Objective To establish a new approach for quantitative detection of VAD1 mRNA in cryptococcus neoformans by RT-FQ-PCR, and evaluate the treatment efficacy of CNM. MethodsThe primers and TaqMan probe were designed according to the published sequence of VAD1 mRNA (GenBank),and RT-FQ-PCR method to detect VAD1 mRNA was established. Cerebrospinal fluid from 25 CNM patients and 30 controls were detected and sensitivity and specificity of the method were evaluated. VAD1 mRNA concentration in cerebrospinal fluid from both acute phase and recovery phase of 25 CNM patients were also detected and significance of CNM treatment efficacy with VAD1 mRNA analysis was evaluated. Results Correlation coefficient of standard curve was - 0. 997 9 in detection of VAD1 mRNA by RT-FQ-PCR, and the detection limit was 101 copies/μl. The intra CV of plasmid standard for high, medium and low concentrations were 0. 65% ,0. 89% and 1.23% respectively, the sensitivity of cryptococcus neoformans detection by RT-FQPCR was 96% (24/25) ,while specificity was 100% (30/30). VAD1 mRNA concentration in acute phase were significant higher than that in recovery phase (3. 042 ±0. 906 vs 2. 187 ±0. 665 ,t =4. 583 ,P ＜0. 01).Conclusions The established RT-FQ-PCR method for the detection of VAD1 mRNA is provided with sound sensitivity, specificity and reproducibility, which might be fit for the detection of VAD1 mRNA. The expression level of VAD1 mRNA is relevant with the treatment efficacy of CNM.

Objective To study the role of homeobox A2(HOXA2) gene in cell growth,cellcycle and apoptosis of hepatoma cells, and discuss the feasibility that HOXA2 gene would be a potential therapy target of hepatoma. Methods Real-time quantitative polymerase chain reaction and revers transcriptional polymerase chain reaction were performed to examine HOXA2 expression in tumorous and non-tumorous tissue of patients was matched with liver cancer. siRNAs were chemically synthesized to interference HOXA2 in PLC/PRF/5 and MHCC-97L cells. Growth curve and soft-agar colony formation assay were performed to evaluate cell growth, and cell cycle and apop-tosis were analyzed by flow cytometry. Results HOXA2 was upregulated in HCC samples compared with matched non-tumor tissues(P<0.05). Two siRNAs against HOXA2 gene were designed and synthesized to specially knock-down HOXA2 in PLC/PRF/5 and MHCC-97L cells. HOXA2 knockdown inhibited cellular growth and soft-agar colony formation in PLC/PRF/5 and MHCC-97L cells. And Fowcytometry analysis revealed that HOXA2 knock-down by RNA interference could result in G1 arrest and S decrease and promoted cellular apoptosis. ConclusionHOXA2 gene has an important role in cell growth of hepatoma cells.

Objective:To reveal the behavior characteristics of health delivery system(HDS) and the dynamical mechanism by which HDS promoting population health,so as to provide theoretical and methodological evidences for the decision-making of national macro health system.Methods: The macro model of HDS was constructed by using the theory of complex system and the method of system dynamics.Results: The principle of "high coverage and reasonable quality" was put forward as developmental strategy of national health service.The improvement of population health was taken as the system objective of HDS;meanwhile,HDS was community-centered and city-country integrative.The macro model of HDS was constructed based on the above objectives.Conclusion: Modeling based on system dynamics is effective to reveal the complexity of health delivery system.The HDS complex system model constructed in this article can be used to analyze and interpret the behavior characteristics of HDS and the dynamic mechanism by which HDS promoting population health.

Objective To investigate the effect of immunocompetent cells and immune regulator on the apoptosis of human mesenchymal stem cells ( MSCs) and on mRNA expression of heme oxygenase-1. Methods MSCs were cultured by density gradient centrifugation and then identified by flow cytometry. RT-PCR was used to detect HO-1 mRNA expression and flow cytometry was used to analyze cell apoptosis after the stimulation of IFN-? and PHA-activated T cells. Results The mRNA expression of Heme oxygenase-1 was observed in MSCs and decreased after the stimulation of IFN-? and activated T cells. IFN-?,znpp-Ⅸ and combined these two caused obvious cell apoptosis in MSCs,with an apoptotic rate of ( 56. 50 ? 0. 16) % ,( 56. 85 ? 2. 27) % ,and ( 82. 53 ? 2. 65) % respectively. All of them had a significant difference compared with the normal MSCs [( 7. 56 ? 1. 43) % ,P

OBJECTIVE:To investigate the effect of aqueous extraction of Lonicerae Flos (SYHW) on anti-influenza A virus H1N1 (H1N1 virus) in vitro. METHODS:Using Madin-Darby canine kid ney (MDCK) cells cultured in vitro by H1N1 virus, half of the tissue culture infection dose(TCID50)was calculated. Culturing MDCK cells for 24 h with different mass concentrations of SYHW,the maximum non-toxic concentration was investigated. And then test was divided into normal cell group,virus control group,SYHW preventive administration group,therapeutic administration group and direct killing group (given SYHW of maxi-mum non-toxic concentration,infecting cells by 100 TCID50 H1N1 virus),and antiviral effective rate (ER) of SYHW was deter-mined. Test was divided into normal cell group,virus control group,SYHW therapeutic group and direct killing group (the same administration and infection as above),changes of cell proliferation index (PI) and cell apoptosis rate were respectively deter-mined. RESULTS:100 TCID50 of H1N1 virus was 1.26×10-7,and the maximum non-toxic concentration of SYHW on MDCK cells was 50 μg/mL(cell survival rate was 91.3%). ERs of preventive administration group,therapeutic administration group and direct killing group were 0,80.3% and 52.7%,respectively. Compared with normal cell group,PI value in virus control group was sig-nificantly reduced (P<0.05),early and late apoptotic rates were significantly increased (P<0.05). Compared with virus control group,PI value in directly killing group was significantly increased(P<0.05),and early apoptotic rate was significantly reduced (P<0.05);early apoptotic rates in therapeutic administration group were significantly reduced (P<0.05). CONCLUSIONS:SY-HW shows anti-H1N1 virus effect in vitro,therapeutic administration and directly killing are preferred in antiviral effect.

