AIM:To observe the inhibitory effect of siRNA targeting to Wip1 gene on the Wip1 gene expression in the colon cancer cells and to investigate the influence of Wip1 gene silencing on the chemotherapy sensitivity of colon cancer cells.METHODS:Wip1-811 siRNA targeting to Wip1 gene was transfected into RKO colon cancer cells with high expression of Wip1 gene.The mRNA expression of Wip1 was measured by real-time PCR.The protein level of Wip1 was detected by Western blotting.The viability of RKO colon cancer cells was measured by MTS assay.The cell apoptosis and cell cycle were analyzed by flow cytometry.RESULTS: Wip1-811 siRNA efficiently inhibited the expression of Wip1 at mRNA and protein levels.The enhanced chemotherapy sensitivity of RKO colon cancer cells was observed after inhibition of Wip1 gene expression.The viability of RKO colon cancer cells was decreased from (89.4 ±6.6)%to (74.7 ±3.9)%af-ter treated with 5-fluorouracil (P<0.05) and decreased from (77.9 ±2.4)%to (66.7 ±2.9)%after treated with oxali-platin ( P<0.05 ) .The cell apoptotic rate was increased from ( 7.7 ±0.5 )% to ( 12.3 ±3.2 )% and from ( 14.7 ± 2.1)% to (34.0 ±2.1)% when RKO colon cancer cells were treated with 5-fluorouracil and oxaliplatin, respectively (P<0.05).CONCLUSION:Wip1 gene silencing enhances chemotherapy sensitivity of colon cancer cells.

OBJECTIVETo investigate the effect of compound Danshen Droplet-pill (DS) combined with trimetazidine (TMZ) in treating senile unstable angina pectoris (SUAP).

METHODSOne hundred and twenty patients with SUAP were eaually and randomly divided into 2 groups, the treated group and the control group. Changes of angina, occurrence of arrhythmia, myocardial infarction and sudden death, myocardial ischemia in ECG and partial indexes of heart function were observed.

RESULTSThe total effective rate in the treated group was 78.3%, while that in the control group was 53.3%, comparison of the two groups showed significant difference (P < 0.05). The incidence rate of arrhythmia in the two groups was 18.2% and 30.0% respectively and that of acute myocardial infarction and sudden death was 0 and 5.0% respectively, also showed significant difference between them (P < 0.05). Condition of myocardial ischemia revealed in ECG and partial indexes of heart function in the treated group were all improved to certain extent. Conclusion DS combined with TMZ is superior in treating SUAP.

Aged ; Aged, 80 and over ; Angina, Unstable ; drug therapy ; Drug Administration Schedule ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Male ; Middle Aged ; Phenanthrolines ; therapeutic use ; Phytotherapy ; Trimetazidine ; therapeutic use ; Vasodilator Agents ; therapeutic use

OBJECTIVETo investigate the vascular activity of extract from mulberry leaves (EML) on rat thoracic aorta and the underlying mechanism.

METHODSIsolated thoracic rings of Sprague-Dawley rats were mounted on the organ bath and the tension of the vessel was recorded.

RESULT(1) EML produced a concentration-dependent vasorelaxation of aorta preconstricted by high K(+) (60 mmol/L) or 10(-6) mol/L phenylephrine (PE) in endothelium-intact and endothelium-denuded arteries. (2) EML at EC(50) concentration reduced the calcium dose-response curve. (3) After incubation of aorta with verapamil, EML induced vasocontraction of aorta preconstricted by PE, which was abolished by ruthenium red.

CONCLUSIONThe vascular effect of EML is biphasic, the vasorelaxation is greater than the vasocontraction. The vasorelaxation induced by EML may be mediated by inhibition of voltage-and receptor-dependent calcium channels in vascular smooth muscle cells, while the vasocontraction is via activation of ryanodine receptor in endoplasmic reticulum.

