The sequences of 5.8S rDNA and the flanking internal transcribed spacers (ITS1 and ITS2) were sequenced from hypha, fruit body and sclerotia of Grifola umbellata and its companion fungus. Their ITS sequences similarity was 99.36%. The results suggested that G. umbellata was closely related to its companion fungus.

This article provides a brief description of systematics on Morchella ,and reviews the advances of molecular systematics on Morchella over the world.

Electrophysiology examination is an important technique in studying amblyopia, which mainly includes electrooculography( EOG), electroretinography ( ERG), visual evoked potential( VEP). This study does not only summarizes the definition, the mechanisms and the meaning of these indexes in the relevant research progress in recent years, but also makes a comment on the controversies among the relevant research conclusions.

Adult ; Body Composition ; Exercise ; physiology ; Female ; Humans ; Lipids ; blood ; Obesity ; blood ; Young Adult

Objective To prospectively evaluate the safety and therapeutic efficacy ofdalteparin in patients with high risk non-ST-elevation acute coronary syndromes (ACS) during percutaneous coronary intervention (PCI). Methods Atotal of 175 patients with high risk non-ST-elevation ACS were randomly assigned to 2 groups [dalteparin group and unfractionated heparin (UFH) group]. The patients in dalteparin group were given dalteparin at a dose of 5,000U subcutaneously soon after diagnosis and then an additional 60U/ kg intravenous bolus ofdalteparin before emergent PCI. Vascular access sheaths were removed immediately after PCI or coronary artery angiography; the patients in UFH group were given UFH intravenously at a dose of 25mgjust before PCI and an additional 65mg bolus was administered if angiographic findings showed that the patients were suitable for percutaneous transluminai coronary angioplasty (PTCA). Sheaths were removed at 4-6 hours after PCI; Results Eighty-three patients in dalteparin group underwent PCI while 82 patients in UFH group underwent PCI; anti-Xa activities of 52 patients in dalteparin group were measured. The average anti-Xa activity was (0.83±0.26) U/ml at 15 minutes after intravenous injection of dalteparin and anti-Xa>0.5U/ml was obtained in 96.1% of the patients; hematomas at puncture sites were significantly fewer in dalteparin group as compared with UFH group (2.3% vs 9. 2%, P < 0.05); none of the patients in 2 groups suffered major bleeding events. No death, acute arterial reocclusion or emergent revascularization events occurred at 30 days after PCI. Conclusions Our study demonstrated that early subcutaneous injection of dalteparin at a dose 5,000U after diagnosis and an additional 60U/kg intravenous bolus ofdalteparin before PCI is safe and efficacious for patients with high risk non-ST-elevation ACS undergoing emergent PCI

Objective To investigate HLA-DPB1, DQB1 gene alleles and their correlation with autoantibodies in type 1 diabetes mellitus. Methods Sequence-based typing was used to determine the alleles of HLA-DPB1 and -DQB1 in 68 patients with type 1 diabetes and 50 healthy controls. Autoantibodies 〔glutamic acid decarboxylase antibody (GADA), islet cell antibody (ICA), insulin autoantibody (IAA)〕were detected by ELISA. Results Compared with the controls, the frequencies of DPB1*0402 and DQB1*0301 were significantly lower in type 1 diabetics (11.76%, 5.15% vs 30.00%, 22,00% respectively, P

BACKGROUND:Human mesenchymal stem cells (hMSCs) become aging and even die after several passages. Some investigations have shown that telomere has a close correlation with life span of the cells. Whether the ectopic expression of human telomerase reverse transcriptase (hTERT) could induce the activity of the telomerase, maintain the length of telomere, and finally prolong the life cycle of MSCs without losing their multipotent differentiation capacity is still uncertain.OBJECTIVE: To observe the influence of the ectopic expression of hTERT on the telomerase activity and cell life cycles of hMSCs.DESIGN: Repetitive measurement trails.SETTING: Research Center of Stem Cell, Zhengzhou University Medical College.MATERIALS: The experiment was conducted in the Research Center of Stem Cell, Zhengzhou University Medical College from October 2003 to December 2005. hMSCs were obtained from 20 healthy donators from the Department of Pediatric Surgery and Outpatient, the Third and First Affiliated Hospitals of Zhengzhou University. Enhanced green fluorescent protein plasmid (pEGFP-C1) and pEGFP-hTERT were provided by Dr. Chantal Autexier of Canada. DH5α strain provided by Dr. Hou Wei-hong, the Key Molecular Medical Laboratory of Zhengzhou University Medical College.METHODS.: Under sterile condition, 2 mL bone marrow of sternum of healthy donors were harvested, and prepared after centrifugalization,dilution and passage.① Transfection of pEGFP-hTERT into hMSCs and the screening and amplification of resistance cloning:The 5th passage cells were seeded in a 24-well plate,and transfected by pEGFP-hTERT with lipofectamine method.The cells were divided into four groups including untransfected group,lipofectamine group,pEGFP-C1 group and pEGFP-hTERT group. Resistance cloning screen and amplification was performed by G418. ②hTERT mRNA expression and detection of telomerase activity:RT-PCR and PCR-ELISA were used to detect the hTERT mRNA expressions of the fifth passage hMSCs transfected with pEGFP-hTERT, and pEGFP-C1, the untransfected tenth passage hMSCs and K562 cells (positive control), and the telomerase activity of the fifth and thirtieth passage hMSCs transfected with pEGFP-hTERT,the fifth pEGFP-C1-transfected cells and the tenth passage untransfected cells. ③Karyotype analysis of hTERT-transfected MSCs: Chromosome analysis was performed by conventional Giemsa staining.④Inducement of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification:The transfected MSCs were cultured in a medium containing epidermal growth factor and basic fibroblast growth factor, which could induce the cells differentiate into neuron-like cells. The culture solution was changed every 3 days, and the changes in cell growth condition and morphologic characteristics were observed under an inverted microscope. The microtubule associate protein (MAP2) and neurofliament subunit M (NF-M) were identified by RT-PCR.MAIN OUTCOME MEASURES:①hMSCs transfection with different kathion liposomes and the screening and amplification of resistance cloning; ②hTERT mRNA expressions of each group and detection of telomerase activity; ③Karyotype analysis of pEGFP-hTERT-transfected MSCs; ④ Induction of differentiation from telomerase-positive MSCs into neuron-like cells and RE-PCR identification.RESULTS: ①With the decrease of G418 concentration, the cells in the untransfected and lipofectamine groups died, and stably EGFP expressed MSCs were obtained; after G418 screening, there was a cell clone undergone 35 passages and continued to proliferate, whose appearance and growth characteristics were similar to the untransfected MSCs observed under inverted microscope. ②The fifth passage pEGFP-C1-transfected hMSCs and tenth passage untransfected hMSCs remained telomerase-negative, but the K562 and fifth passage hTERT-transfected cells showed positive telomerase activity. ③The telomerase activity of the fifth and thirtieth passage hTERT-transfected cells was positive. ④The hTERT-MSCs at passage 10, 20 and 30 had 23 pairs of chromosomes, and two X chromosomes. So they were still normal diploid with normal chromosome appearance and number. ⑤Many hTERT-transfected MSCs had the typical appearance of neuron-like cells. RT-PCR analysis showed that th expressions of MAP2 and NF-M were increased.CONCLUSION:Ectopic expression of the hTERT gene is found in hMSCs,and can induce the telomerase activity of hMSCs.The ectopic expression of the hTERT gene in hMSCs could extend the life spans of cells and maintain their multipotent differentiation capacity.

