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There have been several studies of nuclear factor-κB (NF-κB) and high-mobility group box1 (HMGB1) associated with the pathophysiology of cerebral ischemia. Tricin 7-glucoside, a major bioactive compound extracted from Sedum sarmentosum Bunge. The objectives of this study were to determine the effects of Tricin 7-glucoside on a cultured neuronal cell line, SH-SY5Y in vitro and experimental ischemic stroke in vivo. For oxygen-glucose deprivation (OGD) and tumor necrosis factor-α (TNF-α) stimulated SH-SY5Y cell line in vitro, SH-SY5Y cells were incubated with Tricin 7-glucoside. For in vivo experiment, rats were subjected to middle cerebral artery occlusion (MACO) for 1h, then followed by reperfusion for 23 h. Treatment of SH-SY5Y cells with Tricin 7-glucoside reduced the OGD-induced apoptosis and cytotoxicity, blocked TNF-α-induced NF-κB and IκB-α phosphorylation, and decreased HMGB1 expression. At doses higher than 50mg/kg, Tricin 7-glucoside produced a significant neuroprotective potential in rats with ischemia and reperfusion (I/R). Tricin 7-glucoside (100mg/kg) demonstrated significant neuroprotective activity even after delayed administration at 2h and 4h after I/R. Tricin 7-glucoside 100mg/kg attenuated histopathological damage, decreased brain edema, inhibited NF-κB activation and reduced HMGB1 expression. These data show that Tricin 7-glucoside protects brain against I/R injury with a favorable therapeutic time-window by alleviating cerebral I/R injury and attenuating blood-brain barrier (BBB) breakdown, and its protective effects may involve HMGB1 and NF-κB signaling pathway., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Stroke is the third leading cause of death in industrialized countries and the most important cause of acquired adult disability. Many evidences suggest that inflammation accounts for the progression of cerebral ischemic injury. Arctigenin, a phenylpropanoid dibenzylbutyrolactone lignin isolated from certain plants, has shown anti-inflammatory activity against diabetes and Alzheimer's disease. In this study, we tested whether arctigenin can protect middle cerebral artery occluded (MCAO) rats. Male Sprague-Dawley rats were pretreated with arctigenin or vehicle for 7 d before being subjected to transient occlusion of middle cerebral artery and reperfusion. Rats were evaluated at 24 h after MCAO for neurological deficit scoring. Furthermore, the mechanism of the anti-inflammatory effect of arctigenin was investigated with a focus on inflammatory cells, proinflammatory cytokines, and transcriptional factors. Arctigenin significantly reduced cerebral infarction and improved neurological outcome. Arctigenin suppressed the activation of microglia and decreased the expression of interleukin (IL)- 1β and tumor necrosis factor (TNF)-α. These results revealed that arctigenin has a promising therapeutic effect in ischemic stroke treatment through an anti-inflammatory mechanism.

Objective: To explore the association of urinary podocyte excretion and renal expression of podocyte-specific marker podocalyxin (PCX) with clinicopathological changes in immunoglobulin A nephropathy (IgAN)., Methods: Morning urine samples from IgAN patients and healthy controls were collected. The expression of glomerular PCX was quantified in 50 IgAN patients diagnosed by renal biopsy. IgAN was classified based on the Lee's Grading system and scored according to the Katafuchi semiquantitative criteria. Morphological evaluation of podocyte was determined by electron microscopy., Results: The amount of urinary podocytes in the IgAN patients was significantly higher than that in the healthy controls (p < 0.01). Pairwise comparison among Lee's grades of IgAN showed that the median of urinary podocytes in Lee's I-II group was lower than that in Lee's III, IV, and V groups (p < 0.05); group III lower than group V (p < 0.05). The positive rate of urinary podocytes was the highest in Lee's IV and V groups (100%), and lowest in Lee's I-II group (55%). Multiple comparison among groups of Lee's grades of IgAN showed that the glomerular PCX expression in Lee's I-II group was higher than that in Lee's III, IV, and V groups (p < 0.05); groups III and IV higher than group V (p < 0.05). The amount of urinary podocytes in IgAN patients was negatively correlated with PCX expression (r = -0.702, p < 0.01), but positively correlated with 24-h urinary protein (r = 0.465, p < 0.01) and glomerular (r = 0.233, p < 0.01) and renal tubular pathological scores (r = 0.307, p < 0.05). The glomerular PCX expression was negatively correlated with 24-h urinary protein (r = -0.367, p < 0.05) and glomerular (r = -0.560, p < 0.05) and tubular pathological scores (r = -0.377, p < 0.05). Electron microscopy showed significant changes in podocytes of IgAN, especially in the foot process., Conclusion: The amount of urinary podocyte can reflect the loss of podocytes in renal tissue, which may be a marker of IgAN progression.

Background: Excessive use of computed tomography (CT) in emergency departments (EDs) has become a concern due to its expense and the potential risks associated with radiation exposure. Although studies have shown a steady increase in the number of CT scans requested by ED physicians in developed countries like the United States and Australia, few empirical data are available regarding China., Methods and Findings: We retrospectively analyzed a database of ED visits to a tertiary Chinese hospital to examine trends in CT utilization and their association with ED outcomes between 2005 and 2008. A total of 197,512 ED visits were included in this study. CT utilization increased from 9.8% in 2005 to 13.9% in 2008 (P<.001 for trend). The ED length of stay for visits with CT utilization was 0.6 hour longer than those in which CT was not obtained. CT utilization increased the ED cost by an average $48.2. After adjustment for patients' demographics, arrival hours and clinical condition, CT utilization during ED visits was significantly associated with high ED cost (Odds Ratio [OR]: 21.70; 95% confidence interval [CI], 17.00-27.71), long ED length of stay (OR: 1.22; 95%CI, 1.12-1.34), and more likely to receive emergency operations (OR: 2.31; 95%CI, 1.94-2.76). However, there was no significant correlation between CT use and the possibility to be admitted to inpatient wards (OR: 0.82; 95%CI, 0.65-1.04). With respect to the time-related trends, CT utilization during ED visits in all study years was significantly associated with high ED cost and more likely to receive emergency operations., Conclusion: CT utilization was associated with higher ED cost, longer ED length of stay and more likely to receive emergency operations, but did not correlate with a significant change in the admission rate.

Triple reuptake inhibitors (TRIs) that block the dopamine transporter (DAT), norepinephrine transporter (NET), and serotonin transporter (SERT) are being developed as a new class of antidepressant that may have better efficacy and fewer side effects compared with traditional antidepressants. The purpose of this study was to characterize a new chemical entity, 4-[2-(dimethylamino)-1-(1-hydroxycyclohexyl)ethyl] phenyl 4-methoxybenzoate hydrochloride (TP1). TP1 was designed as a prodrug of desvenlafaxine. Competitive radioligand binding assays were performed using cells expressing the human dopamine (DA) transporter (hDAT), the human serotonin (5-HT) transporter (hSERT), and the human norepinephrine (NE) transporter (hNET) with K(i) values for TP1 of 190 nM, 2076 nM, and 1023 nM, respectively. Uptake assays were performed with IC(50) values for TP1 of 712 nM, 521 nM, and 628 nM, respectively. TP1 (0.06 mmol/kg, orally) rapidly penetrated rat brain and hypothalamus, translated into desvenlafaxine within 1 h, and demonstrated higher bioavailability and better pharmacokinetic properties than desvenlafaxine succinate (DVS). TP1 (0.06 mmol/kg, orally) significantly increased extracellular levels of DA, NE, and 5-HT compared with baseline in the rat hypothalamus by microdialysis assay. In dose-response assays, oral administration of TP1 reduced the time of immobility in a dose-dependent manner during tail suspension test and forced swimming test (FST). This antidepressant-like effect manifests in the absence of significant increases in motor activity even at doses of up to 32 mg/kg. The ability of TP1 to inhibit the reuptake of three biogenic amines closely linked to the etiology of depression may result in a therapeutic profile different from antidepressants that inhibit the reuptake of serotonin and/or NE., (Copyright © 2011 IBRO. Published by Elsevier Ltd. All rights reserved.)

