A study on UHPLC-MS/MS analyses of DNA and RNA oxidative damage metabolites in patients with cervical carcinoma: 8-oxoG in urine as a potential biomarker of cervical carcinoma.

Objective: The purpose of this study was to compare the levels of 8-oxoG and 8-oxodG in urine of patients with cervical carcinoma and healthy women to evaluate their influences on cervical carcinoma. Methods: In this study, urine samples were collected from 70 patients with cervical carcinoma, 24 patients with one-year follow-up, and 100 healthy women. The contents of 8-oxodG and 8-oxoG in urine were assayed by ultra-high performance liquid chromatography tandem mass spectrometry (UHPLC-MS/MS). Results: The levels of 8-oxoG and 8-oxodG were higher in patients with cervical carcinoma (P < 0.000), while AUC of 8-oxoG and 8-oxodG was higher than 0.7. Specifically, the high-risk HPV (HR-HPV) positive group had higher 8-oxoG levels (P < 0.000), but there was no difference in 8-oxodG levels. Yet, 8-oxoG level was associated with lymphatic metastasis, lymph-vascular space infiltration (LVSI) and stromal infiltration, while 8-oxodG level was affected by the differentiation degree and stromal infiltration. According to statistics, the distinct cut-off index of lymphatic metastasis was 7.282 nmol/mmol creatinine. After operation, the concentrations of 8-oxoG and 8-oxodG dropped significantly (8-oxoG P < 0.000, 8-oxodG P = 0.004). Except for chemotherapy group, the urinary 8-oxoG dose of all treatment groups and 8-oxodG dose of chemo-radiotherapy group declined obviously. Conclusions: 8-oxoG may be a potential biomarker for cervical carcinoma.

Metabolic potential of microbial community and distribution mechanism of Staphylococcus species during broad bean paste fermentation.

Although the microbial diversity and structure in bean-based fermented foods have been widely studied, systematic studies on functional microbiota and mechanism of community forms in multi-microbial fermentation systems were still lacking. In this work, the metabolic pathway and functional potential of microbial community in broad bean paste (BBP) were investigated by metagenomics approach, and Staphylococcus, Bacillus, Weissella, Aspergillus and Zygosaccharomyces were found to be the potential predominant populations responsible for substrate alteration and flavor biosynthesis. Among them, Staphylococcus was the most abundant and widespread functional microbe, and closely related Staphylococcus species were diverse and ubiquitously distributed, with the opportunistic pathogen S. gallinarum being the most abundant Staphylococcus specie isolated from BBP. To explain the dominance status of S. gallinarum and species distributions of Staphylococcus genus, we tested the effects of abiotic and biotic factors on three Staphylococcus species using a tractable BBP model, demonstrating that adaptation to environmental conditions (environmental parameters and other functional microbes) led to the dominant position and species coexistence of Staphylococcus, and congeneric competition among Staphylococcus species further shaped ecological distributions of closely related Staphylococcus species. In general, this work revealed the metabolic potential of microbial community and distribution mechanism of Staphylococcus species during BBP fermentation, which could help traditional factories to more precisely control the safety and quality of bean-based fermented foods.

Development of a defined autochthonous starter through dissecting the seasonal microbiome of broad bean paste.

Bean-based fermentation foods are usually ripened in open environment, which would lead to inconsistencies in flavor and quality between batches. The physicochemical metabolism and microbial community of seasonal broad bean paste (BBP) were compared to distinguish discriminant metabolites and unique taxa, as well as their specific reasons for different flavor and quality in this study. Here, we found that environmental variables led to the seasonal distribution of microbiota, and differential microorganisms further contributed to the inconsistency of flavor quality, in which Lactobacillales was responsible for the higher titratable acid and amino acid nitrogen concentration in winter pei, while Saccharomycetales benefited the formation of volatile flavor substances in autumn pei. Additionally, we compared the effect of different combinations of Lactobacillales with Zygosaccharomyces rouxii on the quality of BBP, and found that W. confusa was more suitable for BBP fermentation rather than T. halophilus in terms of sensory characteristics and physicochemical metabolites.

