OBJECTIVETo establish a rapid method for testing drug sussceptibility on Mycobacterium tuberculosis.

METHODSTaking absolute Concentration method for drug susceptibility testing of M. tuberculosis as the "gold standard", we examined the drug-resistant of M. tuberculosis strain with nitrate reducrase assay (NRA) and the drug-resistant of M. tuberculosis germ in sputum with NRA.

RESULTSNRA and absolute concentration method was basically comparable with NRA susceptibility as 96.5% and the specificity was 100%, When comparing with traditional absolute concentration method, NRA could shorten the time about 3 weeks. Using NRA to test the drug-resistant of M. tuberculosis germ in sputum, its susceptibility was more than 66.7% and specificity was 100%, within 10-20 days.

CONCLUSIONNRA could be used as a rapid drug susceptibility testing on M. tuberculosis.

Anti-Bacterial Agents ; pharmacology ; Drug Resistance, Bacterial ; Enzyme Assays ; methods ; Humans ; Microbial Sensitivity Tests ; methods ; Mycobacterium tuberculosis ; drug effects ; pathogenicity ; Nitrate Reductase ; metabolism ; Sputum ; microbiology

Objective To observe the changes of renin-angiotensin system (RAS) in cultured mesangial cells by serum from 3/4 nephrectomized rats feeding with low protein diet and α-keto acid. Methods Thirty male SD rats received 3/4 nephrectomy (Nx) were placed on 18%normal protein diet (NPD,n=10),6% low protein diet(LPD,n=10) or 5% low protein plus 1%α-keto acid diet (LK,n=10) flor 12 weeks.Ten male SD sham-operated rats fed with 18% normal protein diet were used as control (sham group).In addition,mesangial cells were cultured in sera (10%) collected from above animals treated with or without losartan (0.02 mmol/L)for 48 hours.ELISA was applied to detect the level of Ang II,TGF-β1 and fibronectin (FN) in cell medium.Westem blotting was used to determine the protein level of ATI receptor (AT1R)and real-time PCR was used to detect the mRNA level of AT1R,TGF-β1 and FN. Results (1) Nutritional indices including body weight,total protein and albumin had no significant difference in each group. (2) Serum creatinine and 24 h pruteinuria were significantly inceased in nephrectomized groups compared to sham group(P<0.05,respectively).24 h proteinuria was greatly lower in LK group than that in NPD and LPD groups(P<0.05,respectively).(3)LK greatly decteased the level of Ang II[NPD(12.70±0.12)mg/g protein;sham(8.04±0.62)mg/g protein]in supernatant as well as the protein and mRNA expression of AT1R in cultured mesangial cells (P<0.05).(4)NPD serum directly induced higher secretion[FN:sham(20.58±0.46)g/g protein,NPD (39.84±0.06)g/g protein;TGF-β1:sham(10.12±O.56)mg/g protein,NPD(83.85±3.58)mg/g protein] and mRNA expression of FN and TGF-β1 compared with sham group (P<0.05).LPD decreased these increment (P<0.05) and LK showed stronger inhibitory effect (P<0.05). (5)Losartan application sharply reduced FN and TGF-β1 production both in supematant and in mRNA expression in NPD serum treated cells (P<0.05,respectively). Conclusion Low protein diet with α-keto acids supplement directly inhibits the RAS in mesangial cells which may contribute to its beneficial effect on the kidney.

Objective To investigate the daily total fluoride intake in relating to the prevalence of skeletal fluorosis in two villages in Jiangsu Province,in order to provide the scientific evidences for the control and prevention of endemic fluorosis.Methods Adults sampled from a high-fluoride Village,Wamiao,and a low-fluoride Village,Xinhuai,were surveyed in this study according to the fluoride concentration in their household shallow well.The average daily total fluoride intake from difierent sources and the skeletal fluorosis were investigated in each subject.Then the subjects from two villages were allocated into five subgroups(＜2.00,2.00～,3.00～,4.00～,≥5.00 mg/d),the relation fluoride intake and prevalence of osteofluorosis was analyzed.Results The prevalence of skeletal fluorosis in Wamiao Village was 31.06%(41/132),but no skeletal fluorosis case(0/35)was found in Xinhuai Village.According to the daily total fluoride intake,subjects with higher daily total fluoride intake tended to associated with a higher prevalence of skeletal fluorosis in a significant dose-response relationship(regression equation:y=2.624-6.855x+3.424x2:r=0.997).The benchmark dose lower limitation of daily total fluoride intake with 95% confidencewas 2.50 mg/d calculated according to this dose-response relationship,the reference dose(RfD)was 2.50 mg/d.In Wamiao Village a significant difference was also found between daily total fluoride intake in 41 subjects[(5.09±1.20)mg/d]with X-ray detectable skeletal fluorosis and in 91 subjects[(3.08±1.12)mg/d]without X-ray detectable skeletal fluorosis(t=-9.32,P＜0.01).Conclusions These findings indicate that the daily total fluoride intake has a significant dose-response relationship with the prevalence of skeletal fluorosis in an endemic fluorosis area associated with high-fluoride drinking water;and the RfD in this study was lower than that in the national standard of"Chinese hygienic standard for daily total fluoride intake(WS/T 87-1996)"(3.50 mg/d).

