OBJECTIVETo explore the clinical effect of combination of acupressure and magnetic sticker on the quality of life (QOL) including appetite, defecation, and sleep in patients with advanced gastroenteric tumor.

METHODSTotally 147 patients with advanced gastroenteric tumor were assigned to 4 groups according to different treatment methods, i.e., the supportive treatment group (A, 20 cases), the acupressure treatment group (B, 41 cases), the magnetic sticker treatment group (C, 40 cases), and a combination of acupressure and magnetic sticker treatment group (D, 46 cases). They were respectively treated with different methods, supportive treatment for group A, acupressure for group B, magnetic sticker for group C, and a combination of acupressure and magnetic sticker for group D. The scores of food intake, defecation frequency, sleep time, Karnofsky, and QOL were compared before treatment and at day 14 after treatment.

RESULTSAfter treatment, the scores of food intake, defecation frequency, and sleep time were obviously improved in B, C and D groups (P < 0.01). There was statistical difference between group D and group A (P < 0.01). In addition, in comparison with A group, both Karnofsky score and QOL score increased in B, C and D groups (P < 0.01).

CONCLUSIONThe assisted therapy of the combination of acupressure and magnetic sticker could ameliorate QOL such as the digestive functions and sleep in patients with advanced gastroenteric tumor.

Acupressure ; methods ; Adult ; Aged ; Aged, 80 and over ; Female ; Gastrointestinal Neoplasms ; therapy ; Humans ; Magnetics ; Male ; Middle Aged ; Quality of Life ; Treatment Outcome

The chromatographic fingerprint was established by eluting with the mobile phase consisted of acetonitrile and 0.2% formic acid water on an Agilent TC-C18 (2) column (4.6 mm x 250 mm, 5 microm). Six chromatographic peaks were identified by HPLC-MS/MS method. Ten batches of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle were determined, and the similarity was arranged from 0.72 to 0.99. Good precision, stability and repeatability were obtained, and this study provides a reference for the quality control of Glycyrrhizea Radix et Rhizoma Praeparata Cum Melle.
China ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Glycyrrhiza uralensis ; chemistry ; Mass Spectrometry ; Quality Control ; Rhizome ; chemistry

OBJECTIVETo study the effects of Saccharomyces albicans (S. albicans) on the cell cycle distribution and proliferation of human umbilical vein endothelial cell ECV304 cells in vitro.

METHODSThe line of ECV304 cultured in vitro were divided into four groups which were treated by S. albicans supernatant, S. albicans inactivated bacilli, supernatant and inactivated bacilli mixture, normal culture medium. The proliferous effect of ECV304 induced by supernatant, inactivated bacilli, supernatant and inactivated bacilli mixture using the methods of MTT, cell count, microscope and flow cytometry were conducted.

RESULTSIn the condition of different times and different culture concentrations, ECV304 cells incubated with 4-fold diluted S. albicans supernatant for 48 h increased the proliferation rate. The S and G2/M population of ECV304 cells increased after incubated with S. albicans supernatant for 40 h, which showed significant increasing cell proliferation index (PI) (P < 0.05). The PI of the cells treated by inactivated bacilli showed no significant change (P > 0.05).

CONCLUSIONS. albicans could induce ECV304 cell proliferation which depends on the release of metabolic products of S. albicans.

Cell Cycle ; Cell Division ; Cell Proliferation ; Human Umbilical Vein Endothelial Cells ; Humans ; Saccharomyces ; Umbilical Veins

OBJECTIVETo prepare and characterize monoclonal antibodies (mAbs) against the recombinant nucleocapsid (N) protein of 3 human coronaviruses SARS-CoV, 229E and OC43 and study the antigenic relationship between the 3 N proteins.

METHODSBALB/c mice were immunized with the recombinant N proteins of SARS-CoV, 229E and OC43 to obtain the mAbs by means of hybridoma. Screening and identification of the mAbs were performed using indirect enzyme-linked immunosorbent assay (ELISA), Western blotting and indirect immunofluorescence assay. Cross-reactivity between the N proteins of the 3 coronaviruses was analyzed with the prepared mAbs.

RESULTSThe mAbs against the recombinant N proteins of SARS-CoV, 229E and OC43 were obtained, which reacted specifically with the corresponding viral N protein as shown by indirect ELISA, Western blotting and indirect immunofluorescence assay. No cross-reactivity was found between the 3 N proteins.

CONCLUSIONThe prepared mAbs against the recombinant N proteins may provide valuable assistance in studying antigenic relationships of N proteins between the 3 human coronaviruses.

Animals ; Antibodies, Monoclonal ; immunology ; Blotting, Western ; Coronavirus 229E, Human ; genetics ; immunology ; Coronavirus OC43, Human ; genetics ; immunology ; Cross Reactions ; immunology ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Mice ; Mice, Inbred BALB C ; Nucleocapsid Proteins ; genetics ; immunology ; Recombinant Proteins ; immunology ; SARS Virus ; genetics ; immunology

OBJECTIVETo clone and express avian influenza A virus [A/Hong Kong/482/97(H5N1)] H5 subtype hemagglutinin in baculovirus-insect cell expression system and investigate the antigenicity and bioactivity of the recombinant protein.

