The present study was aimed at investigating the expression of calbindin-D28k (CaBP-D28k) in human fallopian tube, which were collected from 33 childbearing age women undergoing abdominal hysterectomy with adnexectomy for benign disease in the pelvic cavity. These women had normal menstrual cycle and history of normal pregnancy. Isthmus, ampullary and umbrella segments of fallopian tubes were respectively collected. These specimens were divided into 6 groups based on their menstrual cycles: early-proliferative stage (n=6), mid-proliferative stage (n=5), late-proliferative stage (n=5), early-secretory stage (n=7), mid-secretory stage (n=5) and late-secretory stage (n=5). The expressions of CaBP-D28k protein and mRNA in fallopian tubes were determined by immunohistochemistry and reverse transcription polymerase chain reaction (RT-PCR) methods. Positive expressions of CaBP-D28k protein and mRNA were observed in human fallopian tubes. There was no significant difference in the expression of CaBP-D28k protein among the isthmus, ampulla and umbrella segments in the same phase of menstrual cycle (P>0.05). However, in the menstrual cycle, the expression level of CaBP-D28k protein in the epithelium was the lowest during the early- and mid-proliferative stages and increased in both the late-proliferative and early-secretory stages (P<0.05), and then decreased in the mid- and late-secretory stages (P<0.05). The expressed CaBP-D28k protein was disposed to gobbets or dispersed sheets in cytoplasm in the early- and mid- proliferative stages, and showed concentrated granules on the top of cells in the late-proliferative and early-, mid-secretory stages. Then in the late-secretory stage redistribution renewed as in the early- and mid-proliferative stages. The CaBP-D28k mRNA obviously increased in the late-proliferative and early-secretory stages (P<0.05). These findings indicate that the expressions of CaBP-D28k protein and mRNA exist in human fallopian tubes and exhibit a cyclic change.
Calbindin 1 ; metabolism ; Fallopian Tubes ; metabolism ; Female ; Gene Expression ; Humans ; Menstrual Cycle

OBJECTIVETo investigate the effect of the secretory proteins of the ventral prostate on the glycoproteins in the oviductal fluid of golden hamsters.

METHODSMale golden hamsters were divided into four groups: sham operation (SH), total removal of accessory sex glands (TX), and retainment of the ventral prostate only (VP). Oviductal fluid was collected from female hamsters at 0.5, 2, 4 and 6 h after mating with the males of different operated groups with or without ventral prostate. Glycoproteins were probed with a panel of lectins and their changes in the oviductal fluid were analyzed by Western blot.

RESULTSThe 47 000, 52 000, 81 000 and 128 000 WGA-binding proteins were observed in the oviductal fluid of the 6 h TX group, the 32 000, 35 500, 47 000 and 52 000 WGA-binding glycoproteins noted in the 6 h VP group, the 47 000, 68 000, 95 000 and 128 000 pisum sativum agglutinin (PSA)-binding glycoproteins shown in the 6 h TX and VP groups, two extra 32 000 and 37 500 bands detected in the 6 h VP group, the 47 000 and 52 000 dolichos biflorus agglutinin (DBA)-binding glycoproteins present in the 6 h VP but absent in the 6 h TX group.

CONCLUSIONVentral prostate secretory proteins affect acetylglucosamine, N-acetylgalactosamine/galactose and mannose in the oviductal fluid collected 6 hours after mating. And these glycoproteins may play an important role in the development of embryos.

Animals ; Copulation ; physiology ; Cricetinae ; Fallopian Tubes ; metabolism ; Female ; Glycoproteins ; metabolism ; Male ; Mesocricetus ; Prostatic Secretory Proteins ; physiology

Carcinoma in Situ ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Fallopian Tube Neoplasms ; metabolism ; pathology ; Fallopian Tubes ; metabolism ; pathology ; Female ; Humans ; Neoplasm Invasiveness ; Ovary ; pathology ; Tumor Suppressor Protein p53 ; metabolism

AIMTo establish a method for determineation of the concentration of ofloxacin in human fallopian tube, uterus and serum.

METHODSThe separation was performed on a Spherisob C18 column (Hypersil, 250 mm x 4.6 mm ID, 5 microns) with a mobile phase of acetonitrile-0.01 moL.L-1 potassium dihydrogen phosphate-0.5 mol.L-1 tetrabutylammonium bromide (9:91:4, pH 2.5). The flow rate was 1.0 mL.min-1 and detection was at 294 nm. The samples were homogenated or ground to powder after freezing with liquid nitrogen. 1% triton-100 and certain volume of ethylacetate-isopropanol (10:1) were added, shaken and centrifuged. Then the entire organic layer was transferred to a tube and vacuum dried. The residue was reconstituted in the mobile phase for HPLC.

RESULTSThere was a linear relationship between the peak area ratio and the ofloxacin concentration over the range of 0.2-8.0 micrograms.mL-1. The limits of detection was 40 ng.mL-1. Using this method to determine the ofloxacin concentrations in relevant organs as well as in the plasma of patients of the Department of Gynecology, and achieved satisfactary results.

CONCLUSIONThe method can be applied to assay the ofloxacin concentration in human tissues. Ofloxacin was well distributed in woman fallopian tube, uterus and serum after single oral administration.

Adolescent ; Adult ; Anti-Infective Agents ; analysis ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Fallopian Tubes ; metabolism ; Female ; Humans ; Ofloxacin ; analysis ; blood ; pharmacokinetics ; Tissue Distribution ; Uterus ; metabolism

To examine whether or not the regulatory sequence of chicken ovalbumin gene can drive transgene expression specifically in hen oviduct, the authors constructed an oviduct-specific expression vector (pOV), containing 3.0 kilobases (kb) of the 5'-flanking sequence and 3.0 kb of the 3'-flanking sequence of the chicken ovalbumin gene. Jellyfish green fluorescence protein (EGFP) reporter gene and bacterial LacZ reporter gene were respectively inserted into the downstream of the 5'-regulatory region. The recombinants were named as pOVEGFP and pOVLacZ. Two transfer systems, in vitro and in vivo, were used to verify the function of the vector. In vitro, the plasmid DNA pOVEGFP and pEGFP-N1 were transfected respectively by the polyethyleneimine procedure into the primary chicken oviduct epithelium (PCOE) and fibroblasts cells isolated from laying hens. In vivo, the recombinant vector pOVLacZ was injected into egg-laying hens via wing vein and the tissues were collected for RT-PCR analysis. The results showed that expression of pEGFP-N1 was achieved at low level in oviduct epithelial cells and at high level in fibroblasts, but that the recombinant vector was not expressed in both cells. RT-PCR analysis showed that the LacZ gene was transcribed in the oviduct, but not in the heart, liver, kidney and spleen of the injected hens. Accordingly, the beta-galactosidase activity was only detected in the oviduct magnum (116.7 mU/ml) and eggs (16.47 mU/ml). These results indicated that the cloned regulation regions of chicken ovalbumin gene could drive exogenous gene expression specifically in the oviducts of hens. In vivo gene injection via wing vein may serve as a rapid production system of recombinant proteins in chicken eggs. In addition, the cultured primary oviduct cells from laying hens were not efficient temporary expression systems for analyzing the function of regulating elements of ovalbumin gene.
Animals ; Cells, Cultured ; Chickens ; Cloning, Molecular ; methods ; Fallopian Tubes ; metabolism ; Female ; Organ Specificity ; Ovalbumin ; biosynthesis ; genetics ; Protein Engineering ; methods ; Recombinant Proteins ; biosynthesis ; Transfection ; methods ; Women

OBJECTIVE: To investigate the expression of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and its role in regulating invasion and migration of trophoblasts. METHODS: Immunohistochemistry and Western blotting were used to detect the localization and expression level of Talin1 in the fallopian tube and chorionic villi in patients with tubal pregnancy and in women with normal pregnancy. In the cell experiment, HTR-8/SVneo cells was transfected with Talin1 siRNA and the changes in cell invasion and migration were assessed using scratch assay and Transwell assay. The expressions of MMP-2, MMP-9, N-cadherin and Snail in the transfected cells were detected by qRT-PCR and Western blotting. RESULTS: Positive expression of Talin1 was detected in both normal fallopian tube tissues and tissues from women tubal pregnancy, and its expression was localized mainly in the cytoplasm of cilia cells. The expression level of Talin1 was significantly higher in both the fallopian tube and chorionic villi in women with tubal pregnancy than in normal fallopian tube and chorionic villi samples (P < 0.01). In HTR-8/SVneo cells, transfection with Talin1 siRNA significantly inhibited cell invasion (P < 0.01) and migration (P < 0.05), down-regulated the expression of N-cadherin, MMP-2 and Snail (P < 0.05), and up-regulated the expression of MMP-9 in the cells (P < 0.05). CONCLUSION The expression of Talin1 in the fallopian tube and chorionic villi is significantly increased in women with tubal pregnancy, suggesting the association of Talin1-regulated trophoblast cell invasion with the occurrence of tubal pregnancy.
Cadherins/metabolism* ; Cell Movement ; Chorionic Villi/metabolism* ; Fallopian Tubes/metabolism* ; Female ; Humans ; Matrix Metalloproteinase 2/metabolism* ; Matrix Metalloproteinase 9/metabolism* ; Pregnancy ; Pregnancy, Tubal/metabolism* ; RNA, Small Interfering/metabolism* ; Talin/metabolism* ; Trophoblasts/metabolism*

OBJECTIVETo investigate the binding of secretory proteins in the ventral prostate to the surface of sperm.

METHODSWe used different techniques to demonstrate the possibility of ventral prostate secretory proteins binding to sperm in golden hamsters. Polyclonal antibodies against crude secretion of the ventral prostate cultured in rabbits were used to detect the antigens in hamster epididymal, uterine and oviductal spermatozoa by indirect immunofluorescence technique. The uterine and oviductal spermatozoa were collected after mating with the males with or without ventral prostate glands. The ventral prostate secretory proteins were isolated and transblotted to the membrane, which was incubated with the biotinylated epididymal sperm membrane proteins, and then the biotinylated binding proteins were stained.

RESULTSAn immunoreaction restricted to the middle piece was observed in the sperm incubated with the ventral prostate secretion and ejaculated sperm recovered from the uteri and oviducts. The rate of the epididymal sperm bound with the ventral prostate secretory proteins was (80 +/- 5) %, and the rats of the sperm binding to the ventral prostate secretory proteins were (30.0 +/- 4.6) % from the uterus and (16.0 +/- 3.6) % from the oviduct after mating with the males with ventral prostate glands, significantly higher than after mating with those without prostate glands (P < 0.01). Five bands were identified by Western blot analysis in vitro of the ventral prostate secretory proteins incubated with biotinylated epididymal sperm membrane proteins.

CONCLUSIONThe present data indicate that ventral prostate secretory proteins bind to the middle piece of sperm in golden hamsters.

Animals ; Blotting, Western ; Cricetinae ; Epididymis ; metabolism ; Fallopian Tubes ; metabolism ; Female ; Fluorescent Antibody Technique, Indirect ; Male ; Mesocricetus ; Prostate ; metabolism ; Protein Binding ; Seminal Vesicle Secretory Proteins ; metabolism ; Spermatozoa ; metabolism ; Uterus ; metabolism

OBJECTIVETo study the serous lesions of the fimbria of the fallopian tube in patients with pelvic serous adenocarcinoma and investigate its significance in the serous carcinogenesis.

METHODSTo observe the morphological features of the fimbria of the fallopian tube in 43 cases of pelvic serous adenocarcinoma (31 cases of ovarian carcinoma and 12 cases of peritoneal carcinoma). Immunohistochemical examination of p53 expression was performed on samples of 69 fallopian tubes of 40 cases.

RESULTSFimbria carcinoma was identified in 44 tubes in 31 of 43 cases. Fourteen of the carcinoma foci were ≤ 5 mm. In 68.3% of the fimbria carcinomas demonstrated involvement of the mucosa. Twenty eight tubes of 20 cases exhibited intraepithelial carcinoma. Twenty three of 44 tubes of the fibria carcinomas showed fimbria adherence and unclear appearance. The early histological changes of the fimbria epithelium included proliferation of local secretory cells, homogeneity, and straightening of the mucous folds. Clusters of tumor epithelial cells or single gland with atypical features floated between mucosal folds were found in 71.4% of the fimbria with intraepithelial carcinoma. The positive expression rate of p53 in the fimbria carcinomas and the fimbria intraepithelial carcinomas were 86.4% and 60.7%, respectively.

CONCLUSIONSFimbria carcinomas is an important component in pelvic serous adenocarcinomas. The fimbria intraepithelial carcinoma is also very common among the cases of pelvic serous adenocarcinoma. The fimbria may be an important primary site of pelvic serous adenocarcinomas.

Carcinoma in Situ ; metabolism ; pathology ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Fallopian Tube Neoplasms ; metabolism ; pathology ; Fallopian Tubes ; metabolism ; pathology ; Female ; Humans ; Neoplasm Invasiveness ; Neoplasms, Second Primary ; metabolism ; pathology ; Ovarian Neoplasms ; metabolism ; pathology ; Pelvic Neoplasms ; metabolism ; pathology ; Peritoneal Neoplasms ; metabolism ; pathology ; Tumor Suppressor Protein p53 ; metabolism

OBJECTIVESTo study the morphologic changes of fallopian tubal epithelium in patients with ovarian serous epithelial tumors and to explore the relationship between the tubal epithelial changes and tumorigenesis of serous ovarian carcinoma.

