Objective To assess the clinical significance of changes tumor necrosis factor(TNF)-alpha level and the relationship between serum TNF-? and glucose levels in patients with gestational diabetes mellitus(GDM). Methods 71 patients with gestational diabetes mellitus were enrolled.40 normal pregnant women were assigned to be control group.Then the concentrations of blood glucose and TNF-? were determined. Results Blood TNF-? level in GDM patients was signficantly higher than that in normal pregnant women(P

OBJECTIVETo investigate the apoptosis-inducing effect of DMF on the human liver cells (HL-7702) in vitro.

METHODSLiver cells were exposed to different concentrations of DMF (0, 50, 100, 150, 200 mmol/L) for 12 hours. Apoptotic rate, the expression of Bax, Bcl-2 and Caspase-3 in liver cells were measured by FCM and western blotting respectively.

RESULTSThe increase in apoptotic rate of hepatocytes in concentration-manner was shown after DMF treatment for 12 h. After treatment the expression of Bcl-2 was decreased steadily and lower than the control group (P < 0.01), the expression of Bax showed no significant difference among the groups of different dosage by one-factor analysis of variance (P > 0.05), as the increase of the dosage of DMF. The ratio of Bcl-2/Bax dropped with the dosage of DMF increasing, and the ratio in 200 mmol/L of DMF was significantly lower than that of the control (P < 0.01). The new lands of procaspase-3 in 150, 200 mmol/L were observed, which demonstrated that there was active caspase-3.

CONCLUSIONDMF can induce apoptosis of cultured adult normal hepatocytes in vitro, and the mechanism might be related to the decrease of Bcl-2/Bax and the cleavage of Caspase-3.

Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cells, Cultured ; Dimethylformamide ; pharmacology ; Hepatocytes ; drug effects ; metabolism ; pathology ; Humans ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; bcl-2-Associated X Protein ; metabolism

Objective To measure the expression of granulocyte colony-stimulating factor receptor (G-CSFR) in human melanocytes and to evaluate the biologic effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) on human melanocytes.Methods Melanocytes were obtained from circumcision specimens of healthy males,and neutrophils were isolated from heparin-andcoagulated peripheral blood of healthy human followed by a primary culture.Then,the melanocytes in third passage were cultured with or without the presence of various concentrations (200,400,600,800 μg/L) of rhG-CSF for 72 hours.The growth and morphology of melanocytes were observed.Flow cytometry was performed to detect the expression of G-CSFR in untreated human melanocytes,neutrophils and erythroleukemia cells (HEL 92.1.7).Western blot and reverse transcription PCR (RT-PCR) were carried out to measure the expression of G-CSFR protein and mRNA respectively in the neutrophils,HEL 92.1.7 cells,treated or untreated human melanocytes.Methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate the proliferation,and dopa-oxidation assay to estimate the tyrosinase activity,of treated melanocytes.Results The expression rate of G-CSFR was 76.81％ ± 10.70％ in human melanocytes,significantly higher than that in the HEL 92.1.7 cells (2.53％ ± 1.54％,P ＜ 0.01 ),but lower than that in the neutrophils (85.76％ ± 15.71％,P ＜ 0.05).Both G-CSFR protein and mRNA were expressed in melanocytes,and there was no significant differences in the expression level of G-CSFR protein and mRNA among melanocytes treated with different concentrations of rhG-CSF (both P ＞ 0.05).The expression levels of G-CSFR protein and mRNA in the melanecytes were significantly higher than those in the HEL 92.1.7 cells (both P ＜ 0.01 ),but lower than those in the neutrophils (P ＜ 0.05 or ＜ 0.01 ).rhG-CSF at 200-800 μg/L displayed a significantly promotive effect on the proliferation of melanocytes (P ＜ 0.01 or ＜ 0.05 ),and the effect was in a dose-dependent manner when rhG-CSF ranged from 200 to 600 μg/L (P ＜ 0.01 ).The rhG-CSF at 600 μg/L and 12-O-tetradecanoyl-phorbol-13-acetate (TPA) at 20 μg/L showed an equivalent effect on the proliferation of melanocytes (164.04％ ± 13.0％ vs.165.62％ ± 10.6％,P ＞ 0.05).However,rhG-CSF from 200 to 800 μg/L had no significant impact on the tyrosinase activity of melanocytes (all P ＞ 0.05 ).Conclusions G-CSFR is expressed in human melanocytes. rhG-CSF can promote the proliferation of cultured human melanocytes,but has no obvious influence on the tyrosinase activity of melanocytes.