Acetates ; isolation & purification ; pharmacology ; Animals ; Aorta, Thoracic ; drug effects ; physiology ; Dose-Response Relationship, Drug ; In Vitro Techniques ; Male ; Morus ; chemistry ; Plant Extracts ; isolation & purification ; pharmacology ; Plant Leaves ; chemistry ; Potassium ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Ryanodine Receptor Calcium Release Channel ; physiology ; Vasoconstriction ; drug effects ; Vasodilation ; drug effects

OBJECTIVETo study the immuno-tolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells (BMSCs) in the allogeneic transplantation.

METHODSForty female C57BL/6 mice and forty male BALB/C mice were respectively used as donors and recipients in skin allogenic graft model. Forty male BALB/C mice were divided randomly into 4 groups: blank control group, CP group, BMSCs group , CP + BMSCs group, with 10 mice in each group. Before skin graft, high-dose abdominal injection of cyclophosphamide (200 mg/kg, 2 d, q. d.) was performed in recipient mice in CP and CP + BMSCs groups. On the transplantation day, a bonus of 2 x 10(6) BMSCs from the SD rat (SD-BMSCs) were injected through the tail vein in the BMSCs and CP + BMSCs groups. The observation and HE staining of skin grafts were used. The expressions of CD29, CD34, CD45 and CD90 of cells were analyzed by using flow cytometry in order to identify BMSCs. The CD4+, CD25+, Foxp3 and Treg cells of spleen were detected by flow cytometry. Cytokine in peripheral blood of recipient mice were measured by ELISA, including TGF-beta, IL-10 and IFN-gamma. T cells were co-cultured with 60Co-irradiated bone marrow MSCs from different individuals. The proliferative activity of T cells were evaluated with MTT assay.

RESULTSThe skin graft survival time was significantly prolonged in the CP + BMSCs group, as compared with that in the blank control group, the CP group, the BMSCs group, respectively. Cells cultured by whole bone marrow adherent cultivation showed CD29 (99.7%), CD44+ (96.7%), CD34- (1.6%), CD45- (1.3%). Compared with the control group and CP group, the ratio of the CD4+, CD25+, Foxp3+ and Treg cells significantly increased in the SD-BMSCs group and CP + BMSCs group (P < 0.05). Analysis of peripheral blood by ELISA showed significant high level of TGF-beta, IL-10 and low level of IFN-gamma in BMSCs group and CP group,compared with that in control group. When co-cultured with BMSCs from different individuals, T- lymphocytes proliferation decreased apparently in SD-BMSCs group and C57-BMSCs group (P < 0.05), but there was no significant difference between SD-BMSCs group and C57-BMSCs group (P > 0.05).

CONCLUSIONSThe immunotolerance mechanism of the third-party bone marrow-derived mesenchymal stem cells in the allogeneic transplantation might be associated with its effect on the proliferation of Treg cells and increasing expression of TGF-beta and IL-10, decreasing expression of IFN-gamma.

Animals ; Bone Marrow Cells ; immunology ; Female ; Immune Tolerance ; Interferon-gamma ; immunology ; Interleukin-10 ; immunology ; Male ; Mesenchymal Stem Cell Transplantation ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Rats ; Rats, Sprague-Dawley ; Skin Transplantation ; T-Lymphocytes, Regulatory ; immunology ; Transforming Growth Factor beta ; immunology ; Transplantation, Homologous

Objective To investigate the inhibiting effect of acupuncture at points Neiguan(PC6) and Shuigou(GV26) on caspase-3, and PARP-1 mRNA and protein expressions in the cerebral cortex in rats with cerebral ischemia-reperfusion. Method Forty male SD rats were randomized to normal, sham operation, model and acupuncture groups. A model of middle cerebral artery infarction was made by modified Longa thread occlusion. Neurological function was scored using a 6-grade 5-point Bederson's scale. Caspase-3, and PARP-1 mRNA and protein expressions in rat cerebral cortex were determined using real-time PCR and Western Blot at 24 hrs after cerebral ischemia. Result Neurological deficit was reduced in the acupuncture group of rats with cerebral ischemia-reperfusion compared with the model group (P<0.01). Acupuncture could down-regulate caspase-3, and PARP-1 mRNA and protein expressions and there was a statistically significant difference compared with the model group (P<0.05, P<0.01; P<0.05, P<0.05). Conclusion Acupuncture at points Neiguan and Shuigou can down-regulate caspase-3, and PARP-1 mRNA and protein expressions in the cerebral cortex in rats with cerebral ischemia-reperfusion.

