Objective To investigate the effect of penehyclidine (PHCD) pretreatment on Toll-like receptor 4 (TLR4) mRNA expression in the lung following acute lung injury (ALI) induced by hemorrhagic shock in rats. Methods Forty healthy SD rats of both sexes weighing 200-250 g were randomly divided into 5 groups( n = 8 each): group Ⅰ sham operation (group S); group Ⅱ ALI (group ALI); group Ⅲ, Ⅴ, PHCD 0.3,1.0, 3.0 mg/kg were given iv respectively at 30 min before hemorrhagic shock (P1-3). Hemorrhagic shock was induced by exsanguinations. MAP was maintained at 35-45 mm Hg for 60 min. The animals were killed at 4 h after resuscitation. Their lungs were removed for microscopic examination, W/D lung weight ratio and determination of TLR4 mRNA expression in the lung tissue (by RT-PCR). NF-κB p65 protein expression in the lung tissue was determined (by immuno-histochemical staining). Results Hemorrhagic shock significantly increased TLR4 mRNA and NF-κB p65 protein expression in the lung tissue and W/D lung weight ratio. Pretreatment with PHCD 1.0 or 3.0 mg/kg significantly inhibited hemorrhagic shock-induced increase in TLR4 mRNA and NF-κB p65 protein expression in the lung and W/D lung weight ratio. The lung injury was significantly ameliorated in group P2,3 as compared to group ALI. Conclusion PHCD pretreatment can attenuate ALI induced by hemorrhagic shock through down-regulation of TLR4 mRNA expression and decreasing NF-κB activity in the lung.

The concentration of 5 nucleosides, uracil, uridine, guanidine, adenine and adenosine in culture of Paecilomyces hepialid was determined by the developed method of HPLC. The HPLC method was performed on a Waters SunFire C18 (4.6 mm x 250 mm, 5 μm) column with methanol-water gradient elution as the mobile phase. The detection wavelength was 260 nm and the colunmn temperature was controlled at 30 °C. The linear range was 10.00-200.00 mg · L(-1) (r = 0.9994) for uracil, 10.10-202.00 mg · L(-1) (r = 0.9992) for uridine, 10.00-200.00 mg · L(-1) (r = 0.9991) for guanidine, 10.30-206.00 mg · L(-1) (r = 0.9992) for adenine and 10.45-209.00 mg · L(-1) (r = 0.9991) for adenosine, respectively. The RSD of precision was 0.032%, 0.035%, 0.039%, 0.049%, 0.00080%, respectively. The average recoveries of uracil, guanidine, adenine, and adenosine were 97.34%, 99.10%, 101.6%, 98.61% and 100.2% with RSD of 1.3%, 2.1%, 0.96%, 0.95%, and 1.3% respectively. The method showed high sensitivity, good selectivity, linearity and repeatability, which was suitable for the content analysis of 5 nucleosides components in P. hepialid and its extracts.
Chromatography, High Pressure Liquid ; methods ; Nucleosides ; analysis ; Paecilomyces ; chemistry

OBJECTIVETo study the effect of Chinese drugs for invigorating qi and tonifying Shen (IQTS) on expression of CD4+CD25+ regulatory T cells in spleen and maternal-fetal interface of abortion-prone mice during pregnancy.

METHODSCBA female mice were mated with DBA/2 male mice to establish abortion-prone models, which were randomly divided into 4 groups, the negative control group (fed with normal saline), the positive control group (treated with CsA), the Chinese medicine group (treated with IQTS), and the Chinese and Western medicine group (treated with IQTS+CsA). Mice were sacrificed in batches on the 9th and the 14th day of gestation, their splenic and decidual tissues were taken out to analyse CD4+CD25+ regulatory T-cell expression by flow cytometry.

