In order to investigate the affinity of aptamers to Bacillus anthracis spore, a custom synthesized 78 mer random DNA library was subjected to 15 rounds of selection against spores of vaccine strain A.16R by using SELEX method. The selected aptamers were cloned and sequenced. Macaw 2.05 and DNAsis 2.5 package were employed to analyze the conserved sequences and second structure of the aptamers, respectively. Affinities of aptamers to the spores were visualized by biotin streptavidin horseradish peroxidase system. The results showed that affinities of the aptamers were different. The highest OD at 450nm was 1.2, and the lowest was 0.25. The second structure analysis revealed possible stem loops for binding to the spores. The conserved sequences, AGGGG, CCCCG, GGGTT and ACACT, were found and the aptamers having same conserved sequence demonstrated similar affinity to the spores.

Objective To analyze the specific peptide of cystatin C (CysC) and its characteristics by bioinformatics technology,and verify the predicted results by mass spectrometry.Methods Online software was applied to analyze the physicochemical properties and homology of CysC peptides hydrolyzed by trypsin and predict the associated parameters of ionized fragmentation of specific peptide by mass spectrometry.Precursor ion scan and product ion scan were conducted on the samples of synthetic specific peptide.The recombinant human CysC and serum samples were analyzed by mass spectrometry after trypsin digestion.The results of analysis were compared with the outcomes predicted by bioinformatics.Results T3 (ALDFAVGEYNK) was considered as the specific peptide of CysC by software analysis.When selecting[M + 2H] 2 + for product ion scan,almost all the y and b ions of fragmentation were observed using tandem mass spectrometry (MS/MS),showing consistency with Skyline predictions.Moreover,both the peptides from the human recombinant CysC and serum sample following the trypsin digestion were eluted at the same time with the isotope-labeled T3 * under the fixed conditions.Conclusion Bioinformatics technology could be available for picking out the specific peptides of target protein quickly and efficiently and predicting the ionized fragmentation precisely by mass spectrometry scanning.

A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.
Calibration ; Chromatography, Liquid ; Humans ; Liquid-Liquid Extraction ; Sitagliptin Phosphate ; blood ; pharmacokinetics ; urine ; Tandem Mass Spectrometry

OBJECTIVE: To explore a simple speedy specific and sensitive method to detect specific IgM (sIgM) and IgG (sIgG) antibodies of hemorrhagic fever with renal syndrome (HFRS),and to study the therapeutic effects of integrated traditional Chinese and western medicine on HFRS. METHODS: The serum of 559 patients with HFRS were tested with colloidal gold immuno-dot assay (CGIDA) for sIgM and sIgG antibodies and compared with enzyme linked immunosorbent assay (ELISA) or indirect fluorescent antibody test (IFAT). One hundred and one patients with HFRS were randomized into treatment group (n=50),treated with Kuhuang Injection, Shenmai Injection and Huangqi Liquid) and control group (n=51),treated with Ribarvirin and Ganlixin Injection). RESULTS: The positive rate of sIgM detected with CGIDA was 70.8% and the positive rate of sIgG detected with CGIDA was 87.5%. The days for fever decline, symptoms alleviation and sign relief between the treatment group and control group were similar (P>0.05). The days for recovery of kidney function in the control group was less than that in the treatment group (P<0.01). The rate of crossing shock stage in the treatment group was higher than that of the control group (P<0.01). CONCLUSION: CGIDA was more simple, speedy, specific and sensitive than ELISA or IFAT in detecting the sIgM or sIgG antibodies in serum of patients with HFRS. Although the sensitivity of CGIDA was lower than that of ELISA the CGIDA had no false positive reaction the sensitivity of CGIDA was higher than that of IFAT on detecting IgG. The effect of the treatment group was similar to that of the control group. But the crossing shock stage rate in the treatment group was higher than that of the control group while the control group was better than the treatment group in recovering the kidney function.

