Objective To investigate the expression of P2X7 receptor (P2X7R) and the effect of P2X7R agonist 2'-3'-O-(4-benzoyl-benzoyl) ethane adenosine triphosphate three amine salt (BzATP) on cell growth and apoptosis in non-small cell lung cancer A549 cells,and to explore the related mechanism.Methods The expression of P2X7R in A549 cells was detected by immunofluorescence.Cells were treated with different concentrations (150,300,600 μmol/L) of BzATP.Cells untreated with BzATP were used as control group.3-(4,5-dimethyl-2-thiazoly)-2,5-diphenyl-2H-tetrazolium bromide (MTF) assay and Hoest33342 staining were respectively used to detect cell viability and apoptosis.Enzyme-linked immunosorbent assay (ELISA) was uesd to detect the concentration of tumor necrosis factor-α (TNF-α) of cell culture supernatants.The expressions of nuclear factor-κB (NF-κB) p65,inhibitor of α of NF-κB (IκBα) and the phosphorylation of inhibitor of α of NF-κB (phospho-IκBα) were detected by Western blotting.Results P2X7R was expressed on the cell membrane of A549 cells.Survival rate of A549 cell was significantly decreased with the concentrations of BzATP at 300 and 600 μmol/L [(67.87 ± 8.98) ％,(44.73 ± 6.92) ％],compared with the control group (98.60 ± 1.44) ％,the differences were statistically significant (t =4.481,P =0.027;t =3.920,P =0.038).BzATP promoted apoptosis,and increased the concentration of TNF-α of supernatant at 300 and 600 μmol/L [(57.35 ±6.41) pg/ml,(78.63 ± 11.33) pg/ml],compared with the control group (42.56 ±0.37) pg/ml,the differences were statistically significant (t =6.410,P =0.035;t =11.330,P =0.005).In addation,the expressions of NF-κB p65 and IκBα were respectively downregulated and upregulated by BzATP,while the expression of phospho-IκBα was not significantly altered.Conclusion P2X7R is expressed on A549 cell membrane.BzATP can inhibit cell proliferation and induce the apoptosis of A549 cells,and the mechanism of action may be related to promoting the release of TNF-α and inhibition of NF-κB pathway.

Key trial activities include: development of the trial protocol;development of standard operating procedures;development of support systems and tools;generation and approval of trial information documents;selection of trial sites and the selection of properly qualified,trained,and experienced investigators and study personnel;ethics committee review and approval of the protocol;review and approval by applicable regulatory authorities;enrollment of subjects into the study: recruitment,eligibility,and informed consent;the investigational product(s): quality,handling,and accounting;trial data acquisition: conducting the trial;trial data acquisition: conducting the trial; safety management and reporting;monitoring the trial;managing trial data;quality assurance of the trial performance and data;reporting the trial.

Objective Pueraria extract puerarin,HPLC assay puerarin extract and compare different doses of correlation with the hang-over effect of puerarin evaluate different doses of puerarin liver hangover effect. Methods Extracted under optimal conditions obtained in the previous experiment puerarin spare,HPLC method for qualitative and quantitative detection of alcohol extract of kudzu root ( PRE) ,the male Kunming mice were randomly divided into control group,positive control group and puerarin group,each group of 10. Give mice fed pueraria extract,30 min after administration of liquor,drunk mice sobering observation time and the determination of mouse liver ADH,GOT,GPT con-tent in order to investigate the effect of puerarin on drunken mice. Results HPLC fraction was measured at 8 times the volume of 70% etha-nol,60 ℃ constant temperature water bath shaker at 30 min for optimal extraction conditions puerarin extraction. Compared with the positive control groups:low,medium and high doses of alcohol extract of pueraria can significantly shorten the time to sober up drunken mice,the dose of PRE could effectively inhibit the absorption of alcohol,reduce liver tissue ADH,GOT,GPT,the effects of high doses of PRE absorption of alcohol was small. Conclusion HPLC method capable of puerarin extract the qualitative and quantitative determination of puerarin on liver injury caused by acute alcoholism a protective regulatory role,and the hangover effect of puerarin dose showed a good positive correlation.