Interstitial Pregnancy after Ipsilateral Salpingectomy: Analysis of 46 Cases and a Literature Review.

STUDY OBJECTIVE: To investigate the relationship between previous ipsilateral salpingectomy and interstitial pregnancy and report on our experience of laparoscopic cornuostomy for interstitial pregnancy. DESIGN: Single-center, retrospective review. SETTING: University-based hospital. PATIENTS: All patients who had undergone ipsilateral salpingectomy previously, diagnosed with interstitial pregnancy and treated between July 2010 and September 2018. INTERVENTIONS: Laparoscopy or laparotomy as a treatment for interstitial pregnancy after ipsilateral salpingectomy. MEASUREMENTS AND MAIN RESULTS: A total of 414 cases of interstitial pregnancy were identified, of which 46 (11.1%) were after ipsilateral salpingectomy. Of the 46 patients, 20 (43.5%) became pregnant by in vitro fertilization and embryo transfer. Ipsilateral salpingectomy was the result of an ectopic pregnancy in 40 patients, hydrosalpinx in 5 patients, and torsion of an ovarian tumor in 1 patient. The laparoscopic approach was used to treat 78.3% of patients with history of previous salpingectomy. Patients who underwent ipsilateral salpingectomy by laparoscopy previously had a shorter interval from salpingectomy to interstitial pregnancy (24 months vs 60 months; p = .038) compared with patients who underwent ipsilateral salpingectomy by laparotomy. Laparoscopic cornuostomy was performed in 38 patients (82.6%); 12 had fetal cardiac activity, 15 had ruptured masses, and 16 used prophylactic methotrexate (MTX) intraoperatively. The median size of the ectopic mass was 2.5 cm (1.0-5.0 cm). At the time of laparoscopic cornuostomy, more patients with interstitial pregnancies with intact ectopic masses were administered prophylactic MTX (81.3% vs 45.5%; p = .043). Only 1 patient with a ruptured ectopic mass, high preoperative human chorionic gonadotropin levels, and without prophylactic MTX administration experienced a persistent ectopic pregnancy. CONCLUSION: Patients with a history of ipsilateral salpingectomy should be cautioned regarding the possibility of interstitial pregnancy. Laparoscopic cornuostomy appears to be an appropriate treatment for interstitial pregnancy in patients wishing to preserve fertility, and the use of concomitant prophylactic MTX may reduce the risk of persistent ectopic pregnancy, especially in patients with ruptured masses and high human chorionic gonadotropin levels.

Downregulation of lncRNA TUBA4B is Associated with Poor Prognosis for Epithelial Ovarian Cancer.

A host of studies have revealed that long non-coding RNAs (lncRNAs) are critically involved in the development and progression of epithelial ovarian cancer. LncRNA TUBA4B is recently identified to be a critical mediator in non-small cell lung cancer. However, the clinical roles and biological functions of lncRNA TUBA4B in epithelial ovarian cancer have yet to be fully clarified. The present study was conducted to explore the expression of lncRNA TUBA4B in human epithelial ovarian cancer tissues and potential roles of lncRNA TUBA4B in ovarian cancer cells. The matched epithelial ovarian cancer specimens and adjacent normal tissues were employed to detect the expression of lncRNA TUBA4B. The prognostic value of lncRNA TUBA4B for tumor progression and survival rate was investigated. The effects of lncRNA TUBA4B on ovarian cancer cell proliferation and migration were also explored. The expression of lncRNA TUBA4B was significantly decreased in epithelial ovarian cancer tissue specimens. The low lncRNA TUBA4B level was closely related with pathological grade, FIGO stage and lymph node metastases, and serum CA125 level. Enforced expression of lncRNA TUBA4B obviously reduced the proliferation of SKOV3 cells, and attenuated the activation of ERK and Akt signaling pathways. Our data demonstrate for the first time that lower lncRNA TUBA4B may be a novel independent prognostic biomarker for overall survival of epithelial ovarian cancer. Overexpression of lncRNA TUBA4B inhibits the proliferation of ovarian cancer cells. LncRNA TUBA4B may be an important target for therapeutic intervention in ovarian cancer.