Objective: To investigate the value of high frequency and color Doppler ultrasonography in detection of synovitis and the intra-articular vascularization in the knee joint of patients with newly-diagnosed rheumatoid arthritis (RA)., Methods: Forty-one patients (30 women, 11 men) with newly-diagnosed RA were recruited to a cross sectional study (RA group). Forty-one age and gender-matched healthy volunteers were used as control group. The thickness of hydatid fluid, synovium hyperplasia, color flow imaging, peak systolic velocity (PSV), end diastolic velocity (EDV), resistance index (RI), venous blood flow and intra-articular perfusion were evaluated by high frequency and color Doppler ultrasonography., Results: Totally 91.46% knee joints with synovial hyperplasia (> 2 mm) were found in 41 patients with RA (75/82 knee joints), and the thickness of the synovial membrane was 2.2 - 19.7 mm (average 6.3 ± 3.4 mm). In aspect of blood flow, the percentage of 0 to 3 grade were 18.67% (14/75), 29.33% (22/75), 45.33% (34/75) and 6.67% (5/75), respectively; the results of arterial blood were indicated with PSV (10.82 ± 3.71 cm/s), EDV (3.86 ± 1.12 cm/s) and RI (0.61 ± 0.07), while the average of venous blood velocity was 2.72 ± 1.02 cm/s. Joint effusion was found in 69 joints (84.15%) with the anteroposterior diameter 2.4 - 16.1 mm (average 6.9 ± 3.2 mm). The thickness of synovial membrane was 1.2 - 1.8 mm (average 1.4 ± 0.4 mm) and no significant difference were observed in joint effusion, signal of blood flow and thickness of synovial membrane in the control group., Conclusions: High frequency and power Doppler ultrasonography may be a valuable and optimal clinical tool to accurately and objectively detect synovial hyperplasia, vascular pannus formation and joint effusion in the knee joint of patients with RA.

Objective: To investigate the distribution of Mycobacterium tuberculosis (MTB) Beijing genotype, which was isolated from tuberculosis (TB) patients registered in local TB dispensaries in Deqing and Guanyun county respectively within 1 year, as well as its drug-resistant phenotypic and genotypic profiles and genotyping features., Methods: A total of 399 TB patients were enrolled from 2 counties. Of the 351 TB patients with MTB isolates available, 237 were male, and 114 were female; aged from 18 - 82 (46 ± 35) years. The proportion method and DNA sequencing were used to define the susceptibility of the isolates to 4 first line anti-TB drugs and the related mutation. Beijing genotype MTB strains were identified by Spoligotyping, while the "cluster" strains and the "unique" strains were defined by IS6110 restriction fragment length polymorphism (RFLP)., Results: Beijing genotype MTB strains were identified in 243 of the 351 strains isolated, and the proportion of multi-drug resistance, mono-resistance to rifampin and isoniazid among Beijing genotype MTB was 18.5% (45/243), 43.2% (105/243) and 22.2% (54/243) respectively, all being significantly higher than the non-Beijing genotype MTB, 7.4% (8/108), 24.1% (26/108) and 12.0% (13/108) respectively. katG and rpoB mutations were observed more common among Beijing genotype MTB than among non-Beijing genotype MTB, 13.2% (32/243) and 4.6% (5/108) respectively, OR = 2.553, 95%CI: 1.031 - 6.324. The Beijing genotype MTB was more likely to be clustered than non-Beijing genotype MTB, 41.2% (100/243) and 11.1% (12/108) respectively, OR = 5.503, 95%CI: 2.851 - 10.622., Conclusions: In eastern rural China, TB patients infected with the Beijing genotype MTB may have a higher risk to develop isoniazid-or rifampin-resistance and multi-drug resistance. The disease is more likely due to recent transmission.

We investigated whether 8-O-acetyl shanzhiside methylester (ND01) regulates angiogenesis and thereby improves functional outcome after stroke. Adult male rats were subjected to 1 hr of middle cerebral artery occlusion (MCAO) and reperfusion, and treated with or without different doses (5 and 10 mg/kg) of ND01, starting 24 hr after ischaemia and reperfusion (I/R) and by intravenous injection daily for 14 days. Neurological functional tests were performed and cerebral Evans blue extravasation was measured. Angiogenesis and angiogenic factor expression were measured by immunohistochemistry and Western blot, respectively. The results indicated that ND01 significantly promoted angiogenesis in the ischaemic brain and improved functional outcome after stroke. ND01 also significantly increased vascularization compared with vehicle treatment. ND01 increased the expression of VEGF, Ang1, phosphorylation of Tie2 and Akt VEGF. The Ang1/Tie2 axis and Akt pathways appear to mediate ND01-induced angiogenesis., (© 2010 The Authors. Basic & Clinical Pharmacology & Toxicology © 2010 Nordic Pharmacological Society.)

The present study was to investigate the neuroprotective efficacy and mechanism of Forsythoside B. Male Sprague-Dawley rats were subjected to middle cerebral artery occlusion for 1 h followed by reperfusion for 23 h. Rats received an intravenous bolus injection of Forsythoside B at 15 min after reperfusion. The results showed that Forsythoside B at doses higher than 8 mg/kg produced a significant neuroprotective potential in cerebral ischemia and reperfusion rats. Forsythoside B (20 mg/kg) demonstrated significant neuroprotective activity even after delayed administration at 1 h, 3 h and 5 h after cerebral ischemia and reperfusion. Forsythoside B 20 mg/kg attenuated histopathological damage as demonstrated by smaller brain infarct size and brain edema, decreased cerebral Evans blue extravasation and myeloperoxidase (MPO) activity, inhibited cerebral phosphor-IkappaB-alpha and nuclear transcription factors kappaB (NF-kappaB) expression. Meanwhile, NF-kappaB expression with immunohistochemical staining was reduced, while circulating polymorphonuclear leukocytes was increased. All of these findings suggested that Forsythoside B exerted potent neuroprotective effects with a favorable therapeutic time-window, reduce of cerebral ischemia and reperfusion injury degree, attenuating blood-brain barrier (BBB) breakdown, and its protective effects may be due to inhibition of inflammatory response., (Copyright (c) 2010 Elsevier B.V. All rights reserved.)

The present study was undertaken to examine the effect of forsythoside B (FB) on rat myocardial ischemia-reperfusion (I/R) model and elucidate the potential mechanism. Left ventricular systolic pressure (LVSP) and +/-dp/dt(max) were detected. Blood samples were collected to determine serum levels of troponin T (Tn-T), TNF-alpha and IL-6. Hearts were harvested to assess histopathological change and infarct size, determine content of MDA, myeloperoxidase (MPO), SOD and GPx activities, analyze expression of high-mobility group box 1 (HMGB1), phosphor-I kappaB-alpha and phosphor-nuclear factor kappaB (NF-kappaB) in ischemic myocardial tissue by Western blot. Compared with control group, rats treatment with FB showed a significant recovery in myocardial function with improvement of LVSP and +/-dp/dt(max). The myocardial infarct volume, serum levels of Tn-T, TNF-alpha and IL-6, content of MDA and MPO activity in myocardial tissue were all reduced, protein expression of HMGB1, phosphor-I kappaB-alpha and phosphor-NF-kappaB were down-regulated, while attenuated the decrease of SOD and GPx activities. Besides, the infiltration of polymorph nuclear leukocytes (PMNs) and histopathological damages in myocardium were decreased in FB treated groups. These findings suggested that FB rescued cardiac function from I/R injury by limiting inflammation response and its antioxidant properties., ((c) 2009 Elsevier GmbH. All rights reserved.)

