Reduced expression of aquaporin 9 in tubal ectopic pregnancy.

Previous studies demonstrate significant roles for passive water channels (aquaporins, AQPs) in maintaining water homeostasis in cell membranes of endometrial cells during decidualisation and embryo implantation. However, there is little information regarding the role of AQPs in the human fallopian tube, specifically their role in human tubal ectopic pregnancy. In this study we took tissue samples from the site of implantation of tubal ectopic pregnancy (group 1, N = 30, mean age 32 years, range 23-42) and the corresponding non-implantation site in women undergoing salpingectomy for tubal pregnancy (group 2). Ampullary fallopian tubes during mid-secretory phase were collected as control group (group 3, N = 17, mean age 37 years, range 30-50). Thin sections were prepared and stained with anti-AQP9, and, for estrogen and progesterone receptors in each group. Immunohistochemical studies showed that AQP9 proteins localize in the cytoplasm of epithelial cells of Fallopian tube. Expression of AQP9 was significantly reduced during tubal pregnancy compared to controls (group 1 vs. group 3, P = 0.036; group 2 vs. group 3, P = 0.029), and, this reduced expression was not related to estrogen receptor or progesterone receptor status (group 2, ER vs. AQP9, Pearson r = 0.173, P = 0.361; PR vs. AQP9, Pearson r = 0.124, P = 0.514, respectively). Similarly, there is no correlation between AQP9 and estrogen receptor or progesterone receptor status in the normal group (group 3, ER vs. AQP9, Pearson r = -0.026, P = 0.923; PR vs. AQP9, Pearson r = -0.292, P = 0.255, respectively). Reduced expression of AQP9 in human fallopian tube may contribute to aspects of pathophysiology of tubal ectopic pregnancy.

Locally elevated leukemia inhibitory factor in the inflamed fallopian tube resembles that found in tubal pregnancy.

OBJECTIVE: To investigate the association between the expressions of leukemia inhibitory factor and the occurrence of tubal pregnancy. DESIGN: Prospective observational study. SETTING: University-based obstetrics and gynecology hospital. PATIENT(S): Thirty women undergoing salpingectomy for tubal pregnancy and 30 nonpregnant patients with benign uterine or appendix disease. INTERVENTION(S): Oviduct tissues with ectopic gestation were separated into implantation sites and nonimplantation sites. Samples of ampullary fallopian tubes during midsecretory phase were collected as control groups. Immunohistochemical and Western blot analysis were performed. MAIN OUTCOME MEASURE(S): The differences of leukemia inhibitory factor expression between the implantation and nonimplantation sites of oviduct tissues and the normal and chronically inflamed fallopian tubes. RESULT(S): The expression of leukemia inhibitory factor in the implantation group is significantly higher than that in the nonimplantation group or in the normal group. A statistically significant difference was also found for leukemia inhibitory factor between the chronic inflammation group and the normal group by Western blot analysis but no difference between the chronic inflammation group and the implantation group or the nonimplantation group. CONCLUSION(S): Leukemia inhibitory factor might be one of the reasons that cause patients with salpingitis to be more susceptible to tubal pregnancy and might be involved in the implantation process of tubal pregnancy.

[Expression of aquaporin-8 and bcl-2 protein in human cervical carcinoma and their correlations].

