Objective:To explore the application of real-time transrectal ultrasound (TRUS) during seminal vesiculoscopy in infertile men with azoospermia or oligoasthenospermia.Methods:We retrospectively analyzed the clinical data of 25 cases of azoospermia or oligoasthenospermia due to ejaculate ducts obstruction who were treated with real-time transrectal ultrasound-guided seminal vesiculoscopy between September 2011 and December 2015. Patients’ age was(29.4±4.5) years. All patients accepted semen analysis, serum sex hormone, MRI, TRUS and then diagnosed as obstructive azoospermia, and 13 cases had intractable obstructive azoospermia or oligoasthenospermia after the failure of simple seminal vesiculoscopy(the path to the ejaculatory duct and seminal vesicle couldn’t be found). All patients were treated with seminal vesiculoscopy under real-time guidance with TRUS. We assessed the success rate of surgery, surgical time and complications.Results:The scope was successfully inserted into the seminal vesicle in 21 of the 25 cases (success rate, 84%). The median operative time was 75(31, 148) min. None of the patients developed severe complications. Among 4 failure cases (4/25, 16%), 1 was due to abnormal congenital development. In 2 cases, a clear outlet of the dual ejaculatory duct could not be found after it was inserted into the prostatic utricle. One case was considered as a Müllerian tubular cyst, and the seminal vesicle scope was used to assess the cystic side wall. The 21 patients were followed up for 3 to 6 months, semen volume 2.0(0-5.2)ml, total sperm 28(0-832) ×10 6/ejaculate, sperm density 5.6(0-110.3)×10 6/ml, mobility rate of sperm 5.4%(0-63.6%), and the differences were significant as compared to that before the surgery [semen volume 0.4(0-2.8)ml, total sperm 0(0-342)×10 6/ejaculate, sperm density 0(0-90.7)×10 6/ml, mobility rate of sperm 0(0-24.1%), all P<0.05]. Among the 17 patients who underwent follow-up of 5 to 9 years, 3 patients was conceived naturally and 9 patients’ postoperative sperm quality has improved and pregnancy in vitro fertilization by extracting sperm from semen. Conclusions:Intraoperative real-time transrectal ultrasound guidance can improved the success rate of seminal vesiculoscopy and promoted operative safety.

Introduction:The 2010 targets of the China Hepatitis B Prevention Programme were a prevalence of hepatitis B surface antigen (HBsAg) less than 1.0% for children less than five years old and less than 6.0% for the total population. This survey assessed the prevalence of Hepatitis B infection in Lianyungang, Jiangsu province, China in 2009–2010.Methods:Multistage sampling was used with 2372 subjects among 17 selected villages. Blood specimen collection and testing by enzyme-linked immunosorbnet assay (ELISA) were completed using the following markers for hepatitis infection: HBsAg and antibody to HBsAg (anti-HBs); hepatitis B e antigen (HBeAg) and antibody to HBeAg (anti-HBe); and hepatitis B core antibody (total anti-HBc). The data were analysed with Epi Info, version 3.3.2.Results:The prevalence of HBsAg was 2.4% (95% Confidence Interval [CI]: 1.8–3.0; Adjusted Prevalence [AP] 2.9%); anti-HBs prevalence was 51.1% (95% CI: 49.1–53.1; AP 49.2%) and total anti-HBc prevalence was 41.7% (95% CI: 39.8–43.7; AP 45.5%). The prevalence of HBsAg and total anti-HBc positivity increased from young to older age groups, yet the prevalence of anti-HBs positivity decreased from young to older age groups (P< 0.001 for all). There was no difference in the prevalences of HBsAg and anti-HBs among females and males (P= 0.108 and 0.089), but females had a higher prevalence than males for total anti-HBc positivity (P< 0.001). Discussion: This survey showed that in 2010 the prevalence of HBsAg among children aged less than five years was lower than the national target of 1.0% and that the prevalence of HBsAg for the total population was lower than the national target of 6.0%.

OBJECTIVETo analyze the relationship between the expression of FasL, Perforin and Granzyme B and the development of acute graft versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT).

METHODSThe peripheral blood mRNA expression of granzyme B, perforin, fasL from 17 patients after allo-HSCT was detected by competitive quantitative RT-PCR and the relationship between FasL, Granzyme B and Perforin expressions and clinical symptom of aGVHD was analyzed.

