microRNAs (miRNAs) are a class of small non-coding RNAs (length 21 to 25 nt).They can complement and bind to 3' non-coding region of their target genes,directly degrade target genes or inhibit the translation of encoding proteins,and extensively participate in development,metabolism,apoptosis,and a variety of disease processes.Studies have shown that miRNAs play important roles in the occurrence and development of ischemic stroke,and participate in post-stroke protection and regulation of repair mechanisms.

OBJECTIVESTo evaluate the relationship between microdeletion or mutation on the Y chromosome and Chinese patients with idiopathic azoospermia and severe oligozoospermia and to establish a molecular detection method.

METHODSMicrodeletion or mutation detection at the AZFa (sY84 and USP9Y), AZFb, AZFc/DAZ and SRY regions of the Y chromosome. Seventy-three azoospermia and 28 severe oligozoospermia patients were evaluated using PCR and PCR-SSCP techniques.

RESULTSTwelve of 101 patients (12%) with the AZFc/DAZ microdeletion were found, including 8 with azoospermia (11%) and 4 with severe oligozoospermia (14.3%), and 1 patient had a AZFb and AZFc/DAZ double deletion. No deletions in the AZFa or SRY regions were found. No deletions in AZFa, AZFb, AZFc/DAZ or SRY regions were found in 60 normal men who had produced one or more children.

CONCLUSIONSMicrodeletion on the Y chromosome, especially at its AZFc/DAZ regions, may be a major cause of azoospermia and severe oligozoospermia leading to male infertility in China. It is recommended that patients have genetic counseling and microdeletion detection on the Y chromosome before intracytoplasmic sperm injection.

Chromosome Deletion ; Humans ; Male ; Oligospermia ; genetics ; Sperm Injections, Intracytoplasmic ; Y Chromosome

Objective To investigate the effect of expressions of endothelial growth factor (VEGF) and VEGF receptor 2 (VEGFR2) in the ischemic cortex on ischemic cerebral injury in rats with diabetes mellitus.Methods A total of 36 healthy male Sprague-Dawley rats were divided into 3 groups:a shamoperation group,a cerebral ischemic group,and a diabetic cerebral ischemic group according to the random number table method.A diabetes model was induced by injection of streptozocin,and then,a permanent focal cerebral ischemic model was induced by the suture method.At 24 h after ischemia,the neurological deficit scores were conducted.The triphenyl tetrazolium chloride staining was used to measure the infarct volume.TUNEL was used to detect the apoptotic cells.Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression levels of VEGF and VEGFR2 mRNAs.Western blot was used to detect the expression levels of VEGF and VEGFR2 proteins.Results In the sham operation group,there were no neurological deficit and infarcts,and there were only a few apoptotic cells and a few expressions of VEGF,VEGFR2 mRNAs and protein.The neurological function score (4.25 ±0.54 vs.2.86 ±0.73);t =5.303,P＜0.001),infarct volume (51.69 ±2.26 mm3 vs.30.15 ±2.08 mm3;t =23.166,P＜0.001),and the number of apoptotic cells (24.22 ± 1.34/HP vs.13.28 ±0.37/HP;t =27.261,P＜0.001) in the diabetic cerebral ischernia group were significantly increased than those in the cerebral ischemic group,while VEGF,VEGFR2 mRNA,and protein expression level were significantly decerased (VEGF mRNA:4.74 ± 0.54 vs.6.71 ± 0.91,P ＜ 0.001;VEGFR2 mRNA:4.06 ± 0.60 vs.6.16 ± 0.96,P ＜ 0.001,VEGF protein:0.99 ± 0.13 vs.1.55 ± 0.23,P ＜ 0.001;VEGFR2 protein:4.12 ± 0.74 vs.6.23 ± 0.76,P ＜ 0.001) compare with the cerebral ischemic group.Conclusions VEGF/VEGFR2 signal pathway participates in diabetes aggravating ischemic cerebral injury.The downregulating of VEGF/VEGFR2 may be one of the mechanisms of diabetes aggravating ischemic cerebral injury.

AIM:To evaluate the effect of microRNA-155(miRNA-155) on the regulation of angiogenesis in diabetic rats with cerebral ischemic injury .METHODS: Adult male Sprague-Dawley rats were randomly divided into 5 groups:sham group, cerebral ischemia group , diabetic cerebral ischemia group , diabetic cerebral ischemia +miRNA-155 inhibitors group and diabetic cerebral ischemia +scramble group .Diabetes model was made by injection of streptozocin and permanent cerebral ischemic model was developed by suture-occluded method .The scores of neurological deficit and infarct volume were estimated at 24 h after cerebral ischemia .miRNA-155 level was detected by real-time polymerase chain reaction.The expression of platelet endothelial cell adhesion molecule-1 ( PECAM-1/CD31 ) and vascular endothelial growth factor ( VEGF) was detected by Western blotting .RESULTS:miRNA-155 inhibitor significantly reduced miRNA-155 levels in the ischemic cortex (P<0.05), improved the scores of neurological deficit , reduced infarction size and up-regulated the levels of CD31 and VEGF (P<0.05).CONCLUSION:miRNA-155 has a critical role in the regulation of angiogenesis in diabetic rats with cerebral ischemia .Down-regulation of miRNA-155 using miRNA-155 inhibitor attenuates brain infarct injury in diabetic rats .