Objective: To investigate the contribution of recent transmission in the epidemic of drug-resistant Mycobacterium tuberculosis (M.TB) and related factors from biomedical and social-demographic perspectives in the Eastern rural areas of China., Methods: Identified by proportion method of drug susceptibility test, 223 drug resistant M.TB isolates and their hosts were included in the present study. These drug resistant tuberculosis isolates were first genotyped by Mycobacterial Interspersed Repetitive Units (MIRU), and those isolates with identical MIRU genotype were further classified by IS6110 restricted fragment polymorphism (RFLP). 'Cluster' was defined as two patients' M. TB isolates harboring the identical MIRU genotype and IS6110-based RFLP pattern simultaneously. Unique strains denoted those with the unparalleled MIRU genotype in the study collection. Socio-demographic and biomedical characteristics of host patients were compared between the clusters and unique groups through univariate and multivariate logistic regression analysis., Results: Based on the MIRU-IS6110 pattern, there were 52 isolates belonged to the "cluster" group and 171 as the "unique" group. Drug resistant M.TB strain isolated from patients at the age of 30 - 60 year had a higher probability of being clustered, comparing to those from patients below 30 years of age (30.9% vs. 11.9%; OR = 3.297; 95%CI: 1.169 - 9.297). Such finding were also seen in the isolates from patients with previous treatment history compared to newly diagnosed patients (32.9% vs. 18.4%; OR = 2.163, 95%CI: 1.144 - 4.090). The multi-drug resistant M.TB strain was found to have been more frequently clustered when comparing to the mono-drug resistant M.TB (47.2% vs. 15.5%; OR = 4.773; 95%CI: 2.316 - 9.837). The transmission pattern of drug resistant tuberculosis was presented mainly by the sporadic distribution in small group within rural villages., Conclusion: Transmission of drug-resistant tuberculosis was seen in the population living in the Eastern rural areas of China, and causal contact within villages was considered as the main route of recent transmission. Patients at middle age and having previous tuberculosis treatment history might have increased the risk of transmission by patients with drug resistant tuberculosis.

This study was conducted to investigate the efficacy of cornin, an iridoid glycoside, in an experimental cerebral ischemia induced by middle cerebral artery occlusion (MCAO) and reperfusion (I/R), and to elucidate the potential mechanism. Adult male Sprague-Dawley rats were subjected to MCAO for 1 h, then reperfusion for 23 h. Behavioral tests were used to evaluate the damage to central nervous system. The cerebral infarct volume and histopathological damage were assessed to evaluate the brain pathophysiological changes. Spectrophotometric assay methods were used to determine the activities of superoxide dismutase (SOD) and glutathione-peroxidase (GPx). Contents of malondialdehyde (MDA), the generation of reactive oxygen species (ROS) as well as respiratory control ratio and respiratory enzymes of the brain mitochondria were also determined. The results showed that cornin significantly decreased neurological deficit scores, and reduced cerebral infarct volume and degenerative neurons. Meanwhile, cornin significantly increased the brain ATP content, improved mitochondrial energy metabolism, inhibited the elevation of MDA content and ROS generation, and attenuated the decrease of SOD and GPx activities in brain mitochondria. These findings indicate that cornin has protective potential against cerebral ischemia injury and its protective effects may be due to amelioration of cerebral mitochondrial function and its antioxidant property., (Copyright (c) 2009 John Wiley & Sons, Ltd.)

8-O-acetyl shanzhiside methylester (ND01), an iridoid glucoside compound, was isolated from the leaves of Lamiophlomis rotata (Benth.) Kudo. The present study elucidated the effects of ND01 on the cultured rat cortical neuron injury induced by oxygen-glucose deprivation. The results showed that ND01 treatment obviously attenuated apoptosis and ameliorated mitochondrial energy metabolism in rat cortical neurons by increasing cell survival rate, mitochondrial respiratory enzyme activities, mitochondrial respiratory control ratio and adenosine triphosphate (ATP) content, and by attenuating lactate dehydrogenase (LDH) leakage, intracellular Ca(2+) level and caspase-3 activity in a concentration-dependent manner. These findings indicated that ND01 has potential against cerebral ischemic injury, and its protective effect on oxygen-glucose deprivation-induced injury might be due to the suppression of intracellular Ca(2+) elevation and caspase-3 activity, and improvement of mitochondrial energy metabolism., (Copyright (c) 2009 Elsevier B.V. All rights reserved.)

This study was carried out to investigate whether rosmarinic acid (RA) has antifibrotic effect on experimental liver fibrosis in vitro and in vivo and its possible mechanism. Culture of hepatic stellate cells (HSCs) determine proliferation and expression of transforming growth factor-beta1 (TGF-beta1), connective transforming growth factor (CTGF) and alpha-smooth muscle actin (alpha-SMA). In carbon tetrachloride (CCL(4))-induced rat liver fibrosis model, determined biochemical indicator, liver fibrosis grade and histopathological changes, immunohistochemical detected liver TGF-beta1 and CTGF expression. The results indicated that RA could inhibit HSCs proliferation, inhibit TGF-beta1, CTGF and alpha-SMA expression in cultured HSCs. It has marked evident in reducing fibrosis grade, ameliorating biochemical indicator and histopathological morphology, reducing liver TGF-beta1 and CTGF expression in CCL(4)-induced liver fibrosis. These findings suggest that RA has potentially conferring antifibrogenic effects., (Copyright 2009 Elsevier GmbH. All rights reserved.)

Hereditary persistence of fetal hemoglobin (HPFH), often associated with mutations in the beta-globin gene cluster, is normally benign, but a person carrying both HPFH and another beta-thalassemia (beta-thal) mutation will develop serious anemia. These people might be erroneously diagnosed as having homozygous beta-thal with common reverse dot-blot methods. Here we report a 5-year old boy with thalassemia intermedia, who is a compound heterozygote for the rare HPFH-6 deletion with codons 41/42 (-TCTT) beta(0)-thal, who inherited the deletion from his mother and the beta(41/42) mutation from his father.

Objective: Multicentre clinical observation for therapeutic efficacy and clinical safety of anticoagulation and thrombolysis in deep venous thrombosis (DVT)., Methods: 127 patients with DVT in left leg were randomly divided into four groups. Group A: treated with batroxobin. Group B: use Low Molecular Weight Heparin (LMWH). Group C: batroxobin and LMWH. Group D: urokinase and LMWH. Observing the perimeter of thigh and calf periodically, monitoring coagulation function, registering the frequency and degree of the complication., Results: All the treatments in four groups can relieve the swell level of the affected legs (P < 0.05). We got the best efficacy when we use batroxobin together with LMWH (P < 0.05), especially in treating the patients with a long course of treatment. Batroxobin can obviously reduce the level of blood FBG (P < 0.05), and have no significant effect with other index of coagulation function (P > 0.05). Inject ing batroxobin into affected legs with the micro pump got a better efficacy and much safer than intravenously guttae from peripheral vein., Conclusion: Treating DVT with batroxobin got a definite efficacy, especially in cases with a long course of treatment; micro pumping can give a better effect and a high safety.

The present study was to investigate the effects of rosmarinic acid (RA) in cultured RAW264.7 cells and experimental model of sepsis induced by cecal ligation and puncture in rats and the potential mechanism. Results showed that RA concentration dependently down-regulated the levels of TNF-alpha, IL-6, and high-mobility group box 1 protein in LPS-induced RAW264.7 cells, inhibited the IkappaB kinase pathway, and modulated nuclear factor-kappaB. Intravenous injection of RA alone or in combination with imipenem reduced cecal ligation and puncture-induced lethality in rats. In addition, serum levels of TNF-alpha, IL-6, high-mobility group box 1 protein, triggering receptor expressed on myeloid cells, and endotoxin were down-regulated; in contrast, serum level of IL-10 was up-regulated. Amelioration of hemodynamics and decrease in serum enzyme activities and myeloperoxidase in lung, liver, and small intestine were also observed after RA injection. These data indicate that the antisepsis effect of RA was mediated by decreasing local and systemic levels of a wide spectrum of inflammatory mediators. This article provides the first evidence that RA has the capacity to inactivate inflammatory response in sepsis. The anti-inflammatory mechanism of RA may inhibit activation of the nuclear factor- kappaB pathway by inhibiting IkappaB kinase activity.