Objective To observe the activities of serum peroxidase capacity,and lipid peroxidation of children from Kaschin-Beck disease (KBD) areas of Xinghai county in Qinhai province,and to explore the relationship between antioxidant capacity and KBD.Methods Sixty four KBD and forty six health subjects without KBD were chosen from KBD endemic areas,which included primary schools of Tangnaihai,Xialujuan and Qushian of Xinghai county in Qinghai province,and fifty nine age-matched healthy control subjects without KBD were from a non-KBD endemic area,Nanfan primary school of Chang'an county in Shaanxi province.Twenty patients with KBD and twenty control subjects from KBD areas and non-KBD area were extracted by simple random sampling method.2,3-DAN fluorescence technique was used to test the hair and blood selenium.The biochemical techniques were used to test the indicators of oxidative stress including malondialdehyde(MDA),antioxidant enzyme activities,total antioxidant capacity(T-AOC),serum superoxide dismutase(SOD),catalase(CAT) and glutathione peroxidase(GSHPx).ResultsAll patients with KBD had significantly lower serum GSH-Px activities[ (59.53 ± 25.23)kU/L] and selenium levels in hair[ (67.64 ± 17.28)μg/L] and blood[(36.27 ± 13.29)μg/L],respectively,than that of control subjects from KBD areas [ ( 91.88 ± 22.99 ) kU/L,( 153.32 ± 24.31 ) μg/L,( 63.06 ± 13.66) μg/L ] and nonKBD areas[ ( 122.68 ± 41.74)kU/L,(242.35 ± 38.56)μg/L,(98.93 ± 17.18)μg/L,all P ＜ 0.05].Serum MDA levels in KBD patients[ (4.64 ± 1.11 )μmol/L] were significantly higher than that in control subjects from KBD [(3.31 ± 1.22)μmol/L] and non-KBD areas[ (3.43 ± 1.29)μmol/L,all P ＜ 0.05].On the other hand,T-AOC,SOD and CAT activities were significantly higher in both KBD[(19.80 ± 6.64),(55.80 ± 8.14),(16.45 ± 5.61 ) kU/L] and control subjects[ (21.71 ± 8.82),(57.45 ± 6.96),(15.63 ± 9.18)kU/L] from KBD areas than that of control subjects from non-KBD area[ (13.56 ± 5.38),(42.79 ± 8.10),(6.05 ± 2.71 )kU/L,all P ＜ 0.05 ].Hair selenium levels,blood selenium levels and GSH-Px activity of control subjects from KBD areas were,respectively,significantly lower than that in control subjects from non-KBD area(all P ＜ 0.05).Conclusions These findings strongly confirm the evidence that KBD patients are susceptible to oxidative stress.The results also show the increase in antioxidant enzymes,which could probably be due to adaptive response to pro-oxidant in KBD state.Hence,there seems to be an imbalance between oxidant and antioxidant systems in KBD patients.

OBJECTIVETo observe amplitude changes of low frequency fluctuation in brain spontaneous nervous activities induced by needling at Hand Taiyin Lung Channel, and to preliminarily explore the possible brain function network of Hand Taiyin Lung Channel.

METHODSBy using functional magnetic resonance imaging (fMRI), 16 healthy volunteers underwent resting-state scanning (R1) and scanning with retained acupuncture at Hand Taiyin Lung Channel (acupuncture, AP). Data of fMRI collected were statistically calculated using amplitude of low frequency fluctuations (ALFF).

