To study the drug-drug interaction of morinidazole and warfarin and its application, a sensitive and rapid liquid chromatography-tandem mass spectrometric (LC-MS/MS) method was developed for the determination of R-warfarin/S-warfarin in human plasma. In a random, two-period crossover study, 12 healthy volunteers received a single oral dose of 5 mg racemic warfarin in the absence and presence of morinidazole. Blood samples were collected according to a pre-designed time schedule. R-warfarin, S-warfarin and methyclothiazide were extracted with ethylether : methylenechloride (3 : 2), then separated on a Astec Chirobiotic V (150 mm x 4.6 mm ID, 5 microm) column using 5 mmol x L(-1) ammonium acetate (pH 4.0) - acetonitrile as mobile phase at a flow-rate of 1.5 mL x min(-1). The mobile phase was splitted and 0.5 mL x min(-1) was introduced into MS. A tandem mass spectrometer equipped with electrospray ionization source was used as detector and operated in the negative ion mode. Quantification was performed using multiple reaction monitoring (MRM). The resolution of warfarin enantiomers is 1.56. The linear calibration curves for R-warfarin and S-warfarin both were obtained in the concentration range of 5 - 1 000 ng x mL(-1). Intra- and inter-day relative standard deviation (RSD) for R-warfarin and S-warfarin over the entire concentration range across three validation runs was both less than 10%, and relative error (RE) ranged from -4.9% to 0.7%, separately. The method herein described is effective and convenient, and suitable for the study of metabolic interaction between morinidazole and warfarin. The results showed that coadministration of warfarin with morinidazole did not affect the pharmacokinetics of either R-warfarin or S-warfarin.

Objective To observe the effects of exercise on serum adiponectin and adiponectin receptor (AdipoR) level in skeletal muscle of insulin-resistant rats. Methods A total of 30 healthy male rats were randomly divided into a control group ( NC, n = 8) and a high-fat group ( HF, n = 22), fed with normal chow and high fat diet, respectively. Eighteen weeks later, the high-fat group was randomly divided into a high-fat diet control group (HC, n = 10) and an exercise group (HE, n = 12). The HC and HE group were continually fed with high fat diet, while the HE group was administered with swimming training for 6 weeks in addition at the same time. After 24 weeks, the insulin sensitivity index was calculated, and serum adiponectin level was detected by using ELISA. The expressions of AdipoR mRNA in skeletal muscle were detected with real-time fluorescence quantitative polymerase chain reaction (PCR). Results After 18 weeks, compared to NC group, the insulin sensitivity index of HF group decreased significantly. It suggested that insulin resistance appeared in HF group. Twenty-four weeks later, compared to NC group, the ISI of HC group was significantly decreased, meanwhile the level of serum adiponectin, expression of AdipoR1 and AdipoR2 mRNA in skeletal muscle of HC group were 71.9% , 59.9% and 69.2% of those of the NC group, respectively; compared to HC group, the ISI was increased significantly by exercise, meanwhile the expression of AdipoR1 mRNA in skeletal muscle was significantly increased by 1.33 times, however the level of serum adiponectin and the expression of AdipoR2 mRNA in skeletal muscle were not altered in HE group. Conclusion Six weeks of exercise improves insulin sensitivity through increasing the expression of AdipoRI mRNA in skeletal muscle.

Objective To investigate the risk factors associated with cranial nerve impairment in patients with tuberculous meningitis.Methods A total of 121 patients with tuberculous meningitis who were admitted to Huashan Hospital from 2000 to 2011 were reviewed retrospectively.Demographic data (gender,age),course of disease,initial results of cerebral spinal fluid (CSF) tests,occurrence of cranial nerve impairment and prognosis of these patients were collected.All the patients were followed up for at least 3 months,and for those with cranial nerve impairment,the minimum follow-up period was 1 year in order to judge the recovery of cranial nerve impairment.Multivariate analysis was performed to study the associated risk factors.Results Out of 121 patients,22 (18.2 ％)developed cranial nerve impairment.Nerves involved were abducens nerve,oculomotor nerve,optic nerve and auditory nerve,and impairment of single nerve occurred in 9 (40.9 ％),8 (36.4 ％),7(31.8％) and 1(4.5％) patient,respectively.Three cases had more than one group of cranial nerves involved,accounting for 13.6％ of the 22 patients with cranial nerve impairment.The incidence of conscious disturbance was significantly higher in patients with cranial nerve impairment than those without impairment (77 ％ vs 45 ％,P=0.020).Delay in diagnosis (OR =1.017,95 ％ CI:1.001-1.033,P=0.040) and occurrence of conscious disturbance (OR =3.242,95 ％ CI:1.142-9.205,P=0.027) were independent predictive factors of cranial nerve injury.During one-year follow-up,90.9％ of patients were fully recovered from cranial nerve impairment,with a median duration of 1 month (range 0.5-6.0 months).Conclusions Cranial nerve impairment is a common complication in patients with tuberculous meningitis.Delay in diagnosis and occurrence of conscious disturbance were independent predictive factors.Most cranial nerve impairment were reversible,and timely diagnosis and treatment are important ways to reduce complications.

