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

ObjectiveTo investigate the influence of different depths and mediums on tissue stiffness measurement by virtual touch tissue quantification technology.Methods Four kinds of in vitro porcine tissue were recruited,three (fat,muscle and liver) were assigned as medium group and three (kidney,liver and muscle) as sample group.The shear wave velocity (Vs) of the samples was measured under different mediums and at different depths (2 cm,3 cm,4 cm and 5 cm),respectively.ResultsAccording to the depth of measurement,two groups of Vs were defined with one group at the depth of 2 cm and 3 cm while the other at 4 cm and 5 cm.The difference of Vs between the two groups was of statistical significance and with the increase of the depth Vs values decreased.Even at the same depth,the differences of Vs under different mediums were statistically significant.ConclusionsDepths and mediums have great influence on evaluating elasticity measurements by virtual touch tissue quantification technology.

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.

This study was to build a canine portal hypertension model by intra-portal administration of high polymer material polyurethane and organic solvent tetrahydrofuran mixed solutions in order to evaluate the effectiveness of the model. Twelve local crossbreed dogs were selected randomly, with intra-portal administration of 8% (weight/volume) polyurethane- tetrahydrofuran solutions through an incision in the upper abdomen to build the portal hypertension model. We measured the portal vein pressure before modeling, during modeling, and four-, eight-, and twelve- weeks after modeling, respectively. Then we evaluated the effectiveness of the model comparing values of data with those data obtained before modeling started, which were regarded as the normal values. The results showed that the portal vein pressure rose by 2. 5 times after the solution administrated instantly as much as that before modeling, and maintained at 1. 5 times after 4 weeks. This method presents an easy operation, low animal mortality and reliable model of portal hypertension. Its less abdominal adhesions and its ability in keeping normal anatomic structure specially make it suit for surgical research of portal hypertension.
Animals ; Disease Models, Animal ; Dogs ; Furans ; adverse effects ; Hypertension, Portal ; Polyurethanes ; adverse effects ; Portal Vein ; physiopathology

Objective Acrysof IQ Restor multifocal toric intraocular lens ( IOL) is a new product , which allows a single sur-gical procedure for presbyopia correction and corneal astigmatism management .This study was to evaluate the early clinical effects of phacoemulsification cataract surgery with implantation of a diffractive multifocal toric IOL . Methods We retrospectively analyzed 7 cases (9 eyes) of corneal astigmatism ≥1.0 diopter (D) treated by phacoemulsification with implantation of an Acrysof IQ Restor toric IOL.The patients were followed up for 3 months for observation of uncorrected distance visual acuity ( UDVA) , best corrected distance visual acuity ( CDVA) , uncorrected near visual acuity ( UNVA ) , best corrected near visual acuity ( CNVA ) , spherical equivalent (SE) refraction, focal depth, residual astigmatism, rotational stability of the IOL, contrast sensitivity (CS), and spectacle independ-ence preoperatively and at 1 week, 1 month, and 3 months after operation . Results At 3 months after surgery , the UDVA ( log-MAR), CDVA, UNVA, and CNVA were 0.07 ±0.10, 0.02 ±0.11, 0.12 ±0.06, and 0.08 ±0.07, respectively, with an SE re-fraction within ±0.50 D of the attempted spherical correction in 8 eyes (88.9%) and a focal depth of (5.32 ±1.78) D.The residual astigmatism at 3 months was significantly reduced as compared with the baseline ([0.25 ±0.28] vs [1.55 ±0.39] D, P<0.05), but showed no statistically significant differences from the preoperative an-ticipated residual astigmatism (P>0.05).At 3 months, the mean IOL axis rotation was (3.11 ±1.61)°and CS was remarkably im-proved ( P<0 .05 ) , while CS with or without glare was not significantly different from that at 1 month at all spatial frequencies ( P>0.05) except at 18.0 cpd (P<0.05). Conclusion Implantation of the Acrysof IQ Restor multifocal toric IOL provides excellent overall quality of vision, spectacle independence, visual quality, and rotational stability for patients with cataract and corneal astigmatism.

Objective To evaluate the changes to mitochondrial ultrastructure in melanocytes of perilesional skin from patients with vitiligo.Methods Skin specimens were obtained from the perilesional area (0.5-1 cm distal to vitiligo lesions) of 10 patients with progressive vitiligo and 10 patients with stable vitiligo,as well as from the normal skin of 10 healthy volunteers.The morphology of melanocytes was observed by using transmission electron microscopy (TEM).Besides,stereological parameters of mitochondria,such as volume density (Vv),surface density (Sv) and numerical density (Nv),were measured.Results In melanocytes from the healthy controls,there were a large number of melanosomes with the number of melanosomes per melanocyte being 28.57± 3.21,which were mainly at stage Ⅲ and Ⅳ; mitochondria with normal structure and densely packed cristae were regularly arranged; autophagosomes were seen occasionally.Compared with the melanocytes from healthy controls,there was an obvious decrease in the number of melanosomes (especially stage Ⅲ melanosomes) in melanocytes from the perilesional skin of patients,with the number of melanosomes per melanocyte being 22.00 ± 6.16 (P ＜ 0.05) and 17.43 ± 6.24 (P ＜ 0.05) in patients with progressive vitiligo and stable vitiligo,respectively.TEM also showed disorganized or disrupted mitochondria in various shapes and sizes,most of which were swelling with obscure cristae and vacuolization,in melanocytes from the perilesional skin,and no autophagy was observed.The three stereological parameters were significantly different between the three groups of tissue specimens (all P ＜ 0.05),with the Nv,Vv and Sv of mitochondria being (7.194 ± 1.434) μm-3,(4.8 ± 1.2) ％,(2.42 ± 0.86) μ m-1 respectively in melanocytes from the healthy controls,(4.055 ± 0.906) μm-3,(7.4 ± 2.1)％,(3.58 ± 1.15)μm-1 respectively from patients with progressive vitiligo,(5.311 ± 0.873) μm-3,(6.5 ± 1.4)％,(2.82 ± 0.94) μm-1 respectively from patients with stable vitiligo.Conclusions Mitochondria are injured in melanocytes from perilesional skin of patients with vitiligo,and the degree of injury is more intense in progressive vitiligo than in stable vitiligo.