Actins ; metabolism ; Antigens, CD ; metabolism ; Antigens, Differentiation, Myelomonocytic ; metabolism ; Diagnosis, Differential ; Female ; Granuloma, Plasma Cell ; metabolism ; pathology ; Histiocytoma, Benign Fibrous ; diagnostic imaging ; metabolism ; pathology ; surgery ; Humans ; Lung Neoplasms ; diagnostic imaging ; metabolism ; pathology ; surgery ; Middle Aged ; Neprilysin ; metabolism ; Pneumonectomy ; methods ; Radiography ; Sarcoma ; metabolism ; pathology ; Solitary Fibrous Tumors ; metabolism ; pathology ; Vimentin ; metabolism ; Xanthomatosis ; metabolism ; pathology

Aged ; Carcinoma, Squamous Cell ; metabolism ; pathology ; surgery ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; surgery ; Male ; Membrane Proteins ; metabolism ; Neoplasm Recurrence, Local ; Reoperation ; Tumor Suppressor Protein p53 ; metabolism ; Vimentin ; metabolism

Adult ; Carcinoma, Medullary ; metabolism ; pathology ; Carcinoma, Papillary ; metabolism ; pathology ; Chromogranin A ; metabolism ; Diagnosis, Differential ; Female ; Humans ; Paraganglioma ; metabolism ; pathology ; surgery ; Phosphopyruvate Hydratase ; metabolism ; S100 Proteins ; metabolism ; Synaptophysin ; metabolism ; Thyroid Neoplasms ; metabolism ; pathology ; surgery ; Thyroidectomy ; methods

Objective To analyse the effect of mobilization and collection's time of peripheral blood stem cells(PBSC) from 8 cases of healthy donors. Methods The 10 donors were studied by self-control design.The number of aphereses was two times every donors. Healthy donors received rhG-CSF according to two different PBSC collection starting time: group 1:PBSC collection was starts 2 hours(2 h) after the fourth day or the fifth day of rhG-CSF. group 2:PBSC collection was starts 4 hours(4 h) after the fourth day or the fifth day of rhG-CSF.(The first dose of rhG-CSF was given on day 1, considering day 0 as the day before starting mobilization). In this study we have compared with two groups of apheresis product. Results The MNC count was significantly higher for donors 4 h collection (groups 2) then 2 h. ( groups 1)(P

Transgenic Atropa belladonna with high levels of scopolamine was developed by metabolic engineering. A functional gene involved in the rate limiting enzyme of h6h involved in the biosynthetic pathway of scopolamine was over expressed in A. belladonna via Agrobacterium-mediation. The transgenic plants were culturing till fruiting through micropropogating and acclimating. The integration of the h6h genes into the genomic DNA of transgenic plants were confirmed by genomic polymerase chain reaction (PCR) analysis. Analysis of the difference of plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight was carried out using SPSS software. The content of hyoscyamine and scopolamine in roots, stems, leaves and fruits was determined by HPLC. The investigation of the expression levels of Hnh6h by qPCR. Both Kan(r) and Hnh6h genes were detected in five transgenic lines of A. belladonna plants (A8, A11, A12, C8 and C19), but were not detected in the controls. The plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight of transgenic plants did not decrease by comparison with the non-transgenic ones, and furthermore some agronomic characters of transgenic plants were better than those of the controls. The highest level of scopolamine was found in leaves of transgenic A. belladonna, and the content of scopolamine was also higher than that of hyoscyamine in leaves. The contents of scopolamine of leaves in different transgenic lines were listed in order: C8 > A12 > C19 > A11 > A8, especially, the content of scopolamine in transgenic line C8 was 2.17 mg x g(-1) DW that was 4.2 folds of the non-transgenic ones (0.42 mg x g(-1) DW). The expression of transgenic Hnh6h was detected in all the transgenic plants but not in the control. The highest level of Hnh6h expression was found in transgenic leaves. Overexpression of Hnh6h is able to break the rate limiting steps involved in the downstream pathway of scopolamine biosynthesis, and thus promotes the metabolic flux flowing toward biosynthesis of scopolamine to improve the capacity of scopolamine biosynthesis in transgenic plants. As a result, transgenic plants of A. belladonna with higher level of scopolamine were developed.
Atropa belladonna ; genetics ; metabolism ; Atropine ; metabolism ; Gene Expression ; Mixed Function Oxygenases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Solanaceae ; enzymology ; genetics

OBJECTIVETo investigate biological functions of hepatitis C virus (HCV) non-structural protein 4A (NS4A).

METHODSYeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4A. HCV NS4A bait plasmid was constructed by ligating the NS4A gene with carrier plasmid pGBKT7, then it was transformed into yeast AH109 (alpha type). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing X-alpha-gal for selection two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent E.coli and analyzed by DNA sequencing and bioinformatics methods.

