Objective To study the molecular biology of rifampin-depending M. Tuberculosis. Methods The seguence (a 319-bp DNA fragment) of rpoB gene were analyzed by automated DNA sequencing machine. (2) The fingerprints of genomic DNA were obtained by random amplified polymorphic DNA (RAPD) fingerprinting. (3)The protein electrophoresis of bacterium by SDS-polyacrylamide gel (SDS-PAG).(4) The cases of pulmonary tuberculosis by rifampin-depending strains were retrospectively analyzed. Results (1) rpoB gene sequenced: The point mutationrate of rifampin-depending strainswas 96.7%(29/30) and that of rifampin-residtant strains 81.1%(30/37), P

Background and purpose:Ara-C is one of the most effective and common agents in the treatment of acute nonlyphocytic leukemia. Telomerase is a unique complex of ribonucleoprotein. It plays an important role in the pathogenesis and development of cancer. In this study, we investigate the changes of mRNA expression of telomerase subunits in HL-60 cells induced by Ara-c and try to come up with a theory that could help to assess the efficacy of Ara-C. Methods:The combinations of various Ara-C concentration and the incubation time were used to treat HL-60. The ratios of apoptotic cell to necrosis cell were determined by flow cytometry and the expressions of telomerase subunits mRNA were evaluated by RT-PCR.Results:① There was no influence on transcription of telomerase subunits gene after HL-60 cells was cultured with 0~0.2ug/ml Ara-C for 12 hours;② 2ug/ml and 10ug/ml of Ara-C could down regulate the expression of hTERT from 0.80+0.07 to 0.50+0.04 and 0.39+0.03, not hTR and hTP1;③ with longer incubation with 10ug/ml of Ara-C, the percentage of apoptosis could be increased. The maximal induction of apoptosis (18.16+4.25%) could be reached at 12hrs treatment of Ara-C, then gradually decreased later on. The rate of necrosis increased with time, the maximal percentage(57.94+12.03%) of necrosis was observed at 48hrs of incubation time with drug. The mRNA level of hTERT gene also decreased along with the cultured time , the lowest value (0.18+0.03) has been documented at 48hrs time point, but not hTR、TP1.Conclusions:① Ara-C could down-regulate the expression of hTERT mRNA in a dose-and time-dependent manner, but not hTR、hTP1;② There might be no relationship between the percentage of apoptosis induced by Ara-C apoptosis and the expression of telomerase hTERT gene mRNA, but a close relationship between necrosis and the expression of hTERT mRNA has been found.

OBJECTIVETo explore whether daunorubicin (DNR) combined with cytosine arabinoside (Ara-C) and DNR alone have similar effect on acute promyelocytic leukemia (APL) cell line NB4 and acute myeloblastic leukemia cell line HL-60 in vitro.

METHODSCell morphology, cells viability, and cell apoptosis (Annexin-V by flow cytometry assay) were analysed.

RESULTSAfter incubation with DNR plus Ara-C for 24 hours,NB4 cell viability [(36.75 +/- 3.82)%] (n = 6) and cell apoptosis rate [(21.24 +/- 5.82)%] (n = 3) did not change significantly compared to that treated with DNR alone for 24 hours [(35.73 + 6.28 )%, (22.55 +/- 3.26)%, respectively] (P > 0.05). However, HL-60 cell viability [(67.17 +/- 2.07)%] and cell apoptosis rate [(48.05 +/- 0.92)%] changed significantly in DNR plus Ara-C group compared with DNR alone [(63.31 +/- 1.80)% ,(41.51 +/- 0.89)%, respectively] (P < 0.01 and < 0.05, respectively).

CONCLUSIONDNR plus Ara-C and DNR alone have similar effect on NB4 cells, but have different effect on HL-60 cells.

