Objective To investigate the relationship between the expression of the survivin gene and CpC methylation in exon 1 of the survivin gene in CA tissue, and to study the expression of survivin protein in CA tissue and its modulation mechanism. Methods Tissue samples were obtained from the CA lesions of 30 patients, normal cervix of 10 female controls, and normal foreskin of 10 male controls. Immunohistochemistry was carried out to detect the expression of survivin protein in these specimens, RT-PCR to measure the mRNA expression of survivin, and methylation specific PCR (MSP) to analyze the methylation status of CpG island in the survivin gene exon 1. Results The positivity rate of survivin protein and mRNA was 90% (27/30) and 93.3% (28/30) in CA tissue specimens, respectively, 5% (1/20) and 5% (1/20) in control tissue specimens, respectively; there was a significant difference between the two groups of specimens in both the parameters (x2 = 35.187, 38.437, both P < 0.01). The demethylation of CpG island in the survivin gene exon 1 was observed in 86.7% (26/30) of the CA tissue specimens and in 15% (3/20) of the control tissue specimens (x2 = 10.865, P < 0.01). There was a positive correlation between the demethylation status of CpG island in exon 1 and the mRNA expression of survivin gene (x2 = 13.929, P = 0.014). Conclusions The expression of survivin protein in CA tissues might be associated with the demethylation of CpG island in exon 1 of the survivin gene, and may play a certain role in the development of CA.

Objective To evaluate the effect of serum LH level on the day of superovulation start on the prolonged gonadotropin-releasing hormone ngonist therapy on the outcomes of in vitro fertilization and embryo transfer (IVF-ET) in patents with ovarian endometriomas. Methods 75 patients with ovarian en-dometriomas were treated by laparoscopic cystectomy or laparotomy cystectomy or ultrasound-mediated cysts puncture, and gonadotropin-releasing hormone agonist (GnRH-α) was given for 3 to 4 times every 28 days after the operation. Superovulation started after 14 ～ 84 days of the last injection. All the patients were di-vided into two groups according to the level of serum LH. Group A included 30 patients whose level of LH was less than 0. 5IU/L, and group B included 45 patients whose level of LH was over 0.5IU/L and less than 1.5IU/L. The outcomes of IVF-ET were evaluated. Results The total ampoules of Gn administration and the ampoules of hMG needed in group A[(32.28±7.7) ampoules, ( 12.0±8. 9) ampoules,]were sig-nificantly more than that in group B[( 25.84±7. 1 ) ampoules, ( 6. 19±7.4) ampoules, P < 0.05] . The successful embryo implantation rate in group A( 18. 1% ) was lower than group B(26. 7% ), and the differ-ence has statistical significance ( P <0. 05). Conclusion The low level of serum LH on the superovula-lion day on the prolonged gonadotropin-releasing hormone agonist protocol will increase the ampoules of Gn administration and decrease the successful embryo implantation rate of IVF-ET, thus LH should be a more important reference parameter of superovulation start.

Objective To present the clinical features and to evaluate interventional therapy for Budd-Chiari syndrome in Chinese youth.Methods From January 1990 to April 2012,227 cases who hospitalized at the age ＜ 29 underwent color Doppler ultrasound scan and digital subtraction angiography (DSA).87 cases were with occlusive inferior vena cava (IVC type),105 cases with occlusive hepatic veins (HV type) and 35 cases with occlusive inferior vena cava and hepatic veins (MIX type).The occlusive veins were opened by percutaneous transluminal angioplasty (PTA),endovascular stent placement,intravenous catheter thrombolysis or combination.Postoperative anticoagulation was given to all patients.Results The symptoms and signs of portal hypertension disappeared or were alleviated in successful cases.Technical success was achieved in 210 patients.The success rate was 100％ in IVC type,85.7％ in HV type and 94.3％ in MIX type.IVC pressure decreased from (26.52 ± 8.16) cm H2O to (14.28 ±4.08) cmH2O(P ＜ 0.05) and HV pressure dropped from(35.70 ± 13.26) cm H2O to(18.36 ±8.16) cm H2O (P ＜0.05).Restenosis or occlusion was found in 21.4％ (45/210) patients after a follow-up of 1 month to 15 years.The rate was 13.8％ (12/87) in IVC type,31.1％ (28/90) in HV type and 15.2％ (5/33) in MIX type.These patients were managed by interventional procedures.Technical successwas achieved in 44 cases with restenosis.Conclusions Hepatic vein occlusion was the most common type of BCS in Chinese youth.The symptoms and signs of portal hypertension were the initial clinical manifestations.Postoperative recurrence rate in HV type was higher than that in the other two types.