RESULTSThe expression level of Granzyme B, Perforin and FasL was 4.6 +/- 0.2, 4.5 +/- 0.1, 1.4 +/- 0.1 before aGVHD occurrence respectively, and was 98.7 +/- 2.5, 91.8 +/- 3.4, 61.5 +/- 2.2, after the occurrence in 14 patients (P < or = 0.05). Over expressions of Granzyme B, Perforin, and FasL during acute GVHD were detected in 13 of 14, 12 of 14, and 12 of 14 patients respectively. The upregulated expressions occurred prior to clinical symptom of aGVHD.

CONCLUSIONThe expressions of Granzyme B, Perforin, and FasL were significantly high in patients with acute aGVHD. Monitoring of the expressions, might predict the occurrence of clinical aGVHD and it severity and prognosis.

Acute Disease ; Adult ; Fas Ligand Protein ; genetics ; Female ; Gene Expression ; Graft vs Host Disease ; blood ; diagnosis ; etiology ; Granzymes ; genetics ; Hematopoietic Stem Cell Transplantation ; adverse effects ; methods ; Humans ; Male ; Perforin ; genetics ; Postoperative Complications ; blood ; diagnosis ; etiology ; Prognosis ; RNA, Messenger ; blood ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Transplantation, Homologous

Objective:To evaluate the association between the level of plasma proprotein convertase subtilisin/Kexin 9 (PCSK9) with incidence and severity of calcified aortic valve disease(CAVD).Methods:We prospectively recruited 120 CAVD patients with at least increased echo density and 40 control patients by transthoracic echocardiography.All patients were grouped by CT quantitative scoring system:aortic valve calcification (AVC)1,2,3 and 4.Calcium score of aortic valve were calculated.Total cholesterol (TC),low density lipoprotein cholesterol (LDL-C),high density lipoprotein cholesterol (HDL-C),triglyceride (TG), carrying lipoprotein A1 (apo A1) and apolipoprotein B (apo B),lipoprotein (a) {LP (a)} and hypersensitive C-reactive protein (hsCRP) were detected by biochemical analyzer.Plasma PCSK9 levels were measured by enzyme-linked immunosorbent assay.The re-lationship between AVC and plasma PCSK9 level,blood lipid,Apo A1,Apo B and hsCRP was analyzed.Results:The data indicated that Apo,B Lp(a) and LDL-C levels in AVC2-4 level was significantly higher than that of AVC1(P<0.05),while TG,APO,A1 HDL-C and hsCRP were not different significantly in the four groups.The levels of TC in group AVC3 and AVC4 were significantly higher than those in group AVC1(P<0.05).At the same time,the patients of grade AVC2-4 have higher level of plasma PCSK9 than patients of group AVC1(P<0.05).Correlation analysis was performed and aortic valve calcium score were significantly correlated with TC (r=0.248,P=0.026),LDL-C (r=0.222,P=0.048),Lp(a) (r=0.276,P=0.013),Apo A1(r=0.245,P=0.012),Apo B(r=0.212, P=0.019) and PCSK9(r=0.309,P=0.005) in all study subjects.PCSK9 was positively correlated with TC,LDL,LP (a),Apo A1, Apo,and no correlation with hsCRP (B).Conclusion:The level of PCSK9 in CAVD patients was significantly higher than that in control group.And there is an association of PCSK9 levels with the presence of CAVD,however.

OBJECTIVETo analyze the hematological and genetic characteristics of unstable hemoglobin Rush (Hb Rush) and compound heterozygote of Hb Rush and thalassemia.

METHODSPeripheral blood samples and genomic DNA from three patients (including two ethnic Dai and one Han Chinese) with anemia of undetermined origin were collected. Hematological phenotypes of these patients were determined through red blood cell analysis and hemoglobin electrophoresis. Genotypes of alpha- and beta-globin genes, -158 XmnⅠ polymorphic site ofγ promoter region, and haplotypes of 7 polymorphic restriction sites in the beta-globin gene cluster were determined using PCR-based methods and DNA sequencing.

RESULTSAll patients have presented hypochromic microcytic anemia and hemoglobin fraction with significant increased measurement (30.5%-59.2%) in the region of fetal hemoglobin during alkaline medium electrophoresis. DNA analysis suggested that all patients have carried mutations leading to the unstable hemoglobin Rush (HBB codon 101, GAG>CAG, Glu>Gln). Two of them were compound heterozygotes of Hb Rush and thalassemia mutations of -α,CD17 and Hb E, respectively. Hb Rush mutation was associated with various haplotypes of the β-globin gene cluster. No significant association was found between increased abnormal hemoglobin fraction in the region of Hb F and the polymorphism ofγ promoter or large deletion of the beta-globin gene cluster.

