Granulocyte colony-stimulating factor (G-CSF) is a kind of hematopoietic growth factor which is produced by monocytes, fibroblasts and endothelial cells. G-CSF acts on neutrophilic progenitor cells by binding to specific cell surface receptors, thereby stimulates proliferation, differentiation, commitment, and selected end-cell functional activation including enhanced phagocytic ability, priming of the cellular metabolism associated with respiratory burst, antibody dependent killing and the increased expression of some functions associated with cell surface antigens. G-CSF is effective and safe for treatment of neutropenia. In this paper, structure of G-CSF and its mechanism, recent status of research on G-CSF, pharmacokinetics, clinical application, adverse effects and prospect of G-CSF are mainly reviewed.
Granulocyte Colony-Stimulating Factor ; pharmacokinetics ; pharmacology ; therapeutic use ; Hematopoiesis ; drug effects ; Humans

In order to confirm the role that the 49th amino acid residue plays in enzymatic inactivity of Glutamine 49 phospholipase A2(Gln49-PLA2),site-directed mutagenesis of its 49th amino acid gene codon was conducted using PCR.Aspartic acid 49 phospholipase A2(Asp49-PLA2-Q49D-PLA2),the mutant of Gln49-PLA2 was expressed in E.coli with pET32a+ vector.The fusion protein,expressed as inclusion body,after being denatured,was on-column refolded and purified by immobilized metal affinity chromatography(IMAC),and then cleaved by Factor Xa.The mature Q49D-PLA2 mutant was obtained by Hitrap SP cation exchange and Superdex 75 gel filtration chromatography,with the recovery rate of 1.3%,and the specific activity of the mature Q49D-PLA2 mutant was 72 U/mg.It has been demonstrated that the 49th glutamine amino acid residue is the main reason in enzymatic inactivity of Gln49-PLA2 and the results are helpful for denatured protein refolding,especially in rich disulfide bonds conditions.

Epidermal growth factor receptor (EGFR) is mutated, dysregulated or overexpressed in many epithelial malignancies, and EGFR activation has been found to be important in tumor growth and progression. Anti-EGFR monoclonal antibodies target the extracellular domain of EGFR; and show promising anti-tumor potential at clinical trials without severe side effects. In this article the pharmacokenetics and clinical study of 3 anti-EGFR monoclonal antibodies (cetuximab, panitumumab and nimotuzomab) were reviewed.
Antibodies, Monoclonal ; pharmacokinetics ; therapeutic use ; Antibodies, Monoclonal, Humanized ; Antineoplastic Agents ; therapeutic use ; Cetuximab ; Humans ; Neoplasms ; therapy ; Receptor, Epidermal Growth Factor ; antagonists & inhibitors ; immunology

Objective To investigate the role of overexpression of Smad7,the inhibitory factor of TGF-?/Smads signaling,in epithelial-mesenchymal transition (EMT) of peritoneal mesothelial cells.Methods Peritoneal fibrosis rat model was built by daily intraperitoneal injection with 4.25% Dineal (100 ml/kg) and lipopolysaccharide(LPS) (0.6 mg/kg) at day 8,10,12,22,24,26. Smad7 or control empty vectors was transferred at day 0,14 and was induced by doxycline in the daily drinking water (200 mg/L).Rats were sacrificed on day 28 and the expression of TGF-beta/ Smads,?-SMA and E-cadherin was examined.Results Compared with normal rats,empty vector rats showed higher expression of phosphorylated Smad2/3.?-SMA expression was elevated but E-cadherin was reduced.Under electron microscope,the mesothelial cells removed to submesothelial zone and showed large bundles of actin microfilaments and dense bodies within the cytoplasm. Basement membrane was broken.After induction of Smad7 in peritoneal fibrosis rats,the morphology of mesothelial ceils normalized partly,phosphorylated Smad2/3 was reduced.Moreover,expression of E-cadherin was increased,expression of?-SMA was dramatically reduced.Conclusion Inhibition of TGF-?/Smad signaling by Smad7 overexpression may inhibit the epithelial-mesenchymal transition of mesothelial cell,which may provide a new therapeutic method for peritoneal fibrosis by overexpression of Smad7.

