China ; Clinical Laboratory Techniques ; Laboratories ; statistics & numerical data ; Occupational Health ; Quality Control

OBJECTIVE:To establish a method for the simultaneous determination of acetic acid and succinic acid in Banxia syrup. METHODS:RP-HPLC was performed on the column of GL InterSustain-C18 with mobile phase of 0.03 mol/L ammonium di-hydrogen phosphate buffer solution(pH2.0)-methanol(gradient elution)at a flow rate of 0.8 ml/min,the detection wavelength was 210 nm,column temperature was 30 ℃,injection volume was 10 μl. RESULTS:The linear range was 0.020 98-0.209 8 μg/ml for acetic acid(0.999 9)and 12.04-120.4 μg/ml for succinic acid(r=0.999 9);limits of quantification were 0.15,18.24 ng,limits of detection were 0.045, 5.53 ng;RSDs of precision, stability and reproducibility tests were lower than 2%;recoveries were 97.45%-101.68%(RSD=1.39%,n=9) and 98.31%-101.08%(RSD=1.01%,n=9). CONCLUSIONS:The method is accurate and rapid,and suitable for the simultaneous determination of acetic acid and succinic acid in Banxia syrup.

ObjectiveTo study the mechanism of differentiation of mesenchymal stem cells(MSCs) into neuron-like cells in vitro.MethodsMSCs of Wistar rats were separated and cultured, and then induced with DMSO and BHA in vitro. The specific marking proteins of neurons, glia and neural stem cells were detected before preinduction, at 24h after preinduction, at 6h, 24h, and 48h after neuronal induction.ResultsAfter the inducement, many MSCs turned into bipolar,multipolar and taper,and then intersected as network structure. Nestin was strong positive at 6h after neuronal induction, and decreased at 24h, 48h after the induction. NeuN was present at 6 h after neuronal induction, and increased at 24h, 48h after the induction.ConclusionMSCs can be induced into neural stem cells(NSCs) at first, and then differentiate into neuron-like cells in vitro.

Objective: To explore the managemental essential ,we have Analyzed retrospectively 4635 articles collected by nation for 15 years in our hospital . Methods: According to the article database which collected our hospital's published articles from 1990-2004 provided by the center of article statistics and analyze of china science and technology institute of national department of science and technology, Windows Access 2000 was used to export articles, and Excel was used to sort﹑collect the results. Results: The embodied number was increased year by year , the authors were mainly from key department, clinical department with master or doctor degree and with high title of professional post, the condition of embodied articles is positive related with research level. Conclusion: It has validated the objective and meaning of the article management; and explained that the article is the talents strength expression,the science research platform,the science research management effects,the subject building collection and the symbol of connecting with the world .

Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apoptosis(bacterium ∶ cell =100 ∶ 1,50 ∶ 1,10 ∶1).The infection model was established using the best MOI =50 ∶ 1.The numbers of in vivo bacteria by colony formation units were calculated after macrophages were infected for different times,including 2,4,8,12,24 and 48 h,and the infected cells were collected.The ratios of apoptosis were detected and the regulation of caspases 3,8 and 9 in apoptosis pathway was elucidated by flow cytometry.Results The numbers of 16M in vivo bacteria were 105.4,104.8,105.8,106.5,108.0 and 109.0,respectively and of M5-90 were 106.1,106.2,106.4,106.3,106.1 and 105.0,respectively.The number of in vivo bacteria of 16M was significantly increased than that of M5-90 after infected for 24 h to 48 h.The ratios of apoptosis induced by 16M after infected for 2,4,8,12,24 and 48 h was (2.67 ± 0.09)％,(13.13 ± 0.30)％,(6.56 ± 0.42)％,(6.49 ± 0.28)％,(16.07 ± 0.86)％ and (24.23 ± 1.67)％,respectively,and by M5-90 was (3.62 ± 0.02)％,(32.01 ± 2.59)％,(17.58 ± 0.44)％,(16.09 ± 0.10)％,(62.53 ± 2.70)％ and (85.53 ± 0.15)％,respectively,and by control group was [(1.90 ± 0.20)％,(1.92 ±0.16)％,(1.99 ± 0.03)％,(2.48 ± 0.11)％,(3.56 ± 0.07)％,(5.26 ± 0.33)％].The differences were statistically between groups in same time.The Brucella melitensis vaccine strain M5-90 was more powerful than virulent strain 16M in respect of inducing macrophage apoptosis after infected for 24 to 48 h.Twenty-four hours after infection,the expression of caspases 3,8 and 9 was (1.47 ± 0.05)％,(1.52 ± 0.02)％ and (2.47 ± 0.12)％,respectively,in control group and the expression was (9.70 ± 0.46)％,(6.08 ± 0.56)％ and (35.08 ± 1.64)％,respectively,after infected for 24 h induced by M5-90.The expression of caspases 3,8 and 9 was significantly higher than that control group (P ＜ 0.01).Twenty-four hours after given caspases 3,8 and 9 inhibitor,apoptosis rate in control group was (66.72 ± 1.28)％,in M5-90 group was (22.58 ± 0.55)％,(53.15 ± 1.85)％ and (29.18 ± 0.23)％,respectively,and compared with control group,apoptosis rate of caspases 3,8 and 9 was significantly lower(P ＜ 0.01).Conclusions Apoptosis of macrophage can be induced by Brucella melitensis virulent vaccine strain 16M and attenuated strain M5-90.M5-90 is stronger than that of strain 16M.Caspases 3,8 and 9 can regulate macrophage apoptosis after M5-90 infection.