OBJECTIVE: To investigate the function and gene expression of active efflux transporters in drug-resistant candida albicans. METHODS: The broth microdilution method was performed to determined the minimal inhibitory concentration of 4 antifungal drugs to 20 candida albicans. We compared the efflux of Rhodamine 6G between sensitive and some of fluconazole-resistant candida albicans, and screened out the resistant strains with significantly increased efflux of Rhodamine 6G. Reverse transcriptase-polymerase chain reaction (RT-PCR) was used to measure the mRNA levels of active efflux gene CDR1 and CDR2. RESULTS: The efflux of Rhodamine 6G was significantly increased in some fluconazole-resistant strains when glucose was added, and the gene expressions of CDR1 and CDR2 were also obviously increased, compared with those in the sensitive strains. CONCLUSION The excessive expression of active efflux pump gene is related to the resistance to fluconazole in candida albicans. The measurement of Rhodamine 6G efflux is a useful method for the identification of drug-resistant strains induced by the excessive expression of active efflux pump.
Antifungal Agents ; pharmacology ; Biological Transport ; Candida albicans ; genetics ; physiology ; Drug Resistance, Fungal ; genetics ; Fluconazole ; pharmacology ; Fungal Proteins ; genetics ; physiology ; Gene Expression Regulation, Fungal ; Membrane Transport Proteins ; genetics ; physiology ; Microbial Sensitivity Tests ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Rhodamines ; metabolism

OBJECTIVE: To explore the expression level of class I integrase (intI 1) mRNA in Acinetobacter baumannii from biofilm cells and planktonic cultured cells ,and to analyze the drug-resistance of Class I integron positive strains. METHODS: Acinetobacter baumannii were collected from hospitals,and Class I integron strains were screened by gene amplification. Total RNA of Class I integron positive strains was extracted, and the intI1 mRNA expression in the bioflim cells and planktonic cultured cells was measured by RT-PCR. Susceptibilities to antibiotics of Class I integron positive strains were also examined. RESULTS: The intI1 gene mRNA was expressed under 2 conditions, and the mRNA expressed in the biofilm cells was about 4 times higher than that in the planktonic cultured cells. Among the 64 strains of Acinetobacter baumannii, 46 strains were Class I integron positive strains. The antibiotic resistance of intI1 gene cassette-positive strains was higher than that of gene cassette-negative strains. CONCLUSION The intI1 gene mRNA can be up-regulated in Acinetobacter baumannii biofilm cells.Class I integron plays an important role in drug resistance. It is much easier to capture gene cassettes for bacteria under biofilm condition.
Acinetobacter Infections ; microbiology ; Acinetobacter baumannii ; genetics ; isolation & purification ; Base Sequence ; Biofilms ; Drug Resistance, Multiple ; genetics ; Humans ; Integrases ; biosynthesis ; genetics ; Molecular Sequence Data ; RNA, Messenger ; biosynthesis ; genetics

Although mesenchymal stem cells (MSCs) are increasingly used to treat graft-versus-host disease (GVHD), their immune regulatory mechanism in the process is elusive. The present study aimed to investigate the curative effect of third-party umbilical cord blood-derived human MSCs (UCB-hMSCs) on GVHD patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and their immune regulatory mechanism. Twenty-four refractory GVHD patients after allo-HSCT were treated with UCB-hMSCs. Immune cells including T lymphocyte subsets, NK cells, Treg cells and dendritic cells (DCs) and cytokines including interleukin-17 (IL-17) and tumor necrosis factor-alpha (TNF-α) were monitored before and after MSCs transfusion. The results showed that the symptoms of GVHD were alleviated significantly without increased relapse of primary disease and transplant-related complications after MSCs transfusion. The number of CD3(+), CD3(+)CD4(+) and CD3(+)CD8(+) cells decreased significantly, and that of NK cells remained unchanged, whereas the number of CD4(+) and CD8(+) Tregs increased and reached a peak at 4 weeks; the number of mature DCs, and the levels of TNF-α and IL-17 decreased and reached a trough at 2 weeks. It was concluded that MSCs ameliorate GVHD and spare GVL effect via immunoregulations.
Adolescent ; Adult ; Cord Blood Stem Cell Transplantation ; methods ; Cytokines ; metabolism ; Dendritic Cells ; metabolism ; Female ; Graft vs Host Disease ; immunology ; therapy ; Hematopoietic Stem Cell Transplantation ; adverse effects ; Humans ; Immunomodulation ; Killer Cells, Natural ; metabolism ; Male ; T-Lymphocyte Subsets ; metabolism ; Transplantation, Homologous ; adverse effects ; Young Adult

