Objective:To observe the effect of acupoint massage plus acupoint sticking therapy for the stress reaction during postoperative anesthesia recovery period in patients undergoing nasal endoscopic surgery.Methods:A total of 120 patients undergoing nasal endoscopic surgery were included,and all patients were under trachea intubation and general anesthesia.The patients were randomized into an observation group and a control group,with 60 patients in each group.Patients in the control group received conventional anesthesia resuscitation,while patients in the observation group received acupoint massage plus acupoint sticking therapy on the basis of conventional anesthesia resuscitation.Changes in the heart rate (HR),systolic blood pressure (SBP) and diastolic blood pressure (DBP) were observed at three time points including the end of the surgery (TO),the removal of the tracheal tube (T1) and 10 min after the removal of the tracheal tube (T2).The awakening and tube removal time,choking cough and restlessness,and adverse reactions (dizziness,nausea and vomiting) in 24 h post-surgery period were compared.Results:At T1 and T2,the comparisons of HR,SBP and DBP between the two groups showed statistical significance (all P＜0.05).Intra-group comparisons showed that the HR,SBP and DBP of the control group at T1 and T2 were significant different from those at TO (all P＜0.05).There were significant differences in the awakening time and tube removal time between the two groups (both P＜0.05).The incidences of choking cough and restlessness were 8.3％ and 3.3％ respectively in the observation group,versus 53.3％ and 30.0％ in the control group,and the between-group comparisons showed statistical significance (both P＜0.05).The incidences of dizziness,nausea and vomiting in 24 h post-surgery period were 3.3％,5.0％ and 0.0％ respectively in the observation group,versus 43.3％,33.3％ and 25.0％ in the control group,and the between-group comparisons showed statistical significance (all P＜0.05).Conclusion:Acupoint massage plus acupoint sticking therapy can effectively regulate the stress reaction during postoperative anesthesia recovery period in patients undergoing nasal endoscopic surgery,and maintain a stable internal environment.

Adolescent ; Anemia, Aplastic ; drug therapy ; immunology ; Antilymphocyte Serum ; administration & dosage ; adverse effects ; Child ; Child, Preschool ; Drug Hypersensitivity ; etiology ; Female ; Humans ; Immunosuppressive Agents ; administration & dosage ; adverse effects ; Infant ; Male ; Retrospective Studies ; Serum Sickness ; chemically induced ; T-Lymphocytes ; immunology ; Treatment Outcome

OBJECTIVETo construct the recombinant plasmid of RCAS1, to express and purify its fusion protein GST-RCAS1, and to investigate its biological function.

METHODSRCAS1 encoding gene was amplified by RT-PCR from total RNA extract of MCF-7 cells and was ligated with expression plasmid vector pGEX-2T by T4 DNA ligase after digested by the restricted endonucleases BamH I and EcoR I. Then the ligated products were inserted into competence JM109 E. Coli and the positive recombinants were identified by restriction endonuclease digestion assay and DNA sequencing. The GST-RCAS1 fusion protein expression was induced by IPTG in BL21 E. Coli and was purified with GST column and identified by SDS-PAGE and Western blotting with anti-GST monoclonal antibody, anti-RCAS1 (N-18) and anti-RCAS1 (C-20) polyclonal antibody. The apoptosis of activated T cells induced by GST-RCAS1 fusion protein was detected by flow cytometry with Annexin V and propidium iodide (PI) staining.

RESULTA 642 bp product was cloned by RT-PCR and the recombinant plasmid was constructed successfully. The GST-RCAS1 fusion protein was recognized by GST monoclonal antibody and RCAS1(N-18 and C-20) polyclonal antibody. FACS analysis showed that GST-RCAS1 fusion protein induced apoptosis in activated T cells.

CONCLUSIONThe recombinant plasmid of RCAS1 has been successfully constructed and the GST-RCAS1 fusion protein expressed and purified. The apoptosis inducing effect of GST-RCAS1 fusion protein on activated T cells is demonstrated.

Antigens, Neoplasm ; biosynthesis ; genetics ; Base Sequence ; Breast Neoplasms ; genetics ; immunology ; Escherichia coli ; genetics ; metabolism ; Gene Expression ; Glutathione Transferase ; biosynthesis ; genetics ; Humans ; Molecular Sequence Data ; Plasmids ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; isolation & purification ; Tumor Cells, Cultured

OBJECTIVETo observe clinical effect of integrated Chinese medical (CM) treatment (as maintenance therapy) on the progression-free survival (PFS) and overall survival (OS) in patients with advanced non-small-cell lung cancer (NSCLC) after first-line chemotherapy.

