Objective To evaluate the efficacy and feasibility of the coil-and-glue method for the reduction of lung volume in rabbit emphysema model.Methods Sixteen rabbits of emphysema model were divided into the occlusion group(n=10),in which both anterior bronchi were occluded using the coil-and- glue method,and the control group(n=6).The maximal static pressure of airway(P_(max)),peak expiratory flow(PEF),end-expiratory volume(EEV)and pressure of oxygen(PO_2)were measured at ante- emphysema,post-emphysema,1 week and 4 week after occlusion respectively.The expectoration(or migration)of coil and collapse of lung were also investigated.Results P_(max)was(20.0?1.3)and(17.1? 1.4)cm H_2O(1 cm H_2O=0.098 kPa)in the occlusion group at ante-emphysema and post-emphysema respectively.P_(max)was(19.2?1.4)cm H_2O in the occlusion group in the 1 week after the occlusion,while (17.1?1.5)cm H_2O in the control group(F=6.68,P

OBJECTIVETo investigate the effect of the Sanxianxinli capsule against fatigue and viability of mice.

METHODThe mouse shinning, burden swimming, heat-resistant, cold resistant and tolerating anoxia were detected by experiment.

RESULTThe results showed that the Sanxansinli capsule have function of improveing the stamina, cold resistant, heat-resistant and tolerating anoxia,and prolonging were the survival time in cold, high temperature and anoxia.

CONCLUSIONThe Sanxianxinli capsule have function against fatigue and can also improve the resistibility of body.

Animals ; Capsules ; Cold Temperature ; Curculigo ; chemistry ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Epimedium ; chemistry ; Fatigue ; physiopathology ; Hot Temperature ; Hypoxia, Brain ; physiopathology ; Male ; Mice ; Oxygen Consumption ; drug effects ; Physical Endurance ; drug effects ; Plants, Medicinal ; chemistry

Selective markers used in yeast vector for gene manipulation were usually drug resistance or autotrophy. Unfortunately, drug resistance selective marker requires drug sensitive host and most industrial strains were not autotrophy. In this paper, flocculation gene (FLO1) from Saccharomyces cerevisiae ABXL-1D was amplified by PCR, sequenced and cloned to construct an expression vector. The new vector was easy to manipulate and suitable for broad host of yeasts without either autotrophy or drugs. beta-glucosidase gene from Bacillus polymyxa was cloned with the vector and expressed in Saccharomyces cerevisiae. The specific activity of beta-glucosidase of the recombinant yeast cell-free extract was 3.91 u/mg protein. The residue glucose of the recombinant yeast was considerably reduced in mixed fermentation of glucose and cellobiose. It should be favorable for ethanol fermentation when utilize lignocellulosic biomass as raw material.
Bacillus ; enzymology ; genetics ; Flocculation ; Genetic Vectors ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Saccharomyces cerevisiae ; genetics ; metabolism ; beta-Glucosidase ; biosynthesis ; genetics

The intestinal absorption properties of the main effective components (glycyrrhizic acid, isoliquiritigenin, 6-gingerol, ginsenoside Rb1, atractylode-I) in Lizhong decoction (LZD) extracts were investigated with an in situ single-pass intestinal perfusion model in rats. UPLC-TQ-MS was used to determine the concentration of the five components in the intestinal perfusion. Animal welfare and experimental procedures were in accordance with the regulations of the Animal Ethics Committee of Nanjing University of Chinese Medicine. As evaluation indexes for the intestinal absorption characteristics, the absorption rate constant (K_a) and the apparent permeability coefficient (P_eff) of the five main ingredients were analyzed. Results showed that the best absorption sites for glycyrrhizic acid, isoliquiritin and 6-gingerol were the ileum, colon and duodenum, respectively, and the differences between different intestinal segments were statistically significant (P <0.05). There was no notable difference in K_a and P_eff between ginsenoside Rb1 and atractylode-I in the different intestinal segments (P > 0.05), suggesting that they were absorbed throughout. The five components were well-absorbed in the whole intestine (P_eff > 1.0×10^-3 cm·min^-1), indicating that LZD is suitable for preparing sustained, controlled release and enteric-coated preparations.