Apoptosis ; drug effects ; Cytarabine ; pharmacology ; Daunorubicin ; pharmacology ; HL-60 Cells ; drug effects ; Humans ; Leukemia, Promyelocytic, Acute ; pathology

Objective To observe the effects of chloroquine phosphate on apoptosis of leukemic cell line U937, and investigate whether chloroquine phosphate induces leukemic cell apoptosis by normalizing protein PNAS-2's abnormal subcellular location. Methods Chloroquine phosphate of different concentrations were added into culture fluid of leukemic cell line U937 at logarithmic phase. MTr was used to measure cell proliferation, flow cytometry and laser confocal microscopy were applied to detect cell apoptosis, and immunofluorescence technology was employed to observe the effects of chloroquine phosphate on the changes of subcellular location of protein PNAS-2. Results Apoptosis of leukemic cell line U937 was significantly induced by 50 μg/mL chloroquine phosphate, and subcellular location of protein PNAS-2 was changed. Conclusion Chlorequine phosphate can induce apoptosis of leukemic cell line U937, and the mechanism may be related to the normalization of PNAS-2's abnormal subcellular location in U937 cell line. Chloroquine phosphate has the potential to be used in leukemic therapy.

The experiment's aim was to optimize the processing technology of Xanthii Fructus which through comparing the difference of UPLC fingerprint and contents of toxicity ingredient in water extract of 16 batches of processed sample. The determination condition of UPLC chromatographic and contents of toxicity ingredient were as follows. UPLC chromatographic: ACQUITY BEH C18 column (2.1 mm x 100 mm, 1.7 microm) eluted with the mobile phases of acetonitrile and 0.1% phosphoric acidwater in gradient mode, the flow rate was 0.25 mL x min(-1) and the detection wavelength was set at 327 nm. Contents of toxicity ingredient: Agilent TC-C18 column (4.6 mm x 250 mm, 5 microm), mobile phase was methanol-0.01 mol x L(-1) sodium dihydrogen phosphate (35: 65), flow rate was 1.0 mL x min(-1), and detection wavelength was 203 nm. The chromatographic fingerprints 16 batches of samples were analyzed in using the similarity evaluation system of chromatographic, fingerprint of traditional Chinese medicine, SPSS16.0 and SIMCA13.0 software, respectively. The similarity degrees of the 16 batches samples were more than 0.97, all the samples were classified into four categories, and the PCA showed that the peak area of chlorogenic acid, 3,5-dicaffeoylquinic acid and caffeic acid were significantly effect index in fingerprint of processed Xanthii Fructus sample. The outcome of determination showed that the toxicity ingredient contents of all samples reduced significantly after processing. This method can be used in optimizing the processing technology of Xanthii Fructus.
Caffeic Acids ; analysis ; toxicity ; Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; toxicity ; Quinic Acid ; analogs & derivatives ; analysis ; toxicity ; Xanthium ; chemistry ; classification

Adolescent ; Adult ; Aged ; Child ; Cholinesterase Reactivators ; therapeutic use ; Female ; Hemoperfusion ; Humans ; Insecticides ; poisoning ; Male ; Middle Aged ; Organophosphate Poisoning ; Poisoning ; therapy

BACKGROUND: The effect of pituitary adenylate cyclase activating polypeptide (PACAP) during traumatic brain injury (TBI) and whether it can modulate secondary injury has not been reported previously. The present study evaluated the potential protective effects of ventricular infusion of PACAP in a rat model of TBI. METHODS: Male Sprague Dawley rats were randomly divided into 3 treatment groups (n=6, each): sham-operated, vehicle (normal saline)+TBI, and PACAP+TBI. Normal saline or PACAP (1g/5L) was administered intracerebroventricularly 20 minutes before TBI. Right parietal cortical contusion was produced via a weight-dropping method. Brains were extracted 24 hours after trauma. Histological changes in brains were examined by HE staining. The numbers of CD4+ and CD8+ T cells in blood and the spleen were detected via flow cytometry. RESULTS: In injured brain regions, edema, hemorrhage, inflammatory cell infiltration, and swollen and degenerated neurons were observed under a light microscope, and the neurons were disorderly arrayed in the hippocampi. Compared to the sham group, average CD4+ CD8– lymphocyte counts in blood and the spleen were significantly decreased in rats that received TBI+vehicle, and CD4– CD8+ were increased. In rats administered PACAP prior to TBI, damage was attenuated as evidenced by significantly increased CD4+, and decreased CD8+, T lymphocytes in blood and the spleen. CONCLUSION: Pretreatment with PACAP may protect against TBI by influencing periphery T cellular immune function.