METHODSH5 gene of influenza A virus was amplified by PCR. The recombinant bacmid was obtained by cloning the gene to the donor plasmid of pFastBacHTB and transformed into DH10Bac competent cells. The recombinant baculovirus stock was prepared by transfecting the recombinant bacmid DNA into the insect cell line for protein expression after amplification. Immunofluorescene assay (IFA) and Western blotting were performed to identify the antigenicity of the recombinant protein, and hemagglutination assay was used to identify its bioactivity.

RESULTSThe recombinant his-H5 protein was expressed in the insect cells with a relative molecular mass of 64,000, which showed erythrocyte-agglutinating activities with the red blood cells of guinea pig. Western blotting and IFA demonstrated that the recombinant his-H5 could be recognized and bound by standard anti-H5 sera.

CONCLUSIONThe recombinant his-H5 with a post-translation modification is successfully obtained in insect cells, which may provide a potential source for further study of the antigen's biological function and for production of the subunit vaccine or monoclonal antibodies.

Animals ; Baculoviridae ; genetics ; Cell Line ; Erythrocytes ; cytology ; immunology ; Genetic Vectors ; genetics ; Guinea Pigs ; Hemagglutination Tests ; Hemagglutinin Glycoproteins, Influenza Virus ; biosynthesis ; genetics ; immunology ; Influenza A Virus, H5N1 Subtype ; genetics ; immunology ; Recombinant Proteins ; biosynthesis ; immunology ; Spodoptera ; Transfection

Objective A multicenter, randomized, controlled and open-labled clinical trial was performed to compare the efficacy and safety of recombinant human insulin injection ( Yousilin R) and treated with Yousilin R versus Novolin R for 12 weeks respectively. Results Compared with baseline,the levels of glycosylated hemoglobin A1c ( HbA1c ) at the end of 12 weeks treatment decreased from 10. 77% to 7. 72% ( P ＜0. 05 ) in Yousilin R group and from 10. 33% to 7. 62% ( P ＜0. 05 ) in Novolin R group,2-hour postprandial plasma glucose ( 2hPG ) decreased from 15.49 mmol/L to 9. 72 mmol/L ( P ＜ 0. 05 ) in Yousilin R group and from 15.33 mmol/L to 10. 07 mmol/L( P ＜ 0. 05 ) in Novolin R group, and fasting plasma glucose (FPG) decreased from 10. 90 mmol/L to 7. 31 mmol/L( P ＜0. 05 ) in Yousilin R group and from 10. 22 mmol/L to 7.21 mmol/L (P ＜0. 05) in Novolin R group. The changes of HbA1c, 2hPG and FPG from baseline to endpoint in Yousilin R group was similar to those in Novolin R group ( P ＞ 0. 05 ).Furthermore, hypoglycemic events(26. 42% vs 30. 48% ), other adverse events( 13.21%vs 16. 19% ) ,and serious adverse events( 1.89%vs 1.90% )were comparable between Yousilin R and Novolin R groups(P ＞0. 05 ). Conclusions Yousilin R has similar efficacy, safety and compliance profiles to Novolin R group in the treatment of diabetic patients.

To establish a method for simultaneous determination of dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid and pachymic acid in Poria, a RP-HPLC method detected by UV wavelengths switch had been developed, including 210 nm (48-55 min) for pachymic acid and 241 nm (0-48 min) for dehydrotumulosic acid, polyporenic acid C, 3-epi-dehydrotumulosic acid, dehydropachymic acid, separately. The system consisting of a Kromasil C18 column (250 mm x 4.6 mm, 5 microm) and a mixture of acetonitrile and 0.05% phosphate acid as the mobile phase was adopted; The flow rate was 1.0 mL x min(-1). The linear response range was 30.5-610.0 microg x mL(-1) (r = 0.999 6) for dehydrotumulosic acid, 12.66-253.2 microg x mL(-1) (r = 0.999 5) for polyporenic acid C, 2.99-59.7 microg x mL(-1) (r = 0.999 7) for 3-epi-dehydrotumulosic acid, 6.13-122.5 microg x mL(-1) (r = 0.999 5) for dehydropachymic acid and 11.3-226.0 microg x mL(-1) (r = 0.9995) for pachymic acid. The average recoveries of these compounds were 98.5% (RSD = 1.9%), 99.4% (RSD = 1.7%), 97.9% (RSD = 1.2%), 96.7% (RSD = 2.5%) and 97.9% (RSD = 2.3%), respectively. The method is simple, accurate and reproducible for quality control of Poria.
Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Lanosterol ; analogs & derivatives ; analysis ; Plants, Medicinal ; chemistry ; Poria ; chemistry ; Quality Control ; Reproducibility of Results ; Spectrophotometry, Ultraviolet ; methods ; Triterpenes ; analysis

OBJECTIVETo characterize the specific monoclonal antibodies to Aspergillus conidia.

