AIMTo further explore the roles of endogenous nitric oxide (NO) or NO derivatives in complex partial seizures and generalized convulsions.

METHODSThe effect of pretreatment with L-nitroarginine (L-NNA), an inhibitor of nitric oxide synthase (NOS), or L arginine (L-Arg), a precursor of NO on kainic acid (KA)-induced seizure in rats and the changes in the concentration of NO2 -/NO- in the hippocampus were determined.

RESULTSThe rats appeared with wet dog shakes (WDS) at 15 min and then occurred generalized convulsions during 1 h to 3 h after administration of KA (10 mg/kg i.p.). However, the pretreatment of L-NNA (50 mg/kg) so dramatically promoted and enhanced KA-induced behavioral seizures that the latency of generalized convulsion was shorten dramatically, and the mortality was greatly high. In contrast, the pretreatment with L-Arg (40 mg/kg) markedly delayed or weakened KA-induced behavioral changes, such as increasing latency of WDS and generalized convulsion, shortening time o f seizure and none of animal died during observed time. The concentration of NO2- /NO3- in the hippocampus increased immediately at 30 min and remained to 7 d after the administration of KA. Compared with control group (pretreatment with NS), the concentration of NO2- / NO3- in the hippocampus apparently increased at 3 h and 3 d after the administration of KA in the rats with L-Arg pretreatment.

CONCLUSIONThe endogenous NO (NO or NO derivatives) mediators may play an important role against excitotoxin induced seizures in rats.

Animals ; Arginine ; pharmacology ; Kainic Acid ; adverse effects ; Male ; Nitric Oxide ; metabolism ; Nitroarginine ; pharmacology ; Rats ; Rats, Wistar ; Seizures ; chemically induced ; metabolism

AIMTo develop a sensitive and rapid LC/MS/MS method for the determination of wogonin, an active flavonoid shown to have an inhibitory effect on the proliferation of a carcinoma cell line, in rat plasma after an oral administration.

METHODSWogonin and daidzein (internal standard) were extracted from plasma directly with n-hexane-diethyl ether (1:4). After liquid-liquid extraction, the analytes of interest were separated on a Diamonsil C18 column. The mobile phase was acetonitrile-water-formic acid (80:20:1) with a flow rate of 0.8 mL.min-1. A Finnigan TSQ (triple stage quadruple) tandem mass spectrometer equipped with an atmospheric pressure chemical ionization (APCI) source was used as detector and was operated in positive ion mode. Selected reaction monitoring (SRM) was used and transitions selected for quantitation were: m/z 284.8-->269.5 for wogonin and m/z 254.7-->198.5 for daidzein. The mass spectrometric conditions were as follows: The temperatures of the vaporizer and heated capillary were 450 degrees C and 250 degrees C, respectively. The corona discharge current was 4.00 microA. Nitrogen was used as the sheath and auxiliary gas, whose settings were 0.6 MPa and 3 mL.min-1, respectively. Argon was used as the collision gas at a pressure of 1.4 Pa. The collision energy of 35 V was chosen for both wogonin and daidzein.

RESULTSThe calibration curve was linear over the concentration range of 0.25 to 20.0 ng.mL-1. The limit of quantitation was 0.25 ng.mL-1. Within-day and between-day precision expressed by relative standard deviation (RSD) was 2.2% to 13.1% and 5.9% to 7.3%, respectively, and the accuracy expressed by RE was -0.3% to 1.3%.

CONCLUSIONThis method proved to be specific, accurate and sensitive enough to be applied to the pharmacokinetic studies of wogonin in rats after a single dose of 5 mg.kg-1 by oral administration.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; isolation & purification ; metabolism ; pharmacokinetics ; Female ; Flavanones ; blood ; isolation & purification ; pharmacokinetics ; Gas Chromatography-Mass Spectrometry ; Male ; Plants, Medicinal ; chemistry ; Rats ; Rats, Wistar ; Scutellaria ; chemistry

