Aged ; Antigens, CD20 ; metabolism ; Humans ; Leukocyte Common Antigens ; metabolism ; Lymphoma, B-Cell ; complications ; metabolism ; pathology ; surgery ; Male ; Prostate ; pathology ; Prostatectomy ; Prostatic Hyperplasia ; complications ; metabolism ; pathology ; surgery

Adenocarcinoma, Clear Cell ; metabolism ; pathology ; surgery ; Adult ; Carcinoma ; metabolism ; pathology ; Carcinoma, Mucoepidermoid ; metabolism ; pathology ; Carcinoma, Renal Cell ; metabolism ; pathology ; secondary ; Diagnosis, Differential ; Humans ; Keratins ; metabolism ; Male ; Nasal Cavity ; Nose Neoplasms ; metabolism ; pathology ; surgery ; S100 Proteins ; metabolism

The purpose of this study was to evaluate the in vivo viability of human platelets cryopreserved at -80 degrees C by using SCID mouse model and flow cytometry. The fresh human platelets were frozen with 5% DMSO at -80 degrees C for 10 days, thawed, and centrifuged for concentration. A 100 ml aliquot of concentrated platelets was injected into the SCID mouse tail vein by using a 1 ml insulin-syringe fitted with a 29-gauge ultra-fine needle. The whole blood was collected into heparinized capillary tube at 0.5, 2, 4, 6, 12, and 24 hours after infusion via a tail vein and was labelled with CD61-PE. Then the human platelets in mouse whole blood were detected by flow cytometry. The 30 minute time point was used as 100% to calculate the survival time of human platelets. The results showed that the survival time of cryopreserved human platelets were more significantly decreased than that of fresh platelets in SCID mice. Survival rates at 4 hours after transfusion of fresh platelets and cryopreserved platelets in SCID mice were 79.5% +/- 9.1% (n = 8) and 40.6% +/- 6.6% (n = 8) respectively, and a T(1/2) estimated were 7 hours for fresh platelets, but 2.5 hours for the cryopreserved. In conclusion, platelets survival time in SCID mice was shortened after frozen with DMSO at -80 degrees C.
Animals ; Blood Platelets ; Blood Preservation ; methods ; Cell Survival ; Cryopreservation ; Humans ; Mice ; Mice, SCID ; Models, Biological ; Platelet Count ; Platelet Transfusion

Objective To develop the method of 16S rRNA gene clone library for tick bacterial flora analysis, and to analyze the detection effective of pathogens in tick and capacity of bacterial flora diversity. Methods Primers were designed according to the specific gene of Borrelia burgdorferi, Bartonella henselae, Anaplasma phagocytophilum, Ehrlichia chaffeensis and templates were choosen by positive PCR result to amplify the DNA extracted from the ticks. One set of primers targeting 16S rRNA gene conserved region were chosen to amplify certain fragments, DNA extraction, PCR reaction, cloning and sequencing. Nucleotide sequences were compared with GenBank database. Calculated Coverage values of clone library and Shannon-Wiener diversity index. Results Sixteen defined genus-or species-bacteria were detected in 103 valid sequences. Eight species were edge type (Clone No. > 5). Three kinds of pathogens were identified (Borrelia burgdorferi, Bartonella henselae and Rickettsia sp). Three kinds of pathogens were not edge type(Clone No. < 5). Coverage value was 96.11%, and Shannon-Wiener index was 2.40. Analysis results of cloning sequence showed that tick-parasitic bacteria mainly were α and γ deformation mycetes which accounted for 56.25% (9/16). Conclusions The 16S rRNA gene sequences technology could make relative quantitative of bacterial flora, and detect many kinds of pathogens in tick. It's a good method for detection of pathogens and bacterial flora analysis.

AIMTo discover new drugs which may be applied to diseases of the immune system, hemogenesis system diseases and tumors, several high-throughput drug screening cell models based on JAK-STAT signal pathway have been established.

METHODSFour repeats of STAT DNA binding conserved sequences were synthesized, subcloned into pGL-Luc reporter vector and stably transfected into cell lines in vitro. Cell clones with high copy numbers of STAT binding sites and reporter genes were chosen as high-throughput drug screening cell models. The cell models were tested with known anti-allergic drugs and anti-tumor drugs by determining luciferase activity. The reaction was performed in 96 well micro-plates with a final volume of 50 microL.

RESULTSThe cell models by performing rapid fluorescence assay were shown to be highly sensitive and stable after testing with cytokine and drugs. The modification of the expression plasmid simplified this method and made it more practical. It also provided good linear correlation, wide range of assay, highly sensitive and good reproducibility.

CONCLUSIONThe method can be performed by high-throughput drug screening for effective extraction of Chinese traditional herbs.