Objective:To observe the clinical efficacy and safety of cranial suture acupuncture plus paroxetine in treating depression,and to discuss the action mechanism of this acupuncture method.Methods:One hundred depression patients were allocated to an observation group and a control group according to the random number table,with 50 cases in each group.The control group was intervened by oral administration of paroxetine tablets,20 mg each time,once a day for successive 6 weeks;the observation group was additionally given cranial suture acupuncture,once a day for 6 weeks.They were scored by Hamilton depression scale-17 (HAMD-17) before the treatment and respectively after 1-week,2-week,4-week and 6-week treatment.The clinical efficacy and safety were also observed.Results:After 6-week treatment,the total effective rate was 94.0％ in the observation group versus 78.0％ in the control group,and the between-group difference was statistically significant (P＜0.01).The HAMD-17 scores respectively after 1-week,2-week,4-week and 6-week treatment were significantly lower than the score before the treatment in the observation group (all P＜0.05);the HAMD-17 scores respectively after 2-week,4-week and 6-week treatment were significantly different from the score before the treatment in the control group (all P＜0.05).There were significant differences in the HAMD-17 score between the two groups respectively after 4-week and 6-week treatment (both P＜0.05).Conclusion:Cranial suture acupuncture plus paroxetine can ease the symptoms of depression,with faster onset and more significant therapeutic efficacy compared with paroxetine alone.

OBJECTIVETo investigate the protective effects of oxymatrine on chronic heart failure induced by isoproterenol (ISO) and to observe its effects on ADMA metabolism pathway in ISO-induced chronic heart failure in rats.

METHODMale Sprague-Dawley rats were given oxymatrine (100,50 mg kg-1) orally for 14 days. Heart failure was induced in rats by subcutaneous injection of isoproterenol (5 mg kg-1 d-1 ) at the 8th day for 1 week. Serum parameters, haemodynamic parameters, Heart weight, and histopathological variables were analysed. Expression of protein levels were measured by Western blot.

RESULTOxymatrine (100,50 mg kg-1) significantly attenuated serum content of cTn I, improved left ventricle systolic and diastolic function and left ventricular remodeling, reduced the ISO-induced myocardial pathological changes compared with ISO group. In addition, oxymatrine (100,50 mg kg-1) significantly reduced serum level of ADMA (P <0. 01), normalize the reduced dimethylarginine dimethylaminohydrolase 2 (DDAH2) expression (P <0. 01) , but had no effect on the isoproterenol-induced upregulated protein arginine methyltransferases 1 expression.

CONCLUSIONOxymatrine could ameliorate the experimental ventricular remodeling in ISO-induced chronic heart failure in rats and the mechanism involved in reducing serum content of ADMA and increased DDAH2 expression.

Alkaloids ; pharmacology ; therapeutic use ; Amidohydrolases ; metabolism ; Animals ; Arginine ; analogs & derivatives ; blood ; metabolism ; Chronic Disease ; Gene Expression Regulation, Enzymologic ; drug effects ; Heart Failure ; drug therapy ; metabolism ; pathology ; physiopathology ; Hemodynamics ; drug effects ; Isoproterenol ; adverse effects ; Male ; Organ Size ; drug effects ; Quinolizines ; pharmacology ; therapeutic use ; Rats ; Rats, Sprague-Dawley ; Troponin I ; metabolism

OBJECTIVETo make prenatal dignosis of X-linked adrenoleukodystrophy (ALD) for the prevention of the disease.

METHODSEighteen amniocenteses were performed on 17 suspected carriers of X-ALD during 18-30 gestation weeks. The very long chain fatty acids (VLCFAs) levels of cultured amniocytes were tested by gas chromatography-mass spectrometry (GC/MS). The plasma VLCFAs levels were measured in 8 of the 18 prenatal diagnosed children when they were born or after abortion. ABCD1 gene mutation analysis was carried out in 8 cases by PCR and sequencing. ALDP of amniocytes was tested by Western blotting in 2 cases from a family, one female, another male, and the VLCFAs of cultured amniocytes were increased in both of them.

RESULTSAmong the 18 fetuses, 10 were males and 8 were females. The VLCFAs levels of the cultured amniocytes were increased in 3 males and 4 females. The postnatal plasma VLCFAs were normal in 5 cases with normal VLCFAs levels of amniocytes, and increased in 3 cases with high VLCFAs levels of amniocytes. ABCD1 gene mutations were found in 4 cases with high VLCFAs levels of amniocytes, no mutation was found in other 4 cases with normal VLCFAs levels of amniocytes. ALDP of amniocytes could be detected in the female with high VLCFAs levels of amniocytes, and it could not be detected in the male with high VLCFAs levels of amniocytes. Three male fetuses with high VLCFAs levels of amniocytes were aborted. The others who were born were normal clinically so far.

CONCLUSIONThe prenatal diagnosis is very important for the prevention of ALD. Amniocyte VLCFAs level analysis combined with ABCD1 gene mutation analysis and ALDP test could make a proper prenatal diagnosis.

