0.05).Conclusion There are no significant effects on bone metabolism and growth of children with small dose of IGs per day for a longer time.

To study the effect of Gegen Qinlian decoction and its major effective components on five hepatic microsomal CYP450 isozymes in rats. The in vitro hepatic microsomal incubation technique was used to co-culture Gegen Qinlian decoction and its major effective components together with each probe substrate. HPLC-MS/MS was used to establish the analytical method for metabolites of the five isoform probe substrates of CYP450 isozymes, detect the linearity among micoromal protein concentration, incubation time and metabolite formation amount. And HPLC-MS/MS was applied to determine the formation rate (V) of corresponding metabolites (acetaminophen, 4-OH-chlorzoxazone, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone) specific probe substrates of the five isoform probe substrates of CYP450 isozymes (phenacetin, polbutamide, dextromethorphan, chlorzoxazone, testosterone), in order to determine the activity of each isozyme. The result showed good linearity among acetaminophen, 4-OH-tolbutamide, dextrophan, 6-OH-chlorzoxazone and 6β-hydroxytestosterone, satisfactory precision, stability and average recovery, suggesting the method was feasible. The optimized in vitro microsomal incubation conditions conformed to the requirements in the guideline of drug-drug interaction. Gegen Qinlian decoction showed different degrees of inhibitor effect on 5 CYP450 isoforms (CYP1A2, CYP2C11, CYP2D2, CYP2E1, CYP3A1/2). Its major effective component berberine could inhibit each CYP450 isoform at high concentrations (except for CYP1A2, CYP3A1/2).
Animals ; Chromatography, High Pressure Liquid ; methods ; Cytochrome P-450 Enzyme Inhibitors ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Isoenzymes ; antagonists & inhibitors ; Liver ; enzymology ; Rats ; Tandem Mass Spectrometry ; methods

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

Objective To investigate the effects of mercury on T lymphocytes and serum immune indexes of workers with Methods occupational mercury exposure. A total of 45 workers with occupational mercury exposure were selected as the ， mercury exposure group and 47 workers without occupational mercury exposure were selected as the control group using the judgment sampling method. Cold atomic absorption spectrometry was used to detect the urinary mercury level of the two groups. （ ） +， + +， + + - + Flow cytometry was used to detect the proportion of cluster of differentiation CD 3 CD3CD4 CD3CD8 and CD3CD19 ， - （ - ） - （ - ） cells in peripheral blood and the levels of tumor necrosis factor α TNF α and interleukin 8 IL 8 in serum. The levels of （ ） ， Results immunoglobulin Ig A IgG and IgM in serum were measured by immune nephelometry. The urinary mercury level of （ ： vs ，P ） individuals in the mercury exposed group was higher than that of the control group median 92.7 13.2 μg/g Cr <0.01 . The +， + +， - + proportion of CD3 CD3CD4 CD3CD19 cells in peripheral blood and serum IgG level in the mercury exposed group （ P ）， - - （ P ） decreased all <0.05 and the serum TNF α and IL 8 levels increased all <0.01 compared with the control group. Urinary - + mercury level was negatively correlated with the proportion of CD3CD19 cells in peripheral blood and serum IgG level in the ［ （r） ， ， P ］， study subjects Spearman correlation coefficient S were −0.21 and −0.31 respectively all <0.05 and positively - - （r ， ， P ） ， correlated with serum TNF α and IL 8 levels S were 0.36 and 0.39 respectively all <0.05 . However the urinary mercury （ P ）， +， + +， level was neither correlated with IgA and IgM levels in serum all >0.05 nor with the proportion of CD3 CD3CD4 + + （ P ） Conclusion CD3CD8 cells in peripheral blood all >0.05 . Occupational exposure to mercury can lead to abnormal ， changes in peripheral blood T lymphocyte subsets B lymphocytes and serum immune factors in workers. The mercury load of occupational mercury exposure workers may impact their immune function.

Objective· To prepare nanocarriers capable of improving the immunogenicity of cholesteryl ester transfer protein (CETP) hapten. Methods · Prepare CETP polypeptide modified by thiol in Fmoc, then Sulfo-SMCC were used to prepare CETP peptide-ovalbumin (OVA) nanoparticle and CETP peptide - OVA molecule conjugates in a two-step reaction scheme. The particle size and zeta potential of nanovaccine were determined and the morphology was observed. New Zealand White rabbits were vaccinated by subcutaneous injection of vaccine and serum collected from rabbits was detected by ELISA assay for the analysis of antibodies, high density lipoprotein cholesterol(HDL-C) and low density lipoprotein cholesterol (LDL-C). The rabbits were randomly allocated to the PBS group (n=3), traditional vaccine group (n=3), and nanovaccine group (n=3). Results · Nanovaccine targeting CETP were successfully synthesized, with about 70 nm in size and about -8.81 mV in zeta potential, and possessed homogeneous spherical shape under transmission electron microscopy. Compared with traditional vaccine group, rabbits in nanovaccine group got higher antibodies. Conclusion · Nanovaccine improve immunogenicity of CETP haptens, and stimulate experiment rabbits to produce higher antibodies.

