Objective To establish a method for studying molecular mechanism of Rhubarb inhibiting anti-Yersinia pesti based on DNA microarray.Methods A whole genome DN A microarray containing 4005 annotated genes of Yersiniapesti Was used.The minimal inhibitory concentration(MIC)of Rhubarb to Yersiniapestiwas determined by liquid dilution method.The gene expression profile of Yersinia pesti was performed after the exposure to Rhubarb at a concentration of 10×MIC for 30 minutes.The total RNA extracted and purified from Yersinia pesti Was reversely transfected to cDNA and labeled by Cy3-Cy5 dye.The labeled probes were hybridized to the microarray anti the results were obtained by a laser scanner and the microarray data was confirmed by real-time quantitative RT-PCR.Results The platform of the DNA microarray-based bacteria transcriptional profile was established.A total of 498 genes of Yersinia pesti changed significantly in response to Rhubarb.Among them.358 genes were up-regulated,140 down-reguated.Conclusions The whole genome DNA microarray can be used in the studying of molecular anti-Yersinia pesti mechanism of Rhubarb.

OBJECTIVETo study the effects of quercetin, a flavonoid, on the learning and memory ability of 3-day-old neonatal rats with hypoxic-ischemic brain white matter damage (WMD).

METHODSSixty 3-day-old Sprague-Dawley rats were randomly divided into four groups: control, WMD model,and quercetin treatment groups (20 and 40 mg/kg). There were 15 rats in each group. Rats in the WMD model and the two quercetin treatment groups were subjected to right common carotid artery ligation followed by 2 hrs of exposure to 8% O2 to induce periventricular white matter injury. After the operation quercetin was administered daily in the two quercetin treatment groups for 6 weeks. Six weeks later, Morris water maze and open-field tests were carried out to test memory and learning ability as well as behavior and cognition.

RESULTSFrom the second day of training, escape latency in the Morris water maze test was more prolonged in the WMD model group than in the control group (P<0.01). The escape latency in the two quercetin treatment groups was shortened significantly compared with the WMD model group (P<0.05). The WMD model group crossed the original platform fewer times compared with the control and quercetin treatment groups (P<0.05). The open-field test indicated that the number of rearings increased and time spent in the centre was extended in the WMD model group compared with the control group. Compared with the WMD model group, the number of rearings was significantly reduced (P<0.05) and time spent in the centre was significantly shortened in the quercetin treatment groups (P<0.05).

CONCLUSIONSQuercetin treatment can improve memory and learning ability as well as cognitive ability in neonates with WMD, suggesting that quercetin protects against WMD resulting from hypoxia-ischemia.

Animals ; Hypoxia-Ischemia, Brain ; drug therapy ; psychology ; Learning ; drug effects ; Maze Learning ; drug effects ; Memory ; drug effects ; Quercetin ; pharmacology ; Rats ; Rats, Sprague-Dawley

AIMTo prepare lipid microsphere of sodium norcantharidin (NCTD) and then study their characters and pharmacokinetic behavior.

METHODSDynamic Light Scattering, HPLC and retrodialysis technique were used to determine the in vitro characters of the NCTD loaded lipid microsphere (LM), such as the particle size, xi-potential, content, incorporation ratio, release profile and changes after dilute. And the plasma concentration was determined by HPLC-MS, compared with NCTD aqueous solution at the same time.

RESULTSEvery property showed that the LM was preferable. The average diameter was about 200 nm. The xi-potential was - 38 mV. The content was close to 100%. And the incorporation ratio exceeded 80%. After i. v. administration of single dose, the pharmacodynamic parameter of LM AUC was 111.28 microg x mL(-1) x h(-1). The data of plasma concentrations showed that the NCTD LM was of two compartment. There was no obvious difference between in vivo parameters of LM and reference solution.

CONCLUSIONThe NCTD LM was eligible and the character of it in vivo was not changed.