Objective To develop a new multiplex allele-specific PCR(MAS-PCR) assay to detect mutation in codon 315 of katG gene of Mycobacterium tuberculosis , mutation in this codon has been reported to be able to account for the Mycobacterium tuberculosis clinical isolates resistant to isoniazid(INH).Method Based on the sequence of katG gene of Mycobacterium tuberculosis , three specific primers are designed to carry out the MAS-PCR, 84 purified DNA preparation with known katG 315 variation detected by PCR-restriction fragment length polymorphism (PCR-RFLP) are used to optimize PCR.Results 84 Mycobacterium tuberculosis clinical stains are detected by the MAS-PCR and PCR-RFLP, respectively.The sensitivity of detection by MAS-PCR is 77.8%,and the specificity is 95.2%.katG mutation S315N(AGC→AAC), neglected in RFLP, can be detected by MAS-PCR.Conclusion MAS-PCR assay is sensitive, specific, economic and easy to carry out , can be used in clinical laboratories to detect the INH-resistant Mycobacterium tuberculosis strains.

BACKGROUND: Nowadays complex bone defects have become a great challenge to orthopedists. A synergistic contribution of various growth factors and a crosstalk between their signaling pathways have been suggested as determinatives for the overall osteogenic outcome.OBJECTIVE: To develop calcium phosphate cement (CPC) incorporated with γ-polyglutamic acid/carboxymethyl chitosan (PGA/CMCS), and to evaluate its physical and chemical properties and sustained-release function. METHODS: The γ-PGA/CMCS polymer composites were prepared by graft copolymerization and spray freeze drying methods, and then loaded with recombinant human bone morphogenetic protein 2 (rhBMP-2) growth factor. CPC served as control group, and γ-PGA/CMCS-CPC containing different contents of rhBMP-2 as experimental groups. A γ-PGA/CMCS-CPC scaffold with regular blade-like crystalline structure was fabricated by injection compression molding. Before mixed with the liquid phase, the solid additives were properly mixed by wet method of CPC solid and the γ-PGA/CMCS carrier, then the pre-blended mix was freeze-dried. The setting time and compressive strength of bone cement in each group were detected, and the microstructure of the material surface was observed under scanning electron microscopy. In vitro release of rhBMP-2 was investigated. The effect of bone cement extracts on cell proliferation was determined through MTS assay.RESULTS AND CONCLUSION: γ-PGA/CMCS-CPC had the same physicochemical properties to the CPC. Initial and final setting time, compressive strength of bone cement had no significant differences among groups. The scanning electron microscope results showed that the γ-PGA/CMCS-CPC scaffold was covered by regular blade-like crystalline structure and the γ-PGA/CMCS particles were uniformly dispersed in the CPC crystals. A sustained release of rhBMP-2 was observed from the γ-PGA/CMCS-CPC. The cell experiments exhibited that the samples with regular blade-like crystalline structure had better cell response compared to CPC control groups with irregular crystalline structure. These findings indicate that γ-PGA/CMCS-CPC can maintain good physicochemical properties, and release growth factor or drug to promote bone formation.

Objective:To explore the characteristics of human melanoma-specific antigen peptides by HLA-A2 restricted tumor-infiltrating lymphocytes.Methods:The HLA-A2 protein and polypeptides molecules were purified from the three tumor cell lines(624-Mel, Chap-Mel and JY) by immunoaffinity chromatography, after the peptides bound to HLA-A2 protein solution were acidified with acetic acid and boiled by high temperature, and centrifuged through an Ultra-CL filter, then the peptides extracts were fractionated by revered phase high pressure liquid chromatography(RP-HPLC). Individual fractions were assessed for their ability to reconstitute melanoma-specific epitopes by adding to the HLA-A2 Ag-procceing mutant cell, T2. The biological feature of one of three active peptides from RT-HPLC samples was performed by mass spectrometric analysis. The synthetic peptides identical to active peptide sequences were determined in the reconstitute test.Results:Three prominent peaks(P19, P25 and P31) of the fraction from 624-Mel were observed in the reconstitute test, TIL killing rate was 67% for (P31) peptide fraction. The mass spectrometric analysis of one of active peptides (P31) showed that at mass-to-charge ratio(m/z) 948 has been usually nine residues. The sequence is H+ Ala Lue Trp Lue Phe Phe Gly Val Lue OH-. The peptide synthesized comprising epitopes were verified.Conclusion:These results showed the peptides derived from active fractions were related to human melanoma-specific tumor antigen peptides recognized by HLA-A2-restriced TIL. These peptides could develop novel peptide-based an anti-tumor vaccine for immunotherapy of CTL.

