Objective To establish and evaluate a gene microarray for determination katG mutations of Mycobacterium tuberculosis isolates associated with resistance to isoniazid(INH).Methods A panel of probes were designed and gene chips were prepared by dotting.Mycobacterium tuberculosis isolates resistance to 5 drugs was determined by proportional dilution methods.Amplicons of Mycobacterium tuberculosis isolates were detected by our chip and sequenced.Results The drug resistance rate of the isolates to at least one of the anti-tuberculosis drugs was 70.8%(97/137).45 strains out 137 Mycobacterium tuberculosis isolates was resistant to INH(32.8%).katG was successfully amplified from 100% of the susceptible strains and 88.9%(40/45)resistant strains.4 of 45 INH resistant isolates' katG were deleted.27 of 40(67.5%) katG has been detected to have katG 315 codon mutations.The mutations were 315 AAC(Asn,13/40), ACC(Thr,6/40),ACA(Thr,4/40),ATC(Ile,2/40),AGC(Arg,2/40).The mutation rate of katG analyzed by gene chips we prepared were identical to katG sequencing.Conclusion The gene microarray techniques we developed for determination of Mycobacterium tuberculosis resistance to INH are specific, sensitive and may be used as an alternative in clinical laboratory.

BackgroundDiabetic retinopathy (DR) is the result of the cytokine network disorders,the imbalance of angiogenic factor and vascular inhibitory factor is the start factor.ObjectiveTo analyze the levels of CXC chemotatic factors of type 2 diabetes mellitus patients,evaluate the clinical application value of them in different clinical types of DR using receiver operating characteristic (ROC)analysis and to approach the new way of individualized treatment.Methods This was a prospective research.The gold standard was ophthalmolscope and fundus fluorescein angiography.The levels of CXC chemotatic factors and multiplicaiton factors were measured in 96 cases with type 2 diabetes mellitus (66 cases with retinopathy and 30 cases without retinopathy as control).The assessment tasks were performed for these index and courses of DR with ROC curve.Results The expression of age,course of disease has significant difference in different courses of DR ( F =8.507,P =0.001 ; F =28.143,P =0.000).Compared with the control group,the expression of growth-related oncogene-α ( GROα ) ( t =- 2.172,P =0.035,AUC =0.625 ),whole blood viscosity 200 ( t =- 3.724,P =0.001,AUC =0.904 ) and neutrophilic leukocyte (t=-2.562,P =0.013,AUC =0.577 ) has significant difference in the group of mild NPDR.Compared with the control group,the expression of interferon-γ-inducible protein 10 ( IP-10 ) ( t =-3.591,P =0.001,AUC =0.592 ),platelet derivation growth factor-BB ( PDGF-BB ) ( t =- 3.233,P =0.003,AUC =0.735 ),vascular endothelial growth factor(VEGF) ( t =- 3.617,P =0.001,AUC =0.776 ),C peptide ( t =- 3.366,P =0.002,AUC =0.962 ),leukocyte ( t=-3.201,P =0.003,AUC =0.852) and neutrophilic leukocyte(t =-4.201,P=0.000,AUC =0.852) has significant difference in the group of moderate and severe NPDR.ConclusionsCXC chemotatic factors may act as reactivator in the pathogenesis of DR,GROα and IP-10 may be useful for clinical monitoring of the severity of DR,and evaluating the imbalance state of chemotatic factors maybe a new approach to clinical monitoring and prognosis of DR.