RESULTSCompared with the negative control group, the expression of splenic CD4+CD25+ regulatory T all significantly increased on the 9th day of gestation (P < 0.01 or P < 0.05). There was no statistical difference in intergroup comparison of the three treatment groups (P > 0.05). Compared with the negative control group, the expression of splenic CD4+CD25+ regulatory T all significantly increased on the 14th day of gestation (P < 0.01 or P < 0.05). Of them, its expression was the highest in the Chinese and Western medicine group, showing significant difference from that in the Chinese medicine group and the positive group (P < 0.01). The difference between the Chinese medicine group and the positive group was insignificant (P > 0.05). On day 9 of gestation, compared with the negative control group, the expressions of CD4+CD25+ regulatory T in maternal-fetal interface increased in the three treated groups, showing no statistical significance (P > 0.05). Its expression was ordered from high to low in sequence as the Chinese and Western medicine group, the positive control group, the Chinese medicine group, and the negative control group. On day 14 its expression was obviously enhanced in the Chinese and Western medicine group, showing statistical difference from that in the negative control group (P < 0.05). But its expression was obviously enhanced in the Chinese medicine group and the positive group, showing insignificant difference from that in the negative group. The same sequence was found in the percentage of CD4+CD25+ T cells in CD4+ T cells.

CONCLUSIONSChinese drugs for IQTS could up-regulate the expression of CD4+CD25+ regulatory T in spleen of abortion-prone mice in the early and late pregnancy stages. When combined with CsA, it also could up-regulate its expression in maternal-fetal interface in the mid and late pregnancy stages, suggesting that Chinese drugs for IQTS are facilitate to maintain the immune tolerance state in mice during pregnancy.

Abortion, Spontaneous ; drug therapy ; immunology ; metabolism ; Animals ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; Male ; Mice ; Mice, Inbred CBA ; Phytotherapy ; Pregnancy ; Spleen ; cytology ; drug effects ; T-Lymphocytes, Regulatory ; cytology ; drug effects ; metabolism

Objective To investigate the expression of JNK2 in hyperoxic lung injury ,and explore the protective effect of sub-stance P (SP) on hyperoxic lung injury and its mechanism .Methods Sixteen SD rats were divided into four groups with 4 rats in each group :room-air and f 9 g/L saline group (group A) ,room-air and SP group (group B) ,hyperoxia injury group and f 9 g/L sa-line group (group C) ,hyperoxia injury group and SP group (group D) .Rats ingroup B and D were injected with SP 1 × 10-6 mol · L -1 · kg -1 · d-1 intraperitoneally ,group A and group C were injected with an equal volume of 9 g/L saline .The animals were sac-rificed after 14 days of experiment .Lung pathology was examined with light microscopy ,lung wet/dry (W/D) ratio and the level of SP and PCNA and TUNEL in lung were evaluated .The Superoxide dismutase (SOD) ,malondialdehyde (MDA) and glutathione (GSH) level were assayed respectively in lung tissue .The quanlity of JNK2 protein was detected by Western blot analysis .Results Compared with group A ,the high oxygen groups all had different degrees of lung injury ,,while the lung pathological pictures in group D was improved significantly compared with group C .Western blot showed that level of JNK2 in group C was obviously higher than that of group A ;After the intervention ,level of JNK2 in group D was lower than that of group C .The lung W/D retio , TUNEL and PCNA expression and distribution SOD ,MDA and GSH was consistent with the trends of JNK2 protein expression . Conclusion High oxygen stress can activate damage lung tissue JNK 2 activity ;SP protection mechanism of high oxygen lung injury may be induced by cutting high oxygen activation of JNK 2 to inhibit oxidative damage .

In order to explore the clinical hypolipidemic features of Daidai flavone extract, the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats were studied and compared. The study established the quantitative determination method of naringin and neohesperidin in plasma by UPLC-MS. Study compared the pharmacokinetics differences of naringin and noehesperidin in normal and hyperlipemia rats on the basis of establishment of hyperlipemia model. Results indicated that the pharmacokinetics features of characteristic active ingredients of Daidai flavone extract in normal and hyperlipemia rats showed significant differences. The C(max) of naringin and neohesperidin in hyperlipemia rats plasma after oral administration of Daidai flavone extract increased obviously, while t1/2, MRT and AUC0-24 h decreased, compared to normal rats. But t(max) showed no differences to that of normal rats. The results further proved Daidai flavone extract would have better hypolipidemic effect in the hyperlipemia pathological status. And the characteristic active ingredients naringin and noehesperidin were the material base of Daidai flavone extract to express the hypolipidemic effect.
Animals ; Citrus ; chemistry ; Drugs, Chinese Herbal ; isolation & purification ; pharmacokinetics ; Flavones ; chemistry ; Hyperlipidemias ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