As a traditional Chinese medicine, the root of Astragalus membranaceus var. mongholicus (AMM) or A. membranaceus (AM) has been widely used in China and other Asian countries for thousands of years. Till now, the flavonoids, phenolic acids and saponins are considered as the main active components contributing to their therapeutic effect in these plants. In order to clarify the distribution and contents of these compounds in different organs of these plants, a rapid and sensitive analytical method for simultaneous determination of 25 active compounds including seven types (i.e. dihydroflavones, iso-flavane, isoflavones, flavones, pterocarpans, phenolic acid and saponins) within 10 min was established using ultra-pressure liquid chromatography coupled with tandem mass spectrometry (UPLC-MS/MS). Then, the established method was fully validated and successfully applied to the determination of the contents of these analytes in different parts (root, rhizome, stem, leaf and flower) of AMM and AM. The results indicated that the contents of the same type of compounds in two different species plants were significantly different. Moreover, the obvious differences were also found for the distribution and con-tents of different type of compounds in five organs of the same species. The present study could provide necessary information for the rational development and utilization of AMM and AM resource.

Measurement of platelet-associated imunoglobulin (PAIg) has frequently been applied for the diagnosis of idiopathic thrombocytopenic purpura (ITP) and other immune thrombocytopenias. In the present study, a flow cytometry (FCM) analysis has been used to detect and characterize PAIg in 47 patients with ITP and Evans' syndrome, 13 patients with non-immune thrombocytopenia, 10 patients with autoimmune hemolytic anemia (AIHA) whose platelet counts were in normal range, and 31 healthy volunteers. With FCM measurement, mean fluorescence intensity (MFI) of platelets from patients with ITP and Evans' syndrome (2.26 +/- 2.29) was significantly higher than those from non-immune thrombocytopenia (0.33 +/- 0.39), AIHA (0.17 +/- 0.07) and control subjects (0.25 +/- 0.15) (P < 0.01). Meanwhile, the percentage of positive platelets of patients with ITP and Evans' syndrome [(44.1 +/- 29.0)%] was also higher than those of non-immune thrombocytopenia [(17.5 +/- 9.4)%], AIHA [(10.7 +/- 7.5)%] and control subjects [(16.6 +/- 8.4)%] (P < 0.01). In addition, some peak shape abnormality appeared (double peaks and peak tail) in the histogram of fluorescence intensity (log) of 11 patients (23.4%) with ITP and Evans' syndrome either alone or accompanied with quantitative alteration of MFI and/or positive platelet percentage. In seven cases, the peak shape abnormality was the unique characteristic that could be detected and have never been seen in normal platelets. This phenotypic alteration perhaps reflects the existence of different platelet populations and could be of diagnostic value. Totally, the positive result of FCM measurement in patients with ITP and Evans' syndrome was 87.2%, slightly higher than 83.0% positive rate with ELISA method, without statistical difference. The correspondent rate of the results of these two analytical settings was 85.1%. This study shows that FCM assay is a rapid and sensitive method for the measurement of PAIg and seems to be suitable as a novel routine diagnostic technique of immune thrombocytopenia.

Objective:To explore correlation between fibrinogen(Fg)level and coronary stenotic degree in aged coronary heart disease(CHD)people and its possible pathogenesis.Methods:A total of 112 patients,who were >65 years and diag-nosed as CHD by coronary angiography(CAG),were selected as CHD group.Another 108 non-CHD patients hospitalized during the same period were enrolled as non-CHD group.According to CAG results,CHD group was divided into single vessel coronary disease group(n=30,single vessel group)and multi vessel coronary disease group(n= 82,multi vessel group).Serum levels of Fg,D-dimer(D-D),C reactive protein(CRP),homocysteine(Hcy)and brain natriuretic peptide (BNP)were measured and compared between two groups.The correlation between serum Fg level and Gensini score was analyzed in CHD patients.Results:Compared with non-CHD group,there were significant rise in serum levels of Fg,D-D, triglyceride(TG),apolipoprotein B(ApoB),lipoprotein a,CRP,BNP and Hcy in CHD group,P=0.001 all;in CHD group,compared with single vessel group,there were significant rise in serum levels of Fg,D-D,ApoB,uric acid,lipo-protein a,CRP,BNP,Hcy,and significant reductions in serum levels of high density lipoprotein cholesterol,ApoA1 and ApoA1/ApoB in multi vessel group,P<0.05 or <0.01. Serum Fg level was significant positively correlated with coronary stenotic degree(Gensini score),r=0.303,P=0.001. Fg was independent risk factor of CHD(OR=2.74,95% CI:1.95~3.97,P=0.001).Conclusion:Serum Fg level is significant positively correlated with coronary stenotic degree in aged CHD patients.Its mechanism of triggering atherosclerosis may be related to risk factors such as dyslipidemia,hyper-homocysteinemia and inflammations etc.