OBJECTIVETo investigate the relationship between point mutation of penicillin-binding protein gene (pbp) and amoxicillin resistance (AMOgamma) of Helicobacter pylori (H. pylori) as well as to compare the protein profiles under proteomic technology to get the candidate resistance-related proteins.

METHODS(1) AMOgamma strains were selected from the sensitive H. pylori strain 26695 by serial passage technique in vitro. (2) Point mutations of five putative resistance genes (HP0597, HP1565, HP1542, HP1556, and HP0160) were analyzed by denaturing high-performance liquid chromatography (DHPLC) and DNA sequencing. (3) Proteins samples were separated by two-dimensional electrophoresis (2-DE). Protein profiles were compared between the AMOgamma strain obtained in vitro and its sensitive parent strain 26695. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) was performed to identify the proteins of interest. The proteins were searched by software MASCOT and identified by peptide fingerprint map using the program MS-FIT of Protein Prospect.

RESULTS(1) An AMOgamma strain (MIC 8 microg/ml) was obtained. Complete loss of the resistant phenotype was observed after cultivation in the absence of AMO or storage at - 80 degrees C. (2) DHPLC and Sequencing result showed no point mutations in five pbp genes in the AMOgamma strain when compared with the corresponding PCR products from its parent strain 26695. (3) Protein profiling showed that eleven protein spots were differently expressed between 26695 and the AMOgamma strain. Of these protein spots in the AMOgamma strain, two new spots (Spot 1 and Spot 2) were observed with one (Spot 3) was up-regulated three-fold and the remained ones (Spot 4-11) were down-regulated.

CONCLUSIONAMO resistance of H. pylori might be resulted from, unstable phenotype change rather than point mutations of pbp genes. These differentially regulated proteins in AMOgamma strain might play a role in development of resistance to AMO in H. pylori.

Amoxicillin ; pharmacology ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; metabolism ; Chromatography, High Pressure Liquid ; Drug Resistance, Bacterial ; genetics ; Electrophoresis, Gel, Two-Dimensional ; Helicobacter pylori ; drug effects ; genetics ; metabolism ; Point Mutation ; genetics ; physiology ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

OBJECTIVESTo investigate prognostic effect of postoperative resection-margin status for intraoperatively positive resection margin in advanced gastric cancer and discuss the treatment choice for intraoperatively positive resection margins.

METHODSA retrospective study was investigated in 64 advanced gastric cancer patients with positive resection margin after potentially curative resection. The survival between 50 patients who was re-excised to a negative resection margin (NR group) and 14 patients who were left with positive resection margin (PR group) was compared. Prognostic factors were analyzed using univariate and multivariate Cox regression model analysis.

RESULTSThe median survival in the PR group was 17.0 months (95%CI: 11.6 - 22.4) as compared with 23.0 months (95%CI: 20.5 - 25.5) in the NR group (P = 0.045). However, resection-margin status lost significance on multivariate analysis. In the subgroup of D2 lymphadenectomy, the median survival in the PR group and NR group were 17.0 months (95%CI: 12.0 - 22.0) and 24.0 months (95%CI: 19.8 - 28.1) respectively; multivariate analysis further identified resection margin status as an independent prognostic factor.

CONCLUSIONSRe-excision for intraoperatively positive margin to negative margin improves the prognosis of the patients with advanced gastric cancer, and re-excision is the first choice when intraoperative frozen section detects a positive margin. Routine frozen section of resection margin should be mandatory in all advanced gastric cancer undergoing potentially curative surgery.