Objective To study the relationship between chromosomal abnormality and Y chromosome microdeletions and male infertility. Methods Lymphocytes were cultured from peripheral blood of 1975 male infertility patients and stained with Giemsa. The chromosomes were analyzed under microscope. Y chromosome specific sequence tags (STS) were selected, then the Y chromosome microdeletions in AZF regions were screened by polymerase chain reaction (PCR) in azoospermia and oligozoospermia patients. Results There were 305 cases of detected chromosomal abnormalities (15.44%) in the 1975 cases. There were 101 cases (5.11 %) with autosome abnormalities which clinically manifested as oligozoospermia and teratospermia. There were 204 cases (10. 33%) of sexual chromosome abnormalities and the patients were mainly characterized with Klinefelter's syndrome. Y chromosome microdeletions were detected in 109 (14.97 %) of the 728 cases of azoospermia or oligozoospermia. The most common microdeletion of Y chromosome was AZFc (62.39%) and these patients were characterized with azoospermia and oligozoospermia. Five patients (4. 59%) who suffered Y chromosome microdeletion in AZFa region and AZFb region were characterized with azoospermia. Fifteen cases (13.76%) with microdeletion in AZFb region and AZFc region were mainly characterized with azoospermia. There were 6 cases (5. 50 % ) of microdeletion in AZFa, AZFb and AZFc regions,these patients were all characterized with azoospermia. Conclusions Both Chromosome abnormalities and Y chromosome microdeletions are important causes for male infertility.

Objective To investigate the correlation between seminal plasma hepatitis B virus (HBV) DNA copy and semen parameters and sperm DNA fragmentation (SDF).Methods The seminal plasma HBV-DNA was detected by the real-time PCR in 148 infertility males,and those with serum HBV-DNA above (positive) or below (negative) 5.0 × 102U/ml were analyzed respectively by semen parameters,sperm morphology and sperm DNA fragmentation (SDF).Results Of 148 male,60 (40.5％) were seminal plasma HBV-DNA positive,and of 60 positive patients,56 (93.3％) were serum hepatitis B e antigen(HBeAg) positive,which was higher than those of seminal plasma HBV-DNA negative males (31cases,35.2％).Serum HBeAg and HBV-DNA in seminal plasma HBV-DNA positive patients were 845.7(0.2 ～ 1455.0) S/CO and (1.7 ± 1.1) × 108U/ml,which were higher than those of HBV-DNA negative patients [HBeAg:0.1 (0.1 ～ 1374.0) S/CO;HBV-DNA:(2.3 ± 1.1) × 107 U/ml,P ＜ 0.01].Seminal plasma HBV-DNA positive patients exhibited lower semen volume,sperm concentration,the percentage of forward moving sperm and less normal morphology compared to HBV-DNA negative patients [(2.44±1.2)mlvs.(3.07±1.3)ml,(66.8±49.1) ×106/mlvs.(87.1 ±65.4) ×106/ml,(54.3± 16.1)％ vs.(59.1 ±15.3)％,(3.77 ±2.8)％ vs.(6.15 ±4.2)％,P＜0.05].The number of patients with teratozoospermia was significantly higher in seminal plasma HBV-DNA positive patients (56.7％ versus 34.1％,(P ＜ 0.01).The SDF in seminal plasma HBV-DNA positive patients was(18.1 ± 12.3)％,while it was(14.4 ± 8.4)％ in negative patients,and the difference of SDF in these two groups was significantly (t =2.197,P ＜ 0.05).Conclusion Seminal plasma HBV-DNA positive could affect the semen parameters,sperm morphology and SDF.