AIM To establish a liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the simultaneous content determination of matrine,oxymatrine,salvia acid B,tanshinol,protocatechualdehyde,liquiritin,astragaloside,cinnamaldehyde,tanshinone Ⅱ A,ophiopogonin D and ruscogenin in Huxin Oral Liquid (Glycyrrhizae Radix et Rhizoma,Astragali Radix,Ophiopogonis Radix,etc.).METHODS The analysis of methanol extract of this drug was performed on a 25 ℃ thermostatic Ultimate XB-C18 column (4.6 mm × 100 mm,3 μm),with the mobile phase comprising of water-methanol (containing 0.05％ formic acid) flowing at 0.4 mL/min in a gradient elution manner.RESULTS Eleven constituents showed good linear relationships with-in their own ranges (r ＞0.999 0),whose average recoveries were 96.66％-103.2％ with the RSDs of 1.4％-3.4％.CONCLUSION This sensitive,reliable and accurate method can be used for the quality control of Huxin Oral Liquid.

Objective To investigate the modulation effects of mesenchymal stem cells (MSC) implantation on the myofibroblasts congregating in the infarct region after myocardial infarction (MI).Methods MI was induced in SD rats by left anterior descending coronary artery ligation,and the experimental animals were assigned randomly into the sham group,MI + PBS group and MI + MSC group (myocardial injection of 0.1 ml 2 × 107/ml in four locations in the infarct region).Echocardiography,hemodynamic examinations and Masson trichrome staining were performed.Implanted MSC differentiation and myofibroblasts congregating in infarct region were investigated by immunofluorescence staining.TGF-β1-Smad2 signaling pathway was examined by real-time RT-PCR and Western blot.Results (1) Four weeks late,heart-weight/body-weight ratio [(3.04 ± 0.16) mg/g vs.(3.34 ± 0.14) mg/g,P ＜ 0.01] and myocardial infarction size [(38.72 ± 2.38) ％ vs.(46.36 ± 2.81) ％,P ＜ 0.01] were significantly reduced in MI + MSC group than in MI + PBS group,while scar thickness of infarct region was thicker [(0.93 ±0.17) mm vs.(0.65 ±0.16) mm,P=0.01],and LVEF was higher [LVEF:(32.5 ±5.9)％ vs.(26.5 ±4.5) ％,P =0.03] in MI + MSC group than in MI + PBS group.(2) Myofibroblasts congregating in the infarct region was significantly enhanced in MI + MSC group compared with MI + PBS group [(196 ± 20) cells/mm2 vs.(89 ± 25) cells/mm2,P ＜ 0.01],and part of implanted MSC expressed α-SMA +.(3) TGF-β1 expression and the phosphorylating of Smad2 in the infarct region were significantly upregulated in MI + MSC group compared with MI + PBS group (all P ＜ 0.05).Conclusions MSC could improve myocardial function and promote myofibroblasts congregating in the infarct region via activating the TGF-β1-Smad2 signaling pathway in this model.

Objective To study the epidemiological characteristics of Keshan disease(KD) and its fiend so as to provide evidences for further research,prevention and treatment of the disease in Sichuan province.Methods Based on KD related data from 1990 to 2008,descriptive method was used to analyze the epidemiological characteristics of KD.Results 87 KD cases were identified during the 19 years.All cases were children from the countryside,with majority of them were Yi nationality.Age of the patients ranged from 5 months to 18 years,with majority at 2-6 year-olds.The annual incidence rates Were from 0/100 000 to 1.73/100 000 with 1999 the highest(1.73/100 000).A total number of 310 preclinical or chronic KD cases were identified and the total detection rates were between 0.28% and 2.8%.with 1992 the highest.As for levels of blood selenium during the 19 years:1995 appeared the lowest(0.1345 μg/g),followed by 1990-2000(0.1558 μg/g) but all of them fell in to the level in the KD epidemic areas.Conclusion There were 5 stages in the development trend of KD disease in Sichuan province,with 2 ascending and 3 descending.The differences between any of the two stages were statistically significant.The 3 descending stages all appeared right after the selenium supplement intervention was taken.Our data showed that the program of selenium supplement was closely related to the incidence of KD,suggesting that a long term mechanism of Selenium supplement in the epidemic areas should be taking into account.