Background and purpose:Vascular endothelial growth factor(VEGF) is a potent angiogenic mediator and angiogenesis has important effects on tumor growth and metastasis.The present study was to investigate the relationship between genetic polymorphism of VEGF and heredity risk factor of lung cancer.Methods:VEGF genotypes were determined by PCR-RFLP method in 171 patients with lung cancer and 172 healthy controls.Software PHASE 1.0 was used to construct the haplotypes of every individual.Unconditional logistic regression model was used to analyze the statistical association of genotypes or haplotypes in the two groups adjusted by gender and age. Results:Individuals with at least one-2578A allele had a significantly decreased risk of lung cancer compared with those carrying-2578CC genotype.When the analyses were stratified by gender,the combined-2578 CA and AA genotype,were associated with a considerably reduced risk of lung cancer(P=0.001,OR=0.303,95%CI=0.15 3-0.601).The distribution of the two haplotypes(936C/-2578C and 936C/-2578A) among overall lung cancer cases was significantly different from that among the controls(P=0.016,0R=0.317,95%CI=0.124-0.809 and P=0.018,OR=0.547, 95%CI=0.331-0.903).When the cases were categorized by tumor histology,the distribution of C-C haplotype in the adenocarcinoma(AC) group was associated with a substantially lowered risk of AC(P=0.004,0R=0.237,95%CI=0.090- 0.627),compared with the reference haplotypes.Conclusion:VEGF polymorphism may be a critical risk for the genetic risk factor to lung cancer.

OBJECTIVETo compare the clinical effect of acupuncture treatment and western medicine Carbamazepine for thalamic pain.

METHODSCrossover trial design was used, 11 cases diagnosed as thalamic pain were randomly divided into two groups according to the mini-unbalance-index method, group I (with 6 cases received acupuncture first and then western medicine) and group II (with 5 cases received western medicine first and then acupuncture). When the effects were evaluated, the two groups were named as acupuncture group and western medicine group, 11 cases in each group. The method of clearing away the heart fire, regulating the spirit, activating blood and relieving pain was adopted in acupuncture treatment, Ximen (PC 4), Yinxi (HT 6), Xuehai (SP 10) and Zhaohai (KI 6) were selected; the western medicine group was treated with oral administration of Carbamazepine, and one course as well as the eluting period were both 10 days. The effects were evaluated with visual analogue scale (VAS) and evaluation scale of Anderson Cancer Center pain in US (MD Pain Evaluation value) respectively.

RESULTSThe VAS and MD value in two groups were obviously decreased after treatment (both P < 0.05), while there was no significant difference between two groups; the markedly effective rate of pain relieving in acupuncture group was 63.6% (7/11), which was higher than that of 36.4% (4/11) in western medicine group, but there was no significant difference between two groups.

CONCLUSIONAcupuncture treatment of regulating spirit, activating blood and relieving pain has a better therapeutic effect for thalamic pain, and can reach to the same therapeutic effect with western medicine Carbamazepine.

Acupuncture Therapy ; Adult ; Aged ; Blood Circulation ; Carbamazepine ; therapeutic use ; Cross-Over Studies ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Pain ; physiopathology ; Pain Management ; Pain Measurement ; Spirituality ; Thalamus ; physiopathology

OBJECTIVETo investigate the effects of quercetin on cell morphology and VEGF expression of acute myeloblastic leukemia cells NB4 in vitro.

METHODSThe cytomorphology of NB4 cells was assessed by Wright-stain, apoptosis rate by apoptotic marker Annexin V, and VEGF secretion level by ELISA.

RESULTSTypical apoptosis was found in NB4 cells after treatment with quercetin. Apoptotic marker Annexin V analysis showed that the apoptotic rate of NB4 cells was increased after treatment with quercetin. The secretion of VEGF of NB4 cells was significantly decreased after treatment with quercetin.

CONCLUSIONQuercetin can induce apoptosis and inhibit secretion of VEGF in NB4 leukemia cells.

Antineoplastic Agents, Phytogenic ; pharmacology ; Apoptosis ; drug effects ; Cell Line, Tumor ; Humans ; Leukemia, Myeloid, Acute ; metabolism ; pathology ; Quercetin ; pharmacology ; Vascular Endothelial Growth Factor A ; secretion