OBJECTIVETo study the adscription of plasma effective constituents of rat after oral administration of Gegen Qinlian decoction.

METHODThis study was performed by using HPLC-UV to identify the chemical constituents of rat plasma samples obtained after oral administration of the whole and individual herbs of Gegen Qinlian decoction. A reversed phase Globalsil C18 column (4.6 mm x 250 mm, 5 microm) was used. The mobile phase was composed of 20 mmol x L(-1) ammonium acetate (adjusted to pH 4. 5 by acetate acid) and acetonitrile with gradient elution. The flow rate was 1.0 mL x min(-1) and the column maintained at room temperature. The detection wavelength was set at 270 nm.

RESULTThe chromatographic analysis of the rat plasma obtained after oral administration of Gegen Qinlian decoction showed 31 compounds, fifteen of which were the original constituents contained in Gegen Qinlian decoction, thirteen of which were metabolites, while the other three only existed after oral administration of the Decoction. By comparison of the retention times of the unknown peaks with that of standard compounds and, by spiking method with the standard compounds, five peaks were identified as puerarin, baicalin, berberine, jatrorrhizine and palmatine.

CONCLUSIONThis HPLC-UV method is simple and reliable in the analysis of the UV absorbing chemical constituents of rat plasma. The compounds absorbed into blood and its metabolites are the possible effective constituents of Gegen Qinlian decoction because they interact directly within the body. The results obtained in this investigation may provide useful information in the further understanding of the action mechanism of the individual and whole herbs in Gegen Qinlian decoction.

Administration, Oral ; Animals ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Male ; Rats ; Reproducibility of Results

OBJECTIVETo study the effect of Feiji Recipe (FR) intervening indoleamine 2,3-dioxygenase (IDO) induced immune escape on the murine model of Lewis lung carcinoma. Methods Totally 48 C57BL/6 mice inoculated with Lewis lung cancer cells transfected with human (enhanced green fluorescent protein,EGFP)-IDO gene were divided into four groups according to radom digit table, i.e., the model group (administered with normal saline by gastrogavage) , the Chinese medicine group (treated with FR Decoction at the daily dose of 100 mg/g by gastrogavage), the 1-methyl-D-trytaphan (1-MT) group (administered with 1-MT mixed liquor at the daily dose of 100 mg/kg by gastrogavage), and the Paclitaxel group (treated with Paclitaxel at the daily dose of 15 mg/kg by peritoneal injection), 12 in each group. The intervention was started from the 2nd day of modeling. The survival time was observed in 24 of them. Ratios of CD4+ CD25+ FoxP3+ regulatory T cells (Treg) in the spleen were detected in the rest 24 mice by flow cytometry respectively.

RESULTSCompared with the model group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells remarkably decreased in the Chinese medicine group, the 1-MT group, and the Paclitaxel group (P < 0. 01). Compared with the Paclitaxel group, the survival time was significantly prolonged in the Chinese medicine group and the 1-MT group (P < 0.01); ratios of Treg cells decreased significantly in the 1-MT group (P < 0.05).

CONCLUSIONFR could inhibit the proliferation of lung cancer cells and immune eseape, improve the immune function, and prolong the survival of tumor-bearing mice.

Animals ; Antineoplastic Agents ; pharmacology ; therapeutic use ; Carcinoma, Lewis Lung ; drug therapy ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Humans ; Indoleamine-Pyrrole 2,3,-Dioxygenase ; Lung Neoplasms ; Mice ; Mice, Inbred C57BL ; Paclitaxel ; T-Lymphocytes, Regulatory

OBJECTIVETo establish a method for studying efficacious materials of traditional Chinese medicines from an overall perspective.

METHODCarthamus tinctorius was taken the example. Its major components were depleted by preparing liquid chromatography. Afterwards, the samples with major components depleted were evaluated for their antioxidant effect, so as to compare and analyze the major efficacious materials of C. tinctorius with antioxidant activity and the contributions.

RESULTSeven major components were depleted from C. tinctorius samples, and six of them were identified with MS data and control comparison. After all of the samples including depleted materials are compared and evaluated for their antioxidant effect, the findings showed that hydroxysafflor yellow A, anhydrosafflor yellow B and 6-hydroxykaempferol-3, 6-di-O-glucoside-7-O-glucuronide were the major efficacious materials.

CONCLUSIONThis study explored a novel and effective method for studying efficacious materials of traditional Chinese medicines. Through this method, we could explain the direct and indirect contributions of different components to the efficacy of traditional Chinese medicines, and make the efficacious material expression of traditional Chinese medicines clearer.

