Objective:To analyze the infection results of The immunofluorescence detection of respiratory syncytial virus (RSV) in huazhou region from 2009 to 2010.Methods: The N gene sequence of RSV was taken as a reference to design specific primers and TaqMan fluorescent probe and to establish real-time The immunofluorescence method to detect sensitivity and specificity.21548 swab samples of children with acute respiratory infections in huazhou region from 2009 to 2010 were tested with real-time fluorescence. The immunofluorescence method.Results: The specificity and sensitivity of real-time fluorescence. The immunofluorescence method for detection of RSV infection were higher, without cross-reactivity to other respiratory viruses.10,477 cases of clinical specimens in 2009 were detected, with positive RSV infection rate of 9.73%, by real-time The immunofluorescence method detection. Positive detection rate of RSV infection had two peaks, which lied in February and December; the positive rate was 11% and 11% respectively. RSV infection rate was the highest for children under 6 years old.Conclusion: Real-time fluorescence features convenient detection, high specificity, and high sensitivity, which can be used for clinical detection of RSV infection. From 2009 to 2010, RSV infection remains a major pathogen of acute respiratory infections in children in huazhou region.

Objective:To study the effects of enamel matrix proteins (EMPs) on the biological characters of osteoblasts.Methods:MC3T3-E1 osteoblasts were cultured in vitro and exposed to EMPs at 300,400 and 500 ?g/ml.Enzyme kinetic method was used to assay alkaline phosphatase (ALP) synthesis and 3H-proline labelling combining collagenase digestion method was used to assay type I collagen synthesis.Results:The results manifested that ALP level in EMPs treated groups was higher than that in control group (P

Traditional teaching mode of biochemistry experiment couldn’t satisfy the demand of cultivating creative medical talents in new era. Offering designing experiment is the developing direction of experiment teaching reforms. But in the practice,there are various of obstacles which prevent the reforming process.

Objective To observe the effect of Guanxin Ⅱ Hao on plasma NO and NOS in dog with myocardial ischemia. Methods 20 dogs of myocardial ischemia were divided into the control group, isosorbidi mononitratis group and the Guanxin Ⅱ Hao group by random. The NO and NOS of serum were measured, before and after treatment. Results In the control group, the level of NO and NOS were higher than that of the isosorbide mononitrate group. There was significant difference between the control group and the Guanxin II Hao groups (P

Objective To evaluate the long-term molluscicidal effect of suspension concentrate of niclosamide (SCN) spraying in marshland regions of the Yangtze River. Methods A marshland with Oncomelania snails in Yangzhong City, Jiangsu Province, was selected as the study spot, and a dose of 6-8 g/m2 of SCN was sprayed yearly for four successive years, and the status of snails was investigated. Results From 2004 to 2007, the average densities of living snails were 6.00, 4.25, 2.04, 1.95 snails/0.1 m2, respectively before the use of molluscicide, and 0.86, 0.86, 0.23, 0.16 snails/0.1 m2, respectively after that. The average densities of living snails before and after the use of molluscicide reduced year by year, and the reduction rates of average densities of living snails were 85.67%, 79.76%, 88.49% and 91.78%, respectively from 2004 to 2007. The rate of frames with snails after the use of molluscicide was 73.46% in 2007, with a reduction rate of 86.85% compared with 9.66% of that before the use of molluscicide in 2004, and the average density of living snails reduced from 6.00 snails/0.1 m2 to 0.16 snails/0.1 m2, with a reduction rate of 97.33%. Conclusions After the spraying for four successive years, the density of snails reduces significantly and the molluscicidal effect is stable and reliable. SCN is a formulation of molluscicide for long-term application in marshland regions.

OBJECTIVETo summarize and analyze rules for acupoint selection and prescription composition in clinical literature regarding acupuncture for polycystic ovary syndrome (PCOS).

METHODSCHKD, VIP and Wan-fang databases were retrieved. The clinical literature data included in the study was collected. The descriptive statistical analysis was conducted on the main acupoints, main meridians of main acupoints, distribution of main acupoints, application of special points and rules for acupoint composition.