Objective: To establish a rat model of mycoplasma pneumonia (MP) for investigating the pathogenesis of MP and its therapy with drugs., Methods: Thirty Wistar rats were randomly divided into 6 groups (n=5), including a control group, a MP model group, a erythromycin lactobionate group and 3 erythromycin microspheres groups (high, middle, and low dose groups). With the exception of those in the control group, all the rats received intranasal MP administration followed by corresponding treatments administered via tail vein injection. At different time points after inoculation of the pathogen, the lungs of the rats were taken for histopathological scoring., Results: In the MP model group, the lung pathology was characterized by patchy interstitial pneumonitis with predominantly lymphocyte infiltration and mucosal edema. The bronchiolar walls became thickened and the lumens narrowed. In erythromycin lactobionate and erythromycin microspheres treatment (high and middle dose) groups, clear cell boundaries were observed in the lungs where no obvious pathological changes were found. RT-PCR amplification showed positive results of MP RNA in the model group, erythromycin lactobionate group and erythromycin microsphere groups., Conclusion: The approach described is practicable to establish rat models of MP. Erythromycin microspheres can effectively relieve the lung inflammations and has therapeutic effect on MP.

Objective: To describe the drug resistance-related molecular characterization and clustering feature of rifampicin-resistant (RIFr) M.tuberculosis (M.tb) in rural area of eastern China., Methods: All patients diagnosed as RIFr M.tb in Deqing and Guanyun county during one year period from 2004 to 2005 were included in the study. By proportion method of drug susceptibility test, 65 isolates were identified resistant to rifampicin and regarded as the studied strains. Hotspots of rpoB gene and katG gene were detected by direct DNA sequencing. Beijing genotype M.tb strains were identified by spoligotyping. IS6110-RFLP (IS6110 restriction fragment length polymorphism) and clustering analysis were performed on all RIFr M.tb isolates available., Results: The mutations in 81 bp rifampicin-resistance determination region (RRDR) of the rpoB gene were observed among 60 (92%) RIFr M.tb isolates, with mutation in locus 531 observed in the majority of RIFr isolates (37/65). 49 (82%) of the 60 isolates were multidrug resistant TB (MDR-TB), which were referred to as resistant to both RIF and isoniazid (INH). Through spoligotyping, 54(83%) isolates were identified as Beijing genotype strains. In clustering analysis of IS6110-RFLP, 24 isolates were grouped into 11 clusters, suggesting that the recent transmission of M.tb did exist among patients. Regarding the drug resistance profile in clusters, all the isolates in clusters were also MDR-TB. 7 clusters contained isolates carrying different mutations were related to RIF-resistance. Multivariate analysis showed the proportion of new cases in clustered patients is higher than that in the un-clustered patients (new/previously treated: OR = 3.342; 95%CI: 1.081 - 10.32)., Conclusion: The acquisition of rifampicin resistance in M.tb was more likely to be resulted from the selective growth of RIFr M.tb in the specific drug resistant M.tb such as isoniazid-resistant M.tb. Previous elongated irregular treatment might favor the epidemic of RIFr M.tb.

Astilbin, a flavonoid compound, was isolated from the rhizome of Smilax glabra Roxb. This study was conducted to investigate the efficacy of astilbin on experimental diabetic nephropathy (DN) in vivo and in vitro and its possible mechanisms. Astilbin was added in high glucose stimulated HK-2 cells, streptozotocin-induced experimental DN, randomized to receive intragastric ( I. G.) astilbin to observe its anti-renal lesion effect. Results showed that astilbin inhibited high glucose stimulated HK-2 cell production of transforming growth factor-beta1 (TGF-beta1) and connective tissue growth factor (CTGF) in vitro, especially CTGF; analogic results was also found in vivo. I. G. of astilbin 2.5 mg/kg or 5 mg/kg significantly ameliorated renal function, reduced kidney index, while it increased body weight and survival time in animals. In addition there was no significant difference in blood glucose level between the STZ-treated group and the astilbin groups. Furthermore, astilbin ameliorated the pathological progress of renal morphology. Astilbin can exert an early renal protective role to DN, inhibit production of TGF-beta1 and especially of CTGF. We suggest that astilbin inhibition of CTGF may be a potential target in DN therapy. This work provides the first evidence for astilbin as a new candidate of DN therapeutic medicine., (Georg Thieme Verlag KG Stuttgart, New York.)

Objective: To evaluate the performance of MIRUs (mycobacterial interspersed repetitive units) genotyping alone, IS6110-RFLP (IS6110 restriction fragment length polymorphism) genotyping alone and their combination applied in the molecular epidemiological study of Mycobacterium tuberculosis (MTB) isolates circulating in rural China., Methods: A cross-sectional study was designed to collect MTB isolates from the TB patients registered in local TB dispensaries of Deqing county and Guanyun county from 2004 to 2005. The proportion method was used to determine drug susceptibility of MTB isolates to the first line anti-TB drugs (isoniazid, rifampin, ethambutol and streptomycin). The Beijing family of MTB was identified by Spoligotyping. All isolates were genotyped by MIRUs alone, IS6110-RFLP alone and their combination., Results: Of the 351 studied MTB isolates, 243 (69.2%) had the genotypes that belonged to the Beijing family, and 223 (63.5%) were resistant to at least 1 anti-TB drug, including 53 (15.1%) resistant to isoniazid and rifampin simultaneously or multidrug resistant (MDR). The heterogeneity of 12 MIRUs loci differed from 0.76 in MIRU26 to 0.003 in MIRU2. MIRUs alone identified 235 genotypes (HGI = 0.9317), including 46 "clusters" containing 162 isolates and 189 "unique" pattern/isolates. Thirty-eight isolates comprised the largest MIRUs defined cluster and presented MIRUs type 2233 2517 3533. MIRUs based clusters were further identified by IS6110-RFLP into 28 subgroups containing 80 isolates (HGI = 0.9989). In comparison, IS6110-RFLP determined 267 genotypes from all isolates, including 46 "clusters" containing 130 isolates (HGI = 0.9684) and 221 "unique" pattern/isolates. All the 46 IS6110-RFLP defined clusters could also be further identified by MIRUs into 31 subgroups (HGI = 0.9992). The performance of MIRUs prior to IS6110-RFLP combination was comparable to that of IS6110-RFLP prior to MIRUs combination, especially in Beijing genotype strain (HGI: 0.9930 vs 0.9933) and MDR-TB strains (HGI: 0.9965 vs 0.9963)., Conclusion: For feasibility, cost and discriminatory power, MIRUs prior to IS6110-RFLP combination is more suitable for the massive epidemiological investigation of MTB in rural China.

