OBJECTIVETo investigate aquaporin 9 (AQP9) mRNA and protein expression in antrum follicle and luteinizing granulosa cells of polycystic ovarian syndrome (PCOS) ovary, and its relation to follicular fluid steroids hormone levels during IVF cycles.

METHODSAQP9 mRNA expression on luteinizing granulosa cells in IVF cycles was detected by RT-PCR. AQP9 protein expression in antrum follicles of PCOS ovary and luteinizing granulosa cells was measured by immunohistochemistry. The concentrations of estradiol (E2), progesterone (P) and testerone (T) in follicular fluid were measured by radioimmunoassay (RIA).

RESULTThe expression of AQP9 mRNA in luteinizing granulosa cells during IVF cycles was positive by RT-PCR. No significant differences in AQP9 mRNA levels in granulosa cells between PCOS and control group were found during IVF cycles. The expression level of AQP9 mRNA in large follicles was higher than that in small follicles, but not significantly. The immunoreactivity for AQP9 was localized in membrane and cytoplast of granulosa cells in antrum follicles from PCOS ovary and luteinizing granulosa cells during IVF cycles. Multiple regression analysis showed that AQP9 mRNA levels on granulosa cells were not correlated with E2, P and T levels in follicular fluid during IVF cycles.

CONCLUSIONAQP9 may play an important role in the follicle development and antrum formation through water transport and AQP9 may be involved in the mechanism of follicle development in PCOS.

Adult ; Aquaporins ; biosynthesis ; genetics ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Granulosa Cells ; metabolism ; Humans ; Immunohistochemistry ; Infertility, Female ; etiology ; genetics ; metabolism ; Polycystic Ovary Syndrome ; complications ; genetics ; metabolism ; RNA, Messenger ; biosynthesis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo evaluate the levels of bone morphogenetic protein-15 (BMP-15) in human follicular fluid (FF) and its association with response to ovarian stimulation.

METHODSWestern blotting was performed to determine the levels of BMP-15 in FF obtained from follicle aspirates in 70 patients undergoing IVF treatment. According to the response to ovarian stimulation the patients were divided into poor responder group and normal responder group.

RESULTBMP-15 levels in FF of poor responders were significantly higher than those in normal responders (1.01 +/- 0.34 vs 0.77 +/- 0.24, P<0.01).

CONCLUSIONIncreased levels of BMP-15 in FF may be associated with poor response to ovarian stimulation.

Adult ; Blotting, Western ; Bone Morphogenetic Protein 15 ; Female ; Follicle Stimulating Hormone ; administration & dosage ; Follicular Fluid ; drug effects ; metabolism ; Gonadotropin-Releasing Hormone ; administration & dosage ; analogs & derivatives ; Growth Differentiation Factor 9 ; Humans ; Infertility, Female ; metabolism ; Intercellular Signaling Peptides and Proteins ; biosynthesis ; Ovary ; drug effects ; metabolism ; Ovulation Induction

OBJECTIVETo investigate the expression of cathepsin D in ovary of patients with polycystic ovarian syndrome (PCOS).

METHODSWestern blot was performed to detect the expression of cathepsin D and immunohistochemistry was used to detect the protein distribution in ovarian tissue.

RESULTSemi-quantity values of cathepsin D expression in PCOS and control group were 2.06 +/- 0.39 and 4.76 +/- 1.43 (P<0.05), respectively. Immunostaining for cathepsin D was obvious in both follicles and stromal cells, and the strongest immunostaining was seen in granulosa cells of follicles. Immunochemical study showed the protein was mainly located on the cytoplasm and cell membrane.

CONCLUSIONCathepsin D expression is down-regulated in ovaries of PCOS patients, which may provide a clue for the abnormality of follicle development in PCOS.

Adult ; Blotting, Western ; Cathepsin D ; biosynthesis ; Down-Regulation ; Female ; Humans ; Immunohistochemistry ; Ovary ; enzymology ; pathology ; Polycystic Ovary Syndrome ; enzymology

OBJECTIVETo evaluate the outcome of in vitro fertilization embryo transfer (IVF-ET) in treatment of polycystic ovarian syndrome (PCOS) with infertility.

