0.05) respectively in groupC1-C4,while in groups T1-T5,the rates were 21%,28%,36%,40%,37.5% ( P 0.05).However,the differentiation rate of MSCs increased probably with the incubation time prolonged.During the procedure of MSCs differentiation,the expression pattern of some genes such as GATA4 and cTNT may make some difference.

Objective:To observe the tansdifferentiation of bone marrow mesenchymal stem cells(MSCs) into cardiomyocyte in a coculture system with cardiomyocytes.Methods:Newly born Wistar Rats' cardiomyocytes were isolated and cultured and the cells were labeled with DAPI.Prepared MSCs were planted into the cardiomyocytes dishes to establish a co-culture system.Supernatant liquid of the cardiomyocytes or fibroblast was added the MSCs(non-labeled) dishes as the control. After one week of coculture,fluorescence microscope observation and immunocytochemical staining with cTnT was performed to evaluate the phenotype changes in non-labeled MSCs.Results:MSCs and cardiomyocytes were connect with each other by net in culture plate after a few days of coculture;some non-labeled cells were positively stained by cTnT antibody while the MSCs cultured with cardiomyocytes or fibroblast supernatant liquid were stained negatively.Conclusion:To obtain cardiomyocyte phenotype in vitro,direct contact of MSCs with cardiomyocytes is essential.The signaling molecules induce MSCs' transformation into cardiomyocytes are likely some membrane proteins on the surface of cardiomyocytes.

Ocular neovascularization,abnormal formation of new vessels from pre-existing capillaries,is a special pathologic change of many ocular diseases,including diabetic retinopathy and retinopathy of prematurity,etc..Ocular neovascular diseases comprise the common causes of blindness in millions of people.Recent studies showed that kringle domains from many endogenous proteins inhibit the proliferation and migration of endothelial cells in vitro and neovascular formation in vivo.Hence kringle domain with conservative structure has been considered as a potential inhibitor of neovascularization.The update studies on antiangiogenic kringle domains and its inhibition on ocular neovascularization were summarized.

This paper argues that the core idea of evidence-based medicine is to “follow the evidence”,whose essence is a model of clinical medical service,and a guiding ideology for the clinical practice.It has become a proper noun nowadays.The wording “evidence-based medicine teaching method”needs discussion, because “evidence-based medicine” and “teaching method” are lack of logical internal relations in concept.The teaching method of scientific sense has its own characteristics and requirements.To name a teaching method,it is necessary to define its peculiar connotation in teaching method,with specific procedures and steps to form a complete set of scientific method of teaching.

ObjectiveTo test the susceptibilities of Candida albicans to 5 antifungal agents including methylrosanilinium chloride and to estimate their relationship with the genotypes of C.albicans.MethodsThe susceptibilitiesof 67C.albicansisolatestomethylrosaniliniumchloride, fluconazole, amphotericinB,itraconazole and ketoconazole were determined by using a broth dilution method as described in the National Committee for Clinical Laboratory Standards(NCCLS) document M27-A2.DNA was extracted from the C.albicans isolates and subjected to the amplification of 25S ribosomal DNA(rDNA) introns by PCR.The C.albicans isolates were divided into A,B and C genotypes according to the amplicons.The relationship was analyzed statistically by exact probability test between the genotypes and antifungal susceptibility of C.albicans.ResultsOf the 67 C.albicans isolates,8.96％ were resistant to fluconazole,2.98％ to itraconazole,1.49％ to ketoconazole,and no resistance to amphotericin B was observed.The minimum inhibitory concentration (MIC) of methylrosanilinium chloride varied from 0.125 to 4 mg/L.Statistical analysis revealed no significant correlation between the genotypes and autifungal susceptibilities of C.albicans(all P ＞ 0.05).Conclusions More attention should be given to the increasing resistance to fluconazole and itraconazole in some isolates of C.albicans as well as to the favorable antifungal activity of methylrosanilinium chloride.The antifungal resistance shows no apparent correlation to the genotypes of C.albicans.