Effect of Momordica charantia protein on proliferation, apoptosis and the AKT signal transduction pathway in the human endometrial carcinoma Ishikawa H cell line in vitro.

Endometrial carcinoma (EC) is one of the most common female malignancies, and there is an urgent requirement to explore new therapeutic strategies. In the present study, Ishikawa H cells were treated with Momordica charantia protein (MCP30). The cell morphology, growth inhibition rate, cell cycle distribution, and expression of phosphate and tensin homolog, P-AKT and AKT were measured. DNA fragmentation analysis and Annexin V-fluorescein isothiocyanate/propidium iodide double staining assay were used to analyze cell apoptosis. MCP30 decreased the viability of Ishikawa H cells in a dose- and time-dependent manner. The early apoptotic rates of Ishikawa H cells treated with MCP30 at 666.67 pM reached to 16.07±0.15%, following 72 h of treatment. DNA ladder was observed in cells treated with 333.33 and 666.67 pM MCP30 following 72 h of treatment. MCP30 blocks Ishikawa H cells from progressing between the S-phase and the G2/M-phase in a time- and concentration-dependent manner. Western blotting revealed that MCP30 treatment decreased the levels of P-AKT in a dose-dependent manner. It was revealed that MCP30 decreases cell proliferation, and induces apoptosis and S-phase cell cycle arrest through the AKT signaling pathway in Ishikawa H cells.

Nomogram Predicting Overall Survival in Operable Cervical Cancer Patients.

OBJECTIVE: Nomograms are widely used as predictive tools to predict oncological outcomes intuitively and precisely. The aim of our study was to develop a nomogram for predicting overall survival (OS) of patients with early stage cervical cancer. METHODS: In this retrospective study, the clinical, pathological, and hematological data and prognosis of 795 cervical cancer patients were investigated. We identified and incorporated independent significant prognostic factors for OS to develop a nomogram. The predictive accuracy and discriminative ability were measured by concordance index. RESULTS: By univariable analysis and subsequent multivariable analysis, we identified body mass index, albumin, platelet, leukocyte, tumor differentiation, and the status of the pelvic lymph node (PLN) (all P < 0.05) as independent prognostic factors. The concordance index of the nomogram integrating these 6 variables was 0.74. The calibration curves for probability of 3- and 5-year OS also demonstrated ideal agreement between nomogram prediction and actual observation. CONCLUSIONS: We developed a novel nomogram that can provide prediction of OS for patients with early stage cervical cancer individually. Furthermore, studies are required to validate whether it can be applied to other cohorts.

Use of cancer antigen 125, cancer antigen 19-9, and the neutrophil-to-lymphocyte ratio to diagnose mature cystic teratoma with torsion.

OBJECTIVE: To determine the efficacy of serum levels of cancer antigen 125 (CA125) and cancer antigen 19-9 (CA19-9), and the neutrophil-to-lymphocyte ratio (NLR) for diagnosis of mature cystic teratoma (MCT) with torsion. METHODS: A retrospective medical record review was conducted of data for women who had undergone surgery for ovarian MCT at the First Affiliated Hospital of Wenzhou Medical University, China, between January 1, 2008, and January 1, 2015. Patients with torsion and a control group who underwent surgery on the same days were identified. Clinical characteristics and the serum levels of CA125, CA19-9, and the NLR were assessed. RESULTS: The serum levels of CA125, CA19-9, and the NLR were higher in the torsion group (n=68) than in the control group (n=120; P≤0.001 for all). Receiver operating characteristic analysis indicated that the area under the curve for the combined use of CA125, CA19-9, and NLR was 0.978 (95% confidence interval 0.954-1.000; P<0.001). This combination had a diagnostic sensitivity of 93.9% and a specificity of 98.3%. Moreover, levels of these inflammatory markers were significantly increased among patients with large tumor diameters (P<0.01 for all). CONCLUSION: The combined measurement of CA125, CA19-9, and the NLR provided an efficient method for the diagnosis of MCT with torsion.