Aim To screen a more suitable transfection recep-tor,and improve the efficiency of constructing cell lines highly expressing human peptide transporters 1 (hPepT1 ).Methods The recombinant plasmid pcDNA3.1 (+)-hPepT1 was transfect-ed into MDCK cells and HeLa cells by LipofectamineTM 2000 transfection reagent,respectively.The monoclonal cells were se-lected and cultured.Expression of hPepT1 mRNA and protein were determined by qRT-PCR and Western blot,respectively. The uptake capacity of Glysar in transfected cells was examined. Results Compared with wild type cells,the expression of hPepT1 and the uptake of Glysar in transfected MDCK cells and HeLa cells significantly increased (P <0.05).Although the up-take of Glysar in HeLa cells was higher than that of MDCK cells,on the contrary,the expression of hPepT1 and the uptake of Glysar in MDCK-hPepT1 cells was higher than that of HeLa-hPepT1 cells.Conclusion MDCK cells may serve as a more suitable transfected receptor for the construction of a cellular model with high expression of hPepT1 ,which would make the construction of a cell model highly expressing hPepT1 more effi-cient.

Objective To investigate the rat bone structural changes of lower limb following lumbar nerve dorsal roots section.Methods Forty-eight mature female Wistar rats were divided into posterior radi- cotomy(PR)and comtrol groups randomly.The bilateral femoral bone mineral density(BMD)and biome- chanics characteristics were analyzed 2,4 and 8 weeks after the radicotomy.The same operation except the radicotomy was done in the sham group.Results In PR group,2,4,and 8 weeks after the radicotomy,the BMD of femur was(0.221?0.008)g/cm~3,(0.213?0.015)g/cm~3 ,and(0.216?0.105)g/cm~3 ,respective- ly;while that was(0.223?0.005)g/cm~3,(0.218?0.014)g/cm~3 ,and(0.208?0.111)g/cm~3 in control group.No significant difference was observed between the two groups(P＞0.05).In PR group,2,4,and 8 weeks after the operation,the mean maximum load in three-point bending test of femun midshaft was(93.64?8.76)N,(89.77?11.18)N and(93.21?8.74)N,respectively,and was lower than the values of the con- trol group at the same time point(95.94?6.29)N,(91.63?9.43)N,(95.57?8.64)N,However,there was no significant difference between the two groups(P＞0.05).Accordingly,there was no significant difference in the energy absorption in femun midshaft between the two groups(P＞0.05).Conclusion The selective rhizotomies of part lumbar never dorsal roots might not cause the loss of the femur BMD and the change of bio- mechanics property significantly in short period.

Objective To evaluate the efficacy and safety of retroperitoneal laparoscopic pyelolithotomy ( RLP) combined with holmium laser lithotripsy under flexible cystoscopy in the treatment of complicated nephrolithiasis. Methods The retrospective analysis was made on the clinical data of 37 patients who underwent RLP and holmium laser lithotripsy under flexible cystoscopy for complicated nephrolithiasis from January 2013 to January 2014. The clinic parameters involved basic data of patients,operational time,blood loss,post-operative hospital stay,the status of stone-free,perioperative complications,and the follow-up data of patients were observed. Results No patient was converted to open surgery. The mean stone size was (2. 8 ± 0. 9) cm in diameter,operational time was (89 ± 24) min,blood loss was (21. 3 ± 7. 7) mL,post-operative hospital stay was (6. 8 ± 1. 7) d,the stone removal rate in one session was 94. 6%. One case occurred urinary leakage,1 case occurred fever after operation,who were all recovered through conservative treatment. All cases were followed up at the sixth months after operation. Conclusion RLP combined with holmium laser lithotripsy under flexible cystoscopy is effective and safe for the treatment of com-plicated nephrolithiasis.

Abstrate:Aim To construct HEK293 cell line with stable and high expression of sodium taurocholate cotransporting polypeptide (NTCP ) efficiently and rapidly.Method Vector expressing EGFP-NTCP fusion protein was constructed and verified by DNA sequencing.The pEGFP-NTCP expression vector was transfected into HEK293 cells by FuGENE 6 transfection reagent. The transfected cells with high expression of green fluorescent protein were selected using fluorescence microscope for screening of G418 for 14 days to obtain cell lines stably and highly expressing NTCP.NTCP expression was detected by RT-PCR,qRT-PCR, Western-blot and the uptake experiment of taurocholic acid.Re-sult RT-PCR,qRT-PCR,Western-blot and the uptake experi-ment revealed that compared to the control cells,the expression of NTCP was significantly positive (P<0.01)in stable trans-fected cells showing green fluorescence (P<0.05 ).Conclusion The HEK293 cell line with stable and high expression of NTCP has been established efficiently and rapidly,which provides a cellular model for the study of the mechanism of the uptake of bile acid derivatives.

Animals ; Chromatography, Thin Layer ; Dichlorvos ; blood ; Dimethoate ; blood ; Insecticides ; blood ; Methyl Parathion ; blood ; Mice ; Organothiophosphorus Compounds ; blood