Alkalies ; chemistry ; Antioxidants ; chemistry ; Carthamus tinctorius ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; Glucosides ; chemistry ; Mass Spectrometry

UNLABELLEDOBJECTIVE; To evaluate the clinical value of different CT diagnostic criteria for peripancreatic artery and vein invasion in pancreatic carcinoma through comparison with the findings on surgical exploration.

METHODSOf 72 patients of having suspected pancreatic carcinoma were examined by multiplane spiral CT. Among 43 confirmed by surgical pathology; 15 underwent pancreaticoduodenectomy; 28 were found to have unresectable tumors. The peri-pancreatic major vessels including the superior mesenteric artery, celiac artery, hepatic artery, superior mesenteric vein and portal vein were explored carefully during surgical exploration.

RESULTSThe criteria for peri-pancreatic artery invasion was the presence of one of the following signs: artery embeded in tumor, or more than half of the artery circumference involved by tumor with wall irregularity or stenosis. The sensitivity of the above described criteria was 75.0% (12/16). If the criteria of tumor involvement exceeding half of the vessel circumference were adhered to, the sensitivity was 87.5% (14/16), which was high than the former, but the specificity was lower than that of the former one (90.2% versus 95.1%). The criteria for peri-pancreatic vein invasion was presence of any of the following signs: vein obliteration, more than half of the vein circumference involved by tumor, vein wall irregularity, vein stenosis, tear-drop sign of superior mesenteric artery. The sensitivity of the above described criteria was 92.9% (39/42), higher than that of the criteria that more than half of the vessel circumference was involved by the tumor (69.0%, 29/42), but the specificity of both criteria was the same (97.4%, 37/38).

CONCLUSIONFor assessing peri-pancreatic artery and vein invasion, using the combination of different CT diagnostic criteria has higher accuracy than when using only criteria of more than half of vessel circumference involved by tumor.

Adult ; Aged ; Carcinoma, Pancreatic Ductal ; diagnosis ; surgery ; Celiac Artery ; diagnostic imaging ; Female ; Hepatic Artery ; diagnostic imaging ; Humans ; Male ; Mesenteric Artery, Superior ; diagnostic imaging ; Mesenteric Veins ; diagnostic imaging ; Middle Aged ; Neoplasm Invasiveness ; Pancreatic Neoplasms ; diagnosis ; surgery ; Pancreaticoduodenectomy ; Portal Vein ; diagnostic imaging ; Reproducibility of Results ; Sensitivity and Specificity ; Tomography, Spiral Computed ; methods

OBJECTIVETuberculosis is still a public health problem. Host genetic factors, such as polymorphisms in NRAMP1 gene, may play a role in the development of tuberculosis. To clarify the effect of NRAMP1 gene polymorphisms on the development of childhood tuberculosis, the association of NRAMP1 gene polymorphisms with susceptibility to tuberculosis in the ethnic Han Chinese children was investigated.

METHODSFrom January 2005 to March 2008, 130 ethnic Han children with tuberculosis (TB group) were enrolled. Three hundred and ninety hospitalized ethnic Han children for physical examination in the surgery department were used as the control group. The controls were matched with tuberculosis children by age, sex and area. PCR-RFLP analysis was performed on DNA samples to identify allele genotypes of INT4 and D543N in NRAMP1 gene. Genotype frequency differences between tuberculosis patients and controls were analyzed using x2 test.

RESULTSNo statistical difference was found in the genotype frequency of variants G/C and C/C at the INT4 locus between the TB and the control groups. At the D543N locus, the frequency of genotype variants (G/A and A/A) was significantly higher in the TB group (34/130) than that in the control group (66/390) (x2=5.349, P<0.05; OR=1.74, 95%CI=1.08-2.79). When stratified by sex, differences in the genotype distribution were observed only in females at the D543N locus, which the variant genotypes were higher in the TB group (16/52) than in the control group (21/155) (x2=7.866, P<0.05; OR=2.84, 95%CI=1.34-5.99). For males, there was no difference between the TB and the control groups. At the INT4 locus, no difference was observed between the two groups in boys and girls.

CONCLUSIONSGenotypic variation at the D543N locus in NRAMP1 gene may be associated with susceptibility to tuberculosis in ethnic Han Chinese children. Variant genotypes in NRAMP1 gene (G/A and A/A) may be susceptible genotypes to tuberculosis in ethnic Han Chinese children. Girls with variant genotypes were more susceptible to tuberculosis.