The present study was carried out to investigate the effects of paeoniflorin in cultured RAW264.7 cell line as well as in an experimental model of sepsis induced by cecal ligation and puncture, and intraperitoneal injection (i.p.) of lipopolysaccharide in rats. Results showed that paeoniflorin concentration-dependently down-regulated the levels of TNF-alpha, IL-6 and high-mobility group-box 1 protein in lipopolysaccharide-induced RAW264.7 cell, inhibited the IkappaB kinase pathway and modulated NF-kappaB. Intravenous injection (i.v.) of paeoniflorin alone or in combination with imipenem reduced i.p. of lipopolysaccharide or cecal ligation and puncture-induced lethality in rats. In addition, serum levels of TNF-alpha, IL-6, high-mobility group-box 1 protein, triggering receptor expressed on myeloid cells and endotoxin were down-regulated; by contrast, serum levels of IL-10 were up-regulated. Amelioration of hemodynamics, decrease of enzyme levels, decrease of myeloperoxidase in lung, liver, and small intestine were also found after paeoniflorin injection. These data indicate that the anti-sepsis effect of paeoniflorin was mediated by decreasing local and systemic levels of a wide spectrum of inflammatory mediators. This work provides the first evidence that paeoniflorin has the capacity to inactivate inflammatory response in sepsis and the anti-inflammatory mechanism of paeoniflorin may inhibit activation of the NF-kappaB pathway by inhibiting IkappaB kinase activity.

This study was conducted to investigate the efficacy of cornuside, a secoiridoid glucoside compound, in cultured macrophages as well as in an experimental model of sepsis induced by cecal ligation and puncture (CLP) in rats. Cornuside was added to cultured macrophages at different concentrations, and all CLP rats were randomized to receive an intravenous injection of the corresponding drug followed by observation of its antisepsis effect. Our results showed that cornuside downregulated the levels of TNF- alpha, IL-6, and NO production in a dose-dependent manner in activated macrophages, while it upregulated the level of IL-10. Intravenous injection of cornuside or imipenem alone or in combination reduced CLP-induced lethality in rats after CLP. In addition, serum levels of TNF- alpha, IL-6, triggering receptor expressed on myeloid cells, and endotoxin were downregulated. On the other hand, the serum levels of IL-10 were upregulated. Decreased bacterial counts in blood, peritoneum, spleen, liver, and mesenteric lymph nodes and decreased myeloperoxidase in lung, liver, and small intestine also were found after cornuside injection. These data indicate that the antisepsis therapeutic effect of cornuside is mediated by decreased local and systemic levels of a wide spectrum of inflammatory mediators. This work provides first evidence for the clinic use of cornuside as a new immunomodulatory drug that has the capacity to inhibit the inflammatory response in sepsis., (Copyright Georg Thieme Verlag KG Stuttgart. New York.)

Objective: To study the oral chronic toxicity of 97% isopropyl thioxanthone (97% ITX) in rats, determine the no-observed adverse effect levels (NOAEL)., Methods: Four groups of rats were fed with foodstuff containing 97% ITX in the dosage of 1000.0, 250.0, 62.5 mg/kg respectively for 2 years. The general behavior, body weight, food availability ect. were observed during the experiment. At the end of the experiment, blood and urine samples were collected for routine and biochemical assays. The internal organs were taken for calculating their organ coefficients and histopathological examinations., Results: During the experimental period, no obvious abnormality were found in the experimental animals. The body weight and the total food availability rate in the high dosage group of male were lower than that of control (P < 0.05). Hematology examination showed that the quantity of Hb and RBC in high dosage groups of both the male and female and Hb in the male middle group were all lower than the control group (P < 0.01 or P < 0.05). Analysis of correlation indicated that r = -0.433, P < 0.01 in male, r = -0.337, P < 0.01 in female of Hb; r = -0.266, P < 0.05 in male, r = -0.317, P < 0.01 in female of RBC. There were obviously negative correlation. Serum biochemistry examination showed the concentration of CHO in the high and middle dosage treated rats of male and female were higher than that of the control (P < 0.01 or P < 0.05). Analysis of correlation indicated that r = 0.497, P < 0.01 in male, r = 0.417, P < 0.01 in female. No abnormality were found in urine examination. The organ weight and organ coefficient such as liver, were higher than control group (P < 0.01). The result of histopathological examinations displayed that the renal tubule Cast and the tubulointerstitial nephritis in the treated groups were higher than that of control group (P < 0.01)., Conclusion: 97% ITX could obviously interfere with the animals' physical condition, and reduce the number of RBC and the concentration of Hb in the blood, interact metabolism of lipoid and induce the concentration of CHO in the serum. The livers of the treated rats are compensatory enlarged. And kidneys of the poisoning animals are damaged. The 2 years oral NOAEL of 97% ITX in rats are more than 4.63 mg/kg for female rats, and larger than 4.06 mg/kg for male rats.

Cornuside, a secoiridoid glucoside compound, was isolated from the fruit of Cornus officinalis Sieb. et Zucc. The present study elucidates the effects of cornuside on cultured rat cortical neuron damage induced by oxygen-glucose deprivation. The results show that cornuside treatment obviously attenuates apoptosis and ameliorates mitochondrial energy metabolism in rat cortical neurons by increasing the cell survival rate, mitochondrial antioxidant enzyme activities, mitochondrial respiratory enzyme activity, mitochondrial respiratory control ratio and the ATP content, and by decreasing the mitochondrial malondialdehyde content, lactate dehydrogenase leakage rate, intracellular Ca(2+) level and caspase-3 activity in a concentration-dependent manner. These findings indicate that cornuside has protective potential against cerebral ischemic injury, and its protective effects may be due to the suppression of intracellular Ca(2+) elevation and caspase-3 activity, and improvements in mitochondrial energy metabolism and antioxidant properties., (Copyright 2009 S. Karger AG, Basel.)

Objective: To evaluate the antimicrobial effects of erythromycin microspheres against Mycoplasma pneumoniae in rats., Methods: With erythromycin lactobionate as the positive control, erythromycin microspheres at 3 non-toxic doses (0.1, 0.5, and 1.2 g.kg(-1).d(-1)) were administered intragastrically for 6 consecutive days in Wistar rats with Mycoplasma pneumoniae infection. The general condition and lung index of the rats were observed and measured to assess the therapeutic effects of the treatments against Mycoplasma pneumoniae infection., Results: The erythromycin microspheres at 0.1, 0.5, 1.2 g.kg(-1).d(-1) significantly alleviated the symptoms of the rats infected with Mycoplasma pneumoniae and reduced the pulmonary index of the infected rats from 1.75 to 1.45, 1.38 and 1.25, respectively (P < 0.01). An obvious dosage-effect relationship was noted between the dose of erythromycin microsphere and the tissue pathologies due to the infection., Conclusion: Erythromycin microspheres possess strong activity against Mycoplasma pneumoniae in rats.

Aim: To evaluate the efficiency of 3 short hairpin RNA (shRNA) interfering with the herpes simplex virus type 1 (HSV-1) gene coding glycoprotein D (gD) for inhibiting the gD expression and virus replication in vitro., Methods: Vero cells were selected for an in vitro model of infection. Three shRNA sequences (shRNAgD1, -gD2, and -gD3) targeting specifically the gD gene of HSV-1 were selected for evaluating the antiviral effects. The antiviral effects of shRNA in the cells infected with HSV-1 were evaluated by cytopathic effect (CPE) observations and plaque assays. The transcription level of viral RNA and the gD expression were studied by RT-PCR, Western blotting, and flow cytometry., Results: With the 3 shRNA at a final concentration of 120 nmol/L, a significant inhibition of CPE in the HSV-1-infected cells was observed. The ED50 of shRNA-gD1, gD2, and gD3 were 48.74+/-2.57, 57.13+/-3.24, and 114.64+/-5.12 nmol/L, respectively. The gD gene decreased significantly after viral infection in the Vero cells pretreated with shRNA compared to the virus group. The expressions of the gD protein, determined by Western blotting and flow cytometry, were also drastically decreased in shRNA-transfected cells., Conclusion: Exogenous shRNA molecules can suppress the HSV-1 gD expression. They are inhibitors of HSV replication during infection in Vero cells.