RESULTSAmong twenty-two positive colonies there were eleven positive for metallothionein 2A, three for eukaryotic translation elongation factor 1 alpha 1, two for albumin, two for RNA binding motif protein 21, two for myomesin, one for cytochrome C oxidase II, and one for ATPase.

CONCLUSIONSGenes of HCV NS4A interacting proteins in hepatocytes were successfully cloned and the results pave the way for studying the biological functions of NS4A and associated proteins.

Carrier Proteins ; genetics ; Cloning, Molecular ; Hepacivirus ; genetics ; Hepatocytes ; metabolism ; Humans ; Two-Hybrid System Techniques ; Viral Nonstructural Proteins ; Viral Proteins ; genetics

OBJECTIVEDiscussion on utility and purposed value of obesity and abdomen obesity when body mass index (BMI), waist circumference (WC), waist to hip ratio (WHR) used as indexes predicting hypertension, hyper-blood glucose, and both clusters, to provide scientific basis for the decision on the indexes and their cut-off points of obesity and abdomen obesity in Chinese people.

METHODSUsing the data of diabetes mellitus (DM) from epidemiological studies carried out in 11 provinces/autonomous regions/municipalities of China from July 1995 to June 1997. Partial relative analysis, logistic multi-factors regression analysis, interaction analysis were used. Relative risk (RR), attributable risk proportion (ARP) and population attributable risk proportion (PARP) of hypertension, hyper-blood glucose, and the both cluster as BMI, WC, WHR with the different cut off points were analysed.

RESULTS1) The correlations between BMI, WC and blood pressure, blood glucose were better than the WHR. 2) After adjusted by age, sex, occupation leisure physical activity, education degree and the family history of DM, the results suggested that BMI, WC, WHR were important predictive factors, with relative importance as BMI > WC > WHR. 3) There were augment interactions on BMI, WC and WHR with hypertension, hyper-blood glucose, with the interaction of BMI and WC in particular. Their pure attributable interaction proportion were from 5.95% to 29.34%. 4) The values of RR were about 2.5 when BMI >/= 23, >/= 24 and >/= 25, suggesting the relationship with exposure factors and diseases were with medium and high maleficent extent. Their ARP were from 0.580 to 0.623 with PARP from 0.259 to 0.425. The values of RR were from 2.06 to 3.08 as WC >/= 85 cm in males, WC >/= 80 cm in females while WC >/= 90 cm in males, WC >/= 80 cm in females, which suggested that the relationship with exposure factors and diseases were in medium and high maleficent extent. Their ARP were from 0.515 to 0.676 while PARP from 0.241 to 0.431.

CONCLUSIONSSince the maleficent extent of exposure factors to diseases, the acceptability for overweight and obesity in population, and the prevention and care for overweight and obesity were just in the introduction stage in China. The utility value of predicted hypertension, hyper-blood glucose in BMI and WC seemed to be better then in WHR. We suggested that BMI used as the obesity index, with the diagnostic cut-off point BMI >/= 24. WC as the abdomen obesity index. The diagnostic cut-off points are suggested to be WC >/= 85 cm in males, and WC >/= 80 cm in females.

Adult ; Aged ; Blood Glucose ; metabolism ; Blood Pressure ; physiology ; Body Constitution ; Body Mass Index ; Diastole ; Female ; Humans ; Hyperglycemia ; diagnosis ; physiopathology ; Hypertension ; diagnosis ; physiopathology ; Logistic Models ; Male ; Middle Aged ; Obesity ; physiopathology ; Predictive Value of Tests ; Systole

OBJECTIVETo investigate the brain oxidative stress injury induced by nano-alumina particles in ICR mice.

METHODSSixty male ICR mice were randomly divided into 6 groups: control group, solvent control group, 100 mg/kg micro-alumina particles group, 3 groups exposed to nano-alumina particles at the doses of 50, 100 and 200 mg/kg. The mice were exposed by nasal drip for 30 days. Then levels of malondialdehyde (MDA), glutathione (GSH), superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GSH-PX) in brain tissues of mice were detected.

RESULTSThere was no difference of SOD activity in mouse brain between control group [(17.32 +/- 6.23)U/gHb] and 50 mg/kg nano-alumina particles group [(17.89 +/- 1.82) U/gHb]. The SOD activity [(4.93 +/- 2.30)U/gHb] in 200 mg/kg nano-alumina particles group was significantly lower than that in control group (P < 0.05). The MDA levels in 3 nano-alumina particles groups were (0.76 +/- 0.13), (1.00 +/- 0.30) and (1.16 +/- 0.39)nmol/ml, respectively, which were significantly higher than that [( 0.24 +/- 0.09)nmol/ml] in control group (P < 0.05). The GSH levels in 3 nano-alumina particles groups were (0.72 +/- 0.08), (0.55 +/- 0.19) and (0.61 +/- 0.20)mg/gpro, respectively, which were significantly lower than that [(1.55 +/- 0.34)mg/gpro]] in control group (P < 0.05). The CAT activity in 50 and 100 mg/kg nano-alumina particles groups were (10.40 +/- 3.84) and (10.40 +/- 2.00)U/mgpro, respectively, which were significantly higher than that [(5.79 +/- 0.96) U/mgpro] in control group (P < 0.05). The CAT activity [(3.25 +/- 1.04)U/mgpro] in 200 mg/kg nano-alumina particles group was significantly lower than that in control group (P < 0.05 ).