BACKGROUND:Implanted site and direction of miniscrew implants, post-implantation movement of implants, contact between tooth and implant after tooth movement al can cause root damage.
OBJECTIVE:To investigate the changes of the root surface, periodontal tissue and osteoprotegerin expression in the periodontal tissue at periods of time after miniscrew implant loading.
METHODTwo beagle dogs were selected. For each dog, miniscrews were implanted into the labial alveolar bone near the roots of the maxil ary second, third, and fourth premolars and the maxil ary second, third, and fourth premolars and first molar. Fourteen miniscrew implants were in one dog, including six in the maxil ary and eight in the mandible;two miniscrews of the maxil ary acted as control group with no loading, and the rest miniscrews were as experimental group under orthodontics force. Two weeks after miniscrew implantation, 150 g force was loaded between two miniscrews with nickel-titanium closed-coil springs. The dogs were sacrificed after miniscrews loading 4 and 8 weeks respectively, the teeth were cut completely with alveolar bone tissue and decalcified to make the histological specimen. The histological changes of the root and periodontal tissue were observed by hematoxylin-eosin staining and expression of osteoprotegrin in the periodontal ligament was also inspected by using immunohistochemistry.
RESULTS AND CONCLUSION:When the miniscrews were adjacent to the roots, absorption lacuna occurred in the alveolar bone;after horizontal force was loaded opposite to the roots, alveolar resorption became much more animate. When the miniscrews contacted with the pericementum, some areas of the cement absorbed seriously and even reached the dentin;when sustaining pressure from the miniscrews, the whole layer of the cement absorbed and the dentin was exposed and began to be absorbed. The osteoprotegrin in the periodontal ligament of control group expressed sharply and its expression was notable at 8 weeks;while the osteoprotegrin of experimental group weakly expressed at 8 weeks, and there was significant divergence between control group and experimental group at 8 weeks (P<0.01). The roots and periodontal tissue were al damaged variously after miniscrew implantation adjacent to the roots. When horizontal orthodontic force was loaded opposite to the roots, the expression of osteoprotegrin was influenced slightly;as the loading period prolonged, pressure dramatical y restrained the expression of osteoprotegrin. When discovering the miniscrew implant adjacent to roors, the miniscrew implant anchorage should not be used to load orthodontical force opposite to the roots but ought to be removed in order to leave the root repair autonomously and avoid further damage.

ObjectiveTo investigate the efficacy and side effects of telbivudine and lamivudine for treatment of chronic hepatitis B for 1 year. MethodsIn this random and control study,the efficacy of telbivudine and lamivudine treatments were compared in 120 patients who consisted of 60 cases with HBeAg-negative and 60 cases with HBeAg-positive chronic hepatitis B.The patients were randomly assigned to a daily 600mug telhivudine treatment group or daily 100mg lamivudine group for 52 weeks.The clinical efficacy and adverse reactions of the two groups were observed in control after 52 weeks of therapy,and dynamics of serum HBsAg,HBeAg levels were monitored and compared. ResultsAt week 52,mean reductions of serum HBV DNA from baseline and undetectable serum HBV DNA rates among patients with HBeAg-negative and HBeAg-positive chronic hepatitis B were significantly greater in the telbivudine-treated patients than in the lamivudine-treated group(all P＜0.05).Viral resistance and viral breakthrough was significant common in lamivudine compared with telbivudine(all P＜0.05).Among patients with HBeAg-positive chronic hepatitis B,telbivudine was significant superior to lamivudine with respect to loss of HBeAg(P＜0.05).There was no significant difference in side effects between patients treated with tebivudine and Iamivudine. ConclusionTelbivudine was more effective than lamivudine in treatment of patients with chronic hepatitis B,and the drug was well tolerated.