RESULTSFifty-two articles were included, involving 55 main acupoints with a total frequency of 375. The most frequent acupoints of acupuncture for PCOS were Sanyinjiao (SP 6), Guanyuan (CV 4), Zigong (EX-CA 1), Zhongji (CV 3) and Qihai (CV 6). The meridians of main acupoitns were conception vessel, spleen meridian of foot-taiyin and stomach meridian of foot-yangming. The main acupoints were distributed in the lower abdomen, lower limbs and back. In the special points, the use of front-mu points, five-shu points and back-shu points was more frequent. The prescription was usually consisted of 5 to 7 acupoints, (6. 9 ± 3. 6) acupoints in average.

CONCLUSIONIn the modern treatment of acupuncture for PCOS, Sanyinjiao (SP 6), Guanyuan (CV 4), Zigong (EX-CA 1), Zhongii (CV 3) and Qihai (CV 6) are most used. With the theories of meridian and zang-fu as essential references, the acupoint selection is based on disease differentiation and meridian circulation. Additionally, the methods for selecting the adjacent points and the distant points are adopted.

Objective:To investigate the migration and odontogenesis of rat dental pulp cells transfected with adenovirus vector enco-ding cadherin-11.Methods:The dental pulp cells were cultured and transfected with pDC316-mCMV-EGFP-Cadherin-11.Cad-11 protein expression of the cells was examined by immunohistochemistry staining.The odontogenic differentiation of the cells was studied by alizarin red staining and ALP staining.The adhesion and migration of the cells were tested.Results:Transfection of pDC316-mC-MV-EGFP-Cadherin11 promoted Cad-11 protein expression and ALP activity,increased calcified nodule formation(P <0.05),adhe-sion and migration of rat dental pulp cells(P <0.05).Conclusion:Cadherin-11 may promote the odontoblast differentiation and mi-gration of rat dental pulp cells.

Objective To investigate the effect of different intestinal cleaning methods on bowel preparation for laparoscopic peritoneal vaginoplasty.Methods One hundred patients undergoing laparoscopic peritoneal vaginoplasty were divided into experiment group and control group with 50 cases in each group .For bowel preparation,the experiment group was managed with oral sodium phosphate salt solution of 45 mL and the control group with oral senna leaf of 30 g in boiled water.The two groups were compared in terms of intestinal cleaning degree,adverse reactions and postoperative exhaust time.Results The incidence of adverse reactions in the former group was significantly lower than the control group.Degree of bowel cleaning satisfaction was significantly better than that of the control group and the postoperative exhaust was significantly earlier than that in the control group(P<0.05).Conclusion The method of oral sodium phosphate salt solution for bowel preparation for laparoscopic peritoneal vaginoplasty can improves the intestinal cleanliness,reduce incidence of intestinal adverse reaction and promote postoperative exhaust so that it can improve postoperative intestinal restoration of kinetic energy.

BACKGROUND: Whether human embryonic stem cells (hESCs) cultured on different feeder layers can maintain identical or similar characteristics remains unclear.OBJECTIVE: To compare the characteristics of hESCs cultured on human- and mouse- origin feeder layers.DESIGN, TIME AND SETTING: The in vitro cytology observation was performed at the Kunming Institute of Zoology, Chinese Academy of Sciences between September 2007 and February 2009.MATERIALS: Two ICR pregnant mice with 12.5-13.5 embryonic days were provided by Animal Center of Kunming Medical College. Immortalized human adult fibroblast (HAFi) cell line was presented by School of Medicine, Johns Hopkins University (USA). hESCs line was provided by Kunming Institute of Zoology, Chinese Academy of Sciences.METHODS: Mouse embryonic fibroblasts (MEF) were harvested from ICR mice by trypsinization method. HAFi were conventionally cultured. After γ ray treatment, two kinds of cells were incubated on 6-well gelatin-coated plate with density of 2.5×10~4/cm~3. hESCs were cultured on HAFi or MEF feeder cells containing β-mercaptoethanol DMEM/F12 and basic fibroblast growth factor.MAIN OUTCOME MEASURES: Morphology, expressions of specific molecular markers, Oct-4 positive rate, as well as cell doubling time of hESCs cultured by two methods were compared.RESULTS: ①BG02 cells on MEF and HAFi shared the similar morphology and characteristic pluripotency markers, which expressed SSEA-3, SSEA-4, TRA-1-60, TRA-1-81 and Oct-4, but not SSEA-1. However, the proportion of positive Oct-4 cells in hESCs colonies maintained on MEF was lower than that of HAFi (P < 0.05) with shorter doubling time (P < 0.05).CONCLUSION: hESCs cultured on MEF and HAFi represent some differences in the growth and pluripotency characteristics.