Objective To screen the optimized Buyang Huanwu Decoction (BYHWD);To verify it. Methods H2O2 was used to induce PC12 cell oxidative stress models. MTT method was used to determine the prevention effects of BYHWD at different concentrations (0.1, 0.2, 0.5, 1.0, 2.0, 3.5 mg/mL) on in vitro oxidative stress cell models to define the optimized concentration. Orthogonal design was used to divide BYHWD single medicine into decomposed BYHWD groups, control group (only with DMEM), normal group (without H2O2 and medicine processing), and model group, to investigate the protective effects on PC12 cells. Optimized BYHWD was screened to decide the compatibility ratio of each medicine. MTT was used to detect the cell survival rate in each group. Middle cerebral artery occlusion was used to replicate MACO rat models. SD rats were randomly divided into sham-operation group, model group, BYHWD group and optimized BYHWD high-, medium-and low-dose groups. Each medication group was given relevant medicine for gavage. The screened results were verified. Results Compared with other decomposed BYHWD groups, the protective effects of the compatibility of Astragali Radix+Chuanxiong Rhizoma+Pheretima on PC12 cells was the best (P<0.05), which was nearly equaled to BYHWD. Compared with the model group, BYHWD and the optimized one could evidently reduce cerebral cortex infarction area and improve the impaired brain edema (P<0.05), and the medium-dose group was the best. Conclusion The optimized BYHWD ratio is:Astragali Radix:Chuanxiong Rhizoma:Pheretima=10:3:1.

Objective To investigate and analyze the related factors influencing the quality of life in patients with epilepsy, and provide a theoretical basis for taking appropriate measures to improve the quality of life in patients with epilepsy. Methods Sixty-five patients with epilepsy visiting our hospital from July 2007 to December 2008 were chosen in our study. The table-31 for quality of life in patients with epilepsy (Chinese version) was performed on them. The age, gender, education degree,seizure types, course of disease, cognitive function, medication compliance and fear of attack were considered as the independent variable; related factors in the table-31 for quality of life were adopted as dependent variable; multiple linear regression analysis was performed on these 2 variables. Results Education degree and course of disease could affect the memory of patients with epilepsy (standardized regression coefficients were 0.380 and 0.264, respectively). Age could affect the social activities of the patients (standardized regression coefficient was -0.303). Gender could affect the attack of the patients (standardized regression coefficient was 0.332). Conclusion The influencing factors of quality of life in patients with epilepsy are age, gender, education degree, course of disease; and education degree and course of disease enjoy the greatest influence.

BACKGROUNDPlatelet-rich plasma (PRP) as a storage vehicle of growth factors has been successfully used in clinical applications, but in most cases the platelets were autologous. However, the large volume of blood withdrawn has detrimental effects on patients with anemia or poor general health. To overcome these limitations, this study was designed to separate the growth factors in homologous platelet-rich plasma.

METHODSThe gel chromatography with Superdex-75 column was applied to separate PRP supernatants into 4 major fractions. Then the four fractions were vacuumed freeze-dried and re-dissolved in phosphate buffered saline. Proteins concentrations in PRP and in four fractions were detected by bicinchoninic acid protein assay; platelet derived growth factor-AB (PDGF-AB) and transforming growth factor beta1 (TGF-beta1) levels were determined by sandwich enzyme-linked immunosorbent assays. The effects of fractions on the proliferation of human marrow-derived mesenchymal stem cells (MSCs) were determined by 3-(4, 5- dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay.