Objective To evaluate the number and function of peripheral Vγ2Vδ2+T lymphocytes during Mycobacterium tuberculosis(MTB)infection in human immunodeficiency virus (HIV)infected individuals.Methods Seventy-six HIV/acquired immune deficiency syndrome(AIDS) patients co-infected with MTB were divided into active tuberculosis(TB)group(HIV+TB)and latent TB group(HIV+LTB).T cell subsets of peripheral blood lymphocytes were analysed by flow cytometry.Stimulated by protein purified derivative(PPD)and hydroxymethylbutenyl diphosphate (HMBPP),specific interferon(IFN)-γ producing T cells were detected using enzyme-linked immunospot(ELISPOT)and intracellular cytokine staining(ICS).Data were analyzed by t test.Results The absolute number of CD3'T cells and DroDortion of V72VB2'T cells in CD3+T cells in HIV+TB group were both significantly lower than those in HIV+LTB group(t=-3.67,P<0.01;t=-2.06,P<0.05).PPD-specific IFN-y-producing T cells and percentage of PPD-specific CD4+Tcells in CD3+T cells in HIV+LTB group were both similar with those in HIV+TB group.While HMBPP-specific IFN-γ-producing T cells and percentage of HMBPP-specific Vγ2Vδ2+T cells in CD3+Tcells in HIV+LTB group were both higher than those in HIV+TB group(t=2.71 and t=3.003,respectively;both P<0.0 1).Conclusion The number and function of Vγ2Vδ2+T cells were impaired in HIV/AIDS patients coinfected with active MTB infection,which indicates that Vγ2Vδ2+T cells may be the key immune cells against MTB in individuals with impaired CD4+T cells.

Objective:To compare the biomechanical effects between oblique Ban-pulling manipulation and lumbar erection-rotation manipulation in sitting position in treating lumbar intervertebral disc herniation (LIDH). Methods:A three-dimensional finite element model of L3-S1 was developed to carry out a comparative study between oblique Ban-pulling manipulation and lumbar erection and rotation manipulation in sitting position. The disc protrusion was assumed to be on the rear left of L4 disc, and the manipulations were performed on the right side. The loading process was simulated by two steps. In the first step, only the compression loading was imposed, and in the second step, both the compression loading and axial rotation moment were imposed. The displacement and stress distribution in L4 disc were investigated. Results:The values of stress and displacement in the second step were lower than those in the first step in each manipulation. The stress and displacement differences between the two steps were respectively 1.79 times and 3.03 times larger in oblique Ban-pulling manipulation than those in lumbar erection-rotation manipulation in sitting position. Conclusion: Oblique Ban-pulling manipulation may result in a better biomechanical effect than lumbar erection-rotation manipulation in sitting position for LIDH.

Objective To evaluate the effect of acupuncture point stimulation on the urodynamics of spinal cord injury patients.Methods Sixty spinal cord injury cases were divided into a research group and a comparison group.Both groups were given bladder training, but the research group also received acupuncture point stimulation.The subjects' maximum cystometric capacity, maximum detrusor pressure and residual urine were measured, along with renal function and any urinary tract infections.The scale of activity of their daily lives was compared before and 3 months and 6 months after the treatment.Results There were no statistically significant differences between the groups before the treatment.There were statistically significant differences in maximum cystometric capacity, maximum detrusor pressure, residual urine and Barthel index (BI) after 3 and 6 months of treatment.In the research group, maximum detrusor pressure, residual urine and BI scores had all improved after 6 months compared with the 3 month values.In the comparison group only the BI score had improved significantly between 3 and 6 months, and this was not reflected in a significant change in the scale of activity in daily life.After 3 months, only the average BI differed significantly between the two groups.After 6 months all indexes except the BI and maximum cystometric capacity differed significantly.There were also significant differences between the groups in reflex voiding, percussion voiding, and abdominal pressure voiding.Conclusions Acupuncture point stimulation can improve the bladder function of spinal cord injury patients.It can also decrease the incidence of urinary tract infection and the possibility of renal dysfunction.

Objective To examine the expression of typeⅠcollagen in pulmonary arteries of rats with monocrotaline(MCT)-induced pulmonary hypertension(PH) and explore the mechanism of pulmonary vascular structural remodeling.Methods Twelve male Wistar rats were randomly divided into 2 groups: the MCT group(n=6) and the control group(n=6),which received a single intraperitoneal injection of MCT solution(60 mg?kg-1,the first day) or 9 g?L-1saline,respectively.After 3 weeks,mean of pulmonary artery pressure(mPAP),the value of right ventricle/(left ventricle plus septum)[RV/(LV+S)] and body weight were measured.Lung sections(HE stained) were observed under lightmicroscope for changes of the pulmonary arteries.The protein expression of typeⅠcollagen in pulmonary arteries was detected by immunohistochemical technique.Results Three weeks after MCT injection,compared with control group,mPAP and RV/(LV+S) increased significantly in MCT group[mPAP:(10.60?2.06) mmHg(1 mmHg=0.133 kPa) vs(32.40?3.24) mmHg,P