To explore a new method for identification of Mongolian patent medicine (MPM) by PCR amplification of specific alleles. Eight kinds of MPM were used to study the identification of "Digeda" raw materials. The total DNA of Lomatogonium rotatum and Corydalis bungeana samples were extracted through modified CTAB method, psbA-trnH sequence was amplified by PCR and sequenced directionally. Specific primer was designed. The DNA of 8 kinds of MPM also was extracted and purified by the commercial DNA purification kits. The rbcL and two pair of specific primers sequences were amplified. The specific amplified products were sequenced in forward directions. All specific sequences were aligned and were analyzed. The results indicated that L rotatum can be identified by specific primers from Digeda-4 Tang, Digeda-8 San, Digeda-4 San, and C. bungeana medicinal materials can be identified by specific primers from Li Dan Ba Wei San, Yi He Ha Ri-12 and A Ga Ri-35. PCR amplification of specific alleles can stably and accurately distinguish raw medicinal materials in MPM.
Alleles ; DNA Primers ; genetics ; DNA, Plant ; genetics ; Medicine, Mongolian Traditional ; Molecular Sequence Data ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods

To prepare rivastigmine liposome, rivastigmine was loaded into liposome via ammonium sulfate gradient method. Its pharmacokinetic profile in rats was evaluated after intranasal administration. The size, zeta potential, entrapped efficiency and release of rivastigmine from the liposome in vitro were determined. Plasma concentration of rivastigmine was determined by high performance liquid chromatography-tandem mass spectrometry (HPLC/MS) using antipyrine as internal standard. The pharmacokinetic parameters were calculated by DAS 2.0. The entrapped efficiency of rivastigmine liposome was (33.41 +/- 6.58) %, with the mean diameter of 154-236 nm and zeta potential of (-10.47 +/- 2.41) mV. The release behavior of rivastigmine was fitting the first order equation in vitro. The pharmacokinetic studies indicated that the C(max), T(max) and AUC(0-infinity), of rivastigmine liposome were (1.50 +/- 0.15) mg x L(-1), 15 min and (89.06 +/- 8.30) mg x L(-') x min, respectively. Rivastimine liposome was absorbed rapidly, and could reach a certain concentration in rat plasma after intranasal delivery.
Administration, Intranasal ; Animals ; Area Under Curve ; Chromatography, Liquid ; Drug Carriers ; Drug Compounding ; Liposomes ; Male ; Neuroprotective Agents ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Particle Size ; Phenylcarbamates ; administration & dosage ; blood ; chemistry ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley ; Rivastigmine ; Tandem Mass Spectrometry

OBJECTIVEThe B7-H1/PD-1 co-signaling pathway has recently been found to play a pivotal role in the immune evasion of tumor cells from host immune system. The aim of this study was to examine the B7-H1 and PD-1 expression and TILs status in gastric cancer and to elucidate the clinical relevance of B7-H1 and PD-1 to the pathogenesis of gastric carcinoma.

METHODSImmunohistochemistry and ANAE histochemical staining were used to investigate the in situ expression of B7-H1 and PD-1 and TILs status in the gastric tissues. RT-PCR was used to explore B7-H1 and PD-1 expression at the transcriptional level. The B7-H1 expression at protein level was detected by Western blot.

RESULTSExpression of B7-H1 and PD-1 was found to be increased in gastric carcinoma, but absent in normal gastric tissue. B7-H1 expression in gastric carcinoma was inversely correlated with TILs infiltration. B7-H1 but not PD-1 expression in tumor tissue was significantly correlated with some clinicopathhological variables including depth of invasion, lymph node metastasis and distant metastasis.

CONCLUSIONB7-H1 and PD-1 expressions are increased in gastric carcinoma. This signaling pathway may inhibit antitumor immune responses in gastric carcinoma. B7-H1 expression plays a critical role in the pathogenesis of human gastric carcinoma,and might be a promising prognostic marker and therapeutic target in the treatment of this disease.

Adult ; Aged ; Antigens, CD ; genetics ; metabolism ; Apoptosis Regulatory Proteins ; genetics ; metabolism ; B7-H1 Antigen ; CD4-Positive T-Lymphocytes ; immunology ; Female ; Humans ; Lymphatic Metastasis ; Lymphocyte Subsets ; immunology ; Lymphocytes, Tumor-Infiltrating ; immunology ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Neoplasm Staging ; Programmed Cell Death 1 Receptor ; RNA, Messenger ; metabolism ; Stomach Neoplasms ; genetics ; immunology ; pathology

OBJECTIVETo construct pPICZalphaA-soluble interleukin-1 receptor type I (sIL-1RI) recombinant expression vector containing the gene fragment encoding the extracellular domain of sIL-1RI for its expression in Pichia pastoris.