Objective To investigate the impact of hepatitis B virus (HBV) infection on semen parameters,sperm DNA integrity,acrosin activity and sperm-nucleoprotein transition.Methods Semen samples from 527 subjects including 273 hepatitis B surface antigen (HBsAg) positive and 254 HBsAg negative,who sought medical attention and received in-vitro feritilization in reproductive medicine center of First Hospital of Wenzhou Medical University from Jan 2011 to Oct 2012 were collected.Semen parameters,sperm DNA fragmentation index (DFI),sperm-nucleoprotein transition and acrosin activity of both HBsAg-positive and HBsAg-negative subjects were analyzed.Results Semen parameters of both groups were within the normal range,but sperm concentration and percentage of forward moving sperms of HBsAg positive group were significantly lower than those of HBsAg negative group (P=0.000),while percentage of static sperms of HBsAg positive group were significantly higher than that of HBsAg negative group (P =0.000).DFI in HBsAg positive and negative group were (17.85 ± 0.70) ％ and (11.85 ± 0.50) ％,respectively,which was significantly different (t=6.951,P=0.000).Percentage of sperms with normal morphology in both groups were within the normal range,but sperms with neck and tail deformity in the HBsAg positive group was significantly higer than those in HBsAg negative group (all P＜0.05).Acrosin activity of sperms in HBsAg positive group was significantly lower than that in HBsAg negative group (t=3.756,P=0.000).Linear regression analysis indicated that serum HBsAg level was reversely correlated with sperm concentration (r=-0.140,P =0.021),but positively correlated to DFI (r =0.151,P =0.014).Conclusions HBV infection not only affects the routine semen parameters and sperm morphology,but also compromises sperm function including impaired DFI and acrosin activity.However,the impact of anti-HBV agents on sperm quality and male fertility requires further research.

Objective To assess sperm chromatin structure assay (SCSA) and sperm chromatin dispersion test (SCD) for DNA fragmentation evaluation in human infertility, and the correlation between these two methods. Methods We used SCSA and SCD assays to detect DNA fragmentation in sperm from 134 infertile men. The correlation of SCSA and SCD assays was analyzed. The sperm DNA fragmentation index (DFI) was divided into 3 groups (≤15%DFI, >15~≤30%DFI and>30%DFI), and the difference between SCSA and SCD assays was assessed. Results The SCSA assay was strongly correlated with the SCD assay for sperm DNA fragmentation (r=0.915, P<0.001). There was no significant difference between>15~ ≤30%DFI and>30%DFI groups. However, SCD showed higher levels of DNA fragmentation than that measured by SCSA for≤15%DFI group (13.50 4.82 vs 9.79 2.60, P<0.001). Conclusion There is a strong positive correlation between SCSA and SCD assays in detection of DNA fragmentation. SCD assay showed higher levels of DNA fragmentation than that measured by SCSA for≤15%DFI group.

The BCCIP (BRCA2- and CDKN1A-interacting protein) is an important cofactor for BRCA2 in tumor suppression. Although the low expression of BCCIP is observed in multiple clinically diagnosed primary tumor tissues such as ovarian cancer, renal cell carcinoma and colorectal carcinoma, the mechanism of how BCCIP is regulated in cells is still unclear. The human INO80/YY1 chromatin remodeling complex composed of 15 subunits catalyzes ATP-dependent sliding of nucleosomes along DNA. Here, we first report that BCCIP is a novel target gene of the INO80/YY1 complex by presenting a series of experimental evidence. Gene expression studies combined with siRNA knockdown data locked candidate genes including BCCIP of the INO80/YY1 complex. Silencing or over-expressing the subunits of the INO80/YY1 complex regulates the expression level of BCCIP both in mRNA and proteins in cells. Also, the functions of INO80/YY1 complex in regulating the transactivation of BCCIP were confirmed by luciferase reporter assays. Chromatin immunoprecipitation (ChIP) experiments clarify the enrichment of INO80 and YY1 at +0.17 kb downstream of the BCCIP transcriptional start site. However, this enrichment is significantly inhibited by either knocking down INO80 or YY1, suggesting the existence of both INO80 and YY1 is required for recruiting the INO80/YY1 complex to BCCIP promoter region. Our findings strongly indicate that BCCIP is a potential target gene of the INO80/YY1 complex.
Calcium-Binding Proteins ; genetics ; metabolism ; Cell Cycle Proteins ; genetics ; metabolism ; Chromatin Assembly and Disassembly ; physiology ; DNA Helicases ; genetics ; metabolism ; HeLa Cells ; Humans ; Multiprotein Complexes ; genetics ; metabolism ; Nuclear Proteins ; genetics ; metabolism ; Promoter Regions, Genetic ; physiology ; Transcription, Genetic ; physiology ; YY1 Transcription Factor ; genetics ; metabolism