Objective To investigate the potential effect of cetyltrimethyl ammonium bromide(CTAB)separated mannan of cell wall from Candida albicans on the production of IL -6and IL -8in h uman peripheral blood mononuclear cells(PBMC)induced by lipopoly saccharide(LPS).Methods PBMCs were pretreated with differen t concentrations of CTAB mannan(1.000mg /mL?0.100mg /mL?0.010mg /mL)for 24h.LPS(50?g /mL)was added and co-incubated for 24h.And a t 48h,the supernatants were collected.At 24h and 48h,only the super-natants of stimulated by CTABmannan were collected.LPS(50?g /mL)was the positive control,unstimula ted culture medium the negative control.The con tents of IL -6and IL -8in the supernatants were determined by ELISA.Re-sults At 24h and 48h,no IL -6and IL -8were detected in 3different concentration-CTAB mannan groups.LPS could induce IL -6(478.507?24.876ng /mL),IL -8(529.655?53.279ng /mL).The contents of IL -6and IL -8of negative control were not detectable.In 1.000mg /mL CTAB mannan +LPS group the contents of IL -6were(85.620?16.058ng /mL,P=0.004),IL -8were(123.940?20.319ng /mL,P=0.011).In 0.100mg /mL CTAB mannan +LPS group,IL -6(210.086?27.874ng /mL,P=0.007),IL -8(206.798?31.878ng /mL,P=0.022).In 0.010mg /mL CTAB mannan +LPS grou p,IL -6(201.387?32.396ng /mL,P=0.014),IL -8(203.133?36.012ng /mL,P=0.015).Conclusion CTAB mannan of cell wall from Candida albicans could downregulate the production o f IL -6and IL -8from human peripheral blood mononuclear cells induced by LPS.

OBJECTIVETo investigate and establish the gene diagnosis methods for the frequent mutations of iduronate-2-sulphatase(IDS) gene in mucopolysaccharidosis type II patients.

METHODSpolymerase chain- reaction-single strand conformation polymorphism PCR-SSCP) analysis was applied to detect the mutations of exons 3, 8 and 9 which were hot spots in the iduronate-2-sulfatase gene; DNA sequencing was applied to analyze the mutations which had been detected by PCR-SSCP; PCR-restriction fragment length polymorphism (PCR-RFLP) was applied to detect the results of DNA sequencing.

RESULTSObvious and abnormal bands in exon 9 of the IDS gene were found by applying PCR-SSCP; the mutation(C1672T) of exon 9 was found in the patient through DNA sequencing, which led to amino acid replacement(R468W); the PCR-restriction enzyme digestion showed that only one band(554 bp) appeared in the patient, but there were two bands (257 bp and 297 bp) in his parents, and it verified the results of sequencing analysis.

CONCLUSIONPCR-SSCP analysis, DNA sequencing analysis and PCR-restriction enzyme digestion are effective methods for MPS II diagnosis. Combined applications of these methods can verify and complement each other and improve the accuracy of diagnosis.

Amino Acid Substitution ; Base Sequence ; Child ; China ; Codon, Nonsense ; DNA ; chemistry ; genetics ; DNA Mutational Analysis ; Humans ; Iduronate Sulfatase ; genetics ; Male ; Mucopolysaccharidosis II ; enzymology ; genetics ; Mutation ; Point Mutation ; Polymorphism, Single-Stranded Conformational

OBJECTIVETo clone a novel gene which is related to human testis spermatogenesis apoptosis.

METHODSTo rapidly attain human novel gene full-length cDNA sequence from a human testis cDNA library,the gene-specific primers and the vector-specific primers were designed for nested polymerase chain reaction. Sequencing was performed and the result was analysed.

RESULTSThe present authors discovered the TSARG3 gene(GenBank accession number AF419291) from a human testis cDNA library, using a cDNA fragment (GenBank accession number BE644537) as an electronic probe, which was significantly changed in cryptorchidism and represented a novel gene. Furthermore, a mouse homologue of this gene was identified (GenBank accession number AF419292) by using the same method.

CONCLUSIONA novel gene named TSARG3 was cloned. It is considered that the function of the new gene is related to human testis spermatogenesis apoptosis.

Amino Acid Sequence ; Animals ; Apoptosis ; genetics ; Base Sequence ; Cloning, Molecular ; DNA, Complementary ; chemistry ; genetics ; Female ; Gene Expression ; Heat-Shock Proteins ; Humans ; Male ; Mice ; Molecular Sequence Data ; Proteins ; genetics ; Sequence Alignment ; Sequence Analysis, DNA ; Sequence Homology, Amino Acid ; Spermatocytes ; cytology ; metabolism