OBJECTIVE:To study the optimal monitoring indexes of plasma concentration of cyclosporine A(CsA) in patients receiving hematopoietic stem cell transplantation.METHODS:The lowest CsA concentration(C0) and the highest CsA concentration(C2) in 23 hematopoietic stem cell transplant recipients were detected by FPIA.All the data were analyzed statistically.RESULTS:Within 6 months after hematopoietic stem cell transplanttion,C0,C2,C0+C2 and C2/C0 in the recipients were(228.84?142.48) ?g?L-1,(741.50?294.42) ?g?L-1,(970.34?391.18) ?g?L-1 and(3.88?1.94) ?g?L-1,respectively.CONCLUSION:As reasonable monitoring indexes for the plasma concentration of CsA,C0+C2 and C2/C0 can comprehensively reflect the exposure of drug in body and monitor the toxicity of CsA in liver and kidney.

A rapid and sensitive liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) method for quantification of sitagliptin in human plasma and urine had been developed. This method was applied to the pharmacokinetics study of sitagliptin tablet after single- and multiple-dosing in Chinese population. Plasma samples were prepared by a liquid-liquid extracted method, and urine samples were diluted. Compounds were analyzed by multiple reaction monitoring (MRM) mode with a electrospray ionization (ESI) interface. Mobile phase consisted of methanol and water (85 : 15, v/v). The linear concentration range of calibration curve was 0.5-1 000 ng.mL-1. and 0.2-100 µg.mL , intra-run/between-run accuracy was 98.98%-103.69% and 97.63%-102.29%, intra-run/between-run precision was <5.51% and 4.26% for plasma and urine sample, respectively. The stability of sitagliptin stock solution was tested for 55 days at -30 °C. Sitagliptin was stable when stored under the following conditions: 24 hours in the autosampler after sample preparation; 24 hours at room temperature, after 3 freeze and thaw cycles (from -30 °C to room temperature), 40 days at -30 °C for plasma and urine samples. The absolute recovery in plasma was 71.1%, and no matrix effect was founded. This method was proved simple, specific, sensitive, rapid and suitable for pharmacokinetics study of sitagliptin in human being.

Object In order to obtain further evidence of therapeutic efficacy of sophoramine (SA) *, its effect on transplantation rejection was studied with simultaneous exploration of its mechanism of action Methods Number of T lymphocyte and its sub sets of skin allografted rat model were determined by immunofluorescence technique, and proliferation of T lymphocyte was assessed by thiazolyl blue (MTT) assay Results SA reduced the CD 3 and C 4 cell counts, lowered CD 4/C 8 ratio, inhibited T lymphocyte proliferation and prolonged the survival time of skin allografted rat Conclusion SA could inhibit transplantation rejection by inhibiting the function of T lymphocyte with simultaneous reduction of lymphocyte count

Adolescent ; Adult ; Automobile Driving ; China ; Electrocardiography ; Female ; Heart ; physiopathology ; Humans ; Male ; Middle Aged ; Noise, Occupational ; adverse effects ; Occupational Exposure ; adverse effects ; Stress, Psychological ; etiology ; physiopathology ; Vibration ; adverse effects

Methods The clinical data of 4445 cases (5 530 hernias) who underwent LIHR at Ruijin Hospital from Jan 2001 to Dec 2015 were analyzed retrospectively.2 125 cases underwent 2 402 trans-abdominal preperitoneal procedure(TAPP),2 306 cases did 2 907 totally extraperitoneal (TEP),and 21 IPOMs in 20 cases.There were 3 216 indirect hernias (60.3％),1 164 direct hernias (21.8％),399 recurrent hernias (7.5％),479 complex hernias (9.0％),and 72 femoral hernias (1.4％).The median time of follow-up is 51 months with a range between 7 and 187 months.Results The average operation time was 27.1 ± 8.7 min for unilateral hernia repair,and 43.0 ± 11.0 min for bilateral hernia repair.The average hospital stay was 1.4 ± 1.1 d.There were 250 seroma (4.7％),68 urinary retention (1.3％),23 transient neuropraxia (0.4％) and 3 paralytic obstruction of intestines (0.1％).Severe complications included 1 port site hernia,1 intestinal injury,and 1 mechanical intestinal obstruction.After a medium follow-up of 51 months,there were 13 recurrent cases (0.24％),including 5 cases after TAPP,7 after TEP,1 after IPOM.Conclusion LIHR is a safe and efficient technique for hernia repair.