In recent years, foreign countries are gradually implementing broad consent to improve the utilization of medical data and biological samples, but broad consent may face ethical issues such as imperfect notification and affecting the rights of subjects. There are already relevant regulations and practices on broad consent in foreign countries. The concept of broad consent is not clearly defined in China′s laws. At present, the treatment of biological samples can be roughly divided into four categories in practice, and there is potential application space for broad consent. The specific scope of broad consent should be clarified, distinguished from donation behavior, and the implementation of broad consent should be explored on the basis of protecting the rights of subjects.

OBJECTIVETo study the HIV incidence and risk factors among sero-negative spouses of HIV patients in Dehong prefecture of Yunnan province.

METHODSA cohort of sero-negative spouses of the HIV patients had been developed and followed up since November, 2005. HIV new infections and related behaviors had been investigated every six months.

RESULTSBy the end of June, 2008, 790 sero-negative spouses of HIV patients had been recruited, of whom 702 were followed-up for at least one time. During the total 1202.35 person-years, 31 new HIV infections were identified, with an overall incidence of 2.58/100 person-years. The HIV incidence rates were 2.22/100 person-years in 2006, 2.95/100 person-years in 2007 and 2.74/100 person-years in 2008. Data from the Cox proportional hazard regression model indicated that those who resided in Yingjiang county [hazard ratio (HR) = 4.37, 95%CI: 1.48 - 12.90, P = 0.008], ever using drugs (HR = 3.49, 95%CI: 1.09 - 11.18, P = 0.035), or having an HIV-infected spouse who never exposed to antiretroviral treatment (HR = 3.60, 95%CI: 1.41 - 9.16, P = 0.007) were at higher risk for HIV infection.

CONCLUSIONSero-negative spouses of HIV patients in Dehong prefecture of Yunnan province had a relatively high incidence of HIV new infection during 2006-2008. More efforts should put on those people living in these areas, having a history of drug use or having an HIV-infected spouse who had never been exposed to antiretroviral treatment.

Adolescent ; Adult ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; HIV Seronegativity ; Humans ; Incidence ; Male ; Middle Aged ; Prospective Studies ; Risk Factors ; Sexual Behavior ; Spouses ; Young Adult

OBJECTIVETo explore a new method for finding more HIV/AIDS.

METHODSIn September 2009, newly reported HIV-infected individuals from May to August, 2009 in Dehong prefecture were asked to participate in a survey which requested demographic characteristics, history of high-risk behaviors and contact information of individuals with whom they had high risky contacts. People with risky contacts with HIV-infected cases (index cases) were also approached to participate in this survey and HIV testing was provided.

RESULTSA total of 342 HIV-infected individuals were newly reported and served as index cases from May to August, 2009. Among them, 47.1% (161/342) were transmitted by regular sexual partners. Through three-round surveys, 218 contacts were traced and among them, 84.9% (185/218) were traced by regular heterosexual partners. HIV positive rate of the tested was 34.7% (60/173) in the first-round tracing and 12.5% (1/8) in the second one. Among the 560 individuals (index cases and their high-risk contacts), the proportions of having regular heterosexual partners, non-regular and non-commercial sexual partners, commercial sexual heterosexual partners and men having sex men were 87.9% (492/560), 18.9% (106/560), 22.3% (125/560) and 0.3% (1/318), respectively, while the proportion of having never used condoms when having sex with the above four types sexual partners were 73.8% (363/492), 72.6% (77/106), 63.2% (79/125) and 0.0% (0/1), respectively.

CONCLUSIONAs an epidemiological method for HIV/AIDS finding, contact tracing identified a large number of HIV infectors who were traced by newly reported HIV-infected individuals.

