Alternative splicing of the androgen receptor in polycystic ovary syndrome.

Polycystic ovary syndrome (PCOS) is one of the most common female endocrine disorders and a leading cause of female subfertility. The mechanism underlying the pathophysiology of PCOS remains to be illustrated. Here, we identify two alternative splice variants (ASVs) of the androgen receptor (AR), insertion and deletion isoforms, in granulosa cells (GCs) in ∼62% of patients with PCOS. AR ASVs are strongly associated with remarkable hyperandrogenism and abnormalities in folliculogenesis, and are absent from all control subjects without PCOS. Alternative splicing dramatically alters genome-wide AR recruitment and androgen-induced expression of genes related to androgen metabolism and folliculogenesis in human GCs. These findings establish alternative splicing of AR in GCs as the major pathogenic mechanism for hyperandrogenism and abnormal folliculogenesis in PCOS.

[Expression of suppressor of cytokine signaling-3 and caspase-3 in endometriosis and their correlation].

OBJECTIVE: To investigate the expression of suppressor of cytokine signaling(SOCS)-3 and caspase-3 and their correlative significance in endometriosis. METHODS: Immunohistochemical EnVision method was used to detect the SOCS-3 and caspase-3 protein expression in ectopic and eutopic endometrium (n = 32) of patients with endometriosis, as well as normal endometrium (n = 30) of women without endometriosis. RESULTS: SOCS-3 and caspase-3 proteins were expressed in all three groups and not affected by the menstrual cycles. The expression of SOCS-3 in ectopic endometrium (5.54 ± 2.12) was significantly lower than that in eutopic (7.39 ± 1.09, P = 0.001) and control group (7.48 ± 1.26, P < 0.01), but without difference between the eutopic and control group (P = 0.756). SOCS-3 expression in ectopic and eutopic endometrium was significantly lower in III/IV stages than that in I/II stages of endometriosis (P < 0.05). Significantly lower expression of caspase-3 protein was found in ectopic (3.20 ± 1.24) and eutopic endometrium (3.88 ± 1.93) as compared with the control group (6.49 ± 1.85, P < 0.01), however ectopic and eutopic endometrium showed no significant difference (t = 1.66, P = 0.10). There was no significant difference of the expression of caspase-3 in ectopic and eutopic endometrium at different disease stages (P > 0.05). Positive correlation was found between the expression of SOCS-3 and caspase-3 proteins in ectopic endometrium (r = 0.655, P < 0.01). CONCLUSION: SOCS-3 may be involved in the development of endometriosis through inhibition of apoptosis of ectopic endometrial cells.

[Treatment of rheumatoid arthritis arthralgia by xiaoyan zhitong paste: a clinical observation].

OBJECTIVE: To formulate a comprehensive treatment program for rheumatoid arthritis arthralgia by clinical observing the efficacy of Xiaoyan Zhitong Paste (XZP). METHODS: Adopted was stratified, block randomized, double-blinded, placebo parallel controlled method. Subjects were assigned to the treatment group and the placebo group. Those in the treatment group were treated by external application of XZP, one to two pastes each time, covering the painful area, exchange once per 24 h, with one-day interval during a 7-day consecutive medication, two 7-days of treatment consisting of one therapeutic course. XZP placebos were applied for those in the placebo group in the same medication way. Joint pain and VAS were taken as main indices for observing the clinical efficacy of XZP. RESULTS: The improvement of the analgesic effect and the Chinese medical syndrome efficacy of XZP were superior to that of the placebo. CONCLUSION: XZP showed obvious effect in treating rheumatoid arthritis arthralgia with no obvious adverse reaction.

[Clinical effect analysis of ankylosing spondylitis treated by Chinese medical syndrome differentiation].

