We evaluated clinical application effect of gene chip for detection of rifampin (RFP) and isoniazid (INH) resistance in Mycobacterium tuberculosis (MTB).Rifampin and isoniazid drug-resistance gene loci were detected by gene chip with sputum specimens from smear-positive tuberculosis patients and clinical strains,comparing the results of detection.BACTEC MGIT 960 drug susceptibility test results were used as control to evaluate the detection performance of gene chip.The sequences of the polymerase chain reaction products of the rpoB,katG and inhA genes from 999 strains identified as Mycobacterium tuberculosis were determined to confirm the mutations by DNA sequencing.Results showed that 100 cases were identified as nontuberculous mycobacteria by gene chip in the 1 108 cases of smear-positive samples.Among the rest 1 008 samples,there were only 9 cases of microarray results different from BACTEC MGIT960 culture-positive strains,achieving the coincidences of 99.1％.Compared with BACTEC MGIT 960 drug susceptibility test results,the gene chip method displayed a concordance of 98.1 ％ and 94.5 ％ for RFP and INH respectively in the 999 strains.Compared with the DNA sequencing method,the accuracy of gene chip method was 99.6％ for rifampin resistance and 99.8％ for isoniazid resistance.It's concluded that the gene chip technology can quickly and accurately detect rifampin and isoniazid resistance in MTB and can be used directly for the detection of sputum samples.

The content of endogenous anabolic steroids is extremely low in biological matrices.Its chemical structure contains polar functional groups such as hydroxyl and carbonyl, which limits their applicability in GC-MS.The lack of ionized groups in the chemical structure leads to poor sensitivity in LC-MS, which plays a significant role in various physiological activities analysis.It is an effective way to enhance the response of mass spectrometer by modifying the chemical structure of endogenous anabolic steroids through derivatization technology.This review summarizes various derivatization reagents and corresponding derivatization processes of endogenous anabolic steroids analysis in the methods based on different testing instruments and methodologies.The advantage and disadvantage of all kinds of derivatiaztion methods and the prospect of the endogenous anabolic steroids derivatiaztion techniques are also discussed.

Objective To establish a RP-HPLC method for the determination of 3-amide-indole derivative and its related substances. Methods Diamonsil C18(250 mmí4. 6 mm,5 μm) column was adopted. The mobile phase consisted of a mixture of methanol-acetonitrile-water(431245) at the flow rate of 1. 0 mL·min-1 . The wavelength for ultraviolet detection was 224 nm. The injection volume was 20 μL and the column temperature was room temperature. Results 3-amide-indole derivative and related substances could be well separated. The linearity of the 3-amide-indole derivative curve was well correlated (r=0. 999 7) within the range of 0. 04-0. 16 mg·mL-1. The RSD was 0. 52%with good repeatability. The detection limit was 2. 65 ng. Conclusion The method is accurate,reliable,sensitive and specific,which could be used for the determination of 3-amide-indolederivative and related substances.

The culture of umbilical cord-derived mesenchymal stem cells is extremely important for studies on umbilical cord mesenchymal stem cells. Optimization of cellculture technology is crucial for clinical application of mesenchymal stem cells and even celltherapy. Meanwhile, the labeling and tracer technique of umbilical cord-derived mesenchymal stem cells is a hotspot in stem celltransplantation. OBJECTIVE:To review the research and development of the cellmarkers and tracer methods of umbilical cord-derived mesenchymal stem cells. METHODS:A computer-based search of VIP, CNKI, Medline, Highwire and Foreign Journals Integration System databases was performed for articles concerning culture and labeling of umbilical cord-derived mesenchymal stem cells published from January 2001 to October 2013. The keywords were“stem cells, mesenchymal stem cells, umbilical cord-derived mesenchymalstem cells, cellculture, labeling methods”in Chinese and English, respectively. Final y, 35 articles were included in result analysis. RESULTS AND CONCLUSION:Umbilical cord-derived mesenchymal stem cells have not yet been widely used, mainly because of the immature isolation, culture and staining techniques of umbilical cord-derived mesenchymal stem cells. These techniques are worthy of further optimization studies. Although in recent years, cellmarkers and tracer technology of umbilical cord-derived mesenchymal stem cell s have made great progress, there are stil many problems need to be solved.

To investigate chemical constituents from Radix Pittospori, chloroform extract of the roots was subjected to column chromatography with various chromatographic techniques. The structures were elucidated on the basis of physico-chemical property and spectral analysis. Two triterpenoids were identified as 22-acetyl-21-(2-acetoxy-2-methylbutanoyl)-R1-barrigenol(1) and 3alpha-hydroxyl-20-demethylisoaleuritolic-14(15)-ene-28, 30-dioic acid (2). Compound 1 is a new triterpene and compound 2 is isolated from this plant for the first time.