METHODSFlow cytometry was used to examine the reactivity of the specific monoclonal antibodies to Aspergillus conidia.

RESULTSBoth the monoclonal antibodies MA3 and Con2 showed specific reactivity to Aspergillus conidia suspensions. MA3 was capable of binding to the conidia of A.fumigatus, A.flavus, A.niger and A.terreus, while Con2 was reactive only to the conidia of A.fumigatus.

CONCLUSIONTwo specific monoclonal antibodies (MA3 and Con2) to Aspergillus conidia have been obtained.

Antibodies, Fungal ; immunology ; isolation & purification ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Antibody Specificity ; Aspergillus ; immunology ; Flow Cytometry ; Spores, Fungal ; immunology

OBJECTIVETo achieve secretory and extracellular production of recombinant dengue virus serotypes I-IV envelope glycoprotein domain III (DENV-1-4 EDIII) in Pichia pastoris.

METHODSEDIII genes of DENVI-IV were amplified and cloned into vector pPIC9K, respectively. These recombinant plasmids were then linearized and transferred into Pichia pastoris strain GS115. Clones highly produced in 4.0 mg/ml G418 were amplified and induced by methanol to achieve the secreted recombinant proteins. Ni-NTA agarose beads were used for purification, while SDS-PAGE and Western blotting were used for identification.

RESULTSThe recombinant plasmids pPIC9K-DENV-1-4 EDIII were constructed and successfully transferred into Pichia pastoris strain GS115. The recombinant EDIII proteins were expressed in a secretory way with the molecular weight about 12 × 10(3) and specifically identified by anti-His monoclonal antibody and anti-DENVI-IV mice sera.

CONCLUSIONDENVI-IV EDIII proteins are successfully achieved from Pichia pastoris expression system and could be used for development of dengue vaccines, diagnostic reagents and study of biological function of the E protein.

Dengue Virus ; genetics ; Genetic Vectors ; Pichia ; metabolism ; Recombinant Proteins ; genetics ; Viral Envelope Proteins ; secretion

OBJECTIVETo study selective killing effect of herpes simplex virus-thymidine kinase/ganciclovir (Hsv-tk/GCV) driven by human telomerase catalytic subunit (hTERT) promoter on lung cancer cell line A549 in vitro.

METHODS(1) Expression plasmids of Hsv-tk gene driven by hTERT promoter and sv40 promoter respectively (pGL3-hTp-tk and pGL3-sv40-tk) were transfected into telomerase-positive human lung adenocarcinoma cell line A549 and telomerase-negative fetal lung fibroblast cell line MRC-5. Reverse transcription-PCR was performed to detect expression of tk gene in above transfected cell lines; (2) Inhibition effect on proliferation of above transfected cell lines treated with GCV was investigated with MTT method; (3) Influence of GCV on apoptosis and cell cycle of above transfected cell lines was investigated with flow cytometry.

RESULTS(1) tk mRNA expression was detected in both A549 and MRC-5 transfected with pGL3-sv40-tk, also in A549 transfected with pGL3-hTp-tk, but not in pGL3-hTp-tk transfected MRC-5; (2) GCV showed significant inhibition effects on proliferation of pGL3-sv40-tk transfected A549 and MRC-5 in vitro, also on that of pGL3-hTp-tk transfected A549, but not on that of pGL3-hTp-tk transfected MRC-5; (3) Treated with GCV, apoptosis index (AI) of pGL3-sv40-tk transfected A549 and MRC-5 as well as pGL3-hTp-tk transfected A549 (21.58%, 9.35% and 23.19% respectively) increased significantly, compared with A549, MRC-5 transfected with pGL3-hTp (0.78% and 0.55% respectively) and A549, MRC-5 without plasmid transfection as blank control (2.17% and 0.60% respectively); GCV had no influence on AI of pGL3-hTp-tk transfected MRC-5 (0.88%).

CONCLUSIONtk gene driven by hTERT promoter could express selectively in lung cancer cell A549. Hsv-tk/GCV driven by hTERT promoter could selectively inhibit proliferation of lung cancer cell.

Antiviral Agents ; pharmacology ; Apoptosis ; drug effects ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Flow Cytometry ; Ganciclovir ; pharmacology ; Gene Expression Regulation, Neoplastic ; drug effects ; Genetic Therapy ; methods ; Humans ; Lung Neoplasms ; genetics ; pathology ; therapy ; Promoter Regions, Genetic ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Simplexvirus ; enzymology ; genetics ; Telomerase ; genetics ; Thymidine Kinase ; genetics ; Transfection ; Viral Proteins ; genetics