CONCLUSIONThis study has confirmed the distribution of Hb Rush among various Chinese populations and is the third report of its kind. Hb Rush can result in increased measurement of hemoglobin fraction in the region of fetal hemoglobin (Hb F) during routine hemoglobin electrophoresis under alkaline condition. Hb Rush heterozygote alone can lead to hypochromic microcytic anemia and thalassemia-like phenotype. Prenatal diagnosis of Hb Rush is necessary for carriers.

Adult ; Base Sequence ; Blood Protein Electrophoresis ; methods ; Female ; Fetal Hemoglobin ; genetics ; metabolism ; Genotype ; Haplotypes ; Hemoglobins, Abnormal ; genetics ; metabolism ; Heterozygote ; Humans ; Infant ; Mutation ; Phenotype ; Polymorphism, Genetic ; Sequence Analysis, DNA ; methods ; Thalassemia ; blood ; diagnosis ; genetics ; Young Adult ; alpha-Globins ; genetics ; metabolism ; beta-Globins ; genetics ; metabolism

BACKGROUNDThe purpose of this study was to investigate the feasibility of avoiding axillary lymph node dissection (ALND) for patients with only one sentinel lymph node (SLN) metastasis. The characteristics and predictive factors for non-sentinel lymph node (NSLN) metastasis of patients with single positive SLN were also analyzed.

METHODSPatients with no and only one SLN metastasis (0/n and 1/n group, n &ge; 2) were selected from 1228 cases of invasive breast carcinoma, who underwent axillary dissection in Shandong Cancer Hospital between November 1999 and December 2011, to compare the characteristics of NSLN metastasis between them. For the 1/n group, the factors that influenced the NSLN metastasis were analyzed by univariate and multivariate analysis.

RESULTSDifferences of the NSLN metastasis between the 0/n and the 1/n groups were significant (P < 0.001). There was no significant difference between the axillary lymph node metastasis on level III in 1/n group and 0/n group (P = 0.570). When the total SLN number was &ge; 4 and with one positive case, the NSLN metastasis was not significantly different from that in the 0/n group (P = 0.118). In the 1/n group, clinical tumor size (P = 0.012), over-expression of Her-2 (P = 0.003), tumor grade (P = 0.018) and the total number of SLN (P = 0.047) significantly correlated with non-SLN metastasis. Clinical tumor size (P = 0.015) and the expression of Her-2 (P = 0.01) were independent predictive factors for non-SLN metastasis by the Logistic regression model.

CONCLUSIONUnder certain conditions, breast cancer patients with single SLN metastasis could avoid ALND.

Adult ; Aged ; Breast Neoplasms ; complications ; pathology ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Middle Aged ; Sentinel Lymph Node Biopsy

OBJECTIVETo get the full length of human METH1 cDNA and express it steadily in mammalian cell stably.

METHODSMETH1 was amplified by RT-PCR, and cloned into pCDNA3.0 after confirmed by sequence analysis. HepG2 cells were transfected by Lipofectamine reagent and then selected in medium with G418. The expression level of METH1 was detected by RT-PCR and Western blot.

RESULTSMETH1 with expected length was effectively amplified, and completely matched the published sequence of encoding mature peptide [GI:5725505] as shown by sequence analysis. Eukaryotic vector expressing METH1 was obtained by gene cloning, cells expressing METH1 was got by selection with G418 at 3 weeks after transfection. RT-PCR and Western blot showed high level expression of METH1.

CONCLUSIONFull length of human METH1 gene is cloned successfully and expressed in HepG2 steadily, The results set up a basis for the study of effects of METH1 on hypertrophic scar angiogenesis.

ADAM Proteins ; ADAMTS1 Protein ; Angiogenesis Inhibitors ; genetics ; metabolism ; Blotting, Western ; Cell Line, Tumor ; Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Disintegrins ; genetics ; metabolism ; Humans ; Metalloendopeptidases ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection ; methods

OBJECTIVETo investigate the influence of home nurture environment on language development and social emotion in children with developmental language disorder (DLD).

METHODSThe 1-3 Years Child Home Nurture Environment Scale, Gesell Developmental Scale, and Infant-Toddler Social and Emotional Assessment Scale were used for the evaluation of 125 children with DLD. A total of 130 children with normal language development matched for age and sex were enrolled as control group.