OBJECTIVE: To investigate the expression of aquaporin-8 (AQP8) and apoptosis associated bcl-2 protein in human cervical carcinoma and their relationship. METHODS: The expression of AQP8 and bcl-2 protein in 74 cases of cervical carcinoma (46 cases of squamous-cell carcinoma of the uterine cervix, 28 cases of adenocarcinoma of the uterine cervix), 34 cases of cervical intraepithelial neoplasia (CIN) and 15 cases of normal cervices were detected by immunohistochemical technique, and their clinical significance were analyzed. RESULTS: The expression of AQP8 and bcl-2 protein were detected in intracytoplasm of atypia cells in CIN, squamous-cell carcinoma and adenocarcinoma of the uterine cervix. The positive rates of AQP8 and bcl-2 in squamous-cell carcinoma, adenocarcinoma, CIN and normal cervical epithelium were 98%, 74%; 61%, 71%; 71%, 53% ; 53%, 20% respectively. There were significant differences between squamous-cell carcinoma of the uterine cervix and other groups in AQP8 (P < 0.01), but no significant differences were found in any other groups. There were significant differences between squamous-cell carcinoma of the uterine cervix and CIN or normal cervical epithelium in bcl-2, so were between adenocarcinoma of the uterine cervix. The expression of AQP8 was positively correlated with bcl-2 in human cervical carcinoma( r(s) = 0.463, P = 0.000). CONCLUSIONS: There is a close relationship between high expression of AQP8 and development of human cervical carcinoma. The expression of AQP8 protein is positively correlated with bcl-2 protein in human cervical carcinoma. AQP8 protein may have anti-apoptosis function, although the detailed mechanism in human cervical carcinoma remains to be clarified.

[Expression of estrogen receptor, progesterone receptor and leukemia inhibitory factor on endometrium during different ovarian stimulation protocols in mice].

OBJECTIVE: To evaluate the influence of superovulation by GnRHa protocol and pregnant mare's serum gonadotropin (PMSG) alone on the expression of estrogen receptor (ER), progesterone receptor (PR) and leukemia inhibitory factor (LIF) mRNA on endometrium. METHODS: Forty-five female ICR mice were randomly allocated into 3 groups:(1) GnRHa+PMSG group: alarelin was give first for desensitizing the pituitary, then superovulation with PMSG; (2) PMSG group: mice were injected with PMSG only; (3) Natural cycle group: mice were given with same volume of saline. Endometrium samples were taken at 48 hours after given hCG or ovulation (control group). ER and PR in glandular cells were detected with SP immunohistochemistry semiquantitatively. Expression of LIF mRNA on endometrium was detected with reverse transcriptase-polymerase chain reaction (RT-PCR) analysis. RESULT: The positive rate(%) and expression intense (AU) of ER and PR on glandular epithelium cells were significantly lower in GnRHa+PMSG group and PMSG group than those in natural cycle group (all P <0.01). The expression of LIF mRNA was significantly lower in GnRHa+PMSG group and PMSG group than that in natural cycle group (all P <0.01); but the expressions of ER, PR and LIF in GnRHa+PMSG group were higher than those in PMSG group. CONCLUSION: The protocol with GnRHa down regulates the expressions of ER, PR and the LIF mRNA on the mice of secretive phase endometrium, suggesting it may have an adverse effect on the endometrial receptivity in mice, but it may still be better than PMSG alone.

Expression of chemokines CCL5 and CCL11 by smooth muscle tumor cells of the uterus and its possible role in the recruitment of mast cells.