Animals ; Antineoplastic Agents ; administration & dosage ; blood ; pharmacokinetics ; Area Under Curve ; Bridged Bicyclo Compounds, Heterocyclic ; administration & dosage ; blood ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Female ; Lipids ; Male ; Mass Spectrometry ; Microspheres ; Particle Size ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effects of tumor suppressor gene PTEN on apoptosis and protein expression of p53 in HepG2 cells, as well as to explore its mechanisms.

METHODSHepG2 cells were transfected with GFP plasmids containing wild-type PTEN or G129E-PTEN and C124A-PTEN in vitro. Both the expression of wild-type p53 and the phosphorylation of protein kinase B (PKB/Akt) and focal adhesion kinase (FAK) were detected by Western blotting. Flow cytometry and confocal microscopy were used to analyze apoptosis of the transfected cells.

RESULTSCompared with the control, the expression of phosphorylated FAK and phosphoylated Akt were down-regulated in HepG2 cells transfected with wild-type PTEN (-65%, -93%) and G129E-PTEN (-65%, -35%), whereas the apoptosis percentage increased to (19.8+/-1.2)% and (9.2+/-0.6)%, and p53 expression was up-regulated by 120% and 50%, respectively. However, in the cells transfected with C124A-PTEN, neither the phosphorylation of FAK and Akt nor the apoptosis percentage and p53 expression had changed.

CONCLUSIONPTEN can dephosphrylate FAK through its protein phosphatase activity, and suppress phosphorylation of Akt mainly through its lipid phosphatase activity. Consequently, it can induce apoptosis of HepG2 cells and up-regulate p53 expression.

Apoptosis ; drug effects ; Carcinoma, Hepatocellular ; genetics ; pathology ; Gene Expression Regulation, Neoplastic ; Humans ; Liver Neoplasms ; genetics ; pathology ; PTEN Phosphohydrolase ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics ; Up-Regulation

OBJECTIVETo investigate the effect and mechanisms of tumor suppressor gene PTEN on the induction of anoikis of hepatocellular carcinoma SMMC-7721 cells.

METHODSSMMC-7721 cells were transfected with GFP plasmids containing wild-type PTEN or phosphatase inactivating mutant PTEN (C124A-PTEN) in vitro; The PTEN expression and the phosphorylation levels of focal adhesion kinase (FAK) and protein kinase B (PKB/Akt) were detected by Western blotting; Flow cytometry assay and laser scanning confocal microscopy were used to analyze apoptosis in adherent and non-adherent cells.

RESULTSCompared with the control, PTEN expression in the cells transfected with wild-type PTEN increased to 248%, while the phosphorylation level of FAK and Akt decreased 65.2% and 89.1%, respectively; and the anoikis percentage increased from 9.5% to 31.3%. In the cells transfected with C124A-PTEN, neither the phosphorylation of FAK and Akt nor the anoikis percentage had obviously changed, although the PTEN expression enhanced dramatically in comparison with the control.

CONCLUSIONThrough its phosphatase activity, tumor suppressor gene PTEN can suppress the phosphorylation of FAK and Akt, and induce anoikis in hepatocellular carcinoma cells.

Anoikis ; physiology ; Carcinoma, Hepatocellular ; pathology ; Focal Adhesion Protein-Tyrosine Kinases ; metabolism ; Humans ; Liver Neoplasms ; pathology ; PTEN Phosphohydrolase ; biosynthesis ; genetics ; Phosphoric Monoester Hydrolases ; metabolism ; Phosphorylation ; Tumor Cells, Cultured

OBJECTIVETo determine whether there was excessive risk of cancer among workers exposed to chrysotile fiber alone by applying a meta-analysis technique.

METHODSAll data meeting the criteria of cohort studies on cancer mortality among workers exposed only to chrysotile were incorporated into meta-analysis. Pooled standardized mortality ratios (SMRs) and their corresponding 95% confidence intervals (CIs) for main cancer sites were calculated using two approaches of unweighted ratio and random effect model. The heterogeneity and its sources of the results were examined with a Q-statistic and Z-score test. The dose-response effect as reflected in the percentage of all deaths due to mesothelioma served as a proxy measure of chrysotile exposure.

