Objective To establish a HPLC method for determining the serum methotrexate(MTX)concentration in children patients for monitoring the blood drug concentration in clinic.Methods The chromatographic analytical column was the Shimaduz C-18 colum(250 mm×4.6 mm,5 μm),the mobile phase was 0.025 mol/L NaH2 PO4 (pH 5.41)-methanol(76∶24,V/V).The flow rate was 1 .0 mL/min.The detection wavelength was 313 nm.The column temperature was set at 35 ℃.The plasma samples were deposited down the protein with 10% HClO4 ,the supernatant liquid 130 μL added with 1 mol/L NaOH 10 μL was directly injected for determination.Results The mass concentration of MTX in the range of 0.00-4.84 μmol/L and 4.84-10.00 μmol/L showed the good linear relation with the peak area,r 1 =1 ,r2 =1 .The mean recovery rates were 61.67%and 71.83%.Intra-day and inter-day precision were <10% and 12%,respectively(n =3).Conclusion The used method is simple,accurate and reliable with high sensitivity,good reproducibility and wide linearity range,which is suitable for the monitoring blood MTX concentration.

Objective To investigate the behaviors character of men who have sex with men(MSM) in the local place,learn about the prevail situation of STI and HIV/AIDS among MSM group,and provide evidences for the health promotions on AIDS and STI among MSM.Methods Based on a snowball model,survey on MSM were conducted with an anonymous model.3～5 ml vein blood of the MSM were also collected at the same time.Results The awareness rate of HIV/AIDS information among 452 MSM was high.98.45% can give the right answer to question of whether would be infected with HIV if inject blood or blood product which have been infected with HIV.Total understand rate was 92.86%.The rate of condom use in their last anal sex with male was 64.43%.Rate of never using condom and condom using in every anal sex with male in the late 6 months was 10.95% and 34.83% respectively.The 3 services of HIV that most been welcomed by them were IEC(67.92%),condom distribution(62.61%) and lube distribution(57.30%).1.99% has a positive result on anti-HIV testing. 8.85% have a positive result on syphilis testing.Conclusions High dangerous behaviors are prevalent among MSM in Nanning and propaganda on HIV/AIDS information is not sufficient.Multi-types approaches should be adopted to prevent the epidemics of HIV/AIDS among MSM.

Objective The purpose of this study was to investigate the effects of ropivacaine and bupivacaine on energy supply and mitochondrial oxidation phosphorylation of isolated rabbit heart muscle in order to compare the cardiotoxicity of the two local anestheties and explain the mechanism Methods Twenty four New Zealand rabbits weighing (2135?345)g were randomly divided into three groups of 8 animals each: control group(C); ropivacaine group(R) and bupivacaine group (B) The animals were anesthetized with intraperitoneal urethane 2g?kg -1 and heparinized (heparin 4mg?kg -1 ) Heart was quickly removed and connected to Langendoff preparation and perfused with oxygenated Krebs Henseleit buffer solution at normothermia(37℃) via aorta After 15 min perfusion, ropivacaine (group R) or bupivacaine(group B) was added to perfusate(500ng?ml -1 ) The heart was then perfused for another 10 min Myocardial tissue was taken from ventricle for determination of ATP, ADP and AMP content with high performance liquid chromatography Myocardial mitochondria was prepared and its oxidation of pyruvate and palmitoylcarnitine was measured using Clark electrode Results Myocardial ATP and ADP decreased significantly in group R and B as compared with those in group C The decrease in ATP and ADP in group B was more than that in group R (P

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Abstract: Objective To investigate the preventive and therapeutic effects of anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin on neonatal mice infected with human rotavirus (HRV), and to analyze the underlying mechanism. Methods Hybridoma technology was used to prepare Ab2β of lactadherin. One hundred and twenty 7-day-old Kunming mice were randomly divided into groups A, B, C and control, each consisting of 30 mice. Groups A, B, and C were all infected with HRV via oral gavage. Group A received no treatment, group B was orally administered lactadherin for 7 days prior to infection, and group C was orally administered lactadherin for 7 days after infection, the control group was orally administered cell culture medium that did not contain the virus. The clinical manifestations (diarrhea, body weight) at different time points after infection of the neonatal mice in each group were observed, and the content of rotavirus antigen in the feces of neonatal mice was detected by enzyme-linked immunosorbent assay (ELISA). After HRV infection for 7 days, immunohistochemical staining was used to examine the expression level of intercellular adhesion molecule-1 (ICAM-1) in mouse small intestinal tissues in each group. Results No diarrhea occurred in the control group at any time point. Groups A, B, and C showed diarrhea symptoms after HRV challenge for 1 day. The degree of diarrhea in groups B and C was lower than that in group A at 2-4 days after HRV challenge, and the difference was statistically significant (P<0.05). The HRV antigen content in the feces of the neonatal mice in groups B and C was lower than that in group A at 1-5 days after HRV challenge, and the difference was statistically significant (P<0.05). There was no significant difference in the degree of diarrhea and HRV antigen content between groups B and C at each time point (P>0.05). There was no significant difference in the body weight of the neonatal mice in each group before infection and 1 day after infection (P>0.05); the weight of neonatal mice in groups B and C was higher than that in group A at 3, 5 and 7 days after HRV challenge, and the difference was statistically significant (P<0.05), and there was no significant difference in body weight between groups B and C at each time point after HRV challenge (P>0.05). The number of ICAM-1 expressing cells in the small intestine of the three groups A, B, and C was higher than that of the control group after HRV challenge for 7 days, and the difference was statistically significant (P<0.05). The cell number and gray value of ICAM-1 expressing cells in groups B and C were lower than those in group A, and the difference was statistically significant (P<0.05). Conclusions Anti-idiotypic monoclonal antibodies (Ab2β) of lactadherin has a good preventive and therapeutic effects on human rotavirus infection in neonatal mice, and can significantly improve diarrhea symptoms and reduce HRV viral load. Its specific mechanism may be related to the inhibition of ICAM-1.