Anti-Allergic Agents ; isolation & purification ; pharmacology ; Antineoplastic Agents ; isolation & purification ; pharmacology ; Carcinoma, Hepatocellular ; pathology ; DNA-Binding Proteins ; genetics ; metabolism ; Drug Evaluation, Preclinical ; methods ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Humans ; Janus Kinase 2 ; Jurkat Cells ; metabolism ; Liver Neoplasms ; pathology ; Luciferases ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; STAT3 Transcription Factor ; Signal Transduction ; Trans-Activators ; genetics ; metabolism ; Tumor Cells, Cultured

The effects of pretreatment of supercritical carbon dioxide (SC-CO2) on the supramolecular structure of cellulose and the cellulase catalyzed reaction were investigated. The cellulase activity was not affected when it was treated with SC-CO2 at 10MPa and at 50 degrees C for 30 min. But when the cellulase was treated by SC-CO2 in the presence of cellulose, the catalytic activity of the cellulase was lost. The cellulose pretreated with or without cellulase under the same SC-CO2 condition was then hydrolyzed with tresh crude cellulase. The final reducing sugar yield from the hydrolysis of the cellulose pretreated with cellulase was higher than that of the cellulose pretreated without cellulase. It was also found that the improvement of the enzymolysis had a direct relevance with the amount of cellulase used during the SC-CO2 pretreatment. The moisture content of cellulose before SC-CO2 pretreatment had an obvious influence on the subsequent enzymolysis. When the moisture content of cellulose was 60% (W/W), the reducing sugar yield was higher than when the moisture content was over 100% (W/W). The FT-IR spectra showed that the structure of the cellulose pretreated with cellulase under the SC-CO2 condition was different from that of the cellulose pretreated without cellulase. In the presence of the enzyme, the strength of the hydrogen bonds and the I beta phase at 710cm(-1) in the crystalline cellulose was weakened. These results suggest that the change in the cellulose structure induced by the SC-CO2 treatment favous the subsequent enzymolysis.
Carbon Dioxide ; chemistry ; Cellulase ; chemistry ; metabolism ; Chromatography, Supercritical Fluid ; methods ; Spectroscopy, Fourier Transform Infrared

The purpose of this study is to investigate the applicability of a natural swelling matrix derived from boat-fruited sterculia seed (SMS) as the propellant of osmotic pump tablets. The sugar components, static swelling, water uptake and viscosity of SMS were determined and compared with that of polythylene oxide (WSR-N10 and WSR-303). Both ribavirin and glipizide were used as water-soluble and water-insoluble model drugs. Then, the monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide were prepared using SMS as the osmotically active substance and propellant. SMS was mainly composed of rhamnose, arabinose, xylose and galactose and exhibited relatively high swelling ability. The area of the disintegrated matrix tablet was 20.1 times as that at initial after swelling for 600 s. SMS swelled rapidly and was fully swelled (0.5%) in aqueous solution with relative low viscosity (3.66 +/- 0.03) mPa x s at 25 degrees C. The monolayer osmotic pump tablets of ribavirin and the bilayer osmotic pump tablets of glipizide using SMS as propellant exhibited typical drug release features of osmotic pumps. In conclusion, the swelling matrix derived from boat-fruited sterculia seed, with low viscosity and high swelling, is a potential propellant in the application of osmotic pump tablets.
Arabinose ; chemistry ; isolation & purification ; Chemistry, Pharmaceutical ; Delayed-Action Preparations ; Drug Carriers ; Galactose ; chemistry ; isolation & purification ; Glipizide ; administration & dosage ; chemistry ; Osmosis ; Plants, Medicinal ; chemistry ; Rhamnose ; chemistry ; isolation & purification ; Ribavirin ; administration & dosage ; chemistry ; Seeds ; chemistry ; Solubility ; Sterculiaceae ; chemistry ; Tablets ; Technology, Pharmaceutical ; methods ; Viscosity ; Water ; Xylose ; chemistry ; isolation & purification

To investigate a baculovirus insect cell system for expressing an interferon alpha 2b (IFNa2b)/immunoglobulin G-4 (IgG4) Fc fusion protein, which has long-acting antiviral effects. Human IFNa2b and IgG4 Fc cDNAs were generated by molecular cloning and inserted into a baculovirus shuttle vector, which was then transposed into the DH10 Bac strain to form recombinant Bacmid-IFN/Fc. The Bacmid-IFN/Fc was transfected into High five insect cells, and expression of the IFN/Fc fusion protein was detected by Western blotting and its biological activity was assessed by the cytopathic effect inhibition method. The IFNa2b and IgG4 Fc cDNA fragments were successfully amplified by RT-PCR using human peripheral lymphocytes. After cloning into the baculovirus shuttle vector, pFastBac1, and transforming into DH10 Bac competent cells, screening identified positive clones carrying the recombinant Bacmid-IFN/Fc. A Bacmid-IFN/Fc clone was successfully transfected into the High five insect cells and packaged into the baculovirus for expression of the IFN/Fc fusion protein. Western blotting revealed that the fusion protein expression was specific, and yielded a protein of 45 kD in size. The in vitro antiviral activity of the IFN/Fc fusion protein was 580 IU/mL. A novel IFN/Fc fusion protein was successfully generated using a baculovirus insect cell system, which may prove useful for providing future experimental data for development of a new long-acting interferon to treat chronic viral hepatitis.
Animals ; Antiviral Agents ; metabolism ; Baculoviridae ; genetics ; Cell Line ; Cloning, Molecular ; Gene Expression ; Gene Fusion ; Genetic Vectors ; Humans ; Immunoglobulin Fc Fragments ; biosynthesis ; genetics ; Immunoglobulin G ; biosynthesis ; genetics ; Insecta ; Interferon-alpha ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