Objective To investigate DNA methylation markers in the whole blood of patients with systemic lupus erythematosus(SLE),in hope to facilitate the evaluation of SLE severity.Methods Whole blood samples were obtained from 58 patients with SLE(including 14 cases of severe SLE,25 moderate SLE,19 inactive SLE)and 50 healthy controls.Bisulphite sequencing was performed to determine the methylation status of interleukin-2 common receptor gamma chain(IL-2RG)promoter region,and real-time reverse transcriptionPCR to quantify the expression level of IL-2RG mRNA,in these subjects.Results The methylation level of IL2RG promoter region was 0.217 ± 0.140,0.325 ± 0.230,0.342 ± 0.085 and 0.175 ± 0.036 in the patients withsevere,moderate and inactive SLE and healthy controls,respectively.A significant increase was observed in the methylation level of IL-2RG promoter region in the patients with inactive SLE compared with the patients with severe SLE and healthy controls(both P ＜ 0.01),and in the patients with SLE compared with the healthy controls(0.263 ± 0.047 vs.0.175 ± 0.036,P ＜ 0.05).The expression level of IL-2RG mRNA was significantly lower in the patients with SLE than in the healthy controls(2.550 ± 0.823 vs.4.293 ± 1.283,P ＜ 0.05).A negative correlation was observed between the expression level of IL-2RG mRNA and methylation level of IL2RG promoter region in 20 patients with SLE(r =-0.44,P ＜ 0.05).Conclusion The methylation status of IL2RG promoter region is statistically higher in patients with SLE than in healthy controls,and significantly different between patients with active SLE and those with stable SLE.

OBJECTIVETo investigate variation of sperm DNA fragmentation index (DFI) in male partners of infertile couples.

METHODSA total of 539 males between April 2009 and April 2012 were analyzed. At least one repeated routine semen analysis and sperm DNA fragmentation test were performed for each sample by sperm chromatin dispersion (SCD) analysis following World Health Organization guidelines. Coefficient of variation (CV) for DFI was calculated.

RESULTSRespectively, 1, 2, 3 and 4 repeated SCD analyses were carried out on 473, 59, 6 and 1 semen samples. The median interval between the first and repeated SCD measurements was 3.0 (1.0-11.0) months. For the first tested samples, the between-sample coefficient of variation (CVB) for DFI was 71.2%. A significant difference has been found between DFI of the first measurement and DFI of repeated measurement in 0.5 to 3 months, 3 to 12 months and 12 to 34 months (P< 0.01). Compared with the first test, 26.3% of males were on both sides of the cut-off point of 18%. The median within-subject coefficient of variation (CVw) for DFI of 539 men was 26.0% (12.6%-42.8%). And the median CVw DFI was significantly lower compared with CVw of sperm count, concentration, progressive motility and normal morphology (P< 0.01). Significant correlations were found between the CVw DFI and sperm count, concentration and interval among the samples (P< 0.05).

CONCLUSIONDFI of male partners for infertile couples is a parameter with substantial variation, repeated SCD measurements are therefore recommended.

Adult ; DNA Damage ; DNA Fragmentation ; Female ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Sperm Count ; Spermatozoa ; metabolism ; Young Adult