Objective: To investigate the therapeutic efficacy of short course chemotherapy (SCC) on drug resistant tuberculosis (DR-TB) cases and related influencing socioeconomic factors. TB patients registered in local county TB dispensaries of two rural counties were followed up in Deqing and Guanyun of Eastern China, during 2004/2005., Methods: Culture-positive patients (Deqing: 182, Guanyun: 217) were selected as subjects of this study. A cohort of TB patients was established at the beginning of their treatment and each patient was followed-up three times by questionnaires. Proportional method of drug susceptibility test was used to define the resistance to the 1st-line anti-TB drugs. chi2 test Kaplan-Meier method and Cox analysis were applied in multivariate analysis to investigate the negative conversion of smear positive sputum, treatment result of SCC and its socioeconomic influencing factors., Results: The cure rates of multi-drug resistant TB (MDR-TB), other drug resistant TB (ODR-TB) and pan-drug susceptible TB, were 58.3%, 91.0%, 98.7% and 51.3%, 89.5%, 93.5% respectively in Deqing and Guanyun. The liver dysfunction (RR = 0.18, 95% CI: 0.04-0.69) and previous treatment history (RR = 0.26, 95% CI: 0.07-0.93) were associated with treatment result among MDR-TB. Result on treatment in ODR-TB was influenced by previous treatment history (RR = 0.66, 95% CI: 0.44-0.98) and Patient delay (> 2 weeks) (RR = 0.67, 95% CI: 0.46- 0.97)., Conclusion: The priority in treating MDR-TB would include: managing side effect, developing the fast sensitive drug susceptibility test and modifying the treatment regimen corresponding to drug resistance.

Objective: To establish a rapid multiplex PCR (MPCR) detection system of oxacillin and erythromycin resistance genes in Staphylococcus aureus (S. aureus) and evaluate the genotype distribution of the genes associated to mecA, ermA and ermC resistance in Guangzhou., Methods: The S. aureus strains were identified and susceptibility tests were performed using VITEK-60 or PHOENIX-100 system. The inducible resistance to clindamycin of strains with of erythromycin resistance was conducted using D-test, and the MPCR system of for detecting the antibiotic resistance genes was optimized., Results: The MPCR assay for detecting the resistance genes was constructed successfully. According to the results of MPCR, the positivity rates for mecA, ermA and ermC genes among the 124 strains of S. aureus isolated from clinical samples were 56.5%, 50% and 33.9%, respectively. Good correlation was observed between the antibiotic resistance phenotypes and the S. aureus genotypes. mecA were detected in all the methicillin-resistant S. aureus strains, and ermA and/or ermC in 97.7% of the S. aureus strains with erythromycin resistance., Conclusion: This MPCR system allows rapid and reliable analysis of antibiotic resistance genotypes of S. aureus isolated from clinical samples. mecA, ermA, and ermC genes are among the predominant genetic determinants for the resistance to oxacillin and erythromycin in S. aureus isolates in Guangzhou.

Objective: To compare the nephrotoxicity of high- and low-osmolar contrast media (HOCM and LOCM), and to determine the protective role of fosinopril or telmisartan and its possible mechanism., Methods: Forty eight healthy SD rats were randomly divided into 6 groups: a normal control group, a glycerol control group, a low-osmolar contrast media (LOCM) group, a high-osmolar contrast media (HOCM) group, a fosinopril group, and a telmisartan group. Glycerine for inducing kidney damage was given to all rats except the normal control group. Twenty-four hours after the injection of glycerine, the mixed fosinopril suspension (10mg/kg) or telmisartan (5mg/kg) was poured into the stomach in the preventive group. Serum creatinine (SCr) and plasma angiotensin II (AngII) levels were detected by an automatical biochemical analyzer and radioimmunoassay; caspase-3 activity and claudin-1 expression of the renal tissue were detected by fluorometric method and immunohistochemical method. The renal injury was assessed by hematoxylin and eosin (HE) staining and terminal deoxynucleotide mediated nick and labeling (TUNEL) staining, respectively., Results: In diatrizoate-injected rats, SCr and AngII levels were increased (P<0.05). Expression of claudin-1 protein and caspase-3 activity in the renal tissue was upregulated. The histologic changes and percentage of apoptotic cells were milder in the LOCM rats than those in the HOCM rats. In the group pretreated with fosinopril or telmisartan, no increase in the levels of SCr and AngII was discovered. The expression of claudin-1 protein and caspase-3 activity was significantly lower than that in the HOCM group. The renal injuries induced by diatrizoate were alleviated., Conclusion: Both HOCM and LOCM could cause cellular apoptosis in the kidney.LOCM was less toxic to rat kidney than HOCM. Nephrotoxicity induced by HOCM might be related to caspase-3, claudin-1 and AngII. Fosinopril or telmisartan may protect the renal tissue from nephrotoxicity induced by diatrizoate.

Simple sequence repeats or microsatellites have been used as genetic markers in population genetics because of their abundance and length variation between different individuals. This study examined the SSRs in the completely sequenced Sclerotinia sclerotiorum and Botrytis cinerea genomes. The occurrences, relative abundance, relative density, most common motifs, and the longest SSRs in the two species were analyzed, and compared with other plant pathogenic fungal species, such as Fusarium graminearum, Magnaporthe grisea and Ustilago maydis. The results demonstrated that the SSRs are abundant in S. sclerotiorum and B. cinerea genomes, and 6 539 and 8 627 SSRs were obtained from these species. The types and distributions of SSRs have similarities between the two species. In the genomes of S. sclerotiorum and B. cinerea, tetra-, penta- and hexa-nucleotide repeats were more abundant than other species, indicating high mutation rates in these species. Furthermore, the abundance and relative density of SSRs were not influenced by the genome sizes and GC content. The analysis in this study provided useful information on applications of microsatellites in population genetics of S. sclerotiorum and B. cinerea.

Objective: To identify the type of the intermediate filament (IF) protein of Angiostrongylus cantonensis and analyze its tissue localization., Methods: Recombinant pET-IF of antigen IF was expressed in E.coli with IPTG induction, and the expression products were purified by His.Bind column and identified for determining the type of the IF protein by Western blotting. Anti-IF antibody was prepared by multi-spot subcutaneous injection into mouse and used to detect the tissue slices of A. cantonensis by immunohistochemical analysis., Results: The antigen IF were correctly expressed and purified, and identified as a keratin located in the intestine wall and cytoplusma., Conclusion: The antigen IF is distributed in the intestine wall of A. cantonensis.

Objective: To optimize the condition for inducing the differentiation of 3T3-L1 preadipocytes into adipocytes and study the expression of PTEN tumor suppression gene in this process, aiming to understand the regulatory role of PTEN in normal adipocyte differentiation and collect laboratory evidence for developing drugs targeting PTEN., Methods: The differentiation of 3T3-L1 preadipocytes cultured in high-glucose DMEM were induced according to 2 protocols with different combinations of dexamethasone, isobutylmethylxanthine (IBMX) and insulin, and the resultant adipocytes were identified by oil red O staining. The total proteins of 3T3-L1 were extracted and analyzed by Western blotting, and PTEN homology between mice and human was analyzed by bioinformatic method., Results: For optimized 3T3-L1 differentiation, 3T3-L1 cells were initially induced with the combination of 1 micromol/L dexamethasone, 0.5 mmol/L IBMX and 5 microg/ml insulin for 48 h, followed by treatment with 5 microg/ml insulin in 4.5 g/L glucose DMEM for 48 h, which resulted in high differentiation rate of 3T3-L1 cells (up to 90% on the 10th day) with unified morphology and size. PTEN expression varied quantitatively in the process of differentiation, especially low on the 12th day as compared with those measured on days 4, 6 and 9. The mice PTEN mRNA shared 96% homology and PTEN amino acid 100% homology with their human counterparts., Conclusion: Endogenous PTEN expression is down-regulated during 3T3-L1 differentiation, suggesting that PTEN may enhance insulin sensitivity and promote adipogenesis under physiological conditions.

