Objective　To investigate what effects BmKAS-1 (a polypeptide purified from the Chinese scorpion Buthus martensi Karsch ［BmK］ and named as BmK activator of skeletal-muscle ryanodine receptor) and its upstream mixture BmK1-3-2 have on Na+ channels in dorsal root ganglion (DRG) small diameter neurons. Methods　The whole-cell patch-clamp technique was used to investigate the effects of BmKAS-1 and BmK1-3-2 on Na+ current in rat small diameter DRG neurons. Results　About 50% peak Na+ current was suppressed by 10*!μg/ml of BmK1-3-2. 1.62*!μg/ml of BmKAS-1 also blocked 50% peak Na+ current, and there was an obvious dose-dependent relationship. Conclusion　Both BmK1-3-2 and BmKAS-1 have a blocking effect on Na+ channels, and this may one of the mechanisms for the analgetic effect of BmK1-3-2 and BmKAS-1.

BACKGROUND AND OBJECTIVEThe level-Ib lymph node metastasis is rare in nasopharyngeal carcinoma (NPC). When and how this level should be irradiated with precise radiotherapy remains controversial. This study evaluated the prevalence and prognostic significance of level-Ib lymphadenopathy on the prognosis of NPC patients.

METHODSFrom January 1990 and December 1999, 933 newly diagnosed patients with NPC treated at Sun Yat-sen University Cancer Center were randomly selected, examined with computed tomography (CT) imagining for evidence of level-Ib lymphadenopathy before treatment. All patients received radical radiotherapy with or without chemotherapy. The relationship between level-Ib lymphadenopathy and post-treatment outcomes including overall survival (OS), locoregional recurrence-free survival (LRFS), and distant metastasis-free survival (DMFS) were analyzed using Kaplan-Meier methods. The Cox proportional hazards regression model was used to adjust for other prognostic factors.

RESULTSOf the 933 patients, 55 (5.9%) were found to have level-Ib lymphadenopathy, which was associated with carotid sheath involvement, oropharynx involvement and levels, and lateral cervical lymph node involvement. In the subgroup with carotid sheath involvement, with multivariate analysis accounting for all previously known prognostic factors, level-Ib lymphadenopathy was still associated with a risk of decreased OS (RR, 2.124; P<0.001), DMFS (RR, 2.168; P<0.001), and LRFS (RR, 1.989; P=0.001).

CONCLUSIONLevel-Ib lymphadenopathy in the patients with carotid sheath involvement is an independent prognostic factor.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; diagnostic imaging ; drug therapy ; pathology ; radiotherapy ; Chemotherapy, Adjuvant ; Child ; Cobalt Radioisotopes ; therapeutic use ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; radiotherapy ; Neck ; pathology ; Neoplasm Metastasis ; Neoplasm Recurrence, Local ; Particle Accelerators ; Pharynx ; pathology ; Prognosis ; Proportional Hazards Models ; Radiography ; Radioisotope Teletherapy ; Retrospective Studies ; Survival Rate ; Young Adult

BACKGROUNDExhaled nitric oxide (NO) is a noninvasive biomarker of airway inflammation in pulmonary diseases. Hydrogen sulfide (H2S), as the third member of the gasotransmitter family, is involved in the pathophysiological process in lung diseases. H2S also exists in exhaled breath and can be sampled non-invasively. The study investigated the level of exhaled H2S in patients with chronic obstructive pulmonary disease (COPD) and its correlation with exhaled NO.

METHODSLevels of exhaled NO and H2S, lung function, and cell differential counts in induced sputum were studied in 19 patients with acute exacerbation of COPD (AECOPD), 19 patients with stable COPD and seven healthy smoke controls.