Female ; Follow-Up Studies ; Frozen Sections ; Gastrectomy ; methods ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Proportional Hazards Models ; Retrospective Studies ; Stomach Neoplasms ; surgery

Chinese acupuncture and moxibustion has been widely accepted as a useful therapeutics all over the world, but its mechanism has not been fully defined. For this purpose, a reticular framework of whole-body fascia and connective tissues has been established by means of digitized virtual human technique. The virtual acupoints represented three-dimensionally were compared with the sites for stimulation in practice of traditional Chinese medicine (TCM) acupuncture therapy. The results showed that the fascial network constituted by the connective tissues may be the anatomical basis for acupuncture therapy. We found that the acupoints were mainly located where thick connective tissues were present. In this fascial network, sensitive nerve endings, active cells and lymphatic vessels abounded in the sites with thick connective tissue, and needling at these sites induced definite biological effects. In light of biological phylogeny and embryo development, we believe that the connective tissue network may constitute a new functional system in the human body, the Self-supervision and control system (Fasciology), which provides a theoretical base for acupuncture therapy.
Acupuncture ; methods ; Acupuncture Points ; Connective Tissue ; anatomy & histology ; Fascia ; anatomy & histology ; Female ; Humans ; Medicine, Chinese Traditional ; methods ; Meridians ; Models, Anatomic ; Models, Neurological

BACKGROUNDIn our previous study, we found that DAZAP2 was the most significantly down regulated gene when differential screening of complementary DNA (cDNA) chips were used to analyze mRNA isolated from bone marrow mononuclear cells from newly diagnosed multiple myeloma (MM) patients without anticancer treatment. In this study, we observed DAZAP2 mRNA and protein expression in the mononuclear cells from MM bone marrow and investigated its role in the pathogenesis of MM.

METHODSThe full-length cDNA of DAZAP2 was cloned and sequenced from mononuclear cells from human bone marrow. The nucleotide and amino acid sequences of DAZAP2 were analyzed using the ClustalW program. A dendrogram was constructed by multiple sequence alignment using ClustalW and amino acid sequence identity/similarity was derived based on comparisons attained using the MegAlign software. The recombinant pEGFP expression vector was constructed and the confocal microscopy was used for the localization of the DAZAP2 protein in transfected COS7 cells. The expression of DAZAP2 mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) and the expression level of DAZAP2 protein was detected by Western blotting analysis in MM samples.

RESULTSDAZAP2 proteins of vertebrates is highly conserved in evolution. It contains a proline-rich region, several potential SH2 and SH3 domain-binding motifs and a possible protein kinase C (PKC) phosphorylation site. We showed by confocal microscopy that the DAZAP2 protein predominantly resides in the cytoplasm with a discrete pattern of punctuated distribution. The expression of DAZAP2 was not detected in 24 of 36 MM samples by semi-quantitative RT-PCR. In contrast, DAZAP2 expression was detected in all 30 normal controls. The expression level of DAZAP2 protein was assayed by Western blotting analysis, showing a robust down-regulation in MM patients (P < 0.001) that matched with the results of the RT-PCR.

CONCLUSIONSDAZAP2 is downregulated in MM samples and it may be a signal molecule in MM cells. DAZAP2 is involved in the pathogenesis of MM and could be used as a genetic marker for MM.

Adult ; Aged ; Amino Acid Sequence ; Blotting, Western ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Molecular Sequence Data ; Multiple Myeloma ; etiology ; metabolism ; RNA, Messenger ; analysis ; RNA-Binding Proteins ; analysis ; chemistry ; genetics

OBJECTIVETo evaluate the efficiency and safety of dasatinib in Chinese patients (pts) with chronic myelogenous leukemia (CML) in chronic phase (CP), accelerated-phase (AP) or blast-phase (BP) who are resistant or intolerant to imatinib (IM).

METHODS119 CML pts received dasatinib 100 mg once daily for pts in CP or 70 mg twice daily for pts in AP/BP. The hematologic/cytogenetic response, progression-free-survival (PFS), overall survival (OS) and adverse effects (AE) of the pts were assessed.