Acquired Immunodeficiency Syndrome ; epidemiology ; Adolescent ; Adult ; Child ; Child, Preschool ; China ; epidemiology ; Contact Tracing ; methods ; Female ; HIV Infections ; epidemiology ; Humans ; Infant ; Infant, Newborn ; Male ; Risk Factors ; Surveys and Questionnaires ; Young Adult

BACKGROUNDST-elevated acute myocardial infarction (STEAMI) happening in the first month post percutaneous coronary intervention (PCI) is almost related to acute thrombosis or subacute thrombosis in-stents. This study aimed to investigate the possible causes of myocardial infarction one month later.

METHODSPatients who had a history of successful PCI, and received coronary angiography or re-PCI due to STEAMI were included in this study. The AMI-related lesions and previous angiographic findings such as the number of lesions, the degree of the stenosis, the type of stents and acute results of last PCI were recorded. If the AMI-related lesion was localized in-stents or at the edge of stents (distance apart from the edge < or = 5 mm), it was defined to be late thrombosis; otherwise as a new-lesion induced AMI.

RESULTSOne hundred and ninety-two patients aged 40 - 79 years were included in this study. New lesions, as the cause of STEAMI, were found in 144 patients (Group A, 75%), and late thrombosis in 48 patients (Group B, 25%). Almost all newly built thromboses were found at the sites of previous insignificant lesions (diameter stenosis < 50%). There was a significant difference in the average time from previous PCI to AMI ((30.1 +/- 12.4) vs (20.3 +/- 11.9) months) between the two groups. Diabetes mellitus (DM) and drug-eluting stent (DES) utilization were associated with markedly higher morbidity of late thrombosis in adjusted Logistic regression (hazard ratio (HR) 3.4, 95% confidence interval (CI) 1.1 - 10.9 and 5.3, 95% CI 1.1 - 26.5).

CONCLUSIONSSTEAMIs happening 1 month after PCI are more likely to develop from previous insignificant lesion rupture than from late thrombosis in-stents. Moreover, DM and DES are associated with the high incidence of late thrombosis, which may indicate that intensive antiplatelet therapy should be considered in patients with diabetes.

Adult ; Aged ; Angioplasty, Balloon, Coronary ; adverse effects ; Coronary Angiography ; Coronary Thrombosis ; etiology ; pathology ; Drug-Eluting Stents ; adverse effects ; Female ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; etiology ; pathology

In this study, we established a rapid and sensitive multiplex Real-time reverse transcription polymerase chain reaction (mrtRT-PCR) to simultaneously detect the novel human influenza A H1N1 virus, human seasonal influenza A H1N1 and H3N2. This assay had three pairs of primer to target the conserved regions of the HA gene for each of the HA types including novel H1N1, seasonal H1N1 and seasonal H3N2, and one pair of primer designed to detect the internal control-RnaseP gene. This assay was performed in one-step in one tube. To validate the specificity of the multiplex Real-time RT-PCR assay, different human influenza virus including human seasonal influenza A H1N1 and H3N2, human influenza B and reference A/California/07/2009 (H1N1) sw1 was tested. To evaluate the sensitivity of the assay, serial dilutions of RNA from in vitro transcription of the novel human influenza A H1N1 HA gene was tested. Finally this assay was evaluated with clinical samples from 54 fever patients diagnosed with novel influenza A H1N1 or seasonal H1/H3 or HB infection either by real-time PCR recommended by the WHO or HI assay by the National Influenza Center. Our results showed that the assay could achieve a sensitivity of 20 RNA copies of novel influenza A H1N1 with high specificity and could detect potential mixed co-infection. In conclusion, this multiplex real-time RT-PCR assay combines both rapidity and sensitivity for not only detecting the novel human influenza A H1N1 virus, but also monitoring the human seasonal influenza A H1N1 and H3N2 simultaneously.
DNA Primers ; genetics ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; Humans ; Influenza A Virus, H1N1 Subtype ; genetics ; Influenza A Virus, H3N2 Subtype ; genetics ; Influenza, Human ; diagnosis ; virology ; Reproducibility of Results ; Reverse Transcriptase Polymerase Chain Reaction ; instrumentation ; methods ; Seasons ; Sensitivity and Specificity