Pulsed field gel electrophoresis(PFGE)is a new type technique of gel electrophoresis which can be used to separate large DNA molecules.It has been widely applied to the karyotype analysis,identification of species groups,genetic orientation and genetic analysis for fungi.This article describes the principle,development and general manipulative procedure of PFGE,and elaborates the application in the molecular research of fungi.

BackgroundIt has been well-known that integrin linked kinase(ILK) plays an important role in the pathogenesis of neorascularization.But there are few researches to elucidate the relationship between ILK and retinal nevascularization. Objective This study was to explore the expression and significance of ILK on retinal neovascularization and new vessels regression in the oxygen-induced retinopathy(OIR) mouse model. Methods One hundred and twenty-eight 7-day-old C57BL/6J mice were randomly divided into the normal group and model group.The mice were rose in(75±2)％O2 environment with mother mice together for 5 days and then in normal environment for 5 days to establish the OIR models.The mice in normal group were rose in the normal environment for 21 days.The 17-day-old mice were sacrificed and retinal sections were prepared for histopathological examination and the numbers of the cellular nuclei of vascular endotheliumbreaking retinalinternal limiting membranewere calculated.Retinal sections and sheeting were prepared in 12,14,17 and 21 -day-old mice to examine the formation and regression of new blood vessel using ADP histochemistry staining,and then immunochemistry,real-time PCR and Western-blotting were used to detect the expressions of ILK and its mRNA in retina. ResultsThe numbers of the cellular nuclei of vascular endothelium breaking retinal internal limiting membrane were (45.64 ± 12.17 )in OIR group,and those of the normal group were( 0.35±0.14 )with a significant difference between two groups (t =22.85,P＜0.05).Retinal new blood vessel appeared in 14-day mice,and peaked in 17-day mice and then regression in 21-day mice.ILK protein was expressed mainly in retinal ganglion cell layer,inner nuclear layer,inner plexiform layer and photoreceptor layer.Real-time PCR showed that ILK mRNA expressing levels in retina in model group were( 1.00±0.22),(1.85±0.17),(1.58±0.43) and(1.53±0.36) respectively in 12-,14-,17- and 21-day mice.Westernblotting determined that the A value of the ILK/β-actin was increased in 12-,14-,17-day mice and decreased in 21-day mice,and the A values were significantly higher in model group than the control group in various aged mice ( t =2.97,P＜0.05 ;t =11.88,P＜0.01 ;t =16.84,P＜0.01 ;t =13.00,P＜0.01 ). ConclusionsThese results indicate a space-time corresponding relation between the expression of ILK and retinal neovascularization.The obvious positive expression of ILK may be highly correlated with retinal neovascularization.

Objective:To assess whether familial occurrence has an influence on the morphological characteristics of patients with nonsyndromic cleft lip and/or palate (NsCL/P).Methods:A case-control analysis was performed on the morphological characteristics of familial group and sporadic group,using medical records of 1967 patients with NsCL/P treated in the Affiliated Stomatological Hospital of Nanchang University from 2002 to 2014.Results:164 (8.34％) cases presented a positive history of cleft in their families.The cleft types,the positive familial rate of cleft lip only (CLO),cleft lip and alveolar ridge(CLA),cleft lip and palate (CLP) and cleft palate only (CPO) were 8.11％,8.54％,6.19％ and 9.65％ respectively.A positive family history of NsCL/P was associated with 0.66 times risk of CPO (P =0.036,OR =0.66,95％ CI 0.44-0.98) compared to those of CLO,CLA and CLP.In familial group of CLP,the lateral incidence of male patients was different from that of female patients (P ＜ 0.001).There was no significant difference between familial group and sporadic group on birth weights,parental child-bearing age and clinical manifestations of patients.Conclusion:Familial occurrence might have an influence on cleft type,laterality and gender of the patients with NSCL/P.