OBJECTIVETo explore the correlation between the HLA-DR13, basic core promoter (BCP), changes of T lymphocyte subset and clinical Chinese medical syndromes of chronic hepatitis B (CHB).

METHODSTotally 102 CHB patients were syndrome typed as Gan depression Pi deficiency syndrome (GDPDS), Pi-Shen yang deficiency syndrome (PSYDS), Gan-gallbladder dampness heat syndrome (GGDHS), Gan-Shen yin deficiency syndrome (GSYDS), and static blood blocking collaterals syndrome (SBBCS). Besides, 30 healthy subjects were recruited as the normal control group. The blood HBV-DNA level and HLA-DR13 gene were detected with real time fluorescent PCR. The expression of CD4+ and CD8+ in T lymphocytes was detected using flow cytometry. The mutation of serum A1762T/G1764A was detected using PCR sequencing. Hepatitis Be antigen (HBeAg) was detected with ELISA, and correlation between various Chinese medical syndrome types and objective indicators were analyzed.

RESULTSThere was no statistical difference in HBV-DNA quantitative results among various syndrome types (P > 0.05). HBeAg positive rate was higher in GDPDS than in other syndrome types (P < 0.05). It was sequenced as GDPDS > GSYDS > SBBCS > GGDHS > PSYDS. Compared with the normal control group, percentages of CD3+ and CD3+ CD4+ were lower in PSYDS (P < 0.05). The ratio of CD3+ CD4+/CD3+ CD8 was lower in GGDHS and PSYDS than in the normal control group (P < 0.05). There was no statistical difference in the CD3+ CD8+ percentage among various syndrome types (P > 0.05). The quantitation of HLA-DR13 gene was lower in GDPDS and GSYDS than in the normal control group (P < 0.05). The positive rate of BCP mutation was higher in GSYDS than in other syndrome types (P < 0.05).

CONCLUSIONCo-detection results of HLA-DR13 and BCP could be used as reference indices of Chinese medical syndrome typing of CHB.

HLA-DR Serological Subtypes ; genetics ; metabolism ; Hepatitis B, Chronic ; classification ; diagnosis ; genetics ; Humans ; Medicine, Chinese Traditional ; Promoter Regions, Genetic ; Syndrome ; T-Lymphocyte Subsets ; metabolism ; Yang Deficiency ; Yin Deficiency

OBJECTIVETo investigate the levels of blood pressure and serum lipids, and examine the relationship between hypertension and hyperlipidemia in Hei Yi Zhuang Chinese living in Guangxi.

METHODSA total of 1056 people of Hei Yi Zhuang ethnicity were studied. Blood pressure, body height, body weight, and serum levels of lipids and apolipoprotein were measured. The data were compared with those in 925 people of Han ethnicity, who live in the same region.

RESULTSSystolic blood pressure and pulse pressure were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of isolated systolic hypertension and hypertension was also significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). Serum concentrations of total cholesterol, triglycerides, low-density lipoprotein cholesterol, and apolipoprotein (Apo) B, and the prevalence of hypercholesterolemia and hyperlipidemia were significantly lower in Hei Yi Zhuang than Han Chinese (P < 0.05). Serum concentrations of high-density lipoprotein cholesterol and the Apo A1 to Apo B ratio were significantly higher in Hei Yi Zhuang than Han Chinese (P < 0.001). The prevalence of hypertension in Hei Yi Zhuang Chinese was positively associated with triglycerides (r = 0.425, P < 0.05), whereas the prevalence of hypertension in Han Chinese was positively correlated with total cholesterol (r = 0.623, P < 0.001).

CONCLUSIONThe present study revealed a significant difference in blood pressure and serum lipids between Hei Yi Zhuang and Han ethnic groups, and an association between hypertension and hyperlipidemia.

Adult ; Aged ; Asian Continental Ancestry Group ; ethnology ; Blood Pressure ; China ; epidemiology ; Female ; Humans ; Hyperlipidemias ; epidemiology ; ethnology ; Hypertension ; epidemiology ; ethnology ; Lipids ; blood ; Male ; Middle Aged ; Prevalence ; Sampling Studies ; Young Adult