OBJECTIVE: Smooth muscle tumors of uterus have been reported to contain considerable number of mast cells, especially cellular leiomyoma. However, to our knowledge the mechanism by which mast cells increased in them is not known. The purpose of this study was to reveal the different mast cell subsets in smooth muscle tumors of uterus and to investigate the mechanism of local increase of mast cells. METHODS: Tissue sections from 85 uterine smooth muscle tumors were studied using immunohistochemical double labeling techniques, including 40 cases of ordinary leiomyomas, 30 cases of cellular leiomyomas and 15 cases of leiomyosarcomas. The sections were double immunostained for mast cell tryptase and chymase, mast cell tryptase and ki-67, mast cell tryptase and chemokines (i.e., CCL2, CCL5, CCL11, TGFbeta), as well as tryptase and CCR3. RESULTS: MC(TC)-type of mast cells was the predominant type in ordinary leiomyoma and cellular leiomyoma, whereas MC(T)-type was seldom found in them. There was no MC(C) in smooth muscle tumors. The total intratumoral number of mast cells in cellular leiomyoma group was significantly higher than that in both leiomyosarcoma and ordinary leiomyoma (P<0.01). Mast cells proliferation was rarely detected in smooth muscle tumors, as revealed by constant negative labeling of the proliferation marker Ki-67 in mast cells. Almost all mast cells (tryptase positive) in smooth muscle tumors were also CCL2, CCL5, CCL11 and TGFbeta positive. Expressions of CCL5 and CCL11 in tumor cells in cellular leiomyoma were all significantly higher than that in both ordinary leiomyoma and leiomyosarcoma (P<0.01). While the expression of TGFbeta in tumor cells in cellular leiomyoma was not significantly different from that in ordinary leiomyoma, expression of CCL2 was not observed in smooth muscle tumor cells. There were positive correlations between CCL5 and the number of mast cells (r(s)=0.801, P<0.01) and between CCL11 and the number of mast cells (r(s)=0.744, P<0.01) in smooth muscle tumors as well. The vast majority of the mast cells in cellular leiomyoma were CCR3 positive. CONCLUSIONS: Using the monoclonal anti-mast cell tryptase antibody could detect all mast cells in smooth muscle tumor. The increased intratumoral mast cell counts in cellular leiomyoma might be the result of mast cells recruitment from the peripheral blood rather than local mast cells proliferation. CCL5 and CCL11, which are expressed by smooth muscle tumor cells, are possibly responsible for the recruitment of mast cells in uterine cellular leiomyoma. Whether they combine to CCR3 expressed by mast cells need further study.

Expression and localization of aquaporin-5 in the epithelial ovarian tumors.

OBJECTIVE: To investigate the expression and localization of aquaporin-5 (AQP5) in epithelial ovarian tumors and its clinic significance. METHODS: The expression of AQP5 protein and mRNA in 65 cases epithelial ovarian tumors and 13 cases normal tissue were measured by immunohistochemical technique, Western blotting and RT-PCR, respectively. RESULTS: AQP5 is mainly localized in the basolateral membranes of benign tumor cells, the apical and basolateral membrane of borderline cells and scattered in the membrane of malignant cells and almost no or weak staining in normal ovarian epithelium. The AQP5 expression in ovarian malignant and borderline tumors was significantly higher than that of benign tumors (P < 0.05) and normal tissue (P < 0.05). Of all the epithelial ovarian malignant tumors, the AQP5 expression in cases with ascites volume more than 1000 ml was higher than that of ascites volume less than 500 ml (P < 0.05). Increased AQP5 protein level was associated with lymph node metastasis (P < 0.05). There is a positive correlation between ascites amount and the expression of AQP5 protein and mRNA (P < 0.05), as well as lymph node metastasis and the expression of AQP5 protein and mRNA (P < 0.05). The AQP5 expression was not related with FIGO stage, grade and histological type (P > 0.05). CONCLUSION: The data suggest that overexpression of AQP5 play an important role in tumorigenesis of epithelial ovarian tumors, which may be related to the ascites formation of ovarian carcinoma.

[Value of computed tomography in the staging and predicting resectability of primary advanced ovarian carcinoma].