RESULTSUnder R1 significantly enhanced ALFF occurred in right precuneus, left inferior parietal lobule, bilateral superior temporal gyrus, bilateral middle frontal gyrus, left superior frontal gyrus, left inferior frontal gyrus, left medial frontal gyrus. Under AP significantly enhanced ALFF occurred in right precuneus, bilateral superior frontal gyrus, cerebellum, bilateral middle frontal gyrus, right medial frontal gyrus, and so on. Compared with R1, needing at Hand Taiyin Lung Channel could significantly enhance ALFF in right gyrus subcallosum and right inferior frontal gyrus. Significant decreased ALFF appeared in right postcentral gyrus, left precuneus, left superior temporal gyrus, left middle temporal gyrus, and so on.

CONCLUSIONNeeding at Hand Taiyin Lung Channel could significantly change fixed activities of cerebral cortex, especially in right subcallosal gyrus, right inferior frontal gyrus, and so on.

Acupuncture Therapy ; Brain ; physiology ; Brain Mapping ; Humans ; Magnetic Resonance Imaging

OBJECTIVETo follow up the electrocardiographic and cardiac autonomic function changes after percutaneous transluminal septal myocardial ablation (PTSMA) in patients with hypertrophic obstructive cardiomyopathy (HOCM).

METHODSBaseline, 3 days and 3 years post procedure 12-lead electrocardiographic and 24-hour Holter electrocardiographic recordings including PR interval, QRS duration, cardiac conduct block, QT, QTd, QTcd, JT, JTd, JTcd, heart rate variability (HRV) data (SDNN, SDANN, HF, rMSSD, PNN50, LF, HF, LF/HF) were analyzed in 26 patients with HOCM receiving PTSMA.

RESULTThe PTSMA procedure was successful in all 26 patients. One patient developed complete atrioventricular block requiring permanent pacing. The PR interval was significantly prolonged 3 days after ablation and recovered 3 years post procedure. Right bundle branch block was seen in all patients 3 days after post procedure and in 24 patients at 3 years post procedure. The QRS duration was significantly prolonged at 3 days and 3 years post procedure. There was persistent QT interval prolongation up to 3 years and transient QTd, QTcd prolongation (prolonged at 3 days and returned to baseline at 3 years after ablation) while JT, JTd, JTcd were not significantly changed after PTSMA. LF, HF, rMSSD and PNN50 were significantly increased while LF/HF, SDNN, SDANN remained unchanged post procedure.

CONCLUSIONPTSMA is a safe and effective therapy option for HOCM. Right bundle branch block was the main electrocardiographic change post procedure and PTSMA could partly restore the heart sympathovagal balance by improving vagal activity.

Adult ; Autonomic Nervous System ; physiopathology ; Cardiomyopathy, Hypertrophic ; physiopathology ; therapy ; Catheter Ablation ; methods ; Electrocardiography ; Female ; Follow-Up Studies ; Heart Septum ; Humans ; Male ; Middle Aged