ATP Binding Cassette Transporter, Sub-Family D, Member 1 ; ATP-Binding Cassette Transporters ; genetics ; Adrenoleukodystrophy ; diagnosis ; genetics ; metabolism ; Adult ; DNA Mutational Analysis ; Fatty Acids ; metabolism ; Female ; Gas Chromatography-Mass Spectrometry ; Humans ; Infant, Newborn ; Male ; Pregnancy ; Prenatal Diagnosis ; methods

OBJECTIVEX-linked adrenoleukodystrophy (X-ALD) is the most common inherited disorder of peroxisomal metabolism which is characterized by demyelination of central nervous system, impaired adrenal cortex and abnormal accumulation of very long chain fatty acids in body fluid and tissues. In this study, the clinical features and the genotype-phenotype relationship were investigated so as to help early diagnosis.

METHODSClinical data of 89 Chinese patients with X-ALD were analyzed and mutation spectrums in 53 of the patients were investigated by polymerase chain reaction and DNA sequencing.

RESULTSOf the 89 cases, 60 (67.4%) had childhood cerebral ALD (CCALD, mean age of onset was 6.5 years, range 2 - 10 years), 18 (20.2%) had adolescent cerebral ALD (ACALD, mean age of onset 12.1 years, range 11 - 19 years), 7 (7.9%) had adrenomyeloneuropathy (AMN, mean age of onset 23 years, range 6 - 39.5 years), and two cases were asymptomatic and another 2 had simple Addison's disease only. CCALD was the most common and severe phenotype, visual impairment was the most common initial symptom in CCALD and ACALD patients. Twenty four cases (63%) in whom hydrocortisone and ACTH were measured showed adrenal insufficiency. Forty five different mutations were identified in 53 patients. Missense mutations were the most common. No hotspot mutation was found in these patients and 1415delAG, the most frequent mutation found worldwide seemed not to be the real "hotspot" in these Chinese patients. The same phenotype may be due to diverse genomic mutations. A single mutation may result in different phenotypes even within a family.

CONCLUSIONThe phenotype distribution, initial symptom and gene mutation spectrum of Chinese patients may not be completely consistent with those in other countries. The clinical phenotype of the disease had no definite relationship with the nature of gene mutations.

Adolescent ; Adrenoleukodystrophy ; genetics ; Adult ; Age of Onset ; Asian Continental Ancestry Group ; genetics ; Child ; Child, Preschool ; DNA Mutational Analysis ; Female ; Genotype ; Humans ; Male ; Mutation, Missense ; Phenotype ; Young Adult

OBJECTIVEX-linked adrenoleukodystrophy (X-ALD) is the most common peroxisomal disorder characterized by progressive demyelination of the central nervous system, adrenal cortex insufficiency and accumulation of saturated very long chain fatty acids (VLCFAs) in tissues and body fluids due to the impaired beta-oxidation in peroxisomes. X-ALD shows a wide range of phenotypic variation. Childhood cerebral form (CCER) is the most common phenotype with severe neurological symptoms and often the average interval from onset to total disability or death is 3 years. So far no effective treatment is available for the underlying defect. Screening for carriers of mutated relevant gene and prenatal diagnosis are very important for the prevention of the disease. In this study, the authors explored the method of carrier screening and prenatal diagnosis of X-ALD.

METHODSThe plasma VLCFAs levels of 83 suspected carriers for ALD were determined by using GC/MS and ABCD1 gene mutational analysis was performed in 31 of them. Amniocentesis was performed in 9 suspected carriers for ALD during 18 - 30 gestational weeks. The VLCFAs level of cultured amniocytes was tested with GC/MS. ABCD1 gene mutational analysis was performed on two cases (one was a male and the other a female) whose VLCFAs levels of amniocytes were found elevated. The plasma VLCFAs levels were measured in five of the nine prenatally diagnosed children when they were 1 day to 3.5 years old.

RESULTSFifty-one of 83 suspected carriers had high plasma VLCFAs levels; 29 of 31 suspected carriers showed ABCD1 gene mutation. Among the nine fetuses, four were males and five were females. The VLCFAs levels of the cultured amniocytes were high in two cases, one was female and the other a male. ABCD1 gene mutational analysis of these two cases showed a 871G > A (E291K) mutation and a 726G > A (W242X) mutation, respectively, which confirmed the biochemical result. The VLCFAs levels were normal in the rest of cases and five of them were confirmed by postnatal plasma VLCFAs assay.