RESULTS59 pts in CP, 25 in AP and 35 in BP received dasatinib treatment. The median duration of dasatinib treatment were 19.32, 20.99 and 3.22 months respectively. Complete hematologic response (CHR), major cytogenetic response (MCyR) and complete cytogenetic response (CCyR) were achieved by 91.5%, 50.8% and 42.4% of pts in CP respectively. The median times to achieving MCyR was 12.1 weeks. None of the pts in CP achieved MCyR progressed or died till to last follow-up. CHR and major hematologic response (MaHR) were achieved by 52.0% and 84.0% of pts in AP, respectively. The median time to CHR and MaHR were 16.0 and 12.1 weeks, respectively. 10 pts in AP achieved MCyR and 9 of them were CCyR. The median duration of PFS was 25.7 months for pts in AP. For 35 pts in BP, the rates of CHR and MaHR were 17.1% and 31.4% respectively. Both of the median time to CHR and MaHR were 12.1 weeks and median time of duration of MaHR was 11.2 months. 8 pts in BP achieved MCyR and the median time of duration of MCyR was 13.2 months. The median duration of PFS and OS for the pts in BP were 4.3 and 16.7 months respectively. Grade 3-4 of hematologic AEs related to dasatinib were frequent but manageable by dose interruption/reduction or supportive care. 52.5% and 61.0% of pts in CP experienced grade 3-4 of neutropenia and thrombocytopenia. More than 80% pts in AP/BP occurred grade 3-4 cytopenia. The common non-hematologic AEs related to dasatinib were including grade 1-2 pleural effusion, headache, pneumonia and diarrhea. The frequency of non-hematologic AE was higher in pts with AP/BP than in pts with CP.

CONCLUSIONChinese pts with CML resistant or intolerant to IM treated by dasatinib can achieve relatively sustained hematologic and even cytogenetic remission and are well tolerated.

Adolescent ; Adult ; Aged ; Benzamides ; adverse effects ; pharmacology ; Dasatinib ; Drug Resistance, Neoplasm ; Female ; Humans ; Imatinib Mesylate ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; drug therapy ; Male ; Middle Aged ; Piperazines ; adverse effects ; pharmacology ; Pyrimidines ; adverse effects ; pharmacology ; therapeutic use ; Thiazoles ; adverse effects ; therapeutic use ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the expression of fractalkine (FKN) and its receptor CX3CR1 in cardiac allografts and the effect of Cyclosporin A (CsA).

METHODSThree groups of rats underwent heterotopic cardiac transplantation, 45 cases in each group and 5 cases in control group: SD to SD regarded as isograft group (group A), Wistar to SD divided into CsA untreated allograft group (group B) and CsA treated allograft group (group C), normal SD rats as control group. The FKN mRNA expression was detected by one-step RT-PCR method and the expression of FKN and CX3CR1 protein was detected by standard ABC immunohistochemical technique.

RESULTSThe expression of FKN mRNA and protein was weak in both isografts and normal heart specimens. The changes of FKN mRNA expression were correlated with the process of acute allograft rejection. The peak of FKN mRNA expression (0.8 +/- 0.26) appeared on the seventh day after transplantation, which could be down-regulated by CsA significantly (t = 2.390, P < 0.05). FKN protein was located in endothelia cells and its receptor CX3CR1 was located in infiltrating mononuclear cells in allografts.

CONCLUSIONSUpregulation of FKN and its receptor was significantly correlated with the trafficking of mononuclear cells which play an important role in acute allograft rejection. It may be one of the important mechanisms of CsA to intervene the acute rejection by inhibiting the activation of the FKN-CX3CR1 pathway.

Acute Disease ; Animals ; CX3C Chemokine Receptor 1 ; Chemokine CX3CL1 ; Chemokines, CX3C ; genetics ; metabolism ; Cyclosporine ; pharmacology ; Graft Rejection ; immunology ; pathology ; prevention & control ; Heart Transplantation ; immunology ; pathology ; Immunohistochemistry ; Male ; Membrane Proteins ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Receptors, Cytokine ; genetics ; metabolism ; Receptors, HIV ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous