BACKGROUND: Previous studies demonstrated that recombinant human bone morphogenetic pretein-7 (hBMP-7) can effectively promote the synthesis of extracellular matrix, repair of damaged disc matrix, restore of degenerative disc height. It is hoped that BMP-7 can be used to control and reverse the intervertebral disc degeneration. However, because of the short half-life of recombinant protein and low biological activity, it is difficult to maintain BMP-7 high concentrations on degenerative disc. Gene therapy can prevent these defects effectively.OBJECTIVE: To study the effect of hBMP-7 gene transfeotion on biological activity of primary cultured rabbit nucleus pulposus cells in vitro, to determine the feasibility of hBMP-7 gene which will on gene therapy research of intervertebral disc degeneration,and to provide basis for further study.DESIGN, TIME AND SETTING: A randomized and controlled observation was performed at the Institute of Cardiothoracic Surgery of Changhai Hospital from December 2005 to September 2006.MATERIALS: Six healthy New Zealand white rabbits of either gender, averaging 4 weeks old and weighing 500 g, were used in this study, and Ad-hBMP7 was constructed by the Cardiothoracic Surgery Institute of Changhai Hospital.METHODS: After rabbits sacrifice under aseptic condition, the nucleus pulposus was got. After digested with Pronase, type Ⅱcollagenase and type Ⅱ DNAase (4 hours, 37 "(2), the cells were seeded in the Petri dishes and were kept in the incubator. After 7 days and then twice a week, the media were refreshed. The nucleus pulposus cells were infected by adenovirus integrated with hBMP7 gene. The cells which were transfected by adenovirus vector for Lac-Z gene and which were not transfected were adopted as control.MAIN OUTCOME MEASURES: The expression of hBMP-7 was determined by RTopCR and Western blot. The effect of hBMP-7 on cell proliferation was surveyed by MTT. Furthermore, the effect of hBMP-7 gene on synthesis of proteoglycan and type Ⅱcollagen was detected by modified dimethylmethylene blue method and ELISA, respectively.RESULTS: Gene sequencing and PCR showed that hBMP-7 gene was inserted correctly and no mutation happened. The pdmary cultured rabbit nucleus pulposus cells were identical with those reported in literature. After Ad-hBMP7 transduct into the nucleus pulposus cells, high level of hBMP-7 expression was observed and lasted over 3 weeks. Also hBMP-7 gene can promote cell proliferation and synthesis of proteoglycan and type II collagen with significant difference compared with control groups (P < 0.05).CONCLUSION: hBMP-7 gene mediated by adenovirus can be the target gene for the further study on gene therapy of intervertebral disc degeneration.

Objective To study the role of KRX-725,a specific agonist of Sphingosin-1 phosphate receptor 3,S1PR3),in the function and mechanism of S1PR3 in respect of bacterial clearance.Methods Twenty healthy male C57BL/6 mice were randomly (random number) divided into two groups:KRX-725 group and control group.Septic mice model were established by intraperitoneal injection of E.coli (3 × 106),then KRX-725 (10 mg/kg) or the vehicle was administered intratracheally.Forty-eight-hour survival rate (n =12),bacterial colony numbers in peritoneal cavity and blood (n =5),and lung injury (n =3)were compared between two groups.In vitro,the peritoneal macrophages were stimulated by E.coli (cell∶E.coli=1∶10) with KRX-725 or the vehicle treatment.The reactive oxygen species (ROS) levels were detected by CM-H2DCFDA in macrophages.The bacteria clearance function of KRX-725 was observed by gentamicin protection test.Survival rates were analyzed with the Log-rank test.A 2-tailed student's t test was used to compare difference between two independent groups.Results Compared with the control group,the 48-hour survival rate of KRX-725 group was significantly higher (P ＜ 0.05).Bacterialload in the blood and the peritoneal lavage fluid (PLF) was greatly decreased in the KRX-725 group (blood:t =3.17,P ＜0.05;PLF:t =4.07,P ＜0.01).The lung tissues injury was also obviously reduced in the KRX-725 group of 24 hours after the injection of E.coli (lung injury score:KRX-725 group 1.4 ± 0.25;control group 2.4 ± 0.25) (t =2.89,P ＜ 0.05).In vitro,KRX-725 could up-regulate the ROS levels in macrophage at 20 min and 30 min after E.coli injected intra-peritoneally (20 min fluorescent intensity:KRX-725 group 522.9 ± 38.76,control group 385.9 ± 15.90,P ＜ 0.05;30 min fluorescent intensity:KRX-725 group 519.7 ±25.02,control group 384.5 ± 15.28,P ＜0.01).The bacterial load in the KRX-725 treated macrophage were significantly decreased at 3 h and 6 h after E.coli injected intra-peritoneally (3 h:KRX-725 group 286.5 ±98.35,control group 710.8 ± 107.8,P ＜0.05;6 h:KRX-725 group 72.5 ±6.45,control group 205.8 ±66.76,P ＜0.01).Conclusion In vivo,KRX-725 could improve the survival rate of septic mice,decrease the bacterial lioad in the blood and PLF,and reduce the lung injury.In vitro,KRX-725 could up-regulate the ROS level in macrophages and accelerate the bacterial clearance.

Animals ; Female ; Isocyanates ; toxicity ; Male ; Malondialdehyde ; metabolism ; Mice ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; metabolism ; Pregnancy ; Prenatal Exposure Delayed Effects ; Superoxide Dismutase ; metabolism ; Testis ; drug effects ; metabolism

Objective To explore the effect of dimethylformamide(DMF)on the histological structure and enzymes activity in the testis of mice.Methods The KM male mice were treated with DMF by gavage at the doses of 0,0.5,1 and 2 g/kg respectively, once a day,for 30 consecutive days.On day 31,the mice blood samples were collected through eyes and then the mice were killed. Two mice were randomly selected in each group and one testicle was sampled to do the pathological examination,the other one for enzyme activity determination,including succinic acid dehydrogenase(SDH),lactate dehydrogenase(LDH)and acid phosphatase (ACP).Results Compared with the control group,all treated groups showed a significant decrease in body weight(P

With the development of translational medicine,medical equipment has become a booster of clinical and basic research,and the visualization of the Chinese traditional pulse diagnosis apparatus,NX-8-type pulse information acquisition and analysis system achieved the digital pulse diagnosis and sublimated the method of diagnosis.By using the visual sensing technology and image acquisition and processing technology,NX-8 displays the B-type ultra-sound images.With the parameter set and added,not only the pulse beat can be seen clearly,as well as border,thickness and deformation,and hens to perform direct analysis of the blood vessel elasticity,but also the objective observing and study of the dual diagnostic pulse,Anti-off pulse and other special physiological structure can be realized.By measuring pulse wave velocity (PWV) and other indicators,NX-8-type pulse information acquisition and analysis system provides diagnosis and significant prevention of arteriosclerosis,hypertension,high cholesterol,angina and other cardiovascular diseases,and is expected to achieve a digital auxiliary diagnostic equipment of traditional Chinese medicine (TCM) Sizhen,and be applied to telemedicine and the field of space science and technology.It is like a milestone in the development of TCM diagnostic techniques.

· Currently, refresher doctors’training is an important form of continuing medical education in our country, effectively promoting and improving the clinical ability and the service level of practitioners in basic -level medical organization. We combine ophthalmic professional features with refresher doctors’training in practice to explore ophthalmic continuing clinical training specifications.Training on the current status and new changes has been analyzed in order to further improve the training quality. The competent administrative departments and clinical teaching hospital should make full use of its advantages to institutionalize, specialize and standardize the training. We suggest linking the continuing clinical training with job promotion;strengthen core clinical ability and being strict with appraisal system so that after the training the trainees can spread standardized guidelines and technology, alleviating the current medical demand and the difficulties in the reform.

OBJECTIVETo evaluate the method for collecting NO2 in workplace air using a solid adsorbent and determining the concentration of NO2 by ion-exchange chromatography.

METHODSNO2 in workplace air was collected using sampling tubes filled with 13X molecular sieve soaked with triethanolamine, and the samples were desorbed with a certain concentration of triethanolamine solution to obtain NO2(-). NO2(-) was separated with an anion exchange chromatography column and quantified by a conductivity detector. The concentration of NO2 was determined based on the conversion coefficient of NO2 and NO2(-).

RESULTSDifferent concentrations of NO2 (standard gas) was collected using the sampling tubes at different time points, and the sampling efficiency and desorption efficiency could reach more than 90%. The penetrating capacity of sampling tubes was more than 1.1 mg. The comparative test shows that there was no significant difference between the new method and national standard method (solution absorption) (P > 0.05). The samples could be stored at room temperature for more than 16 days. The calibration curve plotted in the new method was linear in the range of 0.1∼20.0 µg/ml, with a correlation coefficient of 0.9998; the detection limit was 0.01 µg/ml; the minimum detectable concentration was 0.05 mg/m(3) (V0 = 3.0 L).

CONCLUSIONThis determination method meets the requirements of Guide for establishing occupational heath standards-Part 4 Determination methods of air chemicals in workplace (GBZ/T 210.4-2008) and shows obvious advantages for determination of NO2 in workplace air.

Air ; analysis ; Air Pollutants, Occupational ; analysis ; Chromatography, Ion Exchange ; methods ; Nitrogen Dioxide ; analysis ; Workplace

OBJECTIVETo explore the surgical approaches, methods and techniques of repair of cerebrospinal fluid (CSF) rhinorrhea via frontal sinus under transnasal endoscopy.