OBJECTIVE: To evaluate the curative effect and safety of Bushen Qiangji Decoction (BQD) and Qingre Qiangji Decoction (QQD) in treating ankylosing spondylitis (AS) patients, and to verify the clinical utility of AS syndrome differentiation and treatment scheme [Shen-deficiency induced stasis obstruction syndrome (SDISOS) and dampness-heat obstruction syndrome (DHOS) being two basic syndrome types, Shen invigorating blood activating method (SIBAM) and heat clearing dampness resolving method (HCDRM) being two basic treatment methods]. METHODS: Totally 354 AS patients of SDISOS and DHOS were randomly assigned to the treatment group and the control group using a multi-center randomized, positive drug parallel-controlled clinical trail. Patients in treatment group were treated by BQD or QQD according to syndrome typing, while those in the control group took Sulfasalazine enteric-coated tablet (SECT), 24 weeks as one therapeutic course. After treatment, the clinical efficacy was evaluated by using ASAS20 standard (set by Asessment in Ankylosing Spondylitis working group), Chinese medical efficacy evaluation standards, and BASDAI, BASFI, BASMI, night-pain index, spinal pain index, PGA, C-reactive protein (CRP), and erythrocyte sedimentation rate (ESR). RESULTS: After 24 weeks of treatment by BQD or QQD, ASAS20 standard rate was 86.75% in the treatment group, and the total effective rate of Chinese medical syndrome was 85.47%. They could significantly reduce patients' integrals of Chinese medical syndrome, BASDAI, BASFI, BASMI, night-pain index, spinal pain index, and PGA (all P < 0.01). CONCLUSIONS: QQD and BQD got confirmable clinical effects in treating AS, providing strong evidence of evidence-based medicine for syndrome differentiation and treatment of AS.

The Impact of Traditional Chinese Medicine QingreHuoxue Treatment and the Combination of Methotrexate and Hydroxychloroquine on the Radiological Progression of Active Rheumatoid Arthritis: A 52-Week Follow-Up of a Randomized Controlled Clinical Study.

Traditional Chinese medicine (TCM) has been used successfully to treat rheumatoid arthritis (RA). QingreHuoxue treatment (QingreHuoxue decoction [QRHXD]/QingreHuoxue external preparation [QRHXEP]) is a Chinese medicine treatment for RA. To date, very few studies have compared the long-term effects of QRHXD with those of conventional disease-modifying antirheumatic drugs on RA disease activity and radiological progression. QRHXD delayed the radiological progression and showed long-term clinical efficacy of RA. In clinical experiments, the clinical evidence of delaying the radiological progression of RA patients was obtained. A portion of the patients who participated in the "Traditional Chinese Medicine QingreHuoxue Treatment vs. the Combination of Methotrexate and Hydroxychloroquine for Active Rheumatoid Arthritis" study were followed up for 52 weeks, and intention-to-treat (ITT) and compliance protocol (PP) analyses were used to collect and compare the clinical indicators and imaging data between baseline and week 52. Two radiologists who were blind to treatment scored the images independently. Of the 468 subjects, 141 completed the 52-week follow-up. There were no significant differences among the three groups: the traditional Chinese medicine comprehensive treatment group, the Western medicine treatment group, and the integrated traditional Chinese and Western medicine treatment group. There were no differences in the total Sharp score, joint space stenosis score, and joint erosion score at baseline or 52 weeks. In the comparison of the estimated annual radiographic progression (EARP) and the actual annual Sharp total score changes among the three groups, the actual changes were much lower than the EARP at baseline. The radiological progress in all three groups was well controlled. Results of the ITT and PP data sets showed that the disease activity score 28 level of the three groups at 52 weeks was significantly lower than that at baseline. During the 52-week treatment period, the clearance of heat and promotion of blood circulation controlled disease activity and delayed the radiological progress of active RA.

[Expression and significance of LMP2 and PPM1A in gestational trophoblastic disease].