OBJECTIVE: To assess the value of computed tomography( CT) in the staging and predicting respectability of primary advanced ovarian carcinoma. METHODS: The data of preoperative abdomen and pelvis CT scan in 64 women with Stage II or IV ovarian carcinoma were collected from tumor registry database. All CT scans were analyzed retrospectively without knowledge of the operative findings, and the stage as based on CT was compared with the surgical and pathological findings. Residual lesion of < or = 2 cm in maximal diameter was considered as an optimal surgical result. Twenty-senven of these 64 patients (42.2%) underwent optimal cytoreduction surgery for residual disease C2 cm in diameter. Based on the ability of each parameter in predicting cytoreductive surgery outcome, 11 radiographic features were selected for the final model. Each predictive parameter was assigned a numeric value (1 to 7). Sensitivity, specificity, positive predictive value( PPV) , negative predictive value( NPV),and accuracy were calculated for each predictive parameter. Receiver operating characteristic( ROC) curve was used to assess the ability of the model to predict surgical outcome. The correlation between CT stage and surgical-pathologic stage was analyzed by Chi-square test and Spearman's rho analysis. RESULTS: The overall accuracy of CT staging for advanced ovarian carcinoma was 87. 5% ; 86. 5% and 91.7% for stage III and IV patients respectively. The correlation between CT stage and surgicopathologic stage was found to be comformable. In the final predictive index model, when a predictive index scoreed > or = 2, the overall accuracy, sensitivity and specificity was 70. 3% , 67.6% and 74. 1% for identifying patients for suboptimal surgery. The PPV and the NPV was 78. 1% and 62. 5% , respectively. The ROC curve was generated with an area under the curve = 0. 792+/-0. 055 using the predictive index scores. CONCLUSION: CT has a high accuracy in staging and a moderate ability to predict resectability for advanced ovarian carcinoma. Therefore, the predictive index model may be useful in the management of ovarian carcinoma patients.

[Protein and mRNA expression of aquaporin-1 in epithelial ovarian tumors and its clinic significance].

OBJECTIVE: To investigate protein and mRNA expression of aquaporin-1 (AQP1) in epithelial ovarian tumors and its clinic significance. METHODS: The protein and mRNA expressions of AQP1 were measured by immunohistochemical technique, western blot and RT-PCR in 65 cases of epithelial ovarian tumors and 13 cases of normal ovary tissue. RESULTS: AQP1 located in microvascular and small vessel epithelial cells. The protein and mRNA expressions of AQP1 in ovarian cancer (0.39 +/- 0.12, 0.93 +/- 0.51, respectively) and ovarian borderline tumors (0.43 +/- 0.21, 0.95 +/- 0.34, respectively) were significantly higher than that of ovarian benign tumors (0.27 +/- 0.13, 0.51 +/- 0.41, respectively; P < 0.05) and normal ovary tissue (0.24 +/- 0.13, 0.34 +/- 0.29, respectively; P < 0.05). Of all ovarian cancers, expression of AQP1 in cases with ascites more than 1000 ml (0.46 +/- 0.13, 1.25 +/- 0.57, respectively) was higher than that of ascites less than 1 approximately 499 ml (0.35 +/- 0.11, 0.75 +/- 0.45, respectively; P < 0.05). CONCLUSION: Over-expression of AQP1 plays an important role in development of epithelial ovarian tumors, and may be related with formation of ascites of ovarian carcinoma.

[Telomerase RNA antisense oligonucleotides inhibit growth of human choriocarcinoma xenograft in nude mice].

OBJECTIVE: To study the inhibitory effect of antisense oligonucleotides against telomerase RNA on the growth of human choriocarcinoma transplant in nude mice. METHODS: Choriocarcinoma xenografts were established by transplanting JAR cells subcutaneously to female nude mice, and were treated with high and low doses of antisense oligonucleotides. Control groups were treated with NS, random sequence and actinomycin D (Act-D). Tumor growth was monitored once every other day. Telomerase relative activity was assayed by TRAP-ELISA. Western blotting was used to detect expression of hTERT. RESULTS: Low and high doses antisense oligonucleotides, and Act-D inhibited tumor growth by 76.6%, 93.8% and 85.4% respectively, which were significantly different when compared with random sequence and NS groups. Expression of telomerase relative activity and hTERT were decreased as well. But the differences among the first three groups had no significance. CONCLUSION: Telomerase RNA antisense oligonucleotide inhibits growth of human choriocarcinoma xenografts in nude mice. It may be a novel approach to the treatment of choriocarcinoma.

[Survey of gestational trophoblastic disease incidence among 3.6 million pregnancies in China].