Objective Queuing theory is the mathematical study of how waiting lines or queues are formed and dissipated o -ver time, which aims to work out the optimal design and optimal control of queuing systems based on the research of probability and regularity of various queuing systems .The aim of paper was to find out the maximum acceptable waiting time for outpatients in stomatol -ogy department and explore the rational allocation of dentists based on queuing theory model . Methods Questionnaires , worktime measurement and queuing theory model were applied to calculate indicators of queuing system in outpatient services of Stomatology De -partment , getting the maximum waiting time accpetable for patients and the reasonable number of dentists . Results The maximum acceptable waiting time for outpatients was 34.02 ±7.07 minutes, and it was reasonable to allocate 25 doctors in the morning and 16 doctors in the afternoon . Conclusion Applying queuing theory helps to optimize dentist number in outpatient services of Stomatology Department and provide scientific reference to improve medical efficiency .

Objective To establish the quantitative detection of capsule associated protein 10 (CAP10)and virulence-associated DEAD-box RNA helicase 1(VAD1)genes in Cryptococcus neoformans (CN) and compare the diagnostic values of single gene test and combined gene test in CN meningitis.MethodsTwenty-three CN meningitis patients with fungal culture or ink staining or CN antigen detection positive in cerebrospinal fluid (CSF) were recruited and patients with craniocerebral trauma were recruited as controls.Standard plasmids were constructed using standard CN strain.Real time fluorescent quantitative polymerase chain reaction (RT-FQ-PCR) was established to detect the mRNA expressions of CAP10 and VAD1 genes in the CSF of patients with CN meningitis,which were compared with the results of CSF ink staining,fungal culture and antigen detection.The diagnostic values of single gene test and combined gene test were compared by chi square test.Results Among 23 CN meningitis patients,22 (95.6％) were CAP10 mRNA positive detected by RT-FQ-PCR,which was significantly higher than both ink staining (16/23,69.6％,x2 =4.167,P＜0.05) and fungal culture (15/23,65.2％,x2=5.143,P＜0.05),respectively; but not significant different from antigen detection (21/23,91.3％,x2=0.500,P＞0.05).There were also no statistical significant differences between combined detection of CAP 10 + VAD1 and CAP 10 or VAD1 single gene test (P＞0.05).ConclusionRT-FQ-PCR detection is successfully established using virulence genes as target,which is superior to the conventional methods.

Objective To explore the treatment effect of potato homogenate on the treatment of drug extravasation during intravenous injection.Methods 320 cases of drug extravasation patients during the intravenous infusion were randomly divided into the potato homogenategroup,named group A; patato slice group named group B and magnesium sulfate group as group C.The group A,B and C were separately treated with the external application of potato homogenate,thin slice of fresh potato and 33％ magnesium sulfate.The therapeutic effects of the 3 groups were compared.Results The therapeutic effect of group A was superior to that of group B and C,and the healing time in group A was much shorter than that in group B and C,and group B was batter than group C,there was significant difference between the above comparison groups.Conclusions The therapeutic effect of external application of fresh potato in the treatment of extravasation injury caused by the drugs is remarkable.Potato homogenate can improve the treatment effect and shorten healing time when compared with potato slice.

Objective:To evaluate the vestibular function of motion sickness. Method:VNG, which tests the vestibular function of horizontal semicircular canal,and CPT, which tests vestibulospinal reflex and judge proprio-ceptive,visual and vestibular status, were performed in 30 motion sickness pations and 20 healthy volunteers(control group). Graybiel score was recorded at the same time. Result:Two groups' Graybiel score (12. 67±11. 78 vs 2. 10±6. 23;rank test P<0. 05),caloric test labyrinth value [(19. 02±8. 59)°/s vs (13. 58±5. 25)°/s ;t test P< 0. 05],caloric test labyrinth value of three patients in motion sickness group exceeded 75°/s. In computerized pos-tourgrphy testing(CPT) , motion sickness patients were central type(66. 7%) and disperse type(23.3%);all of contral group were central type. There was statistical significance in two groups' CTP area, and motion sickness group was obviously higher than control group. While stimulating vestibulum in CPT, there was abnormality (35%-50%)in motion sickness group and none in contral group. Generally evaluating CPT,there was only 2 pro-prioceptive hypofunction,3 visual hypofunction, and no vestibular hypofunction, but none hypofunction in contral group. Conclusions Motion sickness patients have high vestibular susceptible, some with vestibular hyperfunction. In posturography,a large number of motion sickness patients are central type but no vestibular hypofunction, but it is hard to keep balance when stimulating vestibulum.

Objective To investigate the clinical efficacy of nephrotic syndrome in children adjunctively treated by underatured lactalbumin.Methods Thirty-six cases of nephrotic syndrome were studied for about 1 year.The changes in red blood cells of reduced glutathione(GSH) were analyzed in 18 cases of underatured lactalbumin treatment group and 18 cases of control group.The clinical presentations were observed in both groups.Results GSH levels increased in underatured lactalbumin treatment group when compared with control group.The frequency of relapse and upper respiratory tract infection in underatured lactalbumin treatment group were significantly reduced compared with control group.Conclusion Underatured lactalbumin can provide support for the potential use of an antioxidant therapy in these patients.