Objective To determine effects of T-2 toxin and selenium on expression of aggrecanase in human chondrocyte.Methods Human chondrocytes were treated with T-2 toxin(0,1,10,20 μg/L),and/or sodium selenite(final concentration of selenium 0,0.1 mg/L) for 5 days.Aggrecan expression was determined by Western blotting,aggrecanase-1 and aggrecanase-2 mRNA levels were measured by RT-PCR.ResultsSelenium and T-2 toxin had effects on both aggrecan protein levels and its aggrecanases(include aggrecanase-1 and aggrecanase-2 ) mRNA levels(F =0.294,27.71 for aggrecan,F =107.45,362.25 for aggrecanase-l,F =34.68,22.26 for aggrecanase-2,respectively,all P ＜ 0.05),and there was interaction between selenium and T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression(F =79.99,230.76,388.33,all P ＜ 0.05).Furthermore,selenium presented significant antagonism to T-2 toxin on aggrecan,aggrecanase-1 and aggrecanase-2 expression.Aggrecan expression levels(0.278 ± 0.015,0.235 ± 0.029,0.195 ± 0.028,0.399 ± 0.028,0.299 ± 0.020,0.263 ±0.019) induced by both 1,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium were significantly decreased than the levels(0.472 ± 0.0358,0.197 ± 0.018,all P ＜ 0.05) in control group(0 mg/L toxin).Selenium partially blocked the effects induced by 1,10,and 20 μg/L T-2 toxin(all P＜ 0.05).One,10,20 μg/L T-2 toxin and 0,0.1 mg/L selenium increased both aggrecanase-1 mRNA levels(0.535 ± 0.033,1.071 ± 0.043,1.454 ± 0.058,1.057 ±0.048,0.555 ± 0.036,0.902 ± 0.045) and aggrecanase-2 mRNA levels(0.596 ± 0.038,0.656 ± 0.033,0.949 ±0.049,0.600 ± 0.040,0.453 ± 0.031,0.164 ± 0.011),when compared with control(0.481 ± 0.023,0.346 ±0.020 for aggrecanase-1 and 0.387 ± 0.020,1.021 ± 0.046 for aggrecanase-2,respectively,all P ＜ 0.05).Selenium partially blocked 10,20 μg/L T-2 toxins induced upregulation of aggrecanase-1 (all P ＜ 0.05) and aggrecanase-2 (all P ＜ 0.05 ).Conclusions These data suggest a possible molecular mechanism that T-2 toxin could induce cartilage matrix degradation through the upregulation of aggrecanases expression and enzyme activities.Trace element selenium has some protective effect on cartilage proteoglycan degradation induced by T-2 toxins.

Eleven flavonol glycosides were isolated from the ethanol extract of Lysimachia clethroides by a combination of various chromatographic techniques including column chromatography over silica gel, Sephadex LH-20, and reversed-phase HPLC. Their structures were identified as astragalin (1), isoquercitrin (2), isorhamnetin-3-O-β-D-glucopyranoside (3), quercetin-3-O-β-D-6"-acetylglucopyranoside (4), quercetin-7-O-β-D-glucopyranoside (5), prunin (6), 2-hydroxynaringin-5-O-β-D-glucopyranoside (7), kaempferol-3-O-rutinonoside (8), kaempferol-3-O-robinobioside (9), rutin (10) and kaempferol-3,7-di-O-β-D-glucopyranoside (11). Among them, compounds 4, 7 and 11 were obtained from the Lysimachia genus for the first time, while compounds 3, 5 and 9 were firstly reported from this plant. In the preliminary assays, compounds 2, 6 and 8 possessed significant inhibition against aldose reduc- tase, with IC50 values of 2.69, 1.00, 1.80 μmol · L(-1), respectively; none of compounds 1-11 exhibited obvious cytotoxic activity (IC50 > 10 μmol · L(-1)).
Drugs, Chinese Herbal ; chemistry ; Flavonols ; chemistry ; Glycosides ; chemistry ; Molecular Structure ; Primulaceae ; chemistry ; Spectrometry, Mass, Electrospray Ionization

Objective To evaluate the significance of platelet function monitoring in anti -platelet treatment of acute myocardial infarction . Methods The platelet maximum aggregation rates ( MAR ) induced by arachidonic acid , adenosine diphosphate and collagen were detected in 60 cases of acute myocardial infarction .The platelet aggregation function was compared before and after anti -platelet therapy for 24 h and 1 week.The plasma concentration of clopidogrel acid was detected and the correlation between concentration of clopidogrel acid and platelet aggrega-tion was analyzed.Results The MAR induced by arachidonic acid , adenosine diphosphate and collagen were significantly decreased after an-tiplatelet treatment for 24 h( P<0.01 );but after 1 week, no further re-duce in platelet aggregation rate was found , which showed some tole-rance.After 24 h, patients with MAR decreased by more than 30% ac-counted for 36.5%, 40.4% and 21.2%, respectively induced by arachidonic acid , adenosine diphosphate and collagen; patients with MAR decreased by 10% -29% accounted for 30.8%, 25%, and 34.6%,respectively,andthosewithMARdecreasedlessthan10%accountedfor34.6%,36.5%and44.2%,re-spectively.During half year follow -up, 34% of patients showed reoccurrence of cardiovascular events , and 55% of them were found aggregation rate less than 30%.Conclusion The aggregation rates under induction of arachidonic acid, adenosine diphosphate and collagen are all less than 30%when adopting the same anti -palate regime, but the incidence of cardiovascular events increased significantly and the platelet aggregation function and blood concentration of clopidogrel acid has no correlation .