BACKGROUND:FGF2,TGFβ/activin/nodal and IGF signal pathways are necessary for keeping functions of human embryonic stem calls,but there was no report concerning whether addition of exogenous basic fibroblast growth factor,transforming growth factor β and insulin can maintain self-renewal of human embryonic stern cells.OBJECTIVE:To establish a feeder layer-and serum-free culture system of human embryonic stem cells.DESIGN,TIME AND SETTING:The cytological in vitro study was performed at the Kunming Animal Institute of Chinese Academy of Sciences from September 2007 to February 2009.MATERIALS:Pregnancy 12.5 or 13.5-day ICR strain mica (clean grade) were provided by the Animal Center of Kunming Medical College.Human embryonic stem cell line BG02 was purchased from Bresagen,USA.METHODS:BG02 cells were digested,centrifuged,resuspended in feeder layer-free medium,and then incubated for 5-7 days.Differentiated human embryonic stem cells were removed.Dispase was added for digestion.Samples were cut into blocks,centrifuged,resuspended,and then incubated at 1:3 in a 4-well plate coated with laminin in feeder layer-free and serum-free medium,supplemented with 80% DMEM/F12,20% KSR,2 mmol/L glutamine,1% non-essential amino acid,0.1 mmol/L β-mercaptoethanol,insulin-transferrin-selenium (ITS) (×1),10~6 U/L penicillin,100 mg/L streptomycin,4 μg/L basic fibroblast growth factor (bFGF) and 0.12 μg/L transforming growth factor-β1 (TGF-β1).ICR fetus mica were sterilely obtained to culture mouse embryonic flbroblasts by tissue pancreatin digestion method.These cells were incubated in a 6-well plate coated with 0.1% gelatin.Human embryonic stem cell line BG02 cultured on mouse embryonic fibroblasts feeder layers was transferred to laminin-coated plates in serum-free medium containing bFGF,TGFβ1 and ITS.MAIN OUTCOME MEASURES:The morphology of BG02 cells in feeder layer-and serum-free condition was observed.The specific molecular markers of human embryonic stem cells were determined by immunohistochemical staining.The karyotype and differentiation ability in vitro of BG02 cells were analyzed.The differences in the proliferation and the differentiated rate of BG02 clumps in feeder layer-and serum-free condition or mouse embryonic fibroblast feeder layer condition were compared.RESULTS:BG02 calls had been continuously cultured for at least 20 passages in feeder layer-and serum-free culture condition.BG02 clones in this culture system had typical human embryonic stem cell morphology.BG02 cells all expressed SSEA-4,SSEA-3,TRA-1-60,TRA-1-81,Oct-4,but did not express SSEA-1.After 20 days of culture,BG02 cells had the capacity to differentiate into derivatives of embryonic endoderm,mesoderm,and ectoderm.Differentiated cells could express alpha-fetoprotein,nestin,α-actin.At passage 20,call karyotype was normal (46XY).The growth of BG02 cells cultured in feeder layer-and serum-free condition was more slowly compared with mouse embryonic fibroblast feeder,and doubling time was significantly prolonged (P ＜ 0.05).Differentiation rate of the colonies was significantly increased (P ＜ 0.05).CONCLUSION:A feeder layer-and serum-free condition for culture of BG02 cells has been established.The addition of bFGF,TGFβ1 and ITS can maintain the self-renewal of BG02 cells.