RESULTSExhaled H2S levels were similar in patients with AECOPD (10.0 parts per billion (ppb), 8.0-13.0 ppb), stable COPD (10.0 ppb, 9.0-12.0 ppb), and healthy controls (9.0 ppb, 8.0-16.0 ppb) (P > 0.05). Exhaled NO levels were similar in patients with AECOPD (155.0 ppb, 129.0-190.0 ppb), stable COPD (154.0 ppb, 133.0-175.0 ppb) and healthy controls (165.0 ppb, 112.0-188.0 ppb) (P > 0.05). Exhaled H2S levels correlated positively with exhaled NO in all healthy controls and patients with COPD (r=0.467, P < 0.01). No significant correlation was found between the exhaled H2S level and percentage of predicted FEV1 (P > 0.05) and proportion of different cell types in induced sputum (P > 0.05).

CONCLUSIONSThere is a correlation between exhaled H2S and exhaled NO. The role of exhaled H2S in airway inflammation in COPD still needs further investigation.

Aged ; Breath Tests ; Female ; Forced Expiratory Volume ; physiology ; Humans ; Hydrogen Sulfide ; metabolism ; Male ; Middle Aged ; Nitric Oxide ; metabolism ; Pulmonary Disease, Chronic Obstructive ; metabolism ; physiopathology

OBJECTIVETo study the role of Lon gene in tumor cell proliferation, apoptosis and cell stress response.

METHODSSmall interfering RNAs (smRNAs) for Lon gene were designed using Ambion software and synthesized. The recombinant plasmid pSilencer U6 2.1/Lon was constructed with the smRNAs and pSilencer U6 2.1, followed by transfection into MCF7 cells via Lipofectamine(TM) 2000. The positive cLones were detected by RT-PCR 24 h after cell transfection. The transfected MCF7 cells were then subjected to cisplatin treatment, ultraviolet (UV) exposure and heat stress, respectively, after which the cells growth was tested with MTT assay and the measurements were plotted against time or concentration depending on the treatment administered. Apoptosis of MCF7 cells following the treatments was measured with flow cytometry.

RESULTSThe mRNA of Lon gene was downregulated in cells transfected with the recombinant plasmid pSilencer U6 2.1-Lon, and RT-PCR fail to detect the specific band of Lon as could be detected in untransfected and mock-transfected MCF7 cells. MTT assay showed that pSilencer U6 2.1-Lon transfection resulted in reduced cell proliferation capacity. Stress response test revealed that MCF7 cells with Lon gene down-regulation enhanced cell sensitivity for UV and cisplatin, which was not observed for non-transfected or mock transfection group. The same changes were also observed for heat stress exposure at 41 degrees Celsius;, but not at 43 degrees Celsius; or 45 degrees Celsius;. Increased cell apoptosis rate from (1.14-/+0.79)% to (22.47-/+3.15)% occurred following pSilencer U6 2.1-Lon transfection of the cells.

CONCLUSIONSLon gene can be significantly downregulated by introduction of siRNA in MCF7 cells to result in enhanced sensitivity of MCF7 cells to UV, cisplatin and heat stress.

Antineoplastic Agents ; pharmacology ; Apoptosis ; Breast Neoplasms ; genetics ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Cell Survival ; drug effects ; radiation effects ; Cisplatin ; pharmacology ; Dose-Response Relationship, Drug ; Female ; Humans ; Protease La ; genetics ; RNA Interference ; Reverse Transcriptase Polymerase Chain Reaction ; Temperature ; Time Factors ; Ultraviolet Rays

OBJECTIVETo determine the effect of activation of lambda-opioid receptor with U50, 488H, a selective kappa-opioid receptor agonist, on the changes in electrical coupling during prolonged ischemia and to explore the possible mechanism.

METHODSThe isolated rat heart was perfused in a Langendorff apparatus. The effect of U50, 488H on electrical coupling parameters including onset of uncoupling, plateau time, slope and fold increase in r(t) was observed in isolated perfused rat heart subjected to global no-flow ischemia. The effect of U50, 488H on connexin 43 (Cx43) expression of ventricular muscle during ischemia was determined by immunohistochemistry.

