationship between snail density and variables. Results The snail density appeared a fluctuation in Eastern Dongting Lake regions during the period from 2007 to 2014, with the highest density on October, 2010 (52.79 snails/0.1 m²) and the lowest density on January 2009 (2.15 snails/0.1 m²). Linear mixed-model analysis showed that permanganate index, total phosphorus and the date of the start of submergence affected the snail density (t = 6.386, −2.920 and −3.892, all P values < 0.01). Smooth curve analysis revealed that the associations of the snail density with the permanganate index and total phosphorus appeared an approximately quadratic curve. After the end of April, the earlier date of the start of submergence resulted in a higher snail density. Conclusion Permenganate index, total phosphorus and the date of the start of submergence affect the O. hupensis snail density in Eastern Dongting Lake regions.

The coronary artery disease is a frequent severe disease of cardiovascular system in recent years. Meanwhile, lung cancer, with its high morbidity and mortality, is the most frequent malignant tumor of respiratory system in the world. Clinical studies have shown that the incidence of coronary artery disease and lung cancer is high throughout the year, and comorbidities are becoming more common, especially in elderly patients. The incidence of lung cancer and coronary heart disease may be related. This article summarizes the common risk factors (smoking and environmental pollution, fibrinogen, estrogen, and age), and treatment (surgical treatment, neoadjuvant therapy, and targeted therapy) progress of the two diseases, providing a theoretical basis for clinical prevention and treatment.