OBJECTIVE: To survey incidence of gestational trophoblastic disease (GTD) in China and to provide useful information for prevention and better treatment of the disease. METHODS: The survey was retrospectively carried out from 1991 to 2000 and included 143 hospitals in the following seven Chinese provinces: Zhejiang, Jiangsu, Fujian, Anhui, Jiangxi, Shanxi and Henan. RESULTS: Excluding incomplete data, data from 118 hospitals from seven provinces were finally analyzed. The total numbers of pregnancy and GTD were 3,674, 654 and 14,222, respectively. The GTD cases occurred mainly among 20 - 34 year old women, which accounted for 85.5% of total GTD. The incidence of GTD was 3.87 per thousand. There were 9,194 cases (64.6%) of hydatidiform, 3,452 cases (24.3%) of invasive mole, 1521 cases (10.7%) of choriocarcinoma, 55 cases (0.4%) of placenta-site trophoblastic tumor, respectively. CONCLUSIONS: The results of this survey are reliable and representative due to large sampling and hospital-based data collection. The incidences of GTD decreased significantly compared with 1950s'. It is important to take pathological examination for GTD and to diagnose complate mole and partial mole correctly.

[Expressions of aromatase P450 and estrogen receptor in eutopic and ectopic endometrium in endometriosis and their correlation with endometriosis].

OBJECTIVE: To detect the expression of aromatase P450 and estrogen receptor (ER) in eutopic and ectopic endometrium in endometriosis and their correlation with endometriosis. METHODS: Forty patients who had undergone operation because of endometriosis (all were stage III-IV according to the revised American Fertility Society classification) were enrolled and 83 tissue specimens from different locations of disease foci were obtained and divided into five groups according to the location: group I, eutopic endometrium of 25 cases; Group II, ovarian endometriosis tissues of 23 cases; Group III, vagina-rectum endometriosis tissues of 11 cases; Group IV, peritoneum endometriosis tissues of eight cases; and group V, uterine serosa endometriosis tissues of 16 cases. Control group consisted of normal endometrium taken from 20 fertile women who had endometrial curettage before placement of intrauterine device. The routine two-step immunohistochemical technique was used to measure the expression of aromatase P450 and ER in endometrial cells. RESULTS: Expressions of aromatase P450 and ER in group I [histochemistry score (H-score), 2.6 +/- 1.0, 3.8 +/- 0.5] were significantly higher than those in control group (both P < 0.01), group II (1.4 +/- 1.0, 1.9 +/- 1.6) (P < 0.05, P < 0.01), and group III (1.2 +/- 0.9, 1.8 +/- 1.6) (both P < 0.05). Expressions of aromatase P450 and ER had positive correlation in all groups (r = 0.577, P < 0.01). CONCLUSIONS: Aromatase P450 could increase estrogen and estrogen receptor levels in the local ectopic tissues, thus promoting the growth and implantation of endometriosis. Different types of endometriosis focus have different levels of hormone regulation.

[Establishment of two-dimensional electrophoresis of uterine leiomyoma for the proteome analysis].

OBJECTIVE: To establish and optimize the two-dimensional electrophoresis (2-DE) of uterine leiomyoma for the proteome analysis. METHODS: Run immobilized pH gradient (IPG)-isoelectric focusing electrophoresis as the first dimension, then vertical SDS-PAGE electrophoresis as the second dimension. A series of important steps,such as sample solubility, volume of loading, electrophoresis parameters and protocol for staining were optimized. RESULTS: The 2-DE patterns of uterine leiomyoma and myometrium with good quality were obtained. CONCLUSION: With optimal condition the two-dimensional electrophoresis of uterine leiomyoma can be obtained.

[Expression of R2 protein in gestational trophoblastic diseases].