METHODSsIL-1RI gene was amplified by RT-PCR and inserted into the yeast expression vector pPICZalphaA by digestion ligation. The recombinant plasmid pPICZalphaA-sIL1RI was transformed into E.coli Stb13, and the positive clones were analyzed by PCR and DNA sequencing. The pPICZalphaA-sIL1RI recombinant plasmid was electroporated into GS115 cells and the transformants were analyzed by PCR. After phenotype identification, the recombinant strains were induced by methanol to express the target protein, which was analyzed by Western blotting of the cell extract and supernatant.

RESULTSThe recombinant plasmid pPICZalphaA-sIL-1RI was constructed successfully, and the results of Western blotting showed that yeast induced by methanol expressed a protein of about 39 kD.

CONCLUSIONsIL-1RI protein has been successfully expressed in P.pastoris expression system, which provides the basis for further study of sIL-1RI.

Escherichia coli ; metabolism ; Gene Expression ; Genetic Vectors ; Humans ; Pichia ; metabolism ; Plasmids ; Receptors, Interleukin-1 Type I ; biosynthesis ; genetics

OBJECTIVETo investigate the safety and efficacy of intracoronary transplantation of G-CSF mobilized autologous peripheral blood stem cells in patients with acute myocardial infarction (AMI).

METHODSPatients with AMI were randomly assigned to receive intracoronary PBSCs transplantation following bone marrow cells mobilization by granulocyte colony-stimulating factor (300-600 microg/day subcutaneously for 5 days) in addition to standard therapy (standard drug therapy and PCI, PBSCs transplantation group, n = 35) or standard therapy (standard drug therapy and PCI, n = 35). One day after G-CSF treatment was finished the patient's mononuclear cells were harvested by Baxter CS 3000 blood cell separator in a volume of 57 ml and then transferred into the infarct related artery by occluding the over the wire balloon and infusing artery through balloon center lumen. Complications during intervention and left ventricular function at baseline and 6 months thereafter were monitored.

RESULTSNo severe side effects of G-CSF treatment could be observed. Malignant arrhythmias were not observed either. Left ventricular function was significantly improved 6 months after G-CSF mobilized autologous peripheral blood stem cell transplantation compared to baseline (global left ventricular function ejection fraction: 57.1 +/- 7.8 vs. 50.0 +/- 8.2%, P < 0.0001; WMSI: 1.101 +/- 0.118 vs. 1.219 +/- 0.190, P < 0.0001; left end-systolic volume: 52.6 +/- 20.3 vs. 63.8 +/- 23.9 ml, P = 0.01 and left end-diastolic volume: 119.2 +/- 30.3 vs. 134.2 +/- 36.7 ml, P = 0.07) while these parameters remained unchanged in the control group.

CONCLUSIONThe present study demonstrates that G-CSF mobilized autologous intracoronary PBSCs transplantation is a safe and feasible treatment for patients with AMI and global left ventricular function is improved and left ventricular remodeling attenuated at six-month follow-up.

Aged ; Female ; Granulocyte Colony-Stimulating Factor ; therapeutic use ; Hematopoietic Stem Cell Mobilization ; methods ; Humans ; Male ; Middle Aged ; Myocardial Infarction ; surgery ; therapy ; Peripheral Blood Stem Cell Transplantation ; Transplantation, Autologous ; Treatment Outcome

OBJECTIVETo study the high-yield and high-quality cultivating technology of Plantago asiatica, adapt to the demands of GAP.

METHODThe main factors which influenced the yield and quality in the production process of P. asiatica was studied.

RESULTThe standard system cultivating technology of high yield and quality of P. asiatica was established.

CONCLUSIONThe seeds of P. asiatica can reach the objective of high yield and high quality on the basis of this cultivating technology.

China ; Fertilizers ; Pest Control ; Plant Diseases ; Plantago ; growth & development ; Plants, Medicinal ; growth & development ; Quality Control ; Seeds ; Soil

OBJECTIVETo study the expression of KiSS-1, nuclear factor kappa B (NF-kappaB) p50 and matrix metalloproteinase 9 (MMP-9) in breast cancer tissue and the relationship with clinicpathological factors.

METHODSImmunohistochemical staining for KiSS-1, NF-KappaBp50, and MMP-9 protein was performed in 152 cases of human breast tissue [92 cases of BC, 30 cases of epithelial hyperplasia, and 30 cases of peritumoral breast tissue (PMT)] and 54 cases of axillary lymph node metastases. In-situ hybridization for KiSS-1 mRNA was done in 50 cases of breast cancer, and 20 cases of PMT.