CONCLUSIONNano-alumina particles can induce the oxidative stress damage in brain tissues of mice.

Aluminum Oxide ; toxicity ; Animals ; Cerebral Cortex ; metabolism ; Glutathione Peroxidase ; metabolism ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Nanoparticles ; toxicity ; Oxidative Stress ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate biological functions of non-structural protein 4B (NS4B) of hepatitis C virus (HCV), yeast-two hybrid technique was performed to seek proteins in hepatocytes interacting with HCV NS4B.

METHODSHCV NS4B bait plasmid was constructed by ligating the NS4B gene with carrier plasmid pGBKT7 and transformed into yeast cells AH109 (type alpha). The transformed yeast cells were amplified and mated with yeast cells Y187 (alpha type) containing liver cDNA library plasmid pACT2 in 2 x YPDA medium. Diploid yeast cells were plated on synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) and synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selecting two times. After extracting plasmid from blue colonies, plasmid DNA was transformed into competent Escherichia coli and analysed by DNA sequencing and bioinformatics.

RESULTSFive genes in eight positive colonies were obtained. There were one NADH dehydrogenase subunit 3, one cytochrome c oxidase subunit III, one retinol binding protein 4, one reticulon 3-A (RTN3) and one fibrinogen gamma polypeptide (FGG).

CONCLUSIONGenes of HCV NS4B interacting proteins in hepatocytes were successfully cloned and the results paved the way for studying the biological functions of NS4B and associated proteins.

Blotting, Western ; Carrier Proteins ; genetics ; metabolism ; Cell Line, Tumor ; Cloning, Molecular ; Electron Transport Complex IV ; genetics ; metabolism ; Gene Library ; Hepatocytes ; metabolism ; Humans ; Immunoprecipitation ; Membrane Proteins ; genetics ; metabolism ; NADH Dehydrogenase ; genetics ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; Plasmids ; genetics ; Protein Binding ; Retinol-Binding Proteins ; genetics ; metabolism ; Two-Hybrid System Techniques ; Viral Nonstructural Proteins ; genetics ; metabolism

Objective To investigate the effect of B7-H3 protein,a collaborative signal molecule,on macrophage-inflammatory protein 2 (MIP-2) mRNA level in Streptococcus pneumococcal meningitis mouse models.Methods Forty-eight healthy male BALB/C mice at the age of 1.5-2.0 months were randomly divided into 4 groups:9 g/L saline group(NS group),B7-H3 protein group(B7-H3 group),Streptococcus pneumoniae group(SP group),and Streptococcus pneumoniae plus B7-H3 protein group(combination group),12 mice in each group.Mouse models were established by intracerebral ventricular injection with 9 g/L saline,B7-H3 protein,Streptococcus pneumoniae type 3 or Streptococcus pneumoniae type 3 plus B7-H3 protein.Neurobehavior of different groups was evaluated according to loeffler rule after injection for 6 h and 24 h,then the mice were sacrificed and MIP-2 mRNA levels were tested by Real-time PCR.The results were analyzed by SPSS 18.0 software.Results Neurobehavior scoring results showed that there were no significant differences between B7-H3 group and NS group (P ＞ 0.05) after infection for 6 h and 24 h,while the score of SP group was decreased compared with that of NS group (P ＜ 0.05),and the score of combination group was significantly decreased compared with that of SP group (P ＜ 0.05).Real-time PCR results showed that,compared with the NS group,the relative MIP-2 mRNA level in SP group increased after injection for 6 hours (1.210 ±0.932 vs 1.000 ± 0.008),but the difference was not significant (P ＞ 0.05),while at 24 h post infection,the relative MIP-2mRNA expressions in SP group were significantly increased compared with that of NS group(12.880 ± 7.792 vs 1.000 ±0.091),the difference was significant (P ＜ 0.05).At 6 h post infection,SP + B7-H3 treatment enhanced the MIP-2mRNA production compared to SP infection alone,but the difference was not significant [(1.240 ± 0.804) vs (1.210 ± 0.932)] (P ＞ 0.05) ; while at 24 h post infection,the difference was significant (38.760 ± 6.601 vs 12.880 ± 7.792) (P ＜ 0.05).Conclusion Collaborative signal molecule B7-H3 protein may increase MIP-2 mRNA level in Streptococcus pneumococcal meningitis mouse model,and exaggerate the clinical disease condition.