CONCLUSIONThe carrier screening and prenatal diagnosis are very important for prevention of the X-ALD. Only the combined use of plasma VLCFAs level analysis and ABCD1 gene mutational analysis could detect X-ALD carriers correctly. ABCD1 gene mutational analysis and postnatal plasma VLCFAs level test verified that amniocytes VLCFAs level analysis is a reliable prenatal diagnostic method for this disease.

ATP Binding Cassette Transporter, Sub-Family D, Member 1 ; ATP-Binding Cassette Transporters ; genetics ; Adrenoleukodystrophy ; diagnosis ; genetics ; Adult ; Fatty Acids ; analysis ; Female ; Genetic Carrier Screening ; Humans ; Infant, Newborn ; Male ; Mutation ; Pregnancy ; Prenatal Diagnosis

OBJECTIVETo evaluate the efficacy, median time to progression (TTP), quality of life and toxicity in the patients with advanced non-small cell lung cancer (NSCLC), treated with thalidomide plus vinorelbine and cisplatin (NP) or NP alone.

METHODSSixty six patients with advanced NSCLC were divided randomly into two groups, the trial and control groups. The trial group was treated with vinorelbine 25 approximately 30 mg/m(2) i.v. on D1 and D8, cisplatin 70 approximately 80 mg/m(2) i.v. on D1 (NP regimen), and thalidomide 200 mg orally and daily from D1. The control group received vinorelbine and cisplatin as above described.

RESULTSOf 66 assessable patients, the overall response rate was 51.5% in the trial group and 36.4% in the control group (P = 0.22). The median TTP was 6.0 months for the trial group, and 3.6 months for the control group (P < 0.001). The score of quality of life in trial group was higher than that in the control group, but no significant difference was observed between the two groups (P > 0.05). There were no significant differences in toxicities between the two groups (P > 0.05).

CONCLUSIONNP regimen combined with thalidomide can significantly prolong the median time to tumor progression in patients with advanced NSCLC. Thalidomide may have a synergic activity with NP regimen without increased toxicities.

Adult ; Aged ; Antineoplastic Combined Chemotherapy Protocols ; adverse effects ; therapeutic use ; Carcinoma, Non-Small-Cell Lung ; drug therapy ; pathology ; Cisplatin ; administration & dosage ; adverse effects ; Disease Progression ; Drug Synergism ; Feeding and Eating Disorders ; chemically induced ; Female ; Follow-Up Studies ; Humans ; Leukopenia ; chemically induced ; Lung Neoplasms ; drug therapy ; pathology ; Male ; Middle Aged ; Neoplasm Staging ; Quality of Life ; Remission Induction ; Thalidomide ; administration & dosage ; adverse effects ; Thrombocytopenia ; chemically induced ; Vinblastine ; administration & dosage ; adverse effects ; analogs & derivatives ; Vomiting ; chemically induced

OBJECTIVETo study the application value of normal sperm morphology on the outcomes of classic in vitro fertilization and embryo transfer (IVF-ET).

METHODSThis study included 659 infertile couples admitted to our center for IVF-ET. Based on the percentage of morphologically normal sperm (MNS), we divided the patients into groups A (n = 112, MNS < 2%), B (n = 180, MNS > or = 2 - < 4%), C (n = 74, MNS > or = 4 - < 5%), and D (n = 293, MNS > or = 5%), and compared the rates of fertilization, normal fertilization, embryos obtained, biochemical pregnancy, clinical pregnancy, implantation, and live birth among different groups.

RESULTSThe mean fertilization rate was significantly higher in groups C (71.90%) and D (72.89%) than in A (57.97%) and B (63.29%) (P < 0.05), with no remarkable differences either between A and B (P > 0.05) or between C and D (P > 0.05). The normal fertilization rate was also significantly higher in group D (57.16%) than in A (46.52%) and B (50.89%) (both P < 0.05) as well as in C (54.67%) than in A (P < 0.05). The rate of embryos obtained, too, was markedly higher in group D (55.62%) than in B (45.75%) (P < 0.05), but none with remarkable difference from other groups (all P > 0.05). There were no statistically significant differences among the four groups in the rates of biochemical pregnancy, clinical pregnancy, implantation, abortion, and live birth (all P > 0.05).

CONCLUSIONThe rate of MNS had some influence on IVF-ET, and 5% MNS exhibited a higher value than 4% MNS in predicting the outcomes of IVF.

Adult ; Embryo Implantation ; Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Retrospective Studies ; Spermatozoa ; cytology