METHODSThe study was a prospective, randomized, controlled clinical trial. Totally 69 non-progressive advanced NSCLC patients treated with first-line chemotherapy were randomly assigned to the test group (34 cases) and the control group (35 cases). Patients in the control group were treated with one Western drug chemotherapy (Gemcitabine or Alimta or docetaxel). Those in the test group were treated with integrated CM treatment (CM decoction, CM Intravenous preparation, and point application). Each cycle consisted of 21 days. Treatment lasted till the disease progressed, or intolerable toxic/adverse reactions occurred, or patients refused to continue the treatment. Patients' life spans were regularly followed-up.

RESULTS(1) The median cycle of maintenance therapy was 2 cycles for two groups with no statistical difference (P =0.274). The median PFS was 12.43 weeks in the test group and 10.00 weeks in the control group, showing statistical difference (P =0.025). The middle survival time (MST) was 18.8 months in the test group and 16.73 months in the control group, showing no statistical difference (P =0.437).

CONCLUSIONCM treatment (as maintenance therapy) showed quail effect to one Western drug chemotherapy in prolonging patients' life span.

Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; Deoxycytidine ; analogs & derivatives ; therapeutic use ; Disease-Free Survival ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Pemetrexed ; therapeutic use ; Prospective Studies ; Taxoids ; therapeutic use

0.37).Meanwhile a good correlation (Y=0.99X-0.18,r=0.987) was obtained. Though Westergren method correlated preferably with MICROTEST 1 (Y=0.86X+1.27,r=0.906),there was a markedly different (t=3.174,P=0.001).At last different references values were collected, according to sex and age.Male,32.5 mm/1 h(60 years old);Female, 34.03 mm/1 h(50 years old).Conclusions MICROTEST 1 correlated preferably with Westergren method.The examination by MICROTEST 1 needs small quantity of sample and fewer time.Furthermore,it has good repeatability and stability.The factors such as temperature and Hct have little influence on the results.The result suggested that it is suitable to apply MICROTEST 1 to large- scale clinical laboratory or other labs.But the reference value of ESR was influenced by age,which should be considered in clinical usage.

Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) is a promising anti-cancer agent. Epigallocatechin-3-gallate (EGCG) is a polyphenolic constituent of green tea. In this study, inhibitory effect of combined use of EGCG and TRAIL on human melanoma A375 cells was examined and the possible mechanism investigated. The cells were divided into 4 groups: control group, EGCG group (EGCG: 10, 20 mug/mL), TRAIL group (TRAIL: 25 ng/mL) and EGCG+TRAIL group (combined group). The growth inhibition was measured in the A375 cells treated with different concentrations of TRAIL ((25, 50, 75, 100, 125, 150 ng/mL) by MTT assay. The apoptosis was assessed by flow cytometry. The expressions of DR4 and DR5 were detected by flow cytometry and western blotting. The activities of caspase-8 and caspase-3 were determined by colorimetric assay. The results showed that TRAIL could dose-dependently inhibit the growth of A375 cells and the IC(50) of TRAIL was 150 ng/mL. The apoptosis rate was 11.8% in the TRAIL group, 5%-7% in the EGCG group and 48.9%-59.1% in the combined group. Significant difference was found in the apoptosis rate between the combined group and the EGCG or TRAIL group (P<0.05 for each). The expression of DR4 instead of DR5 was significantly increased in the EGCG group. The activity of caspase-3 rather than caspase-8 was substantially enhanced in the EGCG group. These results suggest that EGCG is useful for the TRAIL-based treatment for melanoma.

Objective: To evaluate the effect of NGX6 with 5-Fu on the apoptosis of colon cancer cells. Methods: The NGX6-transfected HT-29 cell line with 5-Fu was used in the test group. HT-29 cell line with 5-Fu and PDTC was used in the control group. The expression of NF-κB was detected by EMSA. The proliferation of HT-29 cell line was assayed by MTT. The effect of NGX6 on the apoptosis was detected by FCM. HT-29 cells were double-stained by PI/Annexin-V and AO/EB and observed by fluorescence microscopy. Results: The expression of NF-κB was inhibited in NGX6 transfected colon carcinoma cell group and in colon carcino-ma cell group treated with PDTC. Treatment with the chemopreventive compounds 5-Fu and PDTC resulted in different responses in the effects of anti-proliferation and induced apoptosis of colon carcinoma cells. There was no significant difference in apoptosis between NGX6-transfected HT-29 call line with 5-Fu and the cells in the control group. NGX6 gene enhanced the effect of 5-Fu on the proliferation and apoptosis of colon carcino-ma cells. Conclusion: NGX6 gene can induce apoptosis and inhibit the proliferation of colon carcinoma cells. NGX6 gene can enhance the effect of 5-Fu on the proliferation and apoptosis of colon carcinoma cells through NF-κB pathway.

OBJECTIVETo study lysosomes involvement in the degradation of ricin A chain.