Objective To compare MR,CT,and radiography in the detection of arthropathies in patients with hemophilia.Methods Forty-one symptomic joint images in the 14 patients with hemophilia, aged from 11 to 24 years,were used in this study.Each joint had the examinations of radiography,CT and MR within one day.The severity of each joint was staged using conventional radiographic classification. Severe HA patients with stage 5 were excluded from the study.Imaging findings of soft tissue swelling, osteoporosis,epiphyseal overgrowth,joint erosion,cyst,joint space narrowing,bone marrow,joint effusion, hemorrhage,synovial hypertrophy,widened intercondylar notch as well as anterior and posterior crueiate ligaments(only for knee joint)were used for the all imaging comparison.Results The 41 symptomatic joints in 14 patients with hemophilia were classified by radiographic criteria into stage 0(n=5),stage 1(n=7),stage 2(n=6),stage 3(n=8)and stage 4(n=15).Soft tissue swelling or joint effusion was observed in 33 joints by radiographs,in 34 joints by both CT and MR.Joint erosions were demonstrated in 34 joints by MR,in 33 joints by CT and 20 joints by radiographs.Joint cysts were shown in 21 joints by MR,in 18 joints by CT and 9 joints by radiographs.Significant differences in detection of erosion and cyst were found between radiography with either CT(P0.05).MR showed improvement for detecting nlore loci of both erosion and cyst than CT and radiography,and also CT showed the improvement than radiography.Bone marrow edema 14 joints, hemon'hage in 34 joints and synovial hypertrophy in 27 joints were revealed on MR images.Conclusion MRI is superior to CT and conventional radiography in detecting the abnormal changes and should be considered as the first choice among the imaging modafities in evaluating hemophilic arthropathies.

OBJECTIVETo summarize the effectiveness of Chinese and Western integrative medicine in treating medium and advanced lung cancer, and to provide guidelines for clinical application.

METHODSFor this metaanalysis, a comparative search of Chinese medicine data in Chinese National Knowledge Infrastructure (CNKI) and Chinese BioMedical Literature Database (CBM) was undertaken to identify articles related to randomized comparative research of Chinese and Western integrative medicine in treating medium and advanced lung cancer between 1996 to 2006. Quality of life (QOL) was estimated using RevMan 4.2 software for data processing, adopting the odd ratio (OR) and the 95% confidence interval (CI).

RESULTSThrough meta-analysis of 10 qualified articles, the results were as follows: the merging effectiveness of QOL [OR=3.80, 95% CI (2.65, 5.47)]; the rate of survival [OR=3.44, 95% CI (2.04, 5.80)]; the tumor response rate [OR=1.88, 95% CI (1.37, 2.58)]; the tumor developing rate [OR=0.33, 95% CI (0.23, 0.48)]. Significant differences existed between the Chinese and Western integrative medicine treatment group and the Western treatment group (P<0.01).

CONCLUSIONSChinese and Western integrative medicine treatment of medium and advanced lung cancer has shown to improve patients' QOL and survival rate; it also can control tumor development in the short term.

Carcinoma ; epidemiology ; pathology ; therapy ; Combined Modality Therapy ; Disease Progression ; Humans ; Integrative Medicine ; methods ; Lung Neoplasms ; epidemiology ; pathology ; therapy ; Medicine, Chinese Traditional ; methods ; Neoplasm Staging ; Publication Bias ; statistics & numerical data ; Randomized Controlled Trials as Topic ; statistics & numerical data ; Treatment Outcome ; Western World

Antibody-drug conjugate (ADC) is a new class of therapeutics composed of a monoclonal antibody and small cytotoxin moieties conjugated through a chemical linker. ADC molecules bind to the target antigens expressed on the tumor cell surfaces guided by the monoclonal antibody component. The binding ADC molecules can be internalized and subsequently the toxin moieties can be released within the tumor cells via chemical and/or enzymatic reactions to kill the target cells. The conjugation combines the merits of both components, i.e., the high target specificity of the monoclonal antibody and the highly potent cell killing activity of the cytotoxin moieties. However, such complexities make the pharmacokinetic and metabolic studies of ADCs highly challenging. The major challenges should include characterization of absorption, distribution, metabolism and excretion, investigation of underlying mechanisms, assessment of pharmacokinetic- pharmacodynamic relationship, and analytical method development of ADC drugs. This review will discuss common pharmacokinetic issues and considerations, as well as tools and strategies that can be utilized to characterize the pharmacokinetic and metabolic properties of ADCs.
Antibodies, Monoclonal ; pharmacokinetics ; Cytotoxins ; pharmacokinetics ; Humans ; Immunoconjugates ; pharmacokinetics ; Neoplasms ; drug therapy

Aim To investigate the roles of chloride channels in the apoptosis and apoptotic volume de-crease (AVD)induced by adriamycin in nasopharyn-geal carcinoma CNE-2Z cells.Methods Apoptotic rates were detected by flow cytometry,and the volume changes were measured by the time-lapse live cell ima-ging technique.The patch clamp technique was used to record whole-cell chloride currents.Results Adria-mycin induced apoptosis of CNE-2Z cells.An early ap-optotic volume decrease was observed in the cell trea-ted with adriamycin.The cell volume was decreased by about 10% in 2 h.Adriamycin activated a chloride current which showed outward rectification.The chlo-ride channel blocker 5-nitro-2-(3-phenylpropylamino)-benzoate (NPPB ) could inhibit the adriamycin-in-duced chloride currents,apoptosis and prevent cell shrinkage.Conclusions Our findings suggest that ad-riamycin causes cell apoptosis by activation of chloride channels.Chloride channels may be involved in the apoptosis and apoptotic volume decrease induced by adriamycin in CNE-2Z cells.

The paper is to report the development of a high-performance liquid chromatographic/tandem mass spectrometry (HPLC-MS/MS) method for the determination of icaritin (ICT) in rat plasma. After precipitated with acetonitrile from the plasma, ICT was isolated chromatographically on a Dikma C18 column. The mobile phase consisted of acetonitrile-water-acetic acid (72 : 28 : 1.5, v/v/v). Electrospray ionization (ESI) source was applied and operated in the positive ion mode. Multiple reaction monitoring (MRM) mode with the transitions of m/z 387 --> m/z 313 and m/z 331 --> m/z 315 were used to quantify ICT and the internal standard, respectively. The linear calibration curve was obtained in the concentration range of 2.5-1,000 ng x mL(-1). The lower limit of quantification was 2.5 ng x mL(-1). The inter- and intra-day precision (RSD) were less than 9.63%, and the accuracy (relative error) was within +/-7.42%. The method was proved to be suitable for the pharmacokinetics of ICT, which offers advantages of high sensitivity and selectivity.
Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; methods ; Epimedium ; chemistry ; Female ; Flavonoids ; administration & dosage ; blood ; isolation & purification ; pharmacokinetics ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Reproducibility of Results ; Sensitivity and Specificity ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tandem Mass Spectrometry ; methods

OBJECTIVETo investigate the effects of hepatitis C virus (HCV) on transcription regulation genes of host cells by gene chip assays in cultured cells with intact HCV genome.

METHODSHuh-7 hepatoma cells were cultured and infected with in vitro constructed HCV. The total RNAs, proteins and cell culture supernatants of HCV infected cells and control cells were isolated. Proteins and cell culture supernatants were used to detect the HCV replication and protein expression in cell culture system. The HCV protein expression was detected with Western blotting. Released HCV from infected cells was analyzed by real-time fluorescence quantitative PCR. Total RNA was qualified using 10 g/L agarose gel electrophoresis. cRNA was synthesized, fluorescence labeled and purified, then hybridized with Agilent oligo microarray (20173 probes). Differential expression of genes related to transcription in cell culture system was analyzed.

RESULTHCV was positive in cell culture supernatants and HCV protein expression was also positive according to Western blotting results. Eleven up-regulated and 11 down-regulated genes related to transcription were found after Agilent gene chip screening.

CONCLUSIONIntact hepatitis C virus cell culture system provides an useful tool for study on the affects of HCV infection on transcription regulation genes in host cells.