Objective: To investigate the effect of an oral preparation of Alternathera philoxeroides Griseb (APG) against respiratory syncytical virus (RSV) in mice., Methods: APG preparation was administered orally in RSV-infected mice at different daily doses (2.5, 4.5 and 6.5 g/kg) to observe the therapeutic effect of the preparation., Results: Distinct differences were observed between the death rate of the mice treated with APG at daily dose of 4.5 and 6.5 g/kg and that of the untreated mice with infection. After AGP treatment of the mice at 6.5 g/kg, the detection rate of the virus was 31.3% in the blood and 37.5% in the lung tissue, significantly lower than that in the untreated mice. The virus detection rate was 43.8% in the lung tissues of mice treated with APG at 4.5 g/kg, also significantly lower than that in the untreated control. APG treatment at the 3 doses resulted in different lung indices from that of the control., Conclusion: APG may be effective for treatment of RSV infection.

Objective: To investigate the association of Coxsackie B virus (CBV) with habitual abortion., Methods: CBV IgM antibody, viral RNA and virions were detected in 86 women with habitual abortion and 40 with induced abortion by means of enzyme-linked immunosorbent assay (ELISA), RT-PCR and virus isolation, respectively., Results: The positivity rate of CBV IgM were 87.2% and 35% in the two groups, respectively, and the detection rate of the viral RNA was 53.5% and 17.5% in blood lymphocytes, and 59.3% and 17.5% in the placentas. The virions were found in the placentas in 41.9% and 6.9% of the women, respectively. The positivity rates of CBV IgM, viral DNA and virions showed significant difference between the two groups (P<0. 01)., Conclusion: CBV might be one of the causes responsible for habitual abortion.

One-photon fluorescence(OPF) and two-photon fluorescence (TPF) of two chromophores 1,4-bis(9-ethyl-vinyl carbazole) naphthalene (abbreviated as BECVN) and 1,4-bis(4-N, N-methylamino styryl) naphthalene (abbreviated as BMABN) were resported. Strong emission at 492-541 nm and at 500-556 nm, respectively, were obtained under the excitation of -370 nm (Xe lamp) and of 730-800 nm femto-second laser pulses, respectively. Although the relative fluorescence quantum yield (phi(f)) of BECVN is as high as 7.4 times that of BMABN, the latter possesses higher TPF intensity and larger two-photon absorption (TPA) cross-section, suggesting that BMABN containing naphthalene as pi-center end-capped with N,N-methylamino styryl group exhibits more effective two-photon process. Lippert equation showed that the difference of dipole momnet (deltamu(ge)) between the ground state and the excited state for BECVN is nearly twice that for BMABN, conforming to that the deltamu(ge) value is not the key for the enhancement of two-photon absorption cross-section.

Although abnormal processing of amyloid precursor protein (APP) leads to early onset of Alzheimer's disease, the normal function of this protein is poorly understood. APP is widely expressed in axons, dendrites, and synapses in both central and peripheral nervous systems. Mice homozygous for APP or its homologue APP-like protein 2 (APLP2) null mutation (KO) are viable, but double mutants for APP and APLP2 deletions (DKO) are early postnatal lethal. To investigate the role of APP in synapse development, we compared the ultrastructure of submandibular ganglion synapses between DKO and littermate APLP2 KO mice at birth. Using serial electron microscopy, we found that the size of presynaptic boutons and the number of active zones per bouton were comparable in both strains of animals. However, the synaptic vesicle density, active zone size, and docked vesicle number per active zone were significantly reduced in DKO compared to those in APLP2 KO. These results indicate that the APP family of proteins plays an important role in regulating the formation and function of inter-neuronal synapses.

Objective: To evaluate the serious response during tilt-table test (TTT) and its prophylactic management., Method: Seventy-six elderly patients were tested at a tilt angle of 70 degrees for a maximum of 45 min and then subjected to isoproterenol-provocative tilt testing. ECG and blood pressure were monitored during the test and patients were kept at normal saline condition through a peripheral intravenous duct., Results: Fifty-one of 76 patients were defined as positive including 23 having serious response; 6 of the 23 patients had arteriosclerosis involving internal carotid arteries and 7 cases had bradycardia, two of which were associated with II degrees -I A-V block and the others with chronic atrial fibrillation. The serious response consisted of cardiac arrest for more than 5 s (6 cases), or serious bradycardia for more than 1 min (7 cases) or serious hypotension for more than 1 min (10 cases). Those with serious response were managed by returning to supine position, thus driving up legs and intravenous atropine, CPR (2 cases with cardiac arrest) and needing oxygen supplementation (11 cases). Only 2 hypotension patients recovered gradually by 10 min after emergency management, while others recovered rapidly with no complications., Conclusion: Although non-invasive, TTT may result in serious response, especially in elderly. Therefore proper patient selection, control of isoproterenol infusion and close observation of vital signs are decisive for a safe consequence.

Objective: To investigate the effects of Alternanthera philoxeroides Griseb extracts against dengue virus in vitro., Methods: MTT assay and observation of cytopathic effect (CPE) were carried out to determine the cytotoxicity of Alternanthera philoxeroides Griseb on C6/36 cell lines and its effects on dengue virus., Results: None of the 4 kinds of Alternanthera philoxeroides Griseb extracts exhibited obvious cytotoxicity on the cells at different concentrations with the exception of that over 320 microg/ml. The 4 extracts all showed inhibitory effects on dengue virus. Statistical analysis of TD(50) and ED(50) by Probit regression method suggested that extracts from coumarin had the lowest toxicity on the cells (TD(50)=535.91), whereas extracts from petroleum ether showed the strongest inhibitory effects on dengue virus (ED(50)=47.43) among the 4 extracts., Conclusion: Alternanthera philoxeroides Griseb possesses antiviral effects on dengue virus in vitro.

Objective: To study the protective effect of ligusticum chuanxiong phthalides on cerebral ischemia in rats and its related mechanism of action., Method: Middle cerebral artery occlusion (MCAO) model, thrombosis formation, platelet aggregation and hemorrheological parameters were measured to evaluate the protective effect of ligusticum chuanxiong phthalides., Result: Ligusticum chuanxiong phthalides could markedly decrease the infarct size and behavior deficits score, inhibit the thrombus formation and platelet aggregation, ameliorate hemorrheological parameters with a dose-dependent manner in rats., Conclusion: Ligusticum chuanxiong phthalides has protective effects on focal cerebral ischemia in rats, and its mechanism may be relevant to its inhibition of platelet-dependent thrombosis and amelioration of hemorrheological parameters.

Aim: To study the effects of hydroxysafflor yellow A (HSYA) on the mitochondrial function of cortex mitochondrial during cerebral ischemia in rats., Methods: Rat focal cerebral ischemia model in rats was established by ligation of middle cerebral central artery. Cortex mitochondria were isolated and prepared for the measurement of membrane fluidity, swelling, respiratory function, activities of mitochondrial respiratory enzymes and superoxide dismutase (SOD), contents of phospholipid, malondial dehyde (MDA) and Ca2+ to evaluate the function of mitochondria., Results: Focal cerebral ischemia resulted in severe neuronal mitochondrial injuries, which could be alleviated by i.v. HSYA (10, 20 mg x kg(-1)), and nimodipine (Nim, 1.0 mg x kg(-1)). The swelling of mitochondria was ameliorated, the decomposability of membrane phospholipid was decreased, the membrane fluidity of mitochondria was increased, HSYA also significantly inhibited the decrease in the activities of respiratory enzymes and SOD of mitochondria, and the increase in MDA and Ca2+ levels caused by cerebral ischemia in rats., Conclusion: HSYA showed a protective action against the cortex mitochondrial injuries in rats induced by cerebral ischemia. The mechanisms may be derived from reducing lipid peroxides, inhibiting Ca2+ overload, scavenging free radicals and improving the energy metabolism.