Objective To explore the relationship between antifungal resistance and expression of a virulence factor phospholipase B1 in Candida albicans. Methods Total RNA was extracted from 15 fluconazole-resistant and 15 fluconazole-sensitive Candida albicans isolates. RT-PCR was performed to detect the expression of phospholipase B1 mRNA. Extracellular and intracellular proteins secreted by Candida albicans cells were concentrated using salting-out method and extracted by P0013B RIPA lysis buffer, respectively followed by Western-blotting analysis to detect the expression of extracellular and intracellular phospholipase B1 protein. Results The relative expression of phospholipase B1 mRNA was 0.6173 ± 0.1090 in fluconazoleresistant Candida albicans isolates, significantly higher than in fluconazole-sensitive isolates (0.2653 ± 0.0935,P ＜ 0.05). Increased expressions of extracellular and intracellular phospholipase B1 protein were noted in fluconazole-resistant Candida albicans isolates compared with fluconazole-sensitive isolates (0.4145 ± 0.2773 vs 0.2720 ± 0.2194, 0.1825 ± 0.1831 vs 0.2653 ± 0.0935, t = 2.703, 3.443, both P ＜ 0.05). Conclusions The expressions of phospholipase B1 mRNA and protein are elevated in fluconazole-resistant Candida albicans isolates, suggesting that the expression of phospholipase B1 may be associated with the occurrence of antifungal resistance in Candida albicans.

Objective To investigate the effect of hypertensive perfusion which induced by norepinephrine on the TNF-α and IL-6 in serum and the activity of Na~+-K~+-ATPase in lung tissue and to observe the morphology of lung tissue. Method Ten domestic pigs were suffered from ventricular fibrillation for 4 minutes and applied cardiopulmonary resuscitation. The pigs they were divided into two groups. Hypertensive perfusion group( n = 5) :the mean arterial pressure, which induced by norepinephrine was maintained as 130% as baseline for 4 hours; nor-real perfusion group( n = 5) : the mean arterial pressure was maintained as baseline for 4 hours, too. Hemodynamic parameters was observed, blood samples were extracted to detect the TNF-α and IL-6 in serum at the time of baseline, 10 min, 2 h and 4 h after successful resuscitation. The expression of Na~+ -K~+ -ATPase in lung tissue and the change of ultrastmeture and pathology in lung tissue was detected at 24 h. All values were analyzed by student' s t test. Results There was no influence on hemodynamic parameters except the blood pressure and heart rate in hypertensive perfusion group. The release of TNF-α and IL-6 were reduced(P < 0.01 ) and the activity of Na~+-K~+ -ATPase was upgraded in hypertensive perfusion group. Hypertensive perfusion could play a protective role on the morphology of alveolar cell. Conclusions Hypertensive perfusion which induced by norepinephrine could reduce the release of inflammatory cytokines, upgrade the activity of the Na~+-K~+-ATPase, decrease the consumption of surfactant and have an important protective effect on lung tissue.

Objective To summarize the treatment experience of acute cerebral infarction (ACI)combined with deep venous thrombosis (DVT) of lower extremities in the elderly. Methods Thirty cases of elder patients with ACI combined with DVT (ACI group) were enrolled in this study, and 31 cases of elder patients with DVT were served as control group. All the patients were immobilized for 3 days and treated with low molecular heparin calcium (100 IU/kg,Q12 hX14 d). On the tenth day,warfarin was used and INR was controlled in 2.0～2.5. Low molecular dextran was infused intravenously per day for 14 days (500mg/d). Results Edema in the lower limbs of patients were apparently reduced. 4 eases had cerebral hemorrhage in ACI group, and none in the control group.Hemorrhage occured from 3 to 7 days during the treatment with low molecular heparin calcium. One of the patient died of massive intraeranial hemorrhage with lung infection. Conclnsions It should be more careful to use the low molecular heparin to treat the elder patients older than 80 years old with ACI combined with DVT.