Background Recombinant hirudin variant Ⅲ(rHV3) can effectively prevent galactose-induced human lens epithelial cells LECs injury,but little is known about the molecular mechanism of its action.Objective The present study was to investigate the effects of rHV3 on the expression of apoptosis-related genes in damaged LECs induced by galactose.Methods The rHV3 was extracted by our research group,and the biological activity of rHV3 was identified by titration of thrombase according to Markwardt's method.Human LECs (SRA01/04) were cultured using 125×10-3 mol/L D-galactose+10% FBS+D/F12 medium to establish the damaged human LECs model.rHV3 was added into the medium of the damaged human LECs model.Human LECs were cultured in D/F12 medium containing 10% FBS as normal control.The expression of apoptosis-related genes,such as aldose reductase (AR),bax,bcl2 and p53,in LECs at the mRNA level was detected using RT-PCR.The abundance ratio of target genes was presented with the absorbance (A) of gene mRNA/GAPDH mRNA.Results Compared to the normal control group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were significantly elevated in model group (t=3.90E-06,t=8.44E-04,t=5.15E-08,P＜0.01).However,in the rHV3-treated group,the A values of AR mRNA/GAPDH mRNA,bax mRNA/GAPDH mRNA and p53 mRNA/GAPDH mRNA were lower than those of model group (t=5.90E-06,t=1.51E-04,t=3.42E-06,P＜0.01).The bcl2 mRNA/GAPDH mRNA was markedly downregulated in the model group when compared with the normal control group (t=1.86E-05,P＜0.01);while after rHV3 addition,bcl2 mRNA/GAPDH mRNA increased in comparison with the model group (t=8.56E-05,P＜0.01).Conclusion 125×10-3mol/L D-galactose induces the damage and apoptosis of human LECs.rHV3 likely plays a protective function on D-galactose-induced damage of human LECs by inhibiting the polyol pathway and mitochondria-mediated pathway.

【Objective】To explore the optimal conditions in fingerprinting (APHDF).【Methods】The human DNA fingerprints were detected by APHDF.A pair of short primers was used for amplification.The experimental conditions including template,Mg2+,deoxyribonucleotides,and parameters of cycle,were optimized.【Results】The template DNA should be abstracted freshly and the concentration should be ranged from 50～550 mg/L.The best concentration of Mg2+was 5.0 mmol/L.The deoxyribonucleotides concentration was optimal at 0.2 mmol/L.The PCR cycling parameters were as follows :The denaturing temperatures,annealing temperatures and extension temperatures were 94 ℃ and 90 ℃ for 30 s,43 ℃ and 48 ℃ for 40 s or 50 s,and 72 ℃ for 1 min or 80 s,respectively.【Conclusion】The optimal conditions of the experiment are obtained,with good reproducibility and high specificity.Therefore,this method can be widely applied in practice.

The maternal components are constantly changing with gestational weight gains during pregnancy. Body composition analysis is a method for evaluating the maternal body composition accurately during pregnancy, models of which consist of fat mass, extracellular water, intracellular water, inorganic salts and protein basically. Among various methods to execute body composition analysis, bioelectrical impedance analysis is an important method for pregnant women. Many studies have reported about clinical applications based on correlations between maternal individual components and gestational diseases. This article reviews the researches on clinical applications of body composition analysis to pregnant women.

Objective To find a proper way for accurate results on completing blood counts.Methods Based on the results from automatic hematology analyzer XE-2100,set up the criteria for blood cell microscopic examination.1368 blood specimen were detected and the results were analyzed according to the criteria.Statistics on the data were made to evaluate the accordance between warnings of analyzer and manual examination,likewise the reliability of the criteria.Results Comparing with microscopic examination,analyzer warning on low PLT has good accordance,Kappa value was 0.95,U value was 35.19,P

The Sanger sequencing techniques, also known as the first-generation sequencing techniques and the gold standard of sequencing, have promoted the completion of "working draft" of the human genome, but the disadvantages of low throughput and high cost limit its large-scale application. The second-generation sequencing techniques, also known as the next-generation sequencing techniques, have widely used in basic research and clinical application because of its high throughput and low cost, but the short reads has always been an unavoidable shortcoming. Then, the emergence of the third-generation sequencing techniques, with the long reads, provides new technology selection for the analysis of complex repetitive regions on genome sequences and the assembly of high-quality genomes. In recent years, the third-generation sequencing techniques have been further developed, and have gradually demonstrated the clinical application value. This article reviewed the research progress and clinical application of the third-generation sequencing techniques.
Genome ; High-Throughput Nucleotide Sequencing/methods* ; Humans ; Sequence Analysis, DNA/methods*