To identify whether a highly repeated GT sequence from DCA1 promoter from Dunaliella salina,which have been proved to be a salt-inducible promoter in our previous study,would be a salt-inducible regulation element,different primers were designed to amplify 6 different-length fragments of DCA1 promoter from D.salina by PCR.After these fragments were respectively inserted into the HindⅢ-BamH I sites of the vector pU?GUS,serial expression vectors containing the gus gene were generated.D.salina cells transformed with these recombinant plasmids by electroporation were grown in liquid media containing different concentrations of sodium chloride respectively.GUS enzyme activity was measured histochemically and fluorometrically.The results revealed that 3 fragments containing GT repeated sequence drove the external gus gene expression and the expression pattern of the gus gene was regulated by the concentrations of sodium chloride.Additionally,the 2 fragments without tandem GT sequence drove the gus gene expression,but the expression pattern of the gus gene wasn't regulated by the concentration of sodium chloride;Also,the upstream fragment of the tandem GT sequence wasn't able to drive the gus gene expression.In conclusion,the highly repeated GT sequence from the DCA1 promoter plays an important role in the salt-inducible regulation of DCA1 promoter from D.salina and might be a novel salt-inducible element.

OBJECTIVE: To test the technical parameters of GlobalFiler® PCR Amplification Kit for its application to forensic application value and to investigate the genetic polymorphisms. METHODS: The validation was conducted in sensitivity, mixed samples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The amplification and detection of the genomic DNA from 373 unrelated individuals from Beijing Han nationality were extracted by automation workstation. RESULTS: Global-Filer® PCR Amplification Kit was adaptive to some mixed, degraded and inhibited samples. The power of sensitivity and adaptability and peak height balance showed well. The distributions of genotype frequencies for 21 STR loci in the population were all in accordance with Hardy-Weinberg equilibrium (P > 0.05). The PIC value of the 21 STR loci was among 0.536 to 0.940; the H value was among 0.558 to 0.933; the DP value was among 0.783 to 0.992; the PE value was among 0.243 to 0.874. CONCLUSION GlobalFiler® PCR Amplification Kit is suitable for criminal cases and DNA database in forensic practice. And 21 STR loci in Beijing Han nationality have high polymorphism, which have application value in forensic practice and population genetics.
Asian People/genetics* ; Beijing ; Databases, Nucleic Acid ; Ethnicity ; Gene Frequency ; Genetic Loci/genetics* ; Genetics, Population ; Genotype ; Humans ; Polymerase Chain Reaction/standards* ; Polymorphism, Genetic ; Reproducibility of Results ; Species Specificity