METHODSA lysosome-targeted singal KFERQ was added to the C terminus of rRTA by DNA recombinant technology. A pKK223.3 expression system in E. coli was used to produce recombinant ricine A chain (rRTA) and rRTA-KFERQ. Recombinant proteins were purified by affinity chromatography using Blue-Sepharose 6B. The cytotoxicity of recombinant proteins was measured by the MTT method.

RESULTSRecombinant RTA-KFERQ was 49.87%, 54.18% and 88.68% less cytotoxic than RTA itself on the three cell lines HEPG2, Hela and A549, respectively.

CONCLUSIONLysosomes can degrade, but not completely inactivate RTA in different cells, suggesting cells may have other degradation pathways for RTA.

Chromatography, Affinity ; Escherichia coli ; genetics ; metabolism ; HeLa Cells ; Humans ; Lung Neoplasms ; pathology ; Lysosomes ; metabolism ; Recombinant Proteins ; genetics ; isolation & purification ; metabolism ; Ricin ; genetics ; metabolism ; Tumor Cells, Cultured

OBJECTIVETo construct a novel kind of nonviral gene delivery vector based on polyethylenimine (PEI) conjugated with polypeptides derived from ligand FGF with high transfection efficiency and according to tumor targeting ability.

METHODSThe synthetic polypeptides CR16 for binding FGF receptors was conjugated to PEI and the characters of the polypeptides including DNA condensing and particle size were determined. Enhanced efficiency and the targeting specificity of the synthesized vector were investigated in vitro and in vivo.

RESULTSThe polypeptides were successfully coupled to PEI. The new vectors PEI-CR16 could efficiently condense pDNA into particles with around 200 nm diameter. The PEI-CR16/pDNA polyplexes showed significantly greater transgene activity than PEI/pDNA in FGF receptors positive tumor cells in vitro and in vivo gene transfer, while no difference was observed in FGF receptors negative tumor cells. The enhanced transfection efficiency of PEI-CR16 could be blocked by excess free polypeptides.

CONCLUSIONThe synthesized vector could improve the efficiency of gene transfer and targeting specificity in FGF receptors positive cells. The vector had good prospect for use in cancer gene therapy.

Animals ; Binding Sites ; Carcinoma ; therapy ; Cell Line, Tumor ; Cell Survival ; drug effects ; Dose-Response Relationship, Drug ; Female ; Fibroblast Growth Factors ; metabolism ; Gene Transfer Techniques ; Genetic Vectors ; chemical synthesis ; chemistry ; pharmacology ; Humans ; In Vitro Techniques ; Ligands ; Liver Neoplasms ; therapy ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Particle Size ; Peptides ; chemistry ; metabolism ; pharmacology ; Polyethyleneimine ; chemistry ; metabolism ; pharmacology ; Prostatic Neoplasms ; therapy ; Receptors, Fibroblast Growth Factor ; drug effects ; genetics ; metabolism ; Structure-Activity Relationship ; Surface Properties ; Transfection ; Transplantation, Heterologous ; Xenograft Model Antitumor Assays

OBJECTIVETo investigate the influencing factors of polyethylenimine (PEI) in gene transfer in vitro.

METHODSCytotoxic effects of PEI on in vitro cultured NIH 3T3 cells were quantified by MTT assay. The interaction between PEI and DNA at different charge ratios was analyzed by agarose gel electrophoresis retardation assay. The expression of gene transfer was monitored in Cos-7 cells using pEGFP and pSV beta plasmids as the reporter gene systems. Influences of chloroquine, albumin, serum, salt ion strength, and Mg(2+) ion and other factors on PEI/DNA transfer efficiency were evaluated.

RESULTThe survival rate of NIH3T3 cells at 6 mg/L of PEI was 64.2% and at 7 mg/L of PEI was 54.4%. Gel electrophoresis retardation assays showed that PEI completely retarded DNA migration at 3.0 PEI nitrogen per DNA phosphate. Chloroquine enhanced the transfection efficiency of PEI. Albumin and serum in the culture medium decreased the transfection efficiency. HBS(HEPES buffered solution) or 150 mmol/L NaCl as the dilution solution of PEI/DNA was superior over 278 mmol/L glucose solution in the transfection efficiency. Mg(2+) in the dilution solution decreased the transfer efficiency of PEI/DNA.

CONCLUSIONPEI is efficient gene transfer agent of eukaryotes in vitro, and can be possibly used in vivo.

Animals ; COS Cells ; Cell Survival ; Chloroquine ; pharmacology ; Culture Media ; Gene Transfer Techniques ; Magnesium ; pharmacology ; Mice ; NIH 3T3 Cells ; Osmolar Concentration ; Polyethyleneimine ; pharmacology