Ketamine (KTM), a N-methyl-D-aspartate (NMDA) receptor antagonist, was found to has an anti-inflammatory effect, but some patients suffered from exacerbated pro-inflammatory reactions after anesthesia with KTM. The present study was aimed to examine the underlying mechanism of pro-inflammatory effects of KTM. In this study, RAW264.7 cells were exposed to KTM and NMDA alone or combined for 30 min before lipopolysaccharide (LPS) stimulation. The expression levels of IL-6 and TNF-α were detected by RT-PCR and ELISA, and those of NMDA receptors by RT-PCR in RAW264.7 cells. Additionally, the TLR4 expression was determined by RT-PCR and flow cytometry, respectively. The results showed that in RAW264.7 cells, KTM alone promoted the TLR4 expression, but did not increase the expression of IL-6 or TNF-α. In the presence of LPS, KTM caused a significantly higher expression of IL-6 and TNF-α than LPS alone. NMDA could neither alter the IL-6 and TNF-α mRNA expression, nor reverse the enhanced expression of IL-6 and TNF-α mRNA by KTM in LPS-challenged cells. After TLR4-siRNA transfection, RAW264.7 cells pretreated with KTM no longer promoted the IL-6 and TNF-α expression in the presence of LPS. In conclusion, KTM accelerated LPS-induced inflammation in RAW264.7 cells by promoting TLR4 expression, independent of NMDA receptor.
Anesthetics, Dissociative ; pharmacology ; Animals ; Cell Survival ; drug effects ; Gene Expression Regulation ; Inflammation Mediators ; pharmacology ; Interleukin-6 ; genetics ; Ketamine ; pharmacology ; Lipopolysaccharides ; pharmacology ; Macrophages ; drug effects ; metabolism ; Male ; Mice ; N-Methylaspartate ; pharmacology ; RAW 264.7 Cells ; Signal Transduction ; drug effects ; Toll-Like Receptor 4 ; genetics ; metabolism ; Tumor Necrosis Factor-alpha ; genetics

BACKGROUNDChromosome 13q14 deletion (del13q14), chromosome 1q21 gain (amp1q21) and chromosome 17p13 deletion (del17p13) are the most frequent chromosomal aberrations in multiple myeloma (MM). They play an important role in prognosis. The aim of this study was to investigate the clinical significance of the chromosomal changes in Chinese MM patients.

METHODSInterphase fluorescence in situ hybridization (FISH) on bone marrow (BM) cells was performed in 72 enrolled MM patients. Relationships between chromosomal abnormalities and clinical features, response to therapies and prognosis were analyzed.

RESULTSAs a result of interphase FISH, 77.8% (56/72) patients had chromosome changes. The incidences of each probe were RB1 51.4% (37/72), D13S319 47.2% (34/72), 1q21 45.8% (33/72) and p53 22.2% (12/72). Osteolytic lesion, BM plasma cells index, serum calcium and serum M component were significantly correlated to del13q14. BM plasma cells and hemoglobin were correlated to amp1q21. Serum lactate dehydrogenase (LDH) was correlated with del17p13. Patients with del13q14 treated with bortezomib had a notably higher overall response rate than the patients treated with traditional chemotherapies (93% vs. 65%, P = 0.048). Patients carrying amp1q21 or/and del17p13 did not achieve satisfactory response to bortezomib. The median progression-free survival (PFS) for patients with amp1q21 was 5 months and patients without amp1q21 got 9-month PFS (P = 0.001). The median PFS for patients with del13q14 was 5 months (vs. 8 months, P = 0.026). The median PFS for patients with del17p13 was 3 months (vs. 8 months, P = 0.002). Patients with β(2)-microglobulin > 5.5 mg/L also had a worse outcome, whose median PFS was 5 months (vs. 8 months, P = 0.016).

CONCLUSIONSThe prevalence of chromosomal abnormalities of MM patients was similar in Chinese and Caucasian people. Genetic changes were associated with patients' responses to therapies and prognosis.

Adult ; Aged ; Asian Continental Ancestry Group ; Chromosome Aberrations ; Chromosome Deletion ; Humans ; In Situ Hybridization, Fluorescence ; Interphase ; Male ; Middle Aged ; Multiple Myeloma ; drug therapy ; genetics ; Prognosis