RESULTS(1) The expression of KiSS-1 gene was significantly higher in well-differentiated breast cancer than in PMT, and this expression progressively decreased with decreasing degree of tumor differentiation, increasing pathological grade, TNM stage and the presence of lymph node metastases. The expression of KiSS-1 gene in lymph node metastasis was markedly lower than the corresponding primary tumor. There was correlation between the expression of KiSS-1 mRNA and KiSS-1 protein in breast cancer group. (2) The expression of NF-kappaKBp50 and MMP-9 increased progressively with decreasing degree of tumor differentiation, increasing TNM stage, large tumor size ( >2 cm) and the presence of lymph node metastases.

CONCLUSIONSThe expression of KiSS-1 protein showed negative correlation with that of NF-kappaBp50 and MMP-9 respectively. MMP-9 protein expression was positively correlated with NF-kappap50 protein expression. These suggest that the genes of KiSS-1, NF-kappaBp50 and MMP-9 could be involved in the progression and metastasis of breast cancer.

Breast Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Kisspeptins ; Lymphatic Metastasis ; physiopathology ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; NF-kappa B ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Statistics as Topic ; Tumor Suppressor Proteins ; genetics ; metabolism

Adolescent ; Adult ; Aged ; Alanine Transaminase ; blood ; Female ; Hepatitis B virus ; growth & development ; Hepatitis B, Chronic ; blood ; virology ; Humans ; Male ; Middle Aged ; Viral Load

OBJECTIVETo investigate the prevalence of three lamivudine-resistant HBV mutants in lamivudine-treated Chinese patients.

METHODSUsing three pairs of of HBV polymerase gene B and C domain fragments were amplified by PCR. The PCR products were then digested by restriction enzyme Nde I and Nla III. The digested products were analyzed by electrophoresis. With this method, the prevalence of the three lamivudine-resistant mutants in lamivudine-treated Chinese patients was investigated.

RESULTSAfter Nde I digestion of p24 and p29 amplified product, HBV wild type could be easily separated from YMDD mutant. At the same time, YIDD could be separated from YVDD mutant after Nla III digestion of p24 and p29 amplified product. By this method, the authors found that these eleven patients were infected with lamivudine-resistant mutants. Six of them were infected with M5501 mutant; five were infected with M550V mutant (one of them had both M550V and L526M mutations).

CONCLUSIONThe method of the present study was demonstrated to be an easy way to detect HBV lamivudine-resistant mutants and can be applied to clinical monitoring of lamivudine resistance.

Adolescent ; Adult ; Antiviral Agents ; pharmacology ; therapeutic use ; DNA Mutational Analysis ; DNA Primers ; genetics ; DNA, Viral ; analysis ; Drug Resistance, Viral ; genetics ; Female ; Hepatitis B ; drug therapy ; virology ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Humans ; Lamivudine ; pharmacology ; therapeutic use ; Male ; Microbial Sensitivity Tests ; Middle Aged ; Point Mutation ; Polymerase Chain Reaction

OBJECTIVETo evaluate the impact of radiofrequency catheter ablation on left atrial (LA) size and function in patients with paroxysmal atrial fibrillation (PAF) and whether there is any difference between segmental pulmonary vein ostial isolation (SPVI) and circumferential pulmonary vein ablation (CPVA).

METHODSSixty-six patients with highly symptomatic atrial fibrillation were assigned to undergo either SPVI or CPVA. Transthorax echocardiography was performed before, 1 day, 1 months and 3 months after the procedure. LA dimension, LA area, late diastolic peak velocity of mitral valve inflow (A) and peak atrial systolic mitral annulus velocity (A') were recorded.

RESULTSOf 66 consecutive patients with symptomatic PAF, 30 patients underwent SPVI and 36 underwent CPVA. After a mean follow-up of (315 +/- 153) days, 21 patients (70%) after SPVI and 28 patients (75%) after CPVA were free of atrial tachyarrhythmia. As compared with the baseline, LA area decreased at 1-month after ablation in SPVI group and at 3-month in CPVA group. LA dimension decreased also in SPVI group, but did not in CPVA group. A velocity and A' velocity declined remarkably 1 day after CPVA, and restored 3 months later. The former went back to the level of baseline, and the latter exceeded it apparently. In SPVI group, A velocity increased at 1-month, and maintained in 3-month after ablation. A' velocity increased at 3-month after ablation. No reduction of A velocity or A' velocity was found after SPVI.

CONCLUSIONSThis study demonstrated a decrease in LA area and an improvement in LA systolic function 3 months after ablation for PAF. The LA damage by CPVA was more than that by SPVI, which was characterized by the reduction of LA function 1 day after procedure and the delayed improvement of LA size and functional parameters.

Adult ; Atrial Fibrillation ; diagnostic imaging ; physiopathology ; therapy ; Atrial Function, Left ; Catheter Ablation ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; Ultrasonography