RESULTSIn the prolonged ischemia model, U50, 488H concentration dependently delayed the onset of uncoupling, increased time to plateau, and decreased the maximal rate of uncoupling during ischemia. The effect of U50, 488H on electrical uncoupling parameters during ischemia was abolished by a selective kappa-opioid receptor antagonist nor-BNI or a PKC inhibitor chelerythrine. The amount of Cx43 immunoreactive signal in ventricular muscle was greatly reduced after ischemia. U50, 488H markedly increased Cx43 expression during ischemia and its effect was also attenuated by nor-BNI or chelerythrine.

CONCLUSIONThese results demonstrated that U50, 488H delayed the onset of uncoupling and plateau time, decreased the maximal rate of uncoupling and increased Cx43 expression of ventricular muscle during ischemia, and these effects of U50, 488H were mediated by kappa-opioid receptor, in which activation of PKC was involved. The effect of U50, 488H on electrical coupling during ischemia was probably correlated with preservation of Cx43 in cardiac muscle.

3,4-Dichloro-N-methyl-N-(2-(1-pyrrolidinyl)-cyclohexyl)-benzeneacetamide, (trans)-Isomer ; pharmacology ; Animals ; Benzophenanthridines ; pharmacology ; Connexin 43 ; metabolism ; Female ; Heart ; drug effects ; In Vitro Techniques ; Myocardial Ischemia ; metabolism ; physiopathology ; Myocardium ; metabolism ; Naltrexone ; analogs & derivatives ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Receptors, Opioid, kappa ; metabolism ; Signal Transduction ; drug effects

China ; epidemiology ; Cross-Sectional Studies ; Female ; Hospitalization ; Humans ; Infusions, Intravenous ; statistics & numerical data ; Male

Short-term closing live poultry markets have been frequently adopted to control H7N9 epidemic.To evaluate the effect of short-term closing live poultry markets,we analyzed the data of environment surveillance of H7 by RT-PCR,and the human H7N9 cases in Zhongshan City from Feb 2014 to May 2017.RRx =The positive rate during the x-th week after closing / the positive rate of the week before closing days * 100％.Three rounds of short-term closing live poultry markets were administered.Round one:from Feb 10 to 23,2014,the neighboring cities didn't synchronize.The H7 positive rate increased since the trade recovered,and RR1 =0.40 (95％CI:0.28-0.59),RR3 =0.63 (95％CI:0.32-1.24),RR4 =0.83 (95％CI:0.48 -1.46).Two human cases were reported in May,2014.Round two:from Feb 19 to 28,2015,the neighboring cities synchronized as the province policy.The H7 positive rate maintained a level lower than 10％ since the trade recovered,and RR1 =0.15 (95％CI:0.07-0.34),RR2=0.21 (95％CI:0.10-0.41),RR3 =0.03 (95％CI:0.00-0.18),RR4 =0.10 (95％CI:0.04-0.27).No more human cases reported in that epidemic season.Round three:from Jan 8 to 21,2017,the neighboring cities didn't synchronize with Zhongshan City.The H7 positive rate had increased since the trade recovered,and RR1 =0.25 (95 ％ CI:0.09-0.68),RR3 =0.37 (95％CI:0.14-1.00),RR4 =1.07 (95％CI:0.54-2.11).Two human cases were reported in Feb,2017.Results indicated that,if the policy of closing live poultry markets was administered in single city,the environment pollution rate would rise shortly and the risk of human infection would increase once the trade recovered.However,if it was synchronously administered in all the cities in one region,the environment pollution rate could maintain at a low level and the risk of human infection would reduce enormously.