Cation Transport Proteins ; genetics ; Child ; Child, Preschool ; China ; ethnology ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Infant ; Male ; Polymorphism, Genetic ; Sex Factors ; Tuberculosis ; ethnology ; genetics

OBJECTIVETo screen anti-c-Met Fab from a phage antibody library and identify its binding activity.

METHODSThe expression of c-Met of HCC lines was identified by Western blot and immunofluorescence. Antibodies against c-Met were screened with immobilized antigen. After five rounds of panning, 30 randomly selected clones were identified by phage ELISA to select specific clones with high affinity. The positive clones were selected for Fab soluble expression in TOP10F and the binding activities were analysed in HCC lines.

RESULTSc-Met expressed in HCC membrane was confirmed by Western blot and immunofluorescence. A Fab fragment named AM2-26 with fine activity to c-Met was selected. AM2-26 binding specificity was confirmed by IP, FACS and immunofluorescence.

CONCLUSIONThe anti-c-Met Fab binding to c-Met in HCC provides a promising candidate for the biotherapy of hepatoma.

Antibodies ; immunology ; isolation & purification ; Cell Line, Tumor ; Cloning, Molecular ; Gene Library ; Humans ; Immunoglobulin Fab Fragments ; immunology ; Immunoglobulin Variable Region ; immunology ; Peptide Library ; Proto-Oncogene Proteins c-met ; immunology ; Recombinant Fusion Proteins ; immunology

BACKGROUNDDiabetic retinopathy (DR) is one of the most common complications of diabetes. Angiotensin-converting enzyme inhibitor is thought to play an important role in preventing and treating retinal diseases in animal models of DR. The aim of the present study was to investigate the role of angiotensin-converting enzyme inhibitor (ACEI, captopril) in the treatment of patients with non-proliferative DR.

METHODSThree hundred and seventeen type 2 diabetic patients (88.05% of participants) without or with mild to moderate non-proliferative retinopathy were randomly divided into captopril group (n = 202) and placebo group (n = 115). All subjects received 24-month follow-up. General clinical examinations, including blood pressure and glycated hemoglobin, as well as comprehensive standardized ophthalmic examinations were performed. Color fundus photography and optical coherence tomography (OCT) were used to grade diabetic retinopathy and detect macular edema respectively.

RESULTSThe levels of blood pressure and glycated hemoglobin in the two groups of patients remained within the normal range during the entire follow-up and no significant difference was found between the initial and last visits, suggesting that ACEI drugs play a protective role on the DR patients independent of its anti-blood pressure role. DR classification showed that 169 eyes (83.66%) remained unchanged and the DR grade of 33 eyes (16.34%) increased in captopril group, while 84 eyes (73.04%) remained unchanged and the grade of 31 eyes (26.96%) increased in placebo group (P = 0.024). Captopril treatment improved macular edema in 55.45% eyes, which was significantly higher than the 37.39% improvement in placebo group (P = 0.002). No significant difference was found in the visual acuity between the two groups (P = 0.271).

CONCLUSIONCaptopril can improve or delay the development of DR and macular edema, which can be used in the early treatment of DR patients with type 2 diabetic mellitus.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Angiotensin-Converting Enzyme Inhibitors ; therapeutic use ; Captopril ; therapeutic use ; Diabetic Retinopathy ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo explore the toxicity of LPS-induced activated microglia to preoligodendrocytes (preOLs) and the effect of 1400W, a selective inhibitor of inducible nitric oxide synthetase (iNOS), on the blockage of the toxicity.

METHODSCo-cultured microglia and preOLs obtained from two-day-old Sprague-Dawley (SD) rats were divided into three groups: co-culture control group, co-culture LPS group and co-culture LPS plus 1400W group. After cultured cells were induced by LPS (100 ng/ml) for 48 hours, the concentration of nitric oxide (NO) was measured by nitric acid-oeoxidize-colorimetry, the level of peroxynitrite (ONOO(-)) was determined by immunocytochemistry, and the synthetic level of iNOS was detected by Western blotting, respectively. The morphologic observation of apoptotic preOLs stained with Hoechst 33342/PI and the apoptotic rate of preOLs detected by flow cytometry were processed simultaneously. Data were analyzed with SPSS 11.0 software.

RESULTSCompared to co-culture control group, there was significant increase in levels of NO [(82.27+/-3.41) micromol/L vs. (167.86+/-9.87) micromol/L, t=8.593, P<0.01], ONOO(-)[(6.14+/-1.27) x 10(7)/L vs. (34.38+/-7.75) x 10(7)/L, t=5.892, P<0.01], and iNOS [(0.18+/-0.027) vs. (0.79+/-0.068), t=9.26, P<0.01] induced by LPS in co-culture LPS group, and with a higher apoptotic rate of preOLs [(6.73+/-1.39)% vs. (24.77+/-2.05)%, t=12.619, P<0.01]. However, all levels of NO [(69.55+/-5.07) micromol/L, t=8.896, P<0.01], ONOO(-) [(10.33+/-3.47) x 10(7)/L, t=14.96, P<0.01] and iNOS (0.35+/-0.042, t=5.506, P<0.01) decreased significantly with the use of 1400W at a dose of 10 micromol/L in co-culture LPS plus 1400W group, and the apoptotic rate of preOLs [(11.8+/-2.06)%, t=7.715, P<0.01] was also reduced evidently.

CONCLUSIONSNO, ONOO(-) and iNOS, etc. play important roles in the death pathway of preOLs induced by LPS. 1400W can block effectively the toxicity of LPS-activated microglia toxicity to preOLs through inhibiting iNOS selectively and reducing the production of NO and ONOO(-), and improve the survival rate of preOLs.

Amidines ; pharmacology ; Animals ; Benzylamines ; pharmacology ; Cells, Cultured ; Lipopolysaccharides ; toxicity ; Microglia ; drug effects ; metabolism ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase ; antagonists & inhibitors ; metabolism ; Oligodendroglia ; drug effects ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo improve the quality of medical instrument clinical trial in China.

METHODSA systematic analysis on current situation of medical instrument clinical trial in China is conducted to find a way out of problems.

RESULTS & CONCLUSIONSThere is a low level of medical instrument clinical trial now in China. There are still many shortcomings in law and regulations of medical instrument clinical trial. The sponsors and s do not know medical instrument clinical trial well, the measures of quality related to medical instrument clinical trial must be improved.

China ; Clinical Trials as Topic ; standards ; Equipment and Supplies ; Humans

In order to investigate the role of non-adrenergic non-cholinergic nerves in regulating mechanical and electrical activity of gastric circular smooth muscle, the effects of ATP and its analogues on gastric motility and electrical activities were observed in guinea-pig. In organ bath system, isometric force of the circular smooth muscle of guinea-pig gastric antrum was measured. Electrical activity of the muscle was recorded by using intracellular microelectrode. Electrical and mechanical activities were recorded by chart recorder. ATP and 2-MeSATP potentiated the mechanical activity but did not affect electrical activity in gastric circular smooth muscle. ATP and 2-MeSATP-induced contraction was effectively blocked by nonselective P2y-purinoceptor antagonist, reactive-blue-2 and suramin, but ATP-induced contraction was not blocked by alpha,beta-MeATP-induced desensitization of P2x-purinoceptors. However, alpha,beta-MeATP, P2x-purinoceptor agonist, attenuated slow waves with membrane hyperpolarization and inhibited contraction. The relaxation by beta,gamma-MeATP was blocked by alpha,beta-MeATP-induced desensitization of P2x-purinoceptors. ATP-induced contraction was blocked by external calcium-free, but not by nicardipine, a L-type calcium channel blocker. Indomethacin, a nonselective cyclooxygenase inhibitor, did not block ATP-induced contraction. The results suggest that: (1) ATP- and analogues-induced contraction is mediated by P2y-purinoceptor, whereas alpha,beta-MeATP-induced relaxation by P2x-purinoceptor in guinea-pig gastric antral circular smooth muscle. (2) ATP-induced contraction is dependent on extracellular calcium, but Ca2+ entry is not mediated by L-type calcium channel. (3) Prostaglandins are not involved in ATP- and analogue-induced contraction of gastric circular smooth muscle in guinea-pigs, and alpha,beta-MeATP-induced relaxation is related to membrane hyperpolarization.
Adenosine Triphosphate ; analogs & derivatives ; pharmacology ; Animals ; Electrophysiology ; Guinea Pigs ; In Vitro Techniques ; Microelectrodes ; Muscle Contraction ; drug effects ; physiology ; Muscle, Smooth ; drug effects ; physiology ; Purinergic Agonists ; Pyloric Antrum ; drug effects ; physiology ; Thionucleotides ; pharmacology