RESULTSA cohort of twenty six workers exposed to chrysotile alone was summarized. The significantly elevated meta-SMRs for all deaths (1.27), all cancers (1.28), cancers of respiratory organs (2.51), cancers of lung (2.35) and cancers of stomach (1.24) were observed. The significantly elevated meta-SMRs for lung cancer within occupational strata were observed among textile workers (3.55), asbestos product manufacturers (3.30), miners and millers (2.24), cement product workers (1.22), and for stomach cancer among asbestos product manufacturers (1.49). Meta-SMRs for cancers at other sites were not significant. Meta-SMR for lung cancer showed an increasing trend with an elevated percentage of all deaths from mesothelioma, but no such trend for stomach cancer.

CONCLUSIONThere are excessive risks of lung cancer and mesothelioma among workers exposed to chrysotile fiber alone, and likely no convincing indication of an etiological association between chrysotile exposure and cancers at other sites.

Asbestos, Serpentine ; adverse effects ; Cohort Studies ; Dose-Response Relationship, Drug ; Humans ; Lung Neoplasms ; etiology ; mortality ; Mesothelioma ; etiology ; mortality ; Neoplasms ; chemically induced ; mortality ; Occupational Exposure ; Occupational Health ; Risk Assessment ; Stomach Neoplasms ; etiology ; mortality

Objective To observe a variety of the toxicity,and the incidence of super-high-dose cyclophosphamide as a main chemotherapy scheme for treating children with high risk neuroblastoma,and to summarize the prevention and cure of the disease.Methods The high risk neuroblastoma patients who were diagnosed in Hematologic Tumor of Beijing Children's Hospital Affiliated to Capital Medical University were selected.And they were received super high dose of cyclophosphamide mainly chemotherapy scheme-CAV (cyclophosphamide,adriamycin and vincristine) more than 2 courses according to BCH-NB-HR(Beijing Children's Hospital-Neuroblastoma-High Risk) scheme.Their clinical symptoms,signs,blood routine and blood biochemistry in 1-2-course of treatment were retrospectively analyzed and summarized.According to WHO anti-cancer drugs in acute and subacute toxicity reaction grading standards,the data were analyzed.Results There were 33 children consistent with the criterion of research,who received 66 times of super-high-dose cyclophosphamide as a main chemotherapy scheme.All of the patients had bone marrow suppression,granulocyte generated Ⅳ grade toxicity 56 (84.8％),white blood cell (＜ 1.0 × 109/L) Ⅳ grade toxicity was 56 (84.8％),and blood platelet(＜25 x 109/L) Ⅳ grade toxicity was 19(28.8％).Hemoglobin(＜65 g/L) Ⅳ grade toxicity was 4 (6.06％).The digestive system toxicity including nausea and vomiting toxicity (100.0％).Urogenital system toxicity:Hemorrhagic cystitis only was 1 (1.52％) ;Cardiac Ⅰ-Ⅲ grade toxicity was 3(4.55％).But skin and nervous system toxicity was not observed.No patients died from chemotherapy.Conclusions There is a high incidence of toxicity with super-high-dose of cyclophosphamide as a main chemotherapy scheme,and the highest incidence ineludes bone marrow suppression,digestive tract response and secondary infection.But side effects like hemorrhagic cys-titis,liver,kidney and cardiac toxicity are not significant.Super-high-dose cyclophosphamide chemotherapy scheme is clinically safe under the perfect supportive treatment and rigorous monitoring.

OBJECTIVETo investigate the clinical therapeutic effect of methylprednisolone combined with cyclophosphamide and Etanercept method on acute paraquat poisoning.

METHODS136 patients with acute paraquat poisoning were divided into the normal therapy group and the intensive therapy group randomly. Methylprednisolone, cyclophosphamide and Etanercept were used in the intensive therapy group. Methylprednisolone 500 mg was given intravenously per day for continuous three days followed by 200 mg intravenous per day. Then methylprednisolone was decreased gradually 14 d or 21 d later according to the patient's condition. Cyclophosphamide 600 mg was given intravenously twice weekly for 2 weeks and Etanercept 25 mg was given hypodermic injection twice weekly for 3 weeks. Curative effect evaluation was done at 7, 14, 21 d and 12 weeks after therapy.