METHODSThe fallopian tubes in 79 cases of high-grade serous ovarian carcinoma, 12 cases of low-grade serous ovarian carcinoma, 16 cases of serous borderline ovarian tumor and 11 cases of non-ovarian benign tumors were serially examined under light microscope. Immunohistochemical study with EnVision method was used to detect the expression of p53 and bcl-2 protein in the fallopian tubal epithelium in all cases. The occurrences of secretory cell outgrowth (SCOUT), p53 signature, serous tubal intraepithelial carcinoma (STIC) and serous invasive carcinoma were analyzed.

RESULTSSCOUT in tubal epithelium was observed in 60.8% (48/79) of the high-grade serous carcinoma group, 4/12 of the low-grade serous carcinoma group, 3/16 of the serous borderline tumor group and 2/11 of the non-ovarian benign tumor group (P = 0.001). P53 signature, STIC and serous invasive carcinoma occurred only in the fallopian tubal epithelium of patients with high-grade serous ovarian carcinoma, with the positive rates being 29.1% (23/79), 15.2% (12/79) and 44.3% (35/79), respectively. Of the 23 cases with p53 signature, 17 cases had solitary lesion and 6 cases involved more than two sites. A total of 33 p53 signature positive foci were found, with 22 foci located at fimbria and 11 at ampulla. Bcl-2 expression was demonstrated in 90.9% of those foci (30/33). Of the 12 patients with STIC, 7 cases were solitary and 5 cases involved more than two sites. A total of 18 STIC foci were found, with 16 foci located at fimbria and 2 at ampulla. All of them were positive for bcl-2.

CONCLUSIONSSCOUT is found in fallopian tubal epithelium in patients with serous ovarian epithelial tumors, especially high-grade serious carcinoma. On the other hand, p53 signature, STIC and invasive serous carcinoma of tubal epithelium are observed only in patients with high-grade serous ovarian carcinoma, with a predilection of fimbrial involvement. Correlation exists between SCOUT, p53 signature, STIC and high-grade serous ovarian carcinomas. Bcl-2 and p53 immunostaining is helpful for demonstrating such lesions.

Adult ; Aged ; Cell Transformation, Neoplastic ; Cystadenocarcinoma, Serous ; metabolism ; pathology ; Epithelial Cells ; pathology ; Epithelium ; pathology ; Fallopian Tube Neoplasms ; metabolism ; pathology ; Fallopian Tubes ; pathology ; Female ; Humans ; Immunohistochemistry ; Middle Aged ; Neoplasm Staging ; Ovarian Neoplasms ; metabolism ; pathology ; Precancerous Conditions ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Tumor Suppressor Protein p53 ; metabolism

Objective: To investigate the clinical significance of sperm acrosin activity detection in selecting the method of assisted reproduction for patients with unexplained infertility (UI). METHODS: This retrospective study included 49 UI couples treated by IVFET (49 cycles) after three failures in intrauterine insemination (IUI) and another 95 couples with uterine tube obstruction (UTO) treated by IVF (131 cycles). We analyzed the laboratory data, clinical outcomes and sperm acrosin activity in the two groups of patients. According to the level of sperm acrosin activity of the males, we further divided the UI patients into two subgroups, a < 36 IU/106 sperm group (20 cycles) and a ≥36 IU/106 sperm group (29 cycles), and compared the fertilization rates between the two groups. RESULTS: Compared with UI couples treated by IVFET, the UTO couples treated by IVF had a significantly lower rate of fertilization (67.0% vs 76.4%, P < 0.05) and a higher rate of remedial intracytoplasmic sperm injection (ICSI) (20.4% vs 6.1%, P < 0.05), but showed no statistically significant differences in the rates of MII oocytes, available embryos, highquality embryos, implantation, and clinical pregnancy from the latter group (P >0.05). The sperm acrosin activity was remarkably lower in the UI than in the UTO patients (36.03 vs 61.98 IU/106, P < 0.01), and so was the fertilization rate in the < 36 IU/106 than in the ≥36 IU/106 sperm subgroup (47.7% vs 80.3%, P < 0.01). CONCLUSIONS The low fertilization rate caused by decreased sperm acrosin activity may be the main cause of infertility and the potential factor of UI. When sperm acrosin activity is < 36 IU/106 sperm, IVF plus shortterm fertilization by remedial ICSI should be preferred to IUI.
Acrosin ; analysis ; metabolism ; Embryo Implantation ; Fallopian Tubes ; Female ; Fertilization in Vitro ; methods ; statistics & numerical data ; Humans ; Infertility, Female ; Infertility, Male ; Male ; Pregnancy ; Pregnancy Rate ; Reproduction ; Retrospective Studies ; Sperm Injections, Intracytoplasmic ; statistics & numerical data ; Spermatozoa ; metabolism