Objective To establish an HPLC method for determination of the content of radiosensitizer YABQ and its related substances.Methods Diamonsil C18 column (250 mm ×4.6 mm,5 μm) was used.The mobile phase consisted of methanol-0.1%formic acid (22∶78) and isocratic elution at a flow rate of 1.0 ml/min.The detection wavelength was 266 nm,the temperature of the column was 30℃, and the injection volume was 10 μl.Results The linear range of YABQ was 7-84 μg/ml, r=0.9992.The detection limit was 8 ng (S/N≥3), and the quantitation limit was 24 ng (S/N≥10). According to destructive sample processing, the separation coefficient between the peaks was above 1.5, indicating that this chromatographic method could meet the YABQ monitoring requirements.Conclusion Method validation and destructive testing show that both the precision and specificity are fine with this method, which could be used for quality control of YABQ and its related substances.

Objective To establish an high performance liquid chromatography method for determination of 2-indole ketone derivative and its related substances. Methods Agilent Eclipse XDB-C18(250 mm×4.6 mm,5 μm) column was adopted.The mobile phase was acetonitrile-water with gradient elution mode at a flow rate of 1. 0 mL?min-1; the column temperature was 35 ℃;the injection volume was 20 μL and the detection wavelength was set at 257 nm. Results 2-indole ketone derivative ID and related substances could be well separated. The 2-indole ketone derivative had good linear correlation ( r=0.999 4) within the range of 40-300 μg?mL-1 . It had a good precision ( RSD<1%) . The limit of detection was 8 ng. Conclusion The method is accurate,simple,sensitive and selective,which can be used for the quality control of 2-indole ketone derivative and related substances.

Objective\ To study whether gonadotropin releasing hormone receptor (GnRHR)mRNA exist in rat submaxillary gland and it's gene sequence. Methods\ The total RNA isolated from rat submaxillary gland was amplified by RT\|PCR, the PCR products were identified by sequencing with Sanger's method. Results\ The specific amplified band of GnRHR mRNA was detected through agarose gel electrophoresis and gene sequence is identical to the sequence of GnRHR which has been reported in rat pituitary. Conclusion\ GnRHR can be produced by submaxillary and response for modulating biological function of submaxillary.\;

BACKGROUND: Angiotensin Ⅱ has been found to induce atrial electrical remodeling, which can be blocked or inhibited by allicin.OBJECTIVE: To study the effects of allicin on angiotensin Ⅱ-induced calcium channel current and intracellular free calcium concentration in human atrial myocytes.DESIGN: A randomized controlled study based on human atrial myocytes freshly isolated.SETTING: Cardiology department of a military medical university of Chinese PLA.METHODS: This study was carried out from June 2003 to June 2004 in the Laboratory of Cardiology Department, Xijing Hospital, the Fourth Military Medical University of Chinese PLA. Ten patients with congenital heart disease who underwent extracorporeal circulation surgery were included in the study. Among them, there were 6 males and 4 females with the average age of 15 ± 6 years. Tissue samples were taken from their right auricle and sent to the lab, where the atrial myocytes were freshly isolated. There were four co-administration of angiotensin Ⅱ (0. 1 μmol/L)and allicin(50 μmol/L).The conventional whole-cell configuration of the patch-clamp technique was used to detect membrane electric current of Ca2 + in L type. Confocal microscope was used with Fluo-3/AM as calcium indicator to detect changes of intracellular free calcium concentration immediately and 15 minutes after drug intervention, respectively.MAIN OUTCOME MEASURES: The peak density of electric current of Ca2 + in L type and alteration of fluoresence intensity of intracellular free calcium concentration.electric current of Ca2 + in L type in human atrial myocytes was significantly increased by angiotensin Ⅱ of 0. 1 μmol/L[( - 12. 77 ± 1. 61) vs ( -5.78affect electric current of Ca2+ in L type in human atrial myocytes group, the peak density of electric current of Ca2 + in L type was significantly lower than that in angiotensin Ⅱ group[ ( - 8.75 ± 0.97) pA/pF, P ＜ 0. 05 ].in angiotensin Ⅱ group was significantly higher than that in control and allicin groups[(2 610.1±112.6, (299.2±27.3)%; 653.9±42.5, 0;simultaneously with angiotensin Ⅱ, the alteration of intracellular fluoresence intensity was much lower than that in angiotensin Ⅱ group[ ( 1284.9 ± 85.2,(96.5±8.4)%;P ＜0.05].CONCLUSION: Allicin antagonizes angiotensin Ⅱ-induced increase in the peak density of electric current of Ca2+ in L type and intracellular calcium overload, which may relieve atrial electrical remodeling.