METHODSA retrospective analysis was performed from 52 patients with PCOS (PCOS group) and 408 cases with tubal infertility (control group). Both groups underwent IVF-ET treatment from 2001 to 2004. The duration of stimulation, amps of gammaFSH, the level of serum E2, P on the day of HCG injection, the number of oocytes retrieved, the rates of fertilization, cleavage, implantation and pregnancy, the incidence of ovarian hyperstimulation syndrome (OHSS) and cancelled rate of ET were compared between the two groups.

RESULTThe duration of stimulation and amps of gammaFSH were not significantly different between the two groups. The concentration of serum E2, P on the day of HCG injection, the numbers of oocytes retrieved and cleavaged embryos were significantly higher in PCOS group (P <0.01, <0.05). Fertilization rate was significantly lower in PCOS group (P <0.01). The implantation, pregnancy and miscarriage rates per ET were not statistically significant. The OHSS rates and cancelled rates of ET were higher in PCOS group (P <0.01).

CONCLUSIONWomen with PCOS have a lower fertilization rate compared with those with tubal-factor fertility during IVF-ET. However, more oocytes are recovered and the preimplanted embryo has a normal chance of implantation leading to similar pregnancy rates. The OHSS rates and cancelled rates of ET are higher in PCOS because of a greater number of oocytes developed and a higher level of E2.

Adult ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Infertility, Female ; etiology ; Ovarian Hyperstimulation Syndrome ; Polycystic Ovary Syndrome ; complications ; Retrospective Studies ; Treatment Outcome

In order to investigate the patterns of FIX gene mutation in 3 unrelated hemophilia B (HB) patients, the activated partial thromboplastin time (APTT) and FIX activity (FIX: C) tests were adopted for phenotype diagnosis. All of the eight exons and their flank of FIX gene were amplified by polymerase chain reaction (PCR), the nucleic acid sequences were detected by dideoxymediated chain-termination method. The results indicated that as compared with normal control, the APTT value significantly increased, FIX: C value obviously decreased, PT value was normal. Sequencing results showed that all of 3 HB patients had the changes of gene sequences, among 3 patients the G22119A point mutation of exon 6 existed in case No.1, the G7932C point mutation of exon 2 was detected in case No.2 and the T32685C point mutation of exon 8 was found in case No.3. In conclusion, the relevant changes of gene sequences in all of 3 HB patients were detected, which provides some evidences for molecular mechanism of gene deficiency in HB patients.
Base Sequence ; DNA Mutational Analysis ; methods ; Factor IX ; genetics ; Hemophilia B ; genetics ; Humans ; Molecular Sequence Data ; Point Mutation

OBJECTIVEGenetic factors are thought to be involved in the development of vitiligo. The aim of this study is to explore the possible genetic model of vitiligo by analyzing the genetic characteristics of 815 patients from Zhejiang province.

METHODSData for 815 patients with vitiligo together with their first- and second-degree relatives were obtained using a standardized questionnaire. All these information was requested to confirm the answers about family history in order to reduce the possibility of 'recall' bias. The 815 probands would include 411 (50.43%) males and 404 (49.57%) females with a varied age from 2 months to 71 years old. Since the information on general prevalence of vitiligo in this area was absent, a control group was set up to facilitate the calculations of heritability degree. 468 persons of the control group were from non-vitiligo population with a sex ratio of 241(male): 227(female) with varied age of 4 months to 80 years old. Both gender and age were comparable between the vitiligo and the control population. The inheritance pattern estimation, heritability calculation and complex segregation analysis were performed with Penrose method, Falconer regression method and SAGE-REGTL program.