RESULTSCompared with the control group, the DLD group had a significantly higher proportion of children in a bad home nurture environment and significantly lower scores of all domains of home nurture environment (P<0.05). In children with DLD, the home nurture environment score was positively correlated with the level of language development (r=0.536, P<0.01) and the score of ability domain in social emotion (r=0.397, P<0.01) and was negatively correlated with the scores of the domains of explicit behavior, covert behavior, and imbalance in social emotion (r=-0.455, -0.438, and -0.390 respectively, P<0.01). Home nurture environment had direct influence on language development in children with DLD and affected their language development via the mediating effect of social emotion.

CONCLUSIONSHome nurture environment influences language development and social emotion in children with DLD, and social emotion has a partial mediating effect between home nurture environment and language development.

Child, Preschool ; Emotions ; Environment ; Female ; Humans ; Infant ; Language Development ; Language Development Disorders ; psychology ; Male ; Social Behavior

OBJECTIVETo explore miRNA expression change of differentiation of mice marrow mesenchymal stem cells (MSCs) into adipocytes, which lay the foundation for further studies on molecular mechanism of miRNA regulating the differentiation of MSCs into adipocytes.

METHODSC57BL/6 mice MSCs were isolated, cultured through the whole bone marrow method, amplified by the differential adherent method. Cell growth was observed by morphology and the expression of superficial antigen CD29, CD44, CD34 were detected through immunohistochemistry. MSCs was induced to differentiation into adipocytes with adipocyte differentiation medium, and adipogenic differentiation of MSCs was analyzed by oil Red O staining. MicroRNA microarray was used to investigate the differentially expressed miRNAs in MSCs and adipocytes.

RESULTS(1) The fifth passage of MSCs had high purity under an inverted m icroscope. Immunohistochemistry staining showed that CD29, CD44 were positive and CD34 was negative in more than 90% MSCs. There were a large number of lipid droplets in cytoplasm after MSCs were induced with adipocyte differentiation medium, Oil O staining was positive. (2) The microarray experiment showed that 75 differentially expressed miRNAs were obtained in adipocytes compared with MSCs, 20 up-regulated and 55 down-regulated miRNAs were observed among them.

CONCLUSIONThere was a expression change of miRNA of differentiation of MSCs into adipocytes, some miRNAs might play important roles in MSCs adipogenic differentiation.

Adipocytes ; cytology ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Male ; Mesenchymal Stromal Cells ; cytology ; Mice ; Mice, Inbred C57BL ; MicroRNAs ; metabolism ; physiology

The aim of this study was to investigate the role of mesenchymal stem cells in the hematopoietic reconstitution of patients who had received haploidentical allogeneic hematopoietic stem cell transplantation (hi-allo-HSCT). 15 patients who underwent treatment with both MSCs and HSCs, were selected as study group, while 20 patients receiving only HSCT were taken as control. Bone marrow samples were obtained from iliac crest aspirates at several times after HSCT for the isolation, purification and expansion of MSCs. The confluent ratio and time were measured and compared with those of the control. The peripheral blood samples were obtained from patients, then absolute neutrophil and platelet counts were assayed. From day 4 before transplantation to day 28 after transplantation, serum was obtained every four days from patients of the two groups, and then 3 cytokines as SDF-1alpha, TPO and IL-11 were detected by ELISA. The results indicated that as compared with the control group, the ratio of primary confluent layer formation of MSCs in study group was obviously higher (27.3%) (p < 0.01), and the confluence time in culture was significantly less (p < 0.05). In the study group, the concentration of SDF-1alpha amounted to peak value (2975.19 +/- 681.56 pg/ml) on the 8th day after HSCT, which was obviously higher than that before HSCT (2403.70 +/- 522.39 pg/ml, p < 0.05), whereas in the control, the concentration of highest point of SDF-1alpha reached to peak valve (2280.60 +/- 701.25 pg/ml) on the 16th day after HSCT, which was less than that before HSCT (2701.46 +/- 483.21 pg/ml, p < 0.05). The concentration of TPO and IL-11 was higher in study group compared with the control from day 16 to 28 after HSCT (p < 0.05). It is concluded that the transfusion of MSCs combined with hi-all-HSCT may improve the injured state of the hematopoietic microenvironment in bone marrow of patients during allo-HSCT.
Adolescent ; Adult ; Bone Marrow ; metabolism ; pathology ; Chemokine CXCL12 ; metabolism ; Child ; Hematopoietic Stem Cell Transplantation ; methods ; Hematopoietic System ; Humans ; Interleukin-11 ; metabolism ; Mesenchymal Stem Cell Transplantation ; methods ; Middle Aged ; Thrombopoietin ; metabolism ; Young Adult