OBJECTIVE: To study the expression of the small subunit ribonucleotide reductase (R2) in gestational trophoblastic diseases (GTD) and to assess its prognostic value. METHODS: The expression of R2 was detected with immunohistochemical method in 15 cases of normal villi, 38 cases of hydatidiform mole (HM), 42 cases of invasive moles (IM) and 18 cases of choriocarcinoma (CC). RESULTS: R2 expression in HM, IM and CC was significantly increased compared with that of normal villi (P=0.000). There were no significant differences in R2 protein expression among HM, IM and CC. Among 38 cases of HM, R2 expression in 8 cases with malignant transformation was significantly higher than in 30 cases of non-malignant transformation mole (P=0.02). Preoperative chemotherapy of gestational trophoblastic tumor including IM and CC did not influence the R2 expression. Compared with patients of stage I (WHO), the R2 protein in gestational trophoblastic tumor (GTT) patients of stage III or stage II was significantly increased (P=0.023 and P=0.038, respectively). The value of R2 in GTT patients with middle or high risk in WHO prognostic scoring system was higher than in the patients with low risk (P=0.018 and P=0.006, respectively). CONCLUSION: R2 expression in GTD is increased, which may be associated with the hyperplasia of trophoblasts, malignant transformation of hydatidiform mole and drug resistance of trophoblastic tumor.

[Effect of antisense oligodeoxynucleotide of small subunit component of human ribonucleotide reductase on human choriocarcinoma cell line in vitro].

OBJECTIVE: To study effect of the antisense oligodeoxynucleotide (ASODN) of small subunit of human ribonucleotide reductase (RRM2) mRNA on cell line of human choriocarcinoma in vitro. METHODS: Two 20-mer gapmer ASODNs with a full phosphorothioate backbone were artificially synthesized, which were complementary to nucleotides 626-645 (a coding region) and 1572-1591 (a 3'untranslated region) of RRM2, respectively. ASODNs were transfected into JAR cells through oligofectamine. The survival rate was assessed by methyl thiazolyl tetrazolium (MMT) assay, and RRM2 expression was detected by immunoblot and reverse transcriptase polymerase chain reaction (RT-PCR) methods. RESULTS: Antisense oligodeoxynucleotide one (ASODN1) targeting the coding region significantly inhibited growth of JAR cells in a dose- and time-dependent manner and downregulated RRM2 expression in a time-dependent manner. ASODN1 at 100 nmol/L could inhibit significantly cell growth (P = 0.000), and the effects of ASODN1 on JAR cell proliferation were enhanced with increase of ASODN1 concentration and reached the peak point at 400 nmol/L concentration (P = 0.000). Cell growth was significantly inhibited by 200 nmol/L of ASODN1 after 24 h of treatment (P = 0.000). The effect of ASODN1 was at the maximum at 48 h (P = 0.000), and began to decrease at 72 h of treatment. RRM2 expression started to reduce after ASOND1 treatment for 12 hours, and was obviously downregulated at 24 h of treatment, and decreased to the lowest level at 48 h (P < 0.05). RRM2 expression began to recover at 72 h of treatment. Antisense oligodeoxynucleotide two (ASODN2) corresponding to 3'untranslated region and control oligodeoxynucleotides had no significant effect on the proliferation and RRM2 expression, but the combination of ASODN2 and ASODN1 was more effective on reducing RRM2 expression and inhibiting cell proliferation than ASODN1 alone. CONCLUSIONS: ASODN1 of RRM2 can inhibit the proliferation of JAR cells and downregulate RRM2 transcription and translation in a selective and specific manner. The combination of multiple ASODNs targeting different regions of RRM2 mRNA would be an important approach to improve antisense effectiveness.

[Hemeoxygenase expression and activity in human placenta of pregnancy induced hypertension].