Objective： The current study was designed to investigate the effect of heat shock protein 70(HSP70) on stability of mutant p53 protein. Methods： Retroviral recombinant expressing antisense HSP70 RNA was constructed and transfected into MCF7/Adr breast cancer cells. The existence of foreign DNA was identified by PCR method and the HSP70 protein level was determined by Western blot analysis. The half-life of mutant p53 protein(mtp53) was measured by p53 stability assay. Results： The stable expressing strain(MAp70) from transfected cells was obtained through G418 selection. The foreign DNA in transfectant cells were confirmed by PCR, and the repression rate of HSP70 protein was 42％ . The half-life of mutant p53 in MAp70 cell was 12 hours, significantly lower than that of the control cells. Conclusion： Antisense HSP70 RNA can decrease the HSP70 protein level and significantly increase instability of mutant p53 protein in MCF7/Adr breast cancer cell.

Objective To explore whether the expression level of heparanase (HPA) and its coagulation proteins on leukemic blast membrane could determine the hemostatic balance on the surface of leukemia cells.Methods Forty patients of leukemia were studied,and 20 patients with iron dificient anemia as the control group.Expression of tissue factor (TF),heparanase (HPA),tissue factor pathway inhibitor (TFPI),and urokinase plasminogen activator receptor (UPAR) on leukemic blast surfaces were analyzed by flowcytometry.Results The expression of TF,UPAR,and HPA in AML,ALL,CML,CLL and CRAL groups were significantly higher compared with the control group (t =.3.289,3.507,2.701,P ＜0.05; t =2.498,0.802,3.090,P ＜0.05; t =2.642,3.308,2.696,P ＜0.05; t =3.417,3.434,2.382,P ＜0.05; t =2.193,2.272,2.263,P ＜0.05).There were no significantly differences between the leukemic cell expression of TFPI and the control group (P ＞0.05).Expression of TF,UPAR,HPA in AML patients were significantly higher than ALL,CML and CLL groups (t =2.463,2.179,2.276,P ＜0.05; t =2.637,2.402,2.095,P ＜0.05; t =2.548,2.425,2.412,P ＜0.05).The levels of TF,UPAR and HPA in M3,M4 and M5 patients were higher than that of M1,M2 groups (P ＜0.05).There were no significantly differences among M3,M4 and M5 (P ＞0.05).Conclusions These results suggest that TF,UPAR and HPA are predominately expressed on leukemic blast surface,particularly in M3and M4,5 subtypes.The expression of coagulation proteins on blast membrane might determine the hemostatic balance on the surface of leukemia cells.

BACKGROUNDTo construct the full-length complementary DNA of HCV genome from an HCV infected patient.

METHODSFour HCV gene fragments (1.6, 3.5, 2.4 kb and 2.6 kb) were amplified by RT-PCR from serum of a Chinese patient and fused and connected together to produce a 9.2 kb subgenomic fragment, which was further cloned into a cassette vector with fixed 5-prime and 3-prime termini of HCV to make the full-length cDNA. The cDNA heterogeneity was analyzed by comparing the sequences of 4 isolated HVR1 regions. The prokaryotic expressed Core, NS3 protease, NS3 helicase were detected for their specific reactivities with patient serum by Western blot analysis. And the protease activity of NS3 was evaluated in a cell-based NS3/4A-SEAP expression system.

RESULTSThe cDNA covered the near full-length of HCV genome from the patient's serum. The difference among HVR1 regions indicated no selection of HCV variants during RT-PCR and the quasi-species characteristic of the amplified cDNA. The prokaryotic expressed viral proteins could be identified by patient serum. In the NS3/4A-SEAP system, NS3 could cleave the 4A-4B site between NS4A and SEAP proteins and resulted in the secretion of SEAP in culture media.

CONCLUSIONThese results suggest that the cloned HCV cDNA encodes a complete and functional open reading frame and will be useful for further construction of infectious cDNA clone.

Base Sequence ; Cloning, Molecular ; DNA, Complementary ; genetics ; Genome, Viral ; Hepacivirus ; genetics ; Hepatitis C ; virology ; Humans ; Molecular Sequence Data ; Reverse Transcriptase Polymerase Chain Reaction