To study the expression and significance of pten gene in patients with myelodysplastic syndrome (MDS) and acute myeloid leukemia (AML), RT-PCR and Western blot were respectively applied to detect pten mRNA and PTEN protein in Jurkat cells (as negative control), in bone marrow nucleated cells of 35 patients with MDS, 45 patients with AML and 20 normal control. The results showed that pten mRNA expression could not be detected in Jurkat cells, and the positive rate in MDS patients (77.1%) was significantly lower than that in normal control group (90.0%) (p > 0.05), while significant difference was found between AML patients and normal control (60.0% vs 90.0%, p < 0.05); the positive rate in MDS-RAEB patients (70.0%) was lower than that in MDS-RCMD (86.7%); positive rate in de novo and relapsed AML patients (53.3%) was lower than that in AML patients in CR (73.3%), but statistics tests did not show significant difference (p > 0.05). The results of relative expression level of pten mRNA in all groups indicated that both relative expression levels in MDS patients and AML patients were definitely lower than that in normal control group (p < 0.005); the relative expression level in MDS-RAEB patients was lower than that in MDS-RCMD patients (p < 0.05); and in de novo and relapsed AML patients was obviously lower than that in AML patients in CR (p < 0.001). However, there was no significant difference between MDS and AML patients (p > 0.05). The positive rate of PTEN protein expression in both MDS (65.7%) and AML (54.8%) patients were lower than that in normal control (90.0%) (p < 0.05), and there was no significant difference when comparing MDS-RCMD patients (80.0%) with MDS-RAEB patients (55.0%) (p > 0.05), but positive rate of PTEN protein expression in de novo and relapsed AML patients (44.4%) was significantly lower than that in AML patients in CR (73.3%) (p < 0.05). It is concluded that the complete loss of pten mRNA in MDS and AML is uncommon, but the relative expression level in both diseases is significantly lower than that in normal people. The positive rates of PTEN protein expression in both MDS and AML patients are lower, compared with normal people, but are not in accordance with the expression of pten mRNA. The abnormalities of pten gene expression may be involved in the pathogenesis of MDS and AML.
Adult ; Aged ; Female ; Gene Expression ; Genes, Suppressor ; Humans ; Jurkat Cells ; Leukemia, Myeloid, Acute ; genetics ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; PTEN Phosphohydrolase ; genetics ; metabolism ; Young Adult

OBJECTIVETo find a kind of quick and effective haemostasis to decrease the mortality of severe bleeding.

METHODS18 severe bleeding patients with different cause received recombinant activated factor VIIa (rFVIIa) were analyzed retrospectively.

RESULTSOf total 18 cases with severe bleeding, 13 cases cured, 3 cases were effective, 2 cases ineffective. The total clinical effective rate is 88.89%. After using rFVIIa, the PT, APTT and fibrinogen level of 6 DIC patients returned to normal within 12 hours; 13 patients whose the amount of bleeding can be evaluated stopped bleeding quickly. The fastest onset time was 10 min.

CONCLUSIONrFVIIa can stanch severe bleeding for a variety of reasons rapidly and effectively, including coagulopathy, thrombocytopenia, and obstetric hemorrhage. Application of rFVIIa may decrease mortality, when conventional treatment is not valid.

Adult ; Aged ; Aged, 80 and over ; Blood Coagulation Disorders ; drug therapy ; Factor VIIa ; therapeutic use ; Female ; Hemorrhage ; drug therapy ; Humans ; Male ; Middle Aged ; Recombinant Proteins ; therapeutic use ; Retrospective Studies ; Treatment Outcome ; Young Adult

Child ; Family ; Humans ; Lymphohistiocytosis, Hemophagocytic ; Nervous System

Child ; Glutamate Dehydrogenase ; genetics ; Humans ; Hyperinsulinism ; genetics ; Male ; Mutation