OBJECTIVE: To investigate the expression of heme oxygenase-1 (HO-1) and heme oxygenase-2 (HO-2) in placenta tissue of pregnancy induced hypertension (PIH), and the relationship between HO protein expression and enzymatic activity. METHODS: Protein expression was analyzed qualitatively and semi-quantitatively by Western blotting in placental tissue of PIH (PIH group, n = 30) and normal late pregnant women (control group, n = 30). The levels of HO enzymatic activity in placental tissue were measured with the double wavelength scanning by spectrophotometer. RESULTS: (1) Western blotting analysis demonstrated that the relative protein levels of HO-1 and HO-2 in placental samples of control group were 0.7 +/- 0.4 and 0.8 +/- 0.4 respectively, there were no significant difference between HO-1 and HO-2 protein levels (P > 0.05). (2) The relative levels of HO-1 were 0.6 +/- 0.4 in PIH group, there was no significant difference from those in the control group (P > 0.05); the relative levels of HO-2 protein were 0.6 +/- 0.3 in PIH group, that were obviously lower than those in the control group (P < 0.01). The levels of HO enzymatic activity of PIH group were (0.3 +/- 0.2) nmol x mg(-1) x h(-1), significantly lower than that of the control group [(0.5 +/- 0.3) nmol x mg(-1) x h(-1)] (P < 0.01). The levels of HO activity correlated with HO-2 protein levels. CONCLUSIONS: The expression levels of HO-2 protein were decreased and HO enzymatic activity reduced in PIH group. HO may play a role in the pathophysiology of poor placental perfusion and tissue damage in placenta of PIH.

[Inhibitory effect of tumor suppressor PTEN on cell growth of endometrial carcinoma].

OBJECTIVE: To evaluate the inhibitory effect of tumor suppressor PTEN on cell growth of endometrial carcinoma. METHODS: The exogenous wild PTEN cDNA via an adenoviral vector (Ad-PTEN) was introduced into Ishikawa cells. The expression of PTEN protein was detected by Western blot. The growth of Ishikawa cells was evaluated by trypan blue exclusion method and MTT. RESULTS: The expression of PTEN protein was induced on day 1, and greatly increasing on day 3 - 5 after Ad-PTEN infection. The expression of PTEN significantly inhibited the growth of Ishikawa cells, and also significantly inhibited the growth of Ishikawa cells induced by IGF-II. CONCLUSION: Adenovirus-mediated introduction of exogenous PTEN into human endometrial carcinoma cells can induce growth suppression. PTEN gene may be a novel therapeutic agent for endometrial carcinoma.

[Mast cells in cellular leiomyoma and endometrial stromal sarcoma of the uterus].

OBJECTIVE: To study the role of mast cells in the differential diagnosis of cellular leiomyoma and endometrial stromal sarcoma of uterus and its mechanism. METHODS: Using SP immunohistochemical technique, the expression of proliferating cell nuclear antigen (PCNA) and mast cells in 25 cellular leiomyoma (CL) and 26 endometrial stromal sarcoma (ESS) of uterus were examined. The expression of estrogen receptor (ER) and CD44v3 in cellular leiomyoma was also studied. RESULTS: The expression of PCNA was not significantly different from CL or ESS (P > 0.05), while mast cell count was statistically different between them (P < 0.01). Using a value of less than 7 mast cells per high power field was useful for the diagnosis of ESS, yielding 100% sensitivity and 92.0% specificity. There was a positive correlation between the mast cell count and CD44v3 in CL (r(s) = 0.589, P < 0.01), though no correlation was observed between mast cell count and PCNA or ER. CONCLUSION: Number of mast cells is valuable for the discrimination of CL from ESS in the uterus. The mechanism and the role of higher quantity of mast cells in CL need further study.

[Expression of RFC2 and PCNA in different gestational trophoblastic diseases].