Objective: To understand the determinants and epidemiology of drug-resistant tuberculosis (TB) in rural area., Methods: All the diagnosed TB patients in a county with directly observed treatment (DOTS) short-course program in 2002 and a sample of patients in another county without DOTS program located in northern Jiangsu province were surveyed with questionnaires. Drug susceptibility testing (DST) for positive cultures were performed by standardized proportion method. Univariable analysis and multivariate nonconditional logistic regression modeling were applied for data analysis., Results: Among the 152 patients with DST results, 32.9% of the cases showed resistance to at least one of the first-line anti-tuberculosis drugs with 26.3% to isoniazid, 18.4% to rifampin and 17.1% to both isoniazid and rifampin respectively. Previous treatments for TB and residence in the county without DOTS program were independent risk factors for isoniazid and rifampin resistance. TB patients showing indifferent to their health and delayed health seeking for more than 1 month were more likely to have rifampin resistance. Independent predictors of multidrug-resistant TB would include delayed health seeking for more than 1 month (OR = 4.66, 95% CI: 1.26 - 17.24), residing in the county without a DOTS program (OR = 3.01, 95% CI: 1.10 - 8.22), indifference to their health condition (OR = 5.13, 95% CI: 1.06 - 24.90) and suffering from chronic diseases (OR = 0.22, 95% CI: 0.05 - 0.87)., Conclusion: Drug-resistant TB was quite serious in this rural areas, mainly associated with man-made factors but partly due to the availability of the transmission.

Objective: To understand the status of coxsackievirus B (CVB) infection in senior subjects and explore the association of the infection with cardiovascular diseases., Method: By means of ELISA and RT-PCR, CVB IgG antibody and nucleic acid were detected in 172 patients with cardiopathy and 58 control subjects., Results: The positivity rate of CVB IgG antibody was 43.61% (75/172) in the patient group and 15.52% (9/58) in the control group, showing obvious difference between the two groups. CVB RNA tests revealed a CVB RNA-positivity rate of 20.93% (36/172) in the former group and 3.45% (2/58) in the latter, with also a significant difference (P<0.01)., Conclusion: CVB infection is prevalent in aged patients with heart disease, which should be given due attention to.

Objective: To understand the state of Chlamydia pneumoniae (Cpn) infection in patients with coronary heart disease (CHD), and explore the relationship between Cpn infection and the gonesis and progressin of CHD., Method: By adopting sandwich method with double antibodies and indirect ELISA method, we detected serum Cpn immune complex and IgG antibody in 160 CHD patients (confirmed by coronary artery angiography) and 40 subjects without CHD., Results: The positivity rate of Cpn IgG type immune complex in the CHD group (63.8%) was significantly higher than that in the control group (40.0%, P<0.05), similar to the results of Cpn IgG antibody detection (65.0% vs 32.5%, P<0.05)., Conclusion: Cpn infection is closely connected with the CHD, but their exact relationship needs further exploration and study.

Background & Objective: Cell proliferation and differentiation are directed by cell cycle mechanism. When tumor cells proliferate abnormally, cyclins, which are positive agents of cell cycle, may be expressed abnormally at the same time. Now many references proved that cyclins are highly expressed in solid tumors. This study was designed to investigate the relationship between expression of cyclins and prognosis of acute leukemia., Methods: Sixty-eight cases of acute leukemia were enrolled. Reverse transcription polymerase chain reaction (RT-PCR) was performed on tumor samples to examine the expression of cyclin A, cyclin D, cyclin E. All samples were divided into three groups: acute leukemia in complete remission (12 cases), newly diagnosed acute leukemia (16 cases) and refractory leukemia (40 case). Samples of benign hemopoietic patients were used as normal control (15 cases)., Results: All the 15 cases in the control group were negative of cyclin mRNA. No difference of cyclin A mRNA expression was shown between control group and the three experiment groups (P >0.05). But expression of cyclin D and cyclin E mRNA was significantly different between them (P< 0.01). There was no difference of expression of cyclin D and cyclin E mRNA among CR patients with acute leukemia(P >0.05), while the expression of cyclin D mRNA is significantly higher than that of cyclin E in refractory leukemia group. Furthermore the expression of cyclin D mRNA in recurrent refractory leukemia patients is significantly higher than that newly diagnosed cases (P< 0.05). All cyclins mRNA positive cases were divided into single cyclin gene expressed group and multiple cyclins gene expressed group. The positive rates of them were counted. No difference was found between complete remission group and refractory leukemia group (P >0.05)., Conclusion: Cyclin D may act as a prognostic marker for acute leukemia. The amount of cyclins expressed cannot be used as a prognostic factor for acute leukemia.

The epidermal growth factor (EGF) is a member of the growth factor superfamily that can stimulate the proliferation of many types of cells. Overexpression of EGF receptor (EGFR) was observed in many types of cancer cells. Anti-EGFR antibodies or antisense nucleic acid sequences of EGFR can suppress the growth of hepatoma cells. In order to further investigate the proteome alterations associated with malignant growth of the human hepatoma cells and the influence of EGFR signal pathway on the cellular proteome, we have comparatively analyzed the proteomes of human hepatoma cells transfected with antisense EGFR sequence (cell strain JX-1) and its control cells (cell strain JX-0) by two-dimensional (2-D) gel electrophoresis and mass spectrometry. Image analysis of silver-stained 2-D gels revealed that 40 protein spots showed significant expression changes in JX-1 cells compared to JX-0 cells. Three of them, including the tumor suppressor protein maspin, changed with tendency to the normal levels. Two protein spots were identified as HSP27 in the same gel, and one of them had a reduced level in JX-1 cells. The apparent alterations of HSP27 in expression level might be the results from their differential chemical modifications, suggesting the effect of dynamic post-translational modifications of proteins on the growth of hepatoma cells. Other proteins such as glutathione peroxidase (GPX-1) and 14-3-3-sigma also exhibited altered expression in JX-1 cells, and their functional implications are discussed.

Aim: To investigate inhibitory effects of recombinant transforming growth factor alpha-Pseudomonas exotoxin 40 fusion protein (TGFalpha-PE40; TP40) on neointimal proliferation after arterial balloon injury., Methods: Forty male rabbits fed a cholesterol rich diet were randomly divided into TP40 15 microg, 30 microg, 60 microg, physiologic saline control, and normal artery groups (n=8). Rabbits in the treatment groups were treated by local administration of TP40 (15 microg, 30 microg, and 60 microg per rabbit) 24 h postinjury, and those in the control group were treated by physiologic saline 24 h postinjury. Remained 8 rabbits in normal artery group were treated by TP40 (60 microg). Optical microscope, electron microscope, and computer image analysis were used to study arterial segments 2 weeks after treatment., Results: Irregular thickening of the arterial intima, large amounts of smooth muscle cells (SMC) within the neointima, and stenosis of the arterial cavity were observed in the physiologic saline control group. Great inhibition of intimal proliferation and prevention of stenosis of the arterial cavity were observed in the TP40 treated groups that were examined 2 weeks postinjury by optical microscope. The uninjured carotids were histologically normal. Lots of destructural and necrotic SMC were observed in media in TP40 60 microg group by electron microscope. Computer image analysis showed that the neointimal area of the TP40-treated groups was markedly smaller than that of the saline control group (P < 0.01)., Conclusion: Recombinant TP40 greatly inhibited neointimal proliferation after arterial balloon injury.

An RP-HPLC method for determination and detection of sotalol hydrochloride is described. The baseline separation was achieved on ODS column within 15 minutes by using 0.1% HAc-acetonitrile (80:20, V/V) as mobile phase at a flow rate 1.5 mL/min, and the wavelength was set at 227 nm. The linear range was 5-45 mg/L (r = 0.9991) and the limit of detection was 1 mg/L(S/N > 3). The intra-day and inter-day RSDs were 0.20% and 0.93% respectively.