OBJECTIVE: To investigate the expression of low molecular mass polypeptide-2 (LMP2) and protein phosphatase 1A (PPM1A) in gestational trophoblastic disease and elucidate their predictive value in malignant transformation of hydatidiform mole. METHODS: The expressions of LMP2 and PPM1A protein in 196 complete hydatidiform moles (in which 28 cases with malignant transformation), 7 invasive moles, 5 choriocarcinomas and 20 normal chorionic villus were detected with the method of EnVision immunohistochemistry. Their clinicopathologic data were retrospectively analyzed. RESULTS: LMP2 and PPM1A protein expressed in cytotrophocytes, syncytiotrophoblast and extravillous trophoblast. The level of LMP2 expression in deteriorative hydatidiform mole was significantly higher than that in non-deteriorative hydatidiform mole or normal chorionic villus (6.79±2.38, 5.26±2.63 and 3.10±1.65, all P<0.01), while there were no difference compared with gestational trophoblastic neoplasms (6.42±2.68, P=0.113). The level of PPM1A expression was highest in normal chorionic villus, and decreased gradually in hydatidiform mole (non-deteriorative and deteriorative) and gestational trophoblastic neoplasms (6.30±2.98, 4.93±2.50, 4.43±2.04 and 3.33±2.06, all P<0.01); the level of PPM1A expression in deteriorative hydatidiform mole was significantly lower than that in non-deteriorative hydatidiform mole (P=0.001). The expression of LMP2 protein was correlated to theca lutein ovarian cyst, the expression of PPM1A protein was related with uterine size (P<0.05). While, there was no correlation between the expressions of the two proteins (P>0.05). CONCLUSIONS: High expression of LMP2 and low expression of PPM1A might play an important role in the motility and invasiveness of trophoblast cells and malignant transformation of hydatidiform mole. Testing the expression of LMP2 and PPM1A in hydatidiform mole tissues of initial uterine evacuation might be have some reference significance in judging outcomes of hydatidiform mole.

[Preliminary study on the relationship between tubal intraepithelial carcinoma of the fimbria and pelvic high-grade serous carcinoma].

OBJECTIVE: To explore the relationship between tubal intraepithelial carcinoma (TIC) of the fimbria and pelvic high-grade serous carcinoma. METHODS: All 34 cases of pelvic high-grade serous carcinoma with clear fimbria were evaluated from January 2009 to June 2010, including ovarian carcinoma (n = 26), tubal carcinoma (n = 7) and peritoneal carcinoma (n = 1). Among of these ovarian carcinomas, 12 cases were surface deposits and the other 14 cases within ovarian parenchyma. All 42 cases of non high-grade serous carcinoma in this period including 13 endometrioid ovary carcinomas, 11 clear cell ovary carcinomas, 11 mucinous ovary carcinomas, 6 low-grade serous ovary carcinomas, 1 low-grade serous tubal carcinoma, were also collected as a reference. The presence of tubal intraepithelial carcinomas was assessed. Based on the presence of TIC, high-grade serous ovary carcinomas were divided into TIC positive(+) and TIC negative(-) groups, and the clinical and pathological features of them were also evaluated. RESULTS: Fifteen cases (44%) were identified TIC in 34 high-grade pelvic serous carcinomas, and all of them were in the fimbria only, while none of TIC was found in control cases. There were significant difference between the two groups (χ(2) = 23.086, P = 0.000). Eleven cases (42%) were identified TIC in all 26 high-grade ovarian serous carcinomas, in which 8 cases with unilateral ovary carcinomas were associated with ipsilateral TIC, 2 cases with bilateral ovary carcinomas associated with unilateral TIC and one case with bilateral ovary carcinoma was associated with bilateral TIC. Four TIC (4/7) were identified in 7 cases with high-grade tubal serous carcinomas, and there was no presence of TIC in the 1 high-grade serous peritoneal carcinoma. Of all 26 high-grade ovarian serous carcinomas, 6/11 cases were surface deposits, and 5/11 were parenchyma tumors in TIC(+) group while 6/15 cases were surface deposits and 9/15 were parenchyma tumors in TIC(-) group, in which there were correlated in distribution of TIC between the two groups (P > 0.05). The average diameter of ovarian cancer were 6.9 and 6.5 cm between the two groups with no significant differences (t = 0.409, P = 0.690). CONCLUSION: TIC is specific to high-grade serous carcinomas and maybe have something to do with the pathogenesis of pelvic serous carcinomas.

Traditional Chinese Medicine Qingre Huoxue Treatment vs. the Combination of Methotrexate and Hydroxychloroquine for Active Rheumatoid Arthritis: A Multicenter, Double-Blind, Randomized Controlled Trial.

Traditional Chinese medicine (TCM) has been used successfully to treat rheumatoid arthritis (RA). Qingre Huoxue treatment (Qingre Huoxue decoction (QRHXD)/Qingre Huoxue external preparation (QRHXEP)) is a therapeutic scheme of TCM for RA. To date, there have been few studies comparing the efficacy and safety of QRHXD and conventional synthetic disease-modifying antirheumatic drugs (csDMARDs) for the treatment of active RA. This was investigated in a multicenter, double-blind, randomized controlled trial involving 468 Chinese patients with active RA [disease activity score (DAS)-28 > 3.2] treated with QRHXD/QRHXEP (TCM group), methotrexate plus hydroxychloroquine [Western medicine (WM) group], or both [integrative medicine (IM) group]. Patients were followed up for 24 weeks. The primary outcome measure was the change in DAS-28 from baseline to 24 weeks. The secondary outcome measures were treatment response rate according to American College of Rheumatology 20, 50, and 70% improvement criteria (ACR-20/50/70) and the rate of treatment-related adverse events (TRAEs). The trial was registered at ClinicalTrials.gov (NCT02551575). DAS-28 decreased in all three groups after treatment (p < 0.0001); the score was lowest in the TCM group (p < 0.05), while no difference was observed between the WM and IM groups (p > 0.05). At week 24, ACR-20 response was 73.04% with TCM, 80.17% with WM, and 73.95% with IM (based on the full analysis set [FAS], p > 0.05); ACR-50 responses were 40.87, 47.93, and 51.26%, respectively, (FAS, p > 0.05); and ACR-70 responses were 20.87, 22.31, and 25.21%, respectively, (FAS, p > 0.05). Thus, treatment efficacy was similar across groups based on ACR criteria. On the other hand, the rate of TRAEs was significantly lower in the TCM group compared to the other groups (p < 0.05). Thus, QRHXD/QRHXEP was effective in alleviating the symptoms of active RA-albeit to a lesser degree than csDMARDs-with fewer side effects. Importantly, combination with QRHXD enhanced the efficacy of csDMARDs. These results provide evidence that QRHXD can be used as an adjunct to csDMARDs for the management of RA, especially in patients who experience TRAEs with standard drugs. Clinical Trial Registration: ClinicalTrials.gov, identifier NCTNCT025515.

[Protein gene product 9.5-immunoactive nerve fibers and its clinical significance in endometriotic peritoneal lesions].

OBJECTIVE: To investigate the association between distribution of protein gene product (PGP) 9.5-immunoactive nerve fibers in peritoneal endometriotic lesions and disease-associated pain symptoms. METHODS: Thirty two peritoneal endometriotic lesions from patients with endometriosis (16 cases with pain and 16 cases without pain) and matched with 20 peritoneal tissues from patients with uterine leiomyoma without endometriosis were stained immunohistochemically for PGP9.5-immunoactive nerve fibers. RESULTS: The positive rate and density of PGP9.5-immunoreactive nerve fibers in peritoneal endometriotic leision were 62% (10/16) and (3.8+/-1.7)/mm2 in endometriosis patients with pain, which were significantly higher than 19% (3/16) and (1.7+/-0.5)/mm2 in endometriosis patients without pain (P<0.05) and 25% (5/20) and (1.3+/-0.6)/mm2 in peritoneal tissues in women without endometriosis (P<0.05). However, no differences were found between endometriosis patients without pain and women without endometriosis (P>0.05). Moreover, the density of PGP9.5-immunoreactive nerve fibers in peritoneal lesions in endometriosis patients with pain was positively correlated with the severity of pain (r=0.855, P<0.05). In addition, the density of PGP9.5-immunoreactive nerve fibers in peritoneal lesions was statistically higher in endometriosis patients with chronic pelvic pain and (or) dysmenorrhea than those in endometriosis patients with other type of pain (P<0.05), which was not associated with active lesion, site and staging (P>0.05). CONCLUSION: It suggested that PGP9.5-immunoreactive nerve fibers might confer the mechanism of pelvic pain with endometriosis.

[Clinical study on active rheumatoid arthritis treated with simiao xiaobi decoction].

OBJECTIVE: To observe the clinical efficacy and safety of Simiao Xiaobi Decoction (SXD) in treating active rheumatoid arthritis (RA) of humid pyretic toxic Bi-Zheng (HPTB) syndrome type. METHODS: One hundred and twenty RA patients were randomly assigned to 2 groups, 60 in the treatment group receiving SXD, and 60 in the control group receiving methotrexate, all were treated for 12 weeks. Clinical efficacy in patients was evaluated, referring to the criteria recommended by European League Against Rheumatoism (EULAR), in terms of effective rate, main symptoms, signs, scoring on symptom/sign by Chinese medicine scale and DAS28, physical and chemical indices, long-term outcome of patients and the average therapeutic effect initiating time. Meantime, the adverse reaction was recorded. RESULTS: The study was completed in 103 patients, 52 in the treated group and 51 in the control group. According to a per-protocol analysis, the effective rate was better in the treatment group than in the control group with marked difference in terms of Chinese and Western medicine respectively (92.3% vs 70.6% and 86.5% vs 62.7%, P<0.05). Superiorities in the treatment group were also seen in the improvements of main symptoms and signs, symptom/sign scores, DAS28 scores, and long-term outcome. Moreover, the average therapeutic effect initiating time was shorter (5.31 +/- 0.36 weeks vs 8.28 +/- 0.45 weeks), while the incidence of adverse reaction was less in the treatment group than in the control group (6.7% vs 43.3%, P<0.05). CONCLUSION: SXD can improve the joint symptoms and general condition of RA patients of HPTB type with shorter initiating time and less adverse reaction.

[Distribution of nerve fibers in endometrium and its clinical significance in adenomyosis].

OBJECTIVE: To investigate nerve fibers distribution in endometrium of adenomyosis and their relationship with dysmenorrhea. METHODS: Endometrial tissue was sampled from 74 hysterectomy specimens including 32 cases with adenomyosis and 42 cases with uterine fibroids. Two-step Envision immunohistochemical staining was used to detect distribution of nerve fibers in endometrium. Highly specific polyclonal rabbit anti-protein gene product 9.5 (PGP9.5) and monoclonal mouse anti-neurofilament protein (NF) were used to demonstrate both myelinated and unmyelinated nerve fibers in endometrium in women with adenomyosis and uterine fibroids. RESULTS: The positive rate of PGP9.5 immunoreactive nerve fibers in the functional layer of endometrium of pain patients were with 64% (14/22) in adenomyosis and 67% (10/15) in uterine fibroids. And their density were 0.6 (0 - 9.4)/mm(2) and 0.6 (0 - 6.0)/mm(2) without reaching statistical difference (P > 0.05). No expression of NF could be detected in the functional layer of endometrium of adenomyosis and uterine fibroids. There were no PGP9.5 immunoreactive nerve fibers in the functional layer of endometrium in non-pain women with adenomyosis and uterine fibroids. Moreover, No NF immunoreactive nerve fibers in the functional layer of endometrium were shown in non-pain patients with adenomyosis and uterine fibroids. PGP9.5 immunoreactive nerve fibers and the nerve density in the basal layer of endometrium were 64% (14/22), 1.1 (0 - 12.0)/mm(2) in pain adenomyosis and 50% (5/10), 0.6 (0 - 3.0)/mm(2) in non-pain adenomyosis. NF immunoreactive nerve fibers and the density in the basal layer of endometrium were 23% (5/22), (0 - 0.6)/mm(2) in pain adenomyosis and 20% (2/10), (0 - 1.0)/mm(2) in non-pain adenomyosis. PGP9.5 immunoreactive nerve fibers and the nerve density in the basal layer of endometrium were 80% (12/15) and 1.6 (0 - 10.0)/mm(2) in pain fibroids and 44% (12/27), 0 (0 - 5.0)/mm(2) in non-pain fibroids. NF immunoreactive nerve fibers and the nerve density in the basal layer of endometrium were 40% (6/15), 0 (0 - 0.4)/mm(2) in pain fibroids and 15% (4/27), 0 (0 - 1.0)/mm(2) in non-pain fibroids. There was no statistical different PGP9.5 and NF immunoreactive nerve fibers distribution in basal layer of endometrium between pain adenomyosis and pain fibroids or between non-pain adenomyosis and non-pain fibroids (all P > 0.05). However, PGP9.5 immunoreactive nerve fibers density in basal layer of endometrium was higher in pain adenomyosis and fibroids when compared with non-pain adenomyosis and fibroids (P < 0.05). CONCLUSIONS: PGP9.5 immunoreactive nerve fibers might confer the occurrence of pelvic pain, however, NF immunoreactive nerve fibers may not involved in the pathogenesis of pain.

Expression of aquaporin-1 in normal, hyperplasic, and carcinomatous endometria.

OBJECTIVE: To explore the relationship between aquaporin-1 (AQP1) and endometrial adenocarcinoma. METHOD: Intratumoral microvessel density (IMD) was assessed as well as AQP1 and vascular endothelial growth factor expression in samples from 117 women, 75 with endometrioid adenocarcinoma, 17 with endometrial hyperplasia, and 25 with normal proliferative endometria. RESULTS: AQP1 was located in the epithelial cells of microvessels and small vessels in all samples. The AQP1/IMD ratio was highest in samples from the first, less in samples from the second, and least in samples from the third group. In samples from endometrioid adenocarcinoma, the AQP1/IMD ratio was significantly correlated with histologic grade, surgical stage, myometrial invasion, and extrauterine metastasis. There was a positive correlation between AQP1 expression and IMD and between AQP1/IMD ratio and VEGF expression. CONCLUSION: AQP1 may be involved in the tumorigenesis and progression of endometrioid adenocarcinoma by promoting angiogenesis, and AQP1 level may be both a tumor indicator and a new therapeutic target.

[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.

The inhibition of UBC13 expression and blockage of the DNMT1-CHFR-Aurora A pathway contribute to paclitaxel resistance in ovarian cancer.

Paclitaxel is widely used as a first-line chemotherapeutic drug for patients with ovarian cancer and other solid cancers, but drug resistance occurs frequently, resulting in ovarian cancer still presenting as the highest lethality among all gynecological tumors. Here, using DIGE quantitative proteomics, we identified UBC13 as down-regulated in paclitaxel-resistant ovarian cancer cells, and it was further revealed by immunohistochemical staining that UBC13 low-expression was associated with poorer prognosis and shorter survival of the patients. Through gene function experiments, we found that paclitaxel exposure induced UBC13 down-regulation, and the enforced change in UBC13 expression altered the sensitivity to paclitaxel. Meanwhile, the reduction of UBC13 increased DNMT1 levels by attenuating its ubiquitination, and the up-regulated DNMT1 enhanced the CHFR promoter DNA methylation levels, leading to a reduction of CHFR expression, and an increased in the levels of Aurora A. Our findings revealed a novel function for UBC13 in regulating paclitaxel sensitivity through a DNMT1-CHFR-Aurora A pathway in ovarian cancer cells. UBC13 could potentially be employed as a therapeutic molecular drug for reversing paclitaxel resistance in ovarian cancer patients.

[Expression of cyclin B1 and cyclin-dependent kanasel in ectopic and eutopic endometrium tissues of patients with endometriosis].

OBJECTIVE: To investigate the possible roles of cyclin B1 and cyclin-dependent kanase1 cyclin-dependent kanase1 play in the pathogenesis and development of endometriosis and their association with the ovarian hormones. METHODS: Twenty-nine specimens of ectopic endometrium tissues and 20 specimens of eutopic endometrium tissues were obtained from 29 patients with endometriosis, aged 24 approximately 46. Thirty specimens of endometrium form 30 women without endometriosis were used as controls. The intracellular location of cyclin B1 and Cdc2 was detected by microscopy. Western blotting, RT-PCR, and immunohistochemistry were employed to examine the mRNA expression and protein expression of cyclin B1 and Cdc2. Serum estrogen and progestogen were detected. RESULTS: The expression level of cyclin B1 in the ectopic endometrium from the women with endometriosis was significantly higher than that in the ectopic endometrium tissues from the women with and without endometriosis (both P < 0.05). No significant difference was found between the expression level of cyclin B1 in the ectopic endometrium tissues from the women with and without endometriosis (both P > 0.05). The Cdc2 expression levels were not significantly different among th3 3 groups and the proliferative and secretary stage of endometrium (all P > 0.05). Cyclin B1 expression level was positively correlated with the serum estrogen level, and negatively correlated with the serum progestogen level, and Cdc2 expression was not correlated with the serum sex hormone levels. CONCLUSION: The expression of cyclin B1 in the ectopic endometrium is higher than normal. Cyclin B1 may be involved in the proliferation of endometrium in endometriosis.

[Expression profile of aquaporin 1 in patients with menorrhagia].

OBJECTIVE: To investigate the expression of aquaporin 1 (AQP1) in endometrium of patients with menorrhagia. METHODS: RT-PCR and immunohistochemistry were carried out in twenty women with normal menstrual cycle to confirm the expression of AQP1 in endometrium and locate it. Then 51 women with menorrhagia and 40 women with normal menstrual cycle were included in the study. RT-PCR and immunohistochemistry were used to examine the expression of AQP1. RESULT: AQP1 mRNA was expressed in the human endometrium throughout menstruation cycle, which was mainly located in the endothelia of the capillaries and small blood vessels. Quantification of the immunostaining revealed higher density during secretary phase than that in proliferative phase (P<0.01). The staining intensity and density of AQP1-positive microvessel decreased significantly in simple hyperplasia group (P<0.01) and then gradually increased in complex hyperplasia and atypical hyperplasia groups (P<0.001). CONCLUSION: Decreased expression of AQP1 may lead to disturbed endometrial vascular remodeling and may be involved in the occurrence of menorrhagia.

Altered expression of cellular membrane molecules of HLA-DR, HLA-G and CD99 in cervical intraepithelial neoplasias and invasive squamous cell carcinoma.

OBJECTIVE: The aim of this study was to investigate the role of HLA-DR, HLA-G and CD99 during cervical carcinogenesis and to examine the prognostic significance of these protein expressions in invasive squamous cell carcinoma (SCC). METHODS: Using specific antibodies for HLA-DR, HLA-G and CD99, we examined protein expressions in 19 normal cervix, 15 mild dysplasia (CIN I), 22 moderate dysplasia (CIN II), 23 severe dysplasia (CIN III), and 34 invasive squamous cell carcinoma by immunohistochemistry. And we detected the expression of Ki67 in the same specimens. RESULTS: None of normal cervix and CINs except three cases of CIN III expressed HLA-DR. HLA-DR expression increased progressively with the grade of the tumor, and significant differences could be observed between grade 1 and grade 2 (P<0.01) and between grade 1 and grade 3 (P<0.05). In all normal epithelial control samples, HLA-G expression was seen in ectocervical squamous and endocervical columnar epithelium and the staining was strong and uniform. Only a small proportion of CINs and SCCs showed reduced expression of HLA-G. Compared with the results in the control samples, CINs and SCCs showed significantly reduced expression of HLA-G (P<0.001). SCCs showed significantly increased expression of CD99 when compared with normal cervix and CINs (P<0.05). Ki67 was expressed in all specimens. Significant differences were observed between CINs and normal cervix (P<0.001) and SCCs and controls (P<0.001), but no significant differences could be observed between SCCs and CINs. None of the expressions of these proteins was associated with any of clinicopathological parameters. CONCLUSIONS: These results indicate that increased expression of HLA-DR and CD99 may be related to the evolution of cervical cancer. All protein expressions were not associated with clinicopathological parameters.

Aquaporin-2 expression in human endometrium correlates with serum ovarian steroid hormones.

The aim of the present study was to examine the expression of aquaporin-2 (AQP2), a member of the water channel family aquaporins (AQPs), in human uterine endometrium and its modulation of ovarian steroid hormone at the proliferative and secretory phases. Western blot, immunohistochemistry, and RT-PCR were employed in the present study. Western blot revealed a 29-kDa band that represented AQP2 in human endometrium. The expression of AQP2 in endometrium was confirmed by RT-PCR and immunohistochemical results. The immunohistochemical analysis demonstrated that AQP2 was prominent in luminal and glandular epithelial cells of endometrium. The levels of endometrial AQP2 expression changed during the menstrual cycle and were higher in the secretory endometrium than in the proliferative endometrium. A significantly high level of AQP2 was detected at the mid-secretory phase. There was a positive correlation between the levels of the endometrial AQP2 expression and the concentrations of the serum 17beta-estradiol (E2) or/and progesterone (P4). These data for the first time corroborate that AQP2 is expressed in human endometrium and that the expression of AQP2 in human endometrium might be regulated by E2 or/and P4. The changed expression of AQP2 at different phases of the menstrual cycle may be essential to reproductive physiology in human. The high level of endometrial AQP2 expression was observed at the mid-secretory phase, the time of embryo implantation, suggesting that AQP2 might play physiological roles in the uterine receptivity.

[Tumor lymphangiogenesis in cervical squamous cell carcinoma and its clinical significance].

OBJECTIVE: To investigate the clinical significance of lymphangiogenesis in cervical cancer. METHODS: Monoclonal podoplanin was used to immunostain the lymphatic microvessels in the paraffin sections of cervical squamous cancer tissues at the I(b) and II(a) stages from 35 cases kept in the archives. Computer-assisted morphometric analysis was used to quantitatively analyze the lymphatic vessels in intratumor and peritumor areas. The association with clinicopathologic data was analyzed. RESULTS: (1) The lymphatic microvessels were larger and denser in the peritumor areas than the intratumor areas. Cancer cells were found in the intratumoral lymphatic vessels of the lymph metastatic patients. (2) The lymphatic vessel density (LVD) of the peritumor areas was (31 +/- 10) vessels/mm(2), significantly greater than that in the intratumor areas [(20 +/- 10) vessels/mm(2), P < 0.01]. The relative lymphatic vessel area (LVA) in the peritumor areas was (0.75 +/- 0.40)%, significantly larger than that of the intratumor areas [(0.19 +/- 0.11)%, P < 0.01]. (3) The intra- and peri-tumoural LVD and LVA in the patients with lymph node metastasis were (29 +/- 7) vessels/mm(2) and (40 +/- 5) vessels/mm(2), and 0.27% +/- 0.10% and 1.23% +/- 0.36% respectively, all greater than those of the patients without lymph node metastasis [(16 +/- 8) vessels/mm(2) and (28 +/- 9) vessels/mm(2), and (0.16 +/- 0.09)%, (0.56 +/- 0.20)% respectively, P < 0.01, < 0.01, < 0.05, and < 0.01]. (4) The peri-tumoral LVA and intra-tumoral LVD and LVA in the patients with histological grade I, II, and III were (0.42 +/- 0.10)%, (9 +/- 3) vessels/mm(2), and (0.06 +/- 0.04)%; (0.77 +/- 0.37)%, (21 +/- 8) vessels/mm(2), and (0.21 +/- 0.09)%; and (0.83 +/- 0.46)%, (22 +/- 11) vessels/mm(2), and (0.21 +/- 0.12)% respectively. There were significant differences among the values of the grade II patients and grade I patients (all P < 0.05) and the grade III patients and grade I patients (all P < 0.05) and there were not significant differences between the values of the grade II patients and the grade III patients (all P > 0.05), III and grade I, no statistic significances between grade II and III. CONCLUSION: Higher lymphangiogenesis in early cervical squamous cell cancer may be associated with the risk of lymph node metastasis.