Overexpression of salusin-ß is associated with poor prognosis in ovarian cancer.

Ovarian cancer is recognized as one of the worst gynecologic malignancies associated with rapid metastasis and poor overall survival rate. The identified valuable molecular biomarkers criticize importance of timely diagnosis for ovarian cancer. Salusin-ß levels are dramatically increased in women with polycystic ovarian syndrome. However, the roles of salusin-ß in ovarian cancer have yet to be fully elucidated. A total of 57 paired ovarian cancer specimens and matched adjacent normal tissues were used to measure the salusin-ß levels. The prognostic value of salusin-ß for tumor progression and survival rate was investigated. The effects of salusin-ß on ovarian cancer cell proliferation and epithelial-mesenchymal transition were also explored. The expression of salusin-ß was significantly increased in ovarian cancer tissue specimens compared with matched normal adjacent tissue (P<0.05). The high salusin-ß level was closely related with FIGO stage and lymph node metastases. The ovarian cancer patients with high salusin-ß had a shorter overall survival (P<0.05). Salusin-ß obviously enhanced the proliferation and epithelial mesenchymal-transition of SKOV3 cells. Furthermore, salusin-ß substantially decreased the expression of p-GSK-3ß and GSK-3ß, but stimulated the ß-catenin expression and downstream genes of wnt/ß-catenin including cyclin D1 and C-myc. Our data demonstrated for the first time that upregulated salusin-ß may be a novel independent prognostic biomarker for overall survival of ovarian cancer. Salusin-ß accelerated the proliferation and epithelial mesenchymal transition of ovarian cancer cells at least partly via activation of Wnt/ß-catenin signaling pathway. Salusin-ß may be an important target for therapeutic intervention in ovarian cancer.

Preoperative platelet count improves the prognostic prediction of the FIGO staging system for operable cervical cancer patients.

BACKGROUND: Increased platelet has been identified as an independent and unfavorable prognostic indicator in various cancers including cervical cancer. In our study, the prognostic value of preoperative platelet count combining with FIGO (International Federation of Gynecology and Obstetrics) stage in patients with operable cervical cancer was investigated. METHODS: A large cohort study including 800 operable cervical cancer patients was conducted from May 2005 to December 2012. Cancer-related biomarkers such as platelet count, hematocrit, hemoglobin, RDW was evaluated together with FIGO staging system in stage IA1-IIA2 cervical cancer patients. The prediction validity of platelet together with FIGO stage was then evaluated by receiver operating characteristic (ROC) curve, and the areas under the curve (AUCs) were compared by Z test. RESULTS: Univariate cox proportional hazard analysis demonstrated that hematocrit, platelet count, hemoglobin, FIGO stage, tumor differentiation, PLN (pelvic lymph node metastasis), LVSI (vascular lymph node invasion) were associated with overall survival (OS) and disease free survival (DFS), instead of RDW (red cell distribution width), age and histological subtype. Multivariate analysis demonstrated that preoperative platelet and FIGO stage were independent predictors for OS and DFS in cervical cancer. Furthermore, significant improvements were found after the combination of platelet count and FIGO stage in predicting OS and DFS for cervical cancer patients (P=0.0128 and P=0.0385, respectively). CONCLUSIONS: Combination of platelet count and FIGO stage improved the prediction performance of FIGO staging and provide additional risk stratification for operable cervical cancer patients.

Assessment of ADMA, estradiol, and progesterone in severe preeclampsia.