RESULTSThe survival rate of the intensive therapy group was obviously higher than that of the normal therapy group (P<0.01) on 7, 4, 21 d and 12 weeks. The cure rate of the intensive group were 94.6% (intake dose<50 ml 20% paraquat solution), 75.0% (intake dose 50 approximately 100 ml 20% paraquat solution), 12.5% (intake dose>100 ml 20% paraquat solution) respectively, while the cure rate of the normal group were 16.7% (intake dose<50 ml 20% paraquat solution), 8.3% (intake dose 50 approximately 100 ml 20% paraquat solution), 0% (intake dose>100 ml 20% paraquat solution) respectively. The total cure rate of the intensive therapy group (78.3%) 12 weeks later was higher than that of the normal group (11.9%).

CONCLUSIONMethylprednisolone combined with cyclophosphamide and Etanercept intensive therapy has the curative effect on acute paraquat poisoning.

Acute Disease ; Adolescent ; Adult ; Cyclophosphamide ; administration & dosage ; therapeutic use ; Drug Therapy, Combination ; Etanercept ; Female ; Humans ; Immunoglobulin G ; administration & dosage ; therapeutic use ; Male ; Methylprednisolone ; administration & dosage ; therapeutic use ; Middle Aged ; Paraquat ; poisoning ; Poisoning ; drug therapy ; Receptors, Tumor Necrosis Factor ; administration & dosage ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo evaluate the efficacy of salvaged allogeneic hematopoietic stem cell transplantation (allo-HSCT) for refractory/recurrent acute myeloid leukemia (AML).

METHODSA total of 45 patients with refractory/recurrent AML were enrolled from September 2006 to April 2010. The median blasts in bone marrow (BM) were 36% (20% to 92%) before conditioning. The donors were identical siblings (6) or unrelated ones (9) or haploidentical family members (30). Conditioning regiments were individualized according to patients' status, the regimen with high-dose cytarabine plus BuCy/CY was mostly used (20). The patients with impaired organ function received above regimen except using fludarabine instead of cyclophosphamide (16). FLAG followed by reduced-intensified BuCy was employed for the recipients with more than 40% blasts in BM (6) to reduce leukemia burden. TBI/CY or TBI/Fludarabine was used for the recipients with extramedullary infiltration of leukemia or multidrug resistant leukemia. G-CSF, MTX, NVT, Vm26, Acla or Thaltipa was added into conditioning regiments according to leukemia character.

RESULTSAll but 2 patients attained durable engraftment. The incidence of grade II to IV aGVHD and cGVHD were 34%, 59.1%, respectively. With median follow-up 30 (0.5 - 57) months, the relapse rate was 29.2%. Twenty-nine of 45 (60.2%) patients remained in complete remission since salvaged HSCT. Three-years disease-free survival and overall survival were 60.2% and 62.6%, respectively.

CONCLUSIONOur results indicated that the combination of salvaged HSCT with prophylactic immunotherapy might be a promising modality for treatment of refractory/recurrent AML, even with high leukemia burden.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Hematopoietic Stem Cell Transplantation ; methods ; Humans ; Leukemia, Myeloid, Acute ; mortality ; therapy ; Middle Aged ; Recurrence ; Survival Rate ; Transplantation Conditioning ; methods ; Treatment Outcome ; Young Adult

Objective To group and characterize natural plague foci in China.Methods A novel two-class typing method as well as a three-indication nomenclature method were established to group and characterize the natural plague foci,on the basis of eco-geographical landscapes of plague foci,genetics of Yersinia pestis,zoology of rodent reservoirs and the entomology of flea vectors.Results A total of 12 distinct natural plague foci (including 19 subtypes) as well as their biological features were characterized.Conclusion Natural plague foci in China were grouped and characterized in this study.