Cell Line, Tumor ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genome, Viral ; Hepacivirus ; genetics ; growth & development ; Hepatocytes ; virology ; Humans ; Liver Neoplasms ; genetics ; pathology ; virology ; Transcription, Genetic

OBJECTIVETo evaluate the diagnostic values of four risk of malignancy indices (RMI) for malignant adnexal masses.

METHODSThe data of 223 women with adnexal masses admitted to the Department of Obstetrics and Gynecology of Peking Union Medical College Hospital for surgical exploration between June 2008 and December 2008 were retrospectively analyzed. The sensitivity, specificity, positive predictive value and negative predictive value of RMI1, RMI2, RMI3, and RMI4 in the diagnosis of malignant adnexal masses were calculated.

RESULTSWhen the cutoff levels of RMI1, RMI2, RMI3 were set at 200 and RMI4 at 450, the sensitivities for diagnosing malignant adnexal masses ranged 59.0%-67.2%, the specificities ranged 94.4%-96.9 %, the positive predictive values ranged 82.0%-87.8%, and the negative predictive values ranged 90.9%-92.6%. The Youdens indexes (YI) of RMI1, RMI2, RMI3, and RMI4 were 0.559,0.606,0.576, and 0.559, respectively. RMI2 was significantly different from RMI1 (P=0.000), RMI3 (P=0.008), and RMI4 (P=0.000) in terms of diagnostic efficiency. RMI1, RMI2, RMI3, and RMI4 at a cutoff level of 75.688.679.1, 177.2 respectively, according to ROC curves, yielded sensitivities of 77.8%-82.5%, specificities of 84.6%-90.1%, positive predictive values of 69.0%-75.4%, and negative predictive values of 90.9%-92.6%; the relevant YI of RMI1, RMI2, RMI3, and RMI4 were 0.635, 0.665, 0.651 and 0.705, respectively. Under this cutoff level, the difference between RMI1, RMI2, RMI3, and RMI4 in diagnosing malignancy had no statistic significant. The primary histological types arising false negative were early stage epithelial ovarian cancer and non-epithelial ovarian cancer. The primary histological types arising false positive were endometriosis masses and degenerative sex cord-stromal tumor.

CONCLUSIONSRMIs are useful indices for the differentiation between benign and malignant pelvic diseases. Meanwhile, their cutoff levels for Chinese populations need further study.

Adnexal Diseases ; diagnosis ; diagnostic imaging ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; CA-125 Antigen ; blood ; Female ; Humans ; Menopause ; Middle Aged ; Retrospective Studies ; Risk Assessment ; methods ; Sensitivity and Specificity ; Ultrasonography ; Young Adult

CD4+ T cells mainly interact with Sphingosine 1-phosphate (S1P) to regulate immune function through Sphingosine 1-phosphate receptor 1 (S1P1). This study was aimed to investigate the effects of recombinant human granulocyte-colony-stimulating factor (rhG-CSF) mobilization on S1P1 expression in CD4+ T cells of donor's peripheral blood. The CD4+T cells of peripheral blood were isolated by magnetic beads from 17 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and at fourth day of mobilization with rhG-CSF. The S1P1 expression was detected by real time quantitative PCR in the RNA extracted from CD4+ T cells collected before and after rhG-CSF mobilization. The results showed that the expression of S1P1 was found in CD4+ cells before and after rhG-CSF mobilization, but the expression level of SIP1 in CD4+ cells after rhG-CSF mobilization was significantly lower than that before rhG-CSF mobilization (p<0.01). It is concluded that the mobilization with rhG-CSF obviously down-regulates the expression of S1P1 in CD4+ T cells of donor's peripheral blood.
CD4-Positive T-Lymphocytes ; drug effects ; metabolism ; Granulocyte Colony-Stimulating Factor ; pharmacology ; Hematopoietic Stem Cell Mobilization ; methods ; Hematopoietic Stem Cell Transplantation ; Humans ; Receptors, Lysosphingolipid ; metabolism ; Recombinant Proteins