BACKGROUND: Decreased nitrous oxide (NO) levels are crucial factors in severe preeclampsia (sPE), and asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of NO synthetase. Steroid hormones are closely related to the vascular endothelium. This study determined the levels of and correlations between ADMA, estradiol (E2), and progesterone (Pg) in sPE to investigate the roles of these factors in this disease. METHODS: Sixty-two sPE patients (sPE group) were divided into the sPE1 subgroup (28(+1)-32(+0) weeks of pregnancy), the sPE2 subgroup (32(+1)-36(+0) weeks), and the sPE3 subgroup (36(+1)-40(+0) weeks) and 75 normal pregnant women (NC group) were divided into the NC1 subgroup (28(+1)-32(+0) weeks of gestation), the NC2 subgroup (32(+1)-36(+0) weeks), and the NC3 subgroup (36(+1)-40(+0) weeks). Serum and placental ADMA levels were measured by enzyme-linked immunosorbent assay (ELISA), and serum E2 and Pg concentrations were determined by the chemilumineseent immunoassay (CLIA). RESULTS: ADMA concentrations in both the placenta and the maternal serum were significantly higher in the sPE group (p < 0.05). Higher ADMA contents were observed in the placenta relative to the maternal serum (p < 0.05). Serum E2 levels were significantly lower in the sPE group (p < 0.05). For Pg, the only significant difference was observed between the sPE1 and NC1 subgroups (p < 0.05). The Pg/E2 ratios in the sPE groups were significantly higher, with a significant high positive correlation between Pg/E2 ratios and serum ADMA levels. CONCLUSION: Increased serum levels of ADMA in sPE may result from increased secretion from the placenta, and the increased Pg/E2 ratio may play a role in the development of sPE by aggravating ADMA.

A novel prognostic inflammation score predicts outcomes in patients with ovarian cancer.

BACKGROUND: Systemic inflammation and host immunological nutritional status play important roles in the tumorigenesis of malignant cancer. A novel prognostic inflammation score (PIS) based on preoperative serum albumin and neutrophil to lymphocyte ratio (NLR) was designed. We explored its prognostic value in ovarian cancer. METHODS: 143 patients with ovarian cancer were enrolled in this retrospective study. The association of the PIS with clinicopathologic parameters was analyzed. The prognostic significance was determined by univariate and multivariate cox survival analyses. RESULTS: Both univariate and multivariate analyses showed that NLR and albumin were independent prognostic factors for overall survival (OS) and progression-free survival (PFS). An inverse correlation was observed between the NLR and serum albumin concentration. The novel prognostic inflammation score (PIS) was shown to be a significant predictor for OS and PFS (both P<0.001) according to multivariate analysis. Additionally, low PIS was associated with advanced tumor stage (P<0.001), metastasis (P<0.001) and preoperative high PLR (P<0.001). CONCLUSIONS: The PIS is a novel but promising prognostic score in ovarian cancer. It is a significant prognostic marker adjusted for clinicopathologic characteristics to further identify patients' survival differences.

Cervical cancer systemic inflammation score: a novel predictor of prognosis.

Inflammation contributes to development and progression in a variety of cancers, including cervical cancer. We developed a novel cervical cancer systemic inflammation score (CCSIS) based on the preoperative platelet-to-lymphocyte ratio (PLR) and serum albumin levels. A retrospective analysis of clinical data from 795 patients with operable cervical cancer was then conducted to investigate the prognostic value of CCSIS and its association with the patients' clinicopathological features, overall survival (OS), and disease-free survival (DFS). CCSIS was predictive of OS and DFS. High CCSIS was correlated with more advanced FIGO stages, poor tumor differentiation, and the presence of PLN and LVSI. Both albumin levels and the PLR were independent prognostic indicators for operable cervical cancer. The use of the CCSIS could improve risk stratification and traditional clinicopathological analysis in cervical cancer.

Overexpression of astrocyte elevated gene-1 (AEG-1) in cervical cancer and its correlation with angiogenesis.

OBJECTIVES: To explore the expression of astrocyte elevated gene-1 (AEG-1) in cervical cancer and analyze its correlation with microvascular density (MVD), nuclear factor kappaB (NF-kB p65) and vascular endothelial growth factor (VEGF). MATERIALS AND METHODS: Immunohistochemical MaxVision method was adopted to detect the expression level of AEG-1, NF-kB p65 and VEGF in 45 samples of invading cervical cancer and 12 samples of cervicitis from The First Affiliated Hospital of Wenzhou Medical University. Tumor microvascular endothelial marker CD34 combined with Weidner was used to determine the MVD in cervical cancer tissue. The positive expression and staining conditions of AEG-1, NF-kB p65 and VEGF in cervical cancer tissues were observed under a light microscope. Correlations between expression of AEG-1 protein and those of NF-Kb p65 and VEGF, as well as MVD, were analyzed using Pearson correlation. RESULTS: The expression levels of AEG-1 were 0.186±0.043 in cervical cancer and 0.051±0.002 in chronic cervicitis (p<0.01). Moreover, expression of AEG-1 was related to vascular invasion and lymphatic metastasis of cervical cancer (p<0.01), but not with age of the patients, differentiation degree, tumour size, pathological type and parametrial infiltration (p>0.05). Pearson correlation analysis showed that the expression of AEG-1 was linked with NF-kB p65 (r=0.501, p=0.000), VEGF (r=0.718, p=0.000) as well as MVD in cervical cancer tissue (r=0.815, p=0.000). CONCLUSIONS: AEG-1 is highly expressed in cervical cancer and promotes angiogenesis, which might be related to the fact that AEG-1 activating the signal pathway of NF-kB could up-regulate the level of VEGF expression.

Piperlongumine induces apoptosis and synergizes with cisplatin or paclitaxel in human ovarian cancer cells.

Piperlongumine (PL), a natural alkaloid from Piper longum L., possesses the highly selective and effective anticancer property. However, the effect of PL on ovarian cancer cells is still unknown. In this study, we firstly demonstrate that PL selectively inhibited cell growth of human ovarian cancer cells. Furthermore, PL notably induced cell apoptosis, G2/M phase arrest, and accumulation of the intracellular reactive oxidative species (ROS) in a dose- and time-dependent manner. Pretreatment with antioxidant N-acety-L-cysteine could totally reverse the PL-induced ROS accumulation and cell apoptosis. In addition, low dose of PL/cisplatin or paclitaxel combination therapies had a synergistic antigrowth effect on human ovarian cancer cells. Collectively, our study provides new therapeutic potential of PL on human ovarian cancer.

Ovarian cancer initially presenting with isolated ipsilateral superficial inguinal lymph node metastasis: a case study and review of the literature.

Isolated superficial inguinal metastases without any extended intra-abdominal spread is a rare event in patients with ovarian carcinoma. Here we report an isolated superficial inguinal metastasis in a patient with primary ovarian cancer. A 54-year-old Chinese patient with primary ovarian cancer, had an isolated painless enlarged right groin swelling (3×2cm) as the only manifestation, preoperative pathology confirmed metastatic adenocarcinoma. Gynecologic examination, transvaginal ultrasonography of the abdominopelvic cavity revealed a 5-cm mixed, right adnexal mass. At exploratory laparotomy, there was little intra-abdominal tumor dissemination but 100 ml of faint yellow peritoneal fluid and a 5-cm right ovarian tumor with intact capsule. Staging operation was performed and postoperative pathology confirmed adenocarcinoma located within right ovarian, with no evidence of involvement of other sites. Then the patient received adjuvant chemotherapy for Stage IVB. Five years later, the patient is currently still alive without evidence of recurrent disease. This case indicate that ovarian carcinoma isn't a disease localized only within the intra-peritoneal cavity, isolated superficial inguinal lymph node metastasis might occur in rare cases via potential lymphatic and (or) hematogenous route under special conditions. We propose the need to investigate the possible mechanisms, risk factors, metastatic patterns, the biology and natural history of such patients in a large-scale and multicenter analysis. Furthermore, efforts should be made for earlier and differential diagnosis and finally prolong survival time for such patients.