This study was aimed to obtain higher efficiency in gene transfection into K562 cells and to study the role of green fluorescence protein (GFP) as a reporter system. Transfection efficiencies with different methods including electroporation and lipofectamine 2000 transfection, were observed and calculated under fluorescent microscopy by using GFP as a reporter system. The results showed that the transfection efficiency with electroporation (10%) was higher than that with lipofectamine 2000 (1%). In conclusion, the electroporation is a more ideal method for introduction of foreign gene into K562 cells. GFP can be used as a reporter system for optimizing transfection of K562 cells.
Electroporation ; Genes, Reporter ; Green Fluorescent Proteins ; genetics ; Humans ; K562 Cells ; Liposomes ; Transfection

OBJECTIVETo determine the methods for establishing an in vivo model of long-term hepatitis B virus (HBV) infection in the Chinese tree shrew (Tupaia belangeri chinensis).

METHODSSeventy-seven neonate (1-3 days old) and 49 young adult (2 weeks to 1 year old) tree shrews were inoculated with different HBV sources (chronic hepatitis B (CHB) human patient serum, single or pooled; HBV-infected tree shrew serum, single only; HBV-infected HepG2.2.15 cells' culture medium supernatant; HBV genome-transfected HepG2.2.15 cells' culture medium supernatant) through various routes of injection (subcutaneous, intraperitoneal, and direct liver via abdominal skin; adults also received intravenous and indirect liver via spleen). Serum and liver biopsies were collected from the animals at various time points post-inoculation for detection of HBV markers by fluorescence quantitative polymerase chain reaction, enzyme-linked immunosorbent assay, time-resolved immunofluorescence, Southern blotting, dot blotting, immunohistochemistry, and microscopy.

RESULTSAmong the neonatal group of tree shrews, six (7.8%) were confirmed as HBV-infected for more than 72 (up to 228) weeks after inoculation and another seven (9.1%) were suspected of persistent infections. None of the young adult tree shrews developed persistent infection. Inoculation with single-source serum from either CHB humans or tree shrews were responsible for the most cases of infections, and the subcutaneous injection produced more infections than the other inoculation routes. The most reliable methods of determining HBV infection status were detection of serum HBV immunoreactive markers and intrahepatic HBV DNA.

CONCLUSIONIn order to establish an in vivo model of CHB in the tree shrew, the animals should be inoculated in the neonatal period using subcutaneous injection.

Animals ; Disease Models, Animal ; Female ; Hep G2 Cells ; Hepatitis B virus ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Tupaia

OBJECTIVETo explore ZO-1 gene expression and methylation in leukemia cells and the involvement of ZO-1 gene in leukemogenesis.

METHODSRestriction landmark genomic scanning (RLGS) was used to identify new leukemia related gene, and methylation specific PCR (MSP) for ZO-1 methylation status. ZO-1 specific siRNA was designed and prepared by in vitro transcription and transfected into K562 cells, the transfected cells were cultured for 48 hours before harvesting. The effect of ZO-1 siRNA was monitored by Northern blot. Cellular proliferation capacity was assayed by CCK-8, cell apoptosis by Annexin V-fluorescence in isothiocyanate (FITC) assay, and cell cycle by phosphatidylinositol (PI).

RESULTSThe intensified spots in RLGS gel were subjected to bioinformatics analysis and one of the candidate spots was proved to be ZO-1 gene. In fresh leukemia cells, Molt4 cells and HL-60 cells, ZO-1 was hypermethylated, causing it reduced or silenced. ZO-1 gene was highly expressed with no methylation in normal peripheral blood MNC and K562 cells. There was a good correlation between promoter methylation and the gene silence. The silenced gene can be re-activated by demethylation treatment with 5-AZA-dC in most leukemia cell lines. RNA interference for ZO-1 gene in K562 cells did not interfere with cell proliferation, cell cycle and apoptosis.