In order to enhance the specificity of TNF-α monoclonal antibody to inflamed site, a bispecific antibody BsDb that targets TNF-α and the extra-domain B (ED-B) of fibronectin (FN) was constructed by covalently linking the anti-TNF-α single chain Fv antibody (TNF-scFv) and the anti-ED-B scFv L19 via a flexible peptide linker deriving from human serum albumin (HSA). ED-B is an antigen specifically expressed at the inflamed site. BsDb is expressed in E. coli, identified by immunoblot, and purified with affinity chromatography. This was followed by further examination of its bioactivities and pharmacokinetics. We demonstrated that BsDb retained the immunoreactivity of its original antibodies as it could simultaneously bind to TNF-α and ED-B and neutralize the biological action of TNF-α. In the collagen-induced arthritis mice model, BsDb selectively accumulate in the inflamed joint with a maximal uptake of (12.2 ± 1.50)% ID/g in a single inflamed paw and retain in the inflamed paw for at least 72 h. In contrast, BsDb showed a short serum half-life of (0.50 ± 0.05) h and a rapid clearance from normal tissues. The findings reported herein indicate that BsDb has good specificity to the inflamed site and low toxicity to normal tissues. BsDb is therefore likely to have greater clinical applications in the treatment of rheumatoid arthritis and other autoimmune diseases. This laid a stable basis for its preclinical study.
Animals ; Antibodies, Bispecific ; chemistry ; Antibodies, Monoclonal ; chemistry ; Arthritis, Experimental ; Escherichia coli ; Fibronectins ; chemistry ; Half-Life ; Humans ; Mice ; Single-Chain Antibodies ; chemistry ; Tumor Necrosis Factor-alpha ; chemistry

OBJECTIVETo investigate the protective effects of Yuyin Ruangan Decoction(YRD, traditional Chinese medicine) on experimental hepatic injury in mice.

METHODSThe mice were randomly divided into control group, model group and YRD low, middle and high dose group(n = 11). By ip injection of D-GalN, CCk or thioacetamide (TAA), three models of hepatic injury mice were established to investigate the effects of YRD through detecting the indexes of liver function in serum and, the content of antioxidant system in the hepatic tissue.

RESULTSYRD could decrease the content of alanine aminotransferase (ALT)and aspartate aminotransferase (AST) in serum and that of malonaldehyde (MDA) in the hepatic tissue, upregulate the activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) in the hepatic tissue. Furthermore, the above effects were dosedependent in a certain degree. CoNCLUSION: YRD has some protection effects on the model of experimental hepatic injury in mouse.

Alanine Transaminase ; blood ; Animals ; Antioxidants ; metabolism ; Aspartate Aminotransferases ; blood ; Chemical and Drug Induced Liver Injury ; drug therapy ; Drugs, Chinese Herbal ; pharmacology ; Glutathione Peroxidase ; metabolism ; Liver ; drug effects ; enzymology ; Malondialdehyde ; metabolism ; Mice ; Superoxide Dismutase ; metabolism