RESULTSPRP supernatants were separated into four major fractions by gel chromatography. The proteins recovery was 96.72%. Of the four fractions, fraction B contained the highest TGF-beta1 and PDGF-AB levels, and the highest proteins concentrations. Cell proliferation curves of MSC demonstrated that fraction B and C induced a remarkable increase of MTT values compared to the untreated culture (P < 0.05), and the effects of fraction B and C showed no significant difference compared to the PRP group (P > 0.05). Fraction A and D showed no significant difference to the negative control group (P > 0.05).

CONCLUSIONSThe growth factors in PRP supernatants could be preliminarily separated into four fractions by gel chromatography, and the freeze-drying fractions retained the biological activity of growth factors. The growth factors were mostly presented in fraction B and C, and they promoted cell proliferation effectively.

Bone Marrow Cells ; cytology ; Cell Proliferation ; drug effects ; Chromatography, Gel ; Humans ; Mesenchymal Stromal Cells ; cytology ; drug effects ; Platelet Count ; Platelet-Derived Growth Factor ; isolation & purification ; pharmacology ; Platelet-Rich Plasma ; chemistry ; Transforming Growth Factor beta1 ; isolation & purification ; pharmacology

In recent studies some urea derivatives have been identified as potent anti-tuberculosis agents by targeting mycobacterial membrane protein large 3 (MmpL3). However, this compound series as exemplified by AU1235 exhibited poor in vitro pharmacokinetic profile. With AU1235 as the lead, we have identified a novel benzimidazole series as potential anti-tuberculosis agents by using scaffold hopping approach. Among these synthesized compounds, 2-aminobenzimidazole derivative 8b showed the potent anti-tuberculosis activity with the MIC value of 0.03 microg x mL(-1). This compound also showed improved metabolic stability compared to AU1235. Our investigation indicated that benzimidazole derivatives are the promising lead for further optimization as anti-tuberculosis agents.
Antitubercular Agents ; pharmacology ; Benzimidazoles ; chemistry ; pharmacology ; Drug Design ; Humans ; Structure-Activity Relationship ; Tuberculosis ; drug therapy

The purpose of this work was to study the characteristics of rat brain abnormalities at two hemispheres at the early stage of electrogenic epilepsy. Experiments were performed on 37 male Sprague-Dawley rats. Chronically repetitive tetanization (60 Hz, 2 s, 0.4 - 0.6 mA) was used to stimulate the right dorsal hippocampus (DHPC) of the rat brain once a day for 2, 4, 6, 8 or 10 d, respectively. The T(2) weighted magnetic resonance image (T(2)-WI) were obtained from each experimental rat at the end of the experiments. Histological sections were obtained after experimentation. The results showed that the main pathologic changes at the early stage of epilepsy included: (1) T(2)-WI hyperintensification, the histological enlargement of lateral ventricle (LV) and pathological hyperplasia of ventricular choroidea plexus occurred. The pathological hyperplasia was symmetric in two hemispheres, but the LV enlargement was not. (2) Histologically enlarged LV area showed a resemblance to T(2)-WI hyperintensive area. Compared with the control rats, large T(2)-WI hyperintensive area (P=0.0259; P=0.0184; P=0.0184; P=0.0404; P=0.0259) and histologically enlarged LV area (P=0.0210; P=0.01; P=0.0100; P=0.0152) were present in chronically tetanized rats. (3) Dynamic characteristics of histologically enlarged LV area resembled to those of T(2)-WI hyperintensity area in chronically tetanized rats at different stimulating day. Lateralization of T(2)-WI hyperintensity was in accordance with that of T(2)-WI abnormal area and of histologically enlarged LV. These abnormalities were severe on the contralateral side on the stimulating day 6, or on the ipsilateral side on the stimulating day 10. These results imply characteristic propagation of brain abnormalities crossing to the opposite hemisphere at the early stage of an electrogenic rat epilepsy.
Animals ; Cerebral Cortex ; pathology ; physiopathology ; Electric Stimulation ; Epilepsy ; etiology ; pathology ; physiopathology ; Hippocampus ; physiopathology ; Magnetic Resonance Imaging ; Male ; Rats ; Rats, Sprague-Dawley ; Time Factors