CONCLUSIONZO-1 gene methylation might be involved in the tumorigenesis of acute leukemia.

Animals ; DNA Methylation ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; genetics ; pathology ; Membrane Proteins ; genetics ; metabolism ; Mice ; Phosphoproteins ; genetics ; metabolism ; Zonula Occludens-1 Protein

OBJECTIVETo understand pathogen patterns of enteric infectious diseases and its impact on this pattern due to aggregation of a great deal of foreign visitors during Beijing Olympic Games.

METHODSThe diarrheal patient's rectal swabs and stool specimens were collected from Olympic stadium and hospitals of four districts, including Dongcheng, Xicheng, Haidian and Chaoyang. Enteric multiple pathogens were detected from the total 45 specimens. The culture method was used for the enteric bacteria, ELISA and RT-PCR for the enteric viruses. Molecular typing of Salmonella Enteritidis isolation was completed by PFGE.

RESULTSIt was found that 26 out of 45 cases were positive with 57.8 percent for pathogen detection, and 24 were identified as enteric pathogenic bacteria, including Vibrio parahaemolyticus, Salmonella, diarrheagenic Escherichia coli and Campylobacter jejuni, two as norovirus. There were mixed infections of two pathogenic bacteria for three cases. Ten kinds of pathogens were detected from foreign cases, while five kinds from Chinese cases. A total of 5 PFGE patterns were identified in 10 Salmonella Enteritidis isolates from national and foreign diarrheal cases, which were concentrative in some extent.

CONCLUSIONVibrio parahaemolyticus, Salmonella, diarrheagenic Escherichia coli and Campylobacter jejuni were found to be the primary bacterial pathogens during the Olympic Games. Enteric virus infection existed in summer diarrhea.

Adult ; Bacterial Typing Techniques ; Campylobacter jejuni ; classification ; isolation & purification ; China ; Diarrhea ; epidemiology ; etiology ; microbiology ; virology ; Enterobacteriaceae ; classification ; isolation & purification ; Enterovirus ; isolation & purification ; Escherichia coli Infections ; microbiology ; Female ; Humans ; Male ; Salmonella ; classification ; isolation & purification ; Shigella ; classification ; isolation & purification ; Sports ; Vibrio parahaemolyticus ; classification ; isolation & purification

BACKGROUNDThe accurate and comprehensive assessment of glycemic control in patients with diabetes is important for optimizing glycemic management and for formulating personalized diabetic treatment schemes. This study aimed to analyze the correlation between 1,5-anhydroglucitol (1,5-AG) and glycemic excursions in type 2 diabetic patients.

METHODSSeventy-one outpatients with type 2 diabetes mellitus were randomly recruited from Chinese People's Liberation Army General Hospital. Using a continuous glucose monitoring system (CGMS), these patients' blood glucose levels were monitored for three consecutive days to obtain mean blood glucose (MBG) data. Intraday glycemic excursions were evaluated using the mean amplitude of glycemic excursions (MAGE), the largest amplitude of glycemic excursions (LAGE), standard deviation of blood glucose (SDBG) and the M-value. Interday glycemic excursion was assessed by absolute mean of daily difference (MODD). Postprandial glycemic fluctuations were evaluated using postprandial glucose excursions (PPGE) and postprandial incremental area under the curve (iAUC). Fasting venous blood samples were collected to measure serum 1,5-AG, whole-blood hemoglobin A1c (HbA1c) and serum glycated albumin (GA). Clinical markers of glycemia and parameters of glycemic excursions from CGMS were analyzed using the Pearson correlation coefficient and multivariate stepwise regression.

RESULTSPearson correlation analysis revealed that 1,5-AG was significantly correlated with MAGE, SDBG, M-value, LAGE, PPGE and iAUC (r values were -0.509, -0.430, -0.530, -0.462, -0.416 and -0.435, respectively, P < 0.01), especially in moderately and well-controlled patients, based on defined HbA1c levels. Multivariate stepwise regression analysis revealed a negative correlation between 1,5-AG and the above parameters, but not HbA1c and GA. Finally, HbA1c and GA were positively correlated with MBG and fasting blood glucose (FBG).