BACKGROUND & OBJECTIVE: High expression of replication factor C (RFC) mRNA in hydatidiform mole and choriocarcinoma were detected previously with DNA microarray. Replication factor C subunit 2 (RFC2) was reported to be associated with DNA duplication, DNA repair, and the function of cellular checkpoint. The aim of current study was to investigate the expression levels of RFC2 and proliferating cell nuclear antigen (PCNA) in gestational trophoblastic diseases (GTD) and to explore the relationship of expressions of two proteins with GTD. METHODS: The expression of RFC2 and PCNA were detected with immunohistochemical method in 15 cases of normal villi, 38 cases of hydatidiform moles (HM), 42 cases of invasive moles (IM), and 18 cases of choriocarcinomas (CC). RESULTS: The expression of RFC2 and PCNA were significantly increased in HM, IM, and CC than in normal villi (P=0.000 for RFC2,P=0.004 for PCNA). There was no significant difference of the expression of RFC2 among HM, IM, and CC. The PCNA expression was significantly higher in CC than in HM (P=0.037). PCNA expression was significantly higher in the patients with malignantly transformed molar pregnancy than in the patients without malignantly transformed molar pregnancy (P=0.039). RFC2 expression in no preoperative chemotherapeutic group of gestational trophoblastic tumors (GTT) including IM and CC was significantly higher than that in the group with the chemotherapy of more than 3 cycles (P=0.028). Compared with patients at stageI, patients at stage III (WHO) had significantly increased expression of RFC2 (P=0.01). The level of RFC2 was higher in the patients with high risk in WHO prognostic scoring system than that in the patients with low risk (P=0.018). The levels of PCNA were significantly higher in the patients with high risk and middle risk than that in the patients with low risk (P=0.036 and P=0.048, respectively). The expression of PCNA protein was not associated with the preoperative chemotherapy and WHO stage of GTT. The RFC2 expression was positively correlated with the PCNA expression (P=0.000). CONCLUSION: The over-expression of RFC2 and PCNA in GTD may be associated with the malignant transformation of HM and the behavior of trophoblastic tumor.

Immunohistochemical markers in differential diagnosis of endometrial stromal sarcoma and cellular leiomyoma.

OBJECTIVE: Distinction of endometrial stromal sarcoma (ESS) from benign smooth muscle proliferations like cellular leiomyoma (CL) is sometimes problematic. The purpose of this study was to evaluate the potential utility of a panel of antibodies in the differential diagnosis of ESS and CL. METHODS: Using a standard streptavidin-biotin method, the expression of desmin, alpha smooth muscle actin (SMA), calponin h1, h-caldesmon, estrogen receptor (ER), progesterone receptor (PR), CD10, CD44v3, proliferating cell nuclear antigen (PCNA), and mast cells (MCs) were evaluated in 26 cases of ESS (21 low grade, 5 high grade), 25 CL (17 common CL, 8 highly CL), 25 myometria, and 25 endometria. RESULTS: Among ESS, 20 of 26, 17 of 26, 9 of 26, 12 of 26, 14 of 26, and 22 of 26 were positive for expression of desmin, SMA, calponin h1, ER, PR, and CD10, respectively, while only 2 of 26 were positive for CD44v3 and all were entirely negative for h-caldesmon. Of CL, all were positive for SMA, calponin h1, PR, and CD44v3; 24 of 25, 24 of 25, and 19 of 25 were positive for desmin, h-caldesmon, and ER, respectively, whereas 1 of 25 focally marked with antibodies to CD10. There was no significant difference of PCNA expression between ESS and CL, although the ESS cases tended to have higher values. The MC counts were significantly higher in the CL group than in the ESS group (P < 0.01). When using the cut-off value of seven MCs per HPF to distinguish ESSs from CLs, the sensitivity and specificity of this cut-off value were 92.9% and 100%, respectively. CONCLUSIONS: A panel of h-caldesmon, CD10, and CD44v3 should be used and will distinguish ESS from CL in most cases. In addition, counting the number of MCs might be useful as part of a multivariate approach to the differential diagnosis of them. But the biological function of MC and CD44v3 in these tumors is worthy of further investigation.