Objective To detect the sealing ability of iRoot as root end retrograde filling material in vitro experiment,and to compare it with mineral trioxide aggregate(MTA) in order to provide reference basis for clinical use of iRoot.Methods The dental crown was removed in 67 single root canal anterior teeth.The routine root canal preparation and filling were performed.The root end was resected by 3 mm for conducting root end retrograde preparation.Sixty-three teeth in the samples were randomly divided into 4 experimental groups.Then the different materials were used for root end filling:iRoot BP (group A,n=17),iRoot BP plus (group B,n=17),MTA (group C,n =17),and group D(iRoot BP + gutta percha,n =12),the residual 4 samples served as the control group.The apical microleakage of 10 teeth in each experimental group was detected by the fluid dynamics.The microleakage situation at the time points of 1,7,14,28 d was observed.In addition,2 teeth selected from each group were performed the filling material ultramicrostructure observation by SEM.Each 5 teeth were remained in the group A,B and C and performed the Micro CT scanning.The porosity rate of filling material was detected.Results No obvious edge gap was found between the filling materials and dentin surface in the group A,B and D by SEM,but obvious gap could be seen in various samples of group C.Micro CT scanning found that the porosity rates in the group A,B and C were 0.020 ±0.023,0.045 ±0.035 and 0.031 ± 0.011 respectively,but showing no statistical difference(P＞0.05).The fluid dynamic experiment showed that the microleakage detection value in each group had no statistical difference(P＞0.05).Conclusion The sealing ability of iRoot and MTA used in root end retrograde sealing has no obvious difference,But iRoot operability is superior to MTA.

A bacterial strain having 3-deoxyglucosone-metabolizing enzyme was selected from 31 marine bacterial stains. The conditions for enzyme production of the strain was examined. The optimal tempreture, initial pH. and cultivate time for enzyme formation were 28℃, pH7.8~8.0, and 96 hours respectively. Composition of the suitable medium was as following (%): Fish peptone 1.0, Sucrose 0.3, Yeast extract 0.2, NaCI 5.0.3-deoxyglucosone can induce formation of enzyme.

10.3969/j.issn.2095-4344.2013.26.010

Toll-like receptor 4 (TLR4) plays an important role in inflammation and immune response.MicroRNAs (miRNAs) are involved in the regulation of TLR4 signaling pathway in multiple levels and various molecules,which play an important role in inflammatory reaction.A variety of miRNAs are involved in the regulation of TLR4 signaling pathway,and the TLR4 signaling pathway can induce a variety of miRNAs.Chronic diseases such as diabetes,Alzheimer's disease and cardiovascular disease are closely related to inflammatory response.The regulatory role of TLR4 signaling related miRNAs has attracted much attention in inflammatory diseases.In this review,the research progress of TLR4 signaling pathway related miRNAs in the regulation of inflammatory response is summarized,which provides a new research direction for the clinical diagnosis and treatment of inflammatory response related diseases.

Objective To discuss CT,X-ray and sonographic features of atypical kidney cyst.Methods CT,X-ray and sonographic findings in 9 cases with pathologically-proved atypical kidney cyst were retrospectively analyzed.Results CT and sonography could diagnose atipical kidney cyst with a high accuracy.Of 9 cases,bloody kidney cyst 3 cases,calcified kidney cyst 2 cases,separate kidney cyst 3 cases.Renal milk of calcium kidney cyst 1 case.CT and ultrasound can display the forms and features of atypical kidney cyst.Conclusion CT and sonography are of great value for the diagnosis of atypical kidney cyst.It is difficult in differentiating the bloody kidney cyst and renal carcinoma with muddy cystic content.Ultrasound guided puncturing biopsy sould be carried out.