CONCLUSIONS1,5-AG was much better than HbA1c and GA as a marker of glycemic excursions in type 2 diabetic patients. Based on these results 1,5-AG is the best metric for assessing postprandial glucose levels in moderately and well-controlled patients, while HbA1c and GA were superior to 1,5-AG for monitoring MBG and FBG.

Aged ; Blood Glucose ; metabolism ; Deoxyglucose ; blood ; Diabetes Mellitus, Type 2 ; blood ; metabolism ; pathology ; Female ; Glycated Hemoglobin A ; metabolism ; Humans ; Male ; Middle Aged ; Postprandial Period ; physiology

BACKGROUNDOver one million soldiers were treated for battle- or training-fatigue during World War II. Of all ground combat troops, 37% were discharged for psychiatric reasons due to fatigue. The neuroendocrinological and immunological systems played important roles in the work-related fatigue of military personnel. The aim of this study was to investigate the characteristics of fatigue associated with military operations, and we observed changes in the regulatory functions of the neuroendocrinological and immunological systems that may provide theoretical support for improving the combat effectiveness of armies.

METHODSA total of 240 soldiers from the Field Artillery regiment were selected as subjects. Researchers and subjects received training before participating in the study. Data of the subjects' medical histories, physical examinations, scores on a fatigue assessment scale, and assessments of pituitary-adrenal hormones (adrenal cortical hormone (ACTH), cortical hormone (F), and 24-hour urine-free cortisol (UFC)), pituitary-gonadal hormones (luteinizing hormone (LH), follicle-stimulating hormone (FSH), testosterone, estradiol (E2), and prolactin (PRL)), pituitary-thyroid hormones (thyroid-stimulating hormone (TSH), thyroxine (TT4), triiodothyronine (TT3), free thyroxine (FT4), and free triiodothyronine (FT3)), and cellular immune parameters (CD3(+), CD4(+), CD8(+), CD4(+)/CD8(+), B, and NK cells) were investigated before and after large-scale and high-intensity field exercises. Data were statistically analyzed with Student's t test using SPSS software (version 13.0), and P values < 0.05 were deemed to be significant.

RESULTSAfter the high-intensity military training, the scores on the fatigue scale reflected significant increases of feeling of unpleasantness among soldiers. Additionally, the symptom checklist showed notable increases in somatization scores and significant decreases in psychoticism scores. After intensive military work, levels of plasma ACTH, F, and UFC of soldiers were decreased (P < 0.01). The level of testosterone decreased significantly after the maneuver ((23.51 ± 6.49) versus (18.89 ± 5.89) nmol/L; P < 0.001), whereas the thyroid function (TT3, FT4, and FT3) was markedly increased after the maneuver (P < 0.01). The number of CD3(+), CD4(+), CD4(+)/CD8(+) cells, and B lymphocytes were decreased (P < 0.05), and NK cells were increased (P < 0.001) after the maneuver.

CONCLUSIONSFollowing high-intensity military operations, the psychological tolerance of soldiers was depressed. And the hypoadrenocorticism (the functional decreases of hypothalamic-pituitary-gonadal and abnormal pituitary-thyroid axis) contributed to the increased levels of fatigue. Hypoimmunity may increase the susceptibility to diseases after high-intensity military operations.

Adolescent ; Adrenal Glands ; secretion ; Adult ; Endocrine System ; metabolism ; Estradiol ; blood ; Follicle Stimulating Hormone ; blood ; Humans ; Hydrocortisone ; blood ; Luteinizing Hormone ; blood ; Male ; Military Personnel ; Pituitary Gland ; secretion ; Pituitary Hormones ; blood ; Prolactin ; blood ; Testosterone ; blood ; Thyroid Hormones ; Thyrotropin ; blood ; Thyroxine ; blood ; Triiodothyronine ; blood ; Young Adult