RESULTSIn 815 vitiligo probands, 128 had and 687 had not family histories, with a heritability rate of 15.7%. The vitiligo prevalence in proband's first degree relatives was 2.580%, higher than the prevalence of 0.618% in second degree relatives, and both of them were higher than general prevalence: 0.192%. By Penrose method, the rates on different catagories were as follows: sibling prevalence rates s = 0.080 18; population prevalence rate q = 0.001 92; s/q = 41.76. The ratio of s/q did not approach 1/2q (260.42) or 1/4q (130.21), but approached 1/square root of q(22.82), suggesting vitiligo was consistent with a mode of polygenic inheritance. Using Falconer's method, heritabilities of vitiligo in first-and second degree relatives of probands were 59.61% (95% confidence interval 65.37-53.84) and 55.20% (95% confidence interval 43.88-66.52), respectively. The weighted average of heritability in all relatives was 58.7% (95% confidence interval 53.56-63.83). The results of complex segregation analysis suggested that major gene model including the Mendelian dominant, recessive and additive hypotheses were not rejected (P > 0.05). Purely environmental model and no transmission model were rejected at a 0. 001 significance level. According to AIC, Mendelian dominant inheritance was the best-fitted hypothesis.

CONCLUSIONGenetic factors played an important role in the occurrence of vitiligo, and the genetic model of vitiligo could serve as the polygenetic or multifactorial inheritance with major gene trait.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Models, Genetic ; Vitiligo ; epidemiology ; genetics ; Young Adult

OBJECTIVETo explore factor IX gene mutations and molecular mechanism of haemophilia B in 3 unrelated families.

METHODSThe activated partial thromboplastin time (APTT) and FIX activity (FIX: C) assay were used for phenotypic diagnosis. The STR loci gene polymorphisms for genetic linkage analysis in the patients and their family members were assayed. All of the 8 exons and the exon-intron boundaries of FIX gene were amplified by polymerase chain reaction (PCR) and direct sequencing.

RESULTS AND CONCLUSIONMutations were found in the FIX gene of the propositi. Proband 1 had a G22119A mutation in exon 6, proband 2 a G7392C mutation in exon 2 and proband 3 a T32685C mutation in exon 8.

DNA Mutational Analysis ; Factor IX ; genetics ; Genetic Linkage ; Hemophilia B ; genetics ; Humans ; Mutation ; Pedigree ; Polymorphism, Genetic

OBJECTIVETo detect the frequencies of anti-GPIIb/IIIa antibody secreting B cells and platelet-specific antibody in patients with idiopathic thrombocytopenic purpura (ITP) and non-immune thrombocytopenia, and to evaluate their roles in the diagnosis of ITP and their clinical significance.

METHODSThe frequencies of circulating B cells secreting anti-GPIIb/IIIa antibody and platelet-specific antibody in 58 ITP patients, 33 non-ITP patients and 31 healthy controls were tested by Enzyme-linked Immunospot Assay (ELISPOT) and modified monoclonal antibody immobilization of platelet antigens assay (MAIPA) respectively.

RESULTSThe frequencies of circulating B cells secreting anti-GPIIb/IIIa antibody in ITP patients \[(6.6 ± 4.2)/10(5) PBMNC\] were significantly increased (P < 0.05) than that of the controls \[(1.3 ± 0.5)/10(5) PBMNC\] and non-immune thrombocytopenic purpura patients \[(2.2 ± 2.0)/10(5) PBMNC\]. However there was no apparent difference between the latter two groups (P > 0.05). ELISPOT had a sensitivity of 70.69%, a specificity of 90.91% for the diagnosis of ITP, the sensitivity being higher than that of modified MAIPA's (43.10%) (χ(2) = 7.03, P < 0.05). The ROC curve showed the discriminative validity of cytometric bead array was 0.886.

CONCLUSIONThe frequencies of circulating B cells secreting anti-GPIIb/IIIa antibody may reflect the pathogenesis of ITP. ELISPOT assay have high sensitivity and specificity than modified MAIPA for the diagnosis of ITP and the guidance for clinical therapy.

Autoantibodies ; immunology ; B-Lymphocytes ; Blood Platelets ; Humans ; Platelet Glycoprotein GPIIb-IIIa Complex ; immunology ; Purpura, Thrombocytopenic, Idiopathic ; immunology

OBJECTIVESTo measure serum and follicular resistin, steroids hormone levels in women with PCOS (polycystic ovary syndrome) (BMI (body mass index)<25 kg/m(2)), to assess possible correlations of resistin to hormonal and metabolic parameters and to analyze the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) in women with PCOS and tubal infertility.

STUDY DESIGNWe analyzed the clinical outcomes of IVF-ET in women with PCOS (BMI<25 kg/m(2)) and tubal infertility during the years 2002 to 2004 and compared the serum and follicular fluid resistin levels, estradiol (E(2)), progesterone (P), testosterone (T) levels in 20 PCOS and 20 healthy, age-matched women without PCOS during IVF-stimulated cycles. The correlations between the resistin levels and the outcomes of IVF-ET were evaluated.

RESULTSNo significant differences in resistin levels of either serum or follicular fluid between PCOS and control group were found. However, resistin levels in serum were higher than that in follicular fluid in both groups. Multiple regression analysis showed that resistin levels in serum did not correlate with BMI, estradiol, LH (luteinizing hormone) and insulin level in fasting blood. No significant correlations were found between follicular fluid reisistin levels and fertilization rate, implantation rate, clinical pregnancy rate or early miscarriage rate in both PCOS and control groups.

CONCLUSIONOur results show that resistin does not have correlation with the hormonal and metabolic parameters as well as the outcomes of IVF. These data suggest that resistin is unlikely to be a local determinant factor in steroidogenesis and growth and maturation of oocytes during IVF-ET in lean women with PCOS.

Adult ; Case-Control Studies ; Embryo Transfer ; Female ; Fertilization in Vitro ; Follicular Fluid ; metabolism ; Gonadal Steroid Hormones ; blood ; metabolism ; Humans ; Infertility, Female ; blood ; etiology ; metabolism ; therapy ; Polycystic Ovary Syndrome ; blood ; complications ; metabolism ; therapy ; Pregnancy ; Pregnancy Outcome

BACKGROUNDHelicobacter pylori (H. pylori) frequently colonizes the stomach. Gastroesophageal reflux disease (GERD) is a common and costly disease. But the relationship of H. pylori and GERD is still unclear. This study aimed to explore the effect of H. pylori and its eradication on reflux esophagitis therapy.

METHODSPatients diagnosed with reflux esophagitis by endoscopy were enrolled; based on rapid urease test and Warth-Starry stain, they were divided into H. pylori positive and negative groups. H. pylori positive patients were randomly given H. pylori eradication treatment for 10 days, then esomeprazole 20 mg bid for 46 days. The other patients received esomeprazole 20 mg bid therapy for 8 weeks. After treatment, three patient groups were obtained: H. pylori positive eradicated, H. pylori positive uneradicated, and H. pylori negative. Before and after therapy, reflux symptoms were scored and compared. Healing rates were compared among groups. The χ2 test and t-test were used, respectively, for enumeration and measurement data.

RESULTSThere were 176 H. pylori positive (with 92 eradication cases) and 180 negative cases. Healing rates in the H. pylori positive eradicated and H. pylori positive uneradicated groups reached 80.4% and 79.8% (P = 0.911), with reflux symptom scores of 0.22 and 0.14 (P = 0.588). Healing rates of esophagitis in the H. pylori positive uneradicated and H. pylori negative groups were, respectively, 79.8% and 82.2% (P = 0.848); reflux symptom scores were 0.14 and 0.21 (P = 0.546).

CONCLUSIONSBased on esomeprazole therapy, H. pylori infection and eradication have no significant effect on reflux esophagitis therapy.

Adolescent ; Adult ; Aged ; Amoxicillin ; therapeutic use ; Esomeprazole ; therapeutic use ; Esophagitis, Peptic ; drug therapy ; etiology ; microbiology ; Female ; Gastroesophageal Reflux ; drug therapy ; etiology ; microbiology ; Helicobacter Infections ; complications ; drug therapy ; Helicobacter pylori ; drug effects ; pathogenicity ; Humans ; Male ; Middle Aged ; Tinidazole ; therapeutic use ; Young Adult