METHODSCerebrospinal fluid rhinorrhea, located at the posterior wall of the frontal sinus (n = 9) and posterior lateral wall of the frontal recess (n = 4) had been repaired surgically. A transnasal endoscopic approach was chosen in 12 patients and combined approach was used in 1 patient during the first procedure. Three patients needed the second surgery. Among them, one patient needed to repair CSF rhinorrhea, 1 patient needed to treat intracranial abscess of frontal lobe via combined approach and another one was treated because of the complication of frontal cyst.

RESULTSTwelve patients were successfully repaired in the first surgery. Only 1 patient needed second surgery. Two patients occurred complications. One was intracranial infection after surgery, external drainage and packing in the frontal sinus was used. Another was obstructive cyst in frontal sinus, transnasal endoscopic frontal sinusotomy was performed.

CONCLUSIONSCSF rhinorrhea which located at the posterior wall of the frontal sinus can be successfully repaired via transnasal endoscopic approach if the leak was visible under endoscopy. The size of the frontal ostium and leak vantage should be considered to prevent the drainage of the frontal sinus which would result in obstructive cyst in frontal sinus, frontal sinusitis and intracranial infection. Combined approach was suggested to the patients that leakage could not be seen in frontal sinus or frontal ostium was difficult to enlarge.

Adolescent ; Adult ; Cerebrospinal Fluid Rhinorrhea ; surgery ; Child ; Endoscopy ; methods ; Female ; Frontal Sinus ; surgery ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo assess factors predisposing to proteinuria in patients with type 2 diabetes mellitus (DM).

METHODSA case-control study was conducted on two groups of patients with type 2 diabetes without proteinuria (urine protein < 300 mg/24 h, n = 106) and with proteinuria (urine protein >or= 500 mg/24 h, n = 106). The two groups were matched by age (+/- 3 years), sex, race and place of residence. Information on these subjects including demography, history of disease, family history of diseases, lifestyle and behavioral variables, were obtained through questionnaire. Variables including blood pressure, fasting blood glucose (FBG), quantity of protein in 24-h urine were measured. Cox's proportional hazards regression analysis was then performed.

RESULTSFactors that were independently associated with the development of proteinuria, would include occupation, diabetic duration, glycemic control, hypertension, duration of hypertension and daily intake of vegetable after diagnosis of DM. Their corresponding odds ratios (OR) with 95% confidence intervals (CI) were 2.429 (1.299 - 4.542), 1.078 (1.029 - 1.131), 2.316 (1.341 - 3.998), 1.810 (1.059 - 3.092), 1.043 (1.012 - 1.074) and 0.551 (0.334 - 0.907), respectively, while the presence of proteinuria was not associated with family history of DM, family history of hypertension, certain dietary habits and smoking habit. Multivariate logistic regression model was then fitted with three variables, including diabetic duration, glycemic control and hypertension.

CONCLUSIONThe prevalence of proteinuria in labor workers was higher than that in intellectuals. Longer history of diabetic duration, poor glycemic control, hypertension and longer duration of hypertension all independently contributed to the development of proteinuria. The risk of proteinuria decreased with increasing daily intake of vegetables after the patients being diagnosed as having DM.

Adult ; Aged ; Case-Control Studies ; China ; epidemiology ; Diabetes Mellitus, Type 2 ; complications ; Female ; Humans ; Male ; Middle Aged ; Prevalence ; Proportional Hazards Models ; Proteinuria ; epidemiology ; etiology ; Risk Factors

OBJECTIVETo investigate the impact of Staphylococcus aureus from infertile men on sperm motility and the relationship between virulence genes and the activity of spermatozoal immobilization.

METHODSWe collected 60 strains of non-repeated Staphylococcus aureus from the semen of 589 infertile males and analyzed the influence of Staphylococcus aureus on sperm motility using the computer-aided sperm analysis system. We selected the strains that apparently decreased sperm motility and detected their virulence genes by PCR.

RESULTSSperm motility was significantly decreased in 17 of the 60 strains of Staphylococcus aureus (P < 0.05). The main virulence genes in these strains were hlg (33.3%), scn (23.3%), cna (20%), hlb (20%), and clfA (18.3%), others including icaA, fnbA, tst, seb, hld, eta and sea. The scn gene carriers accounted for 47.1% in the spermatozal immobilization positive group, significantly higher than 14% in the negative group (P < 0.05). No statistically significant differences were found in the percentages of the carriers of the other virulence genes between the two groups (P > 0.05).

CONCLUSIONInfections of Staphylococcus aureus in male reproductive system can lead to the decrease of sperm motility, which may be associated with the Staphylococcus complement inhibitor encoding gene scn.

Humans ; Infertility, Male ; microbiology ; Male ; Polymerase Chain Reaction ; Semen ; microbiology ; Species Specificity ; Sperm Motility ; Staphylococcal Infections ; Staphylococcus aureus ; pathogenicity ; Virulence ; genetics