Objective To explore characteristics of sleep structure and the correlation with cognitive function in cerebral infarction combined with obstructive sleep apnea hypopnea syndrome (CI-OSAHS).Methods The patients with CI-OSAHS and OSAHS in Department of Neurology and Breathing Sleep Monitoring Room of Tianjin Medical University General Hospital from December 2009 till March 2011 were collected All the patients completed polysomography(PSG).Sixty patients were selected and divided into 3 groups based on PSG.These 3 groups were combined group 20 persons (CI-OSAHS),OSAHS group 20persons (OSAHS) and control group 20 persons (without cerebral infarction obstructive sleep apnea hypopnea syndrome).All the patients completed image examinations ( CT and MRI ) evaluation of the cognitive function by Mini-Mental State Examination(MMSE) and Montreal Cognitive Assessment(MoCA).Results Sleep structure:the awake time,non-rapid eye movement sleep (NREM) 1,NREM 2 and NREM periods in combined group and OSAHS group were significantly longer,the NREM3 + 4 and rapid eye movement(REM) periods were shorter than the control group.The NREM and NREM 1 periods in combined group were longer,the NREM 3 +4 and REM periods were shorter than the OSAHS group.The correlation analysis of cognitive function and breathing disorders and low oxygen related index:there was negative correlation between the total scores of cognitive function (MMSE and MoCA)and apnea hyponea index,oxygen desaturation index (ODI) ( MMSE r =-0.450,-0.671,MoCA r =-0.486,- 0.494,all P ＜0.05) while,was positive correlation between them and noctumal average hypoxemia and minimum hypoxemia ( MMSE r =0.477,0.485,MoCA r =0.507,0.482,all P ＜0.05) in the OSAHS group.There was negative correlation between ODI,arousal index and the total scores of MoCA in the combined group (MoCA r=-0.463,0.480,both P＜0.05),there was correlation between the total scores of MMSE and the other sleep parameters,but,there was no difference in statistics.The correlation analysis of cognitive function and sleep stages:There was positive correlation between the total scores of cognitive function ( MMSE and MoCA) and the NREM 3 + 4 periods ( r =0.521,0.474,both P ＜ 0.05 ) while,there was negative correlation between the total scores of MMSE and the N REM 1 + 2 periods (r =-0.458,P ＜ 0.05 )in the OSAHS group.There was positive correlation between the REM period and the total scores of MoCA (r =0.472,P ＜ 0.05 ).There was correlation between the total scores of MMSE and the sleep structure,but,there was no difference in statistics in combined group.Conclusions Patients with OSAHS have obvious sleep structure disorder.The awake time and light sleep periods are significantly longer than the control group,while,the deep sleep and REM periods are significantly shorter than the control group.The NREM 1 of the patients with CI-OSAHS is longer than the patients with OSAHS.The higher the AHI,the lower the night blood oxygen,the more obvious cognitive dysfunction The longer the awake time,the longer the light sleep,the shorter the deep sleep and REM periods,the more serious cognitive dysfunction.The correlation between the cognitive impairment and low oxygen is more apparent than sleep structure.There is apparent correlation among the total scores of MoCA,the degree of hypoxia and sleep structure in the patients with CI-OSAHS.The total scores of MoCA are more sensitivity than MMSE in mild vascular cognitive impairment.

Sinisan is a widely used traditional Chinese medicine (TCM) in treating various diseases; however, the in vivo metabolic profile of its multiple components remains unknown. In this paper, ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) was applied to identify the metabolites of Sinisan extract in rat plasma, urine, feces and bile after intragastric administration. Using MS(E) and mass defect filter techniques, 41 metabolites of 10 parent compounds (naringin, naringenin, hesperidin, neohesperidin, liquiritin, liquiritigenin, glycyrrhizic acid, glycyrrhetinic acid, saikosaponin a and saikosaponin d) were detected and tentatively identified. It was shown by our results that these compounds was metabolized to the forms of hydroxylation, glucuronidation, sulfation, glucuronidation with sulfation and glucuronidation with hydroxylation in vivo.

A 37-year-old female was admitted to the hospital for an itching and painful subcutaneous nodule with ulceration on the extensor aspect of her left forearm for more than 6 months.The pain was severe,continuous and localized.Systemic and local treatment with antibiotics resulted in no obvious improvement.The lesion had gradually increased in size over the past 6 months and the ulcer had enlarged for 1 month.On examination,a hard infiltrative plaque measuring about 5.5 cm × 4.0 cm with a well-defined margin was seen on the extensor aspect of her left forearm,along with ulceration and some dirty discharge on the surface.The diagnosis of fibrosarcoma,grade Ⅱ was eventually made by a biopsy of the lesion,which revealed increased pigmentation in the basal layer,and tumor tissue was tightly adherent to the epidermis.Dermis and subcutaneous fat layer were infiltrated with various sizes of spindle cells with fine collagen fiber bundles between the cells.Obvious atypia and mitotic figures were easily observed in some of the cells.Immunohistochemical analysis showed moderately positive staining for fibronectin,but negative staining for human melanoma black-45 (HMB45),S100,smooth muscle actin (SMA),Melan-a,high molecular weight cytokeratin (HCK),CD34,CD68 or cytokeratin.Some diseases should be differentiated from this case,including dermatofibrosarcoma protuberans,cutaneous spindle cell squamous carcinoma,atypical fibroxanthoma,malignant fibrous histiocytoma,and so on.