OBJECTIVETo study the low back pain(LBP) and its cause on female workers in flat-grained veneer wood industry.

METHODSBending posture was analyzed by observation and the prevalence of low back pain was investigated by physical examination and questionnaire among 299 female workers.

RESULTSThe prevalence of fatigue compliant in selecting, remending and sticking workers was 68.8%, 66.7% and 59.0%, respectively, which mainly involved in the part of low back. The prevalence of low back pain in selection (53.8%) and remending (58.7%) workers was higher than that in sticking workers (30.1%), which was in accordance with the tenderness between L4/L5 or L5/L6 and on the psoas major. Posture analysis indicated that the biggest bending range of selecting and remending workers (80 degrees ) was larger than that of sticking workers (60 degrees ), as well as the daily bending times[(4396+/-817), (1696+/-286), (1094+/-476)] and the time they kept bending[(6.5+/-0.6), (6.2+/-1.3), 4.5+/-0.9) h].

CONCLUSIONBending posture is common among female workers especially those who work in selecting and remending and might be the major causes for the high prevalence of LBP in flat-grained veneer wood industry.

Adult ; Female ; Humans ; Low Back Pain ; epidemiology ; etiology ; Occupational Diseases ; epidemiology ; etiology ; Posture ; Prevalence ; Risk Factors ; Surveys and Questionnaires ; Wood

OBJECTIVETo study the promoter hypermethylation of several tumor suppressor genes in gastric carcinoma (GC) tissue and adjacent normal gastric foveolar epithelium (GFE).

METHODSMethylation specific PCR (MSP) was used to examine the promoter methylation of tumor suppressor genes E-cadherin, hMLH1, APC and MGMT in paraffin-embedded gastric cancer tissue and adjacent normal foveolar epithelium in 106 cases.

RESULTSThe positive rate of genes promoter methylation was 44.3% (47/106 cases) and 72.6% (77/106 cases) at one or more genes tested in the normal GFE and GC tissue, respectively. There was a significant difference in the positive rates of gene promoter methylation between normal GFE and GC tissue (P = 0.0001). There was a significant association with Laurén classification, degree of differentiation and pTNM staging in GC (P < 0.05), but no significant association with Ming's classification (P > 0.05).

CONCLUSIONTumor suppressor genes promoter methylation is frequently present in GC and adjacent normal gastric foveolar epithelium, especially in Laurén diffuse type GC, poorly differentiated GC, mucus-secreting (signet ring) cell GC and pTNM stage III and IV GC. Our findings indicate that the gene promoter methylation is a common and early event in GC carcinogensis.

Adaptor Proteins, Signal Transducing ; genetics ; metabolism ; Adenomatous Polyposis Coli Protein ; genetics ; metabolism ; Adult ; Aged ; Aged, 80 and over ; Cadherins ; genetics ; metabolism ; DNA Methylation ; DNA Modification Methylases ; genetics ; metabolism ; DNA Repair Enzymes ; genetics ; metabolism ; Epithelium ; metabolism ; pathology ; Female ; Gastric Mucosa ; metabolism ; pathology ; Genes, APC ; Genes, Tumor Suppressor ; Humans ; Male ; Middle Aged ; MutL Protein Homolog 1 ; Neoplasm Staging ; Nuclear Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Suppressor Proteins ; genetics ; metabolism ; Young Adult