Triterpenoids as reversal agents for anticancer drug resistance treatment.

Overexpression of ATP-binding cassette (ABC) transporters in cancer cells results in multidrug resistance (MDR), which is one of the major obstacles in the treatment of cancer patients. None of the strategies to overcome MDR has been successfully applied in the clinic until now. Plenty of evidence shows that some triterpenoids function as reversal agents of MDR for anticancer drug resistance treatment. Here, we review the latest findings of reversing cancer MDR with triterpenoids. Findings are summarized showing that triterpenoids are MDR modulators and potential chemosensitizers. Finally, we contemplate future prospects of modulating MDR in the clinic.

Non-suture technique for rabbit oviduct anastomosis with 2-octyl cyanoacrylate: a histopathologic and biomechanical analysis.

AIM: The aim of this study was to investigate histological and biomechanical properties of oviduct anastomosis with 2-octyl cyanoacrylate (OCA) in the rabbit. MATERIAL AND METHODS: Sixty female rabbits were randomly divided equally into three groups: A (control), B (traditional catgut suture), and C (non-suture technique using OCA). After suture or OCA anastomosis, gross examination (adhesion formation) and histopathology (hematoxylin-eosin), ultrastructure (transmission electron microscopy), and biomechanics (bursting pressure) on para-anastomotic site were investigated on oviduct taken at 1 (A1, B1, C1) and 4 (A2, B2, C2) weeks, respectively. RESULTS: Adhesion score in group B was more severe than that in groups A and C at 1 and 4 weeks. Histopathology showed that acute endosalpingitis in group B was the most intense at 1 week, followed by significantly more tissue stimulation induced by catgut and foreign-body giant cells in group B than in group C at 4 weeks. Ultrastructural damage of ciliated cells was reversed partly (B2) and completely (C2) at 4 weeks. Bursting pressure in C1 was weaker than that in B1, followed by no significant difference at 4 weeks. CONCLUSION: Non-suture using OCA for oviduct anastomosis can be accepted as a new-perspective technique.

[Effects of Brucea javanica oil emulsion on human papilloma virus type 16 infected cells and mechanisms research].

OBJECTIVE: To explore the in vitro inhibitive effect and underlying mechanisms of Brucea Javanica oil emulsion (BJOE) on human papilloma virus (HPV) type 16 infected cells. METHODS: The HPV16 E61E7 immortalized human ectocervical Ect1/E6E7 cell line and the CaSki cell line were selected as the in vitro models of premalignant cervical lesion and cervical cancer respectively. After treated with BJOE at different concentrations (5, 10, 20, and 40 microg/mL) at the operation time points (24, 48, and 72 h), the effects of BJOE on proliferative activities were measured by MTT assay. The morphologic changes of cell apoptosis stained with Hochest 33,258 were observed by fluorescence microscope. The effect on the cell apoptosis rate was analyzed by Annexin V-FITC/PI double-labeled flow cytometry. The mRNA expressions of HPV16 E6 and E7 were determined by semi-quantitative RT-PCR. The protein expressions of HPV16 E6, E7 oncogene, and specifically interacted p53, Rb antioncogene were stained by immunocytochemical staining (Elivison two-step procedure). RESULTS: (1) The proliferative activities of the Ect1/E6E7 cell and the CaSki cell treated with BJOE at different concentrations (5, 10, 20, and 40 p g/mL) at the operation time points (24, 48, and 72 h) were obviously inhibited, showing dose- and time-dependent manners (P <0.05). (2) Typical changes of apoptosis were observed in both HPV 16 positive cell lines after treated with BJOE. The cell apoptosis rates increased markedly after being cultured with BJOE at different concentrations (5, 10, and 20 microg/mL) for 48 h (P < 0.05). (3) After treated with BJOE at different concentrations (5, 10, and 20 microg/mL) for 48 h, the HPV16 E6 and E7 mRNA relative expressions in both HPV 16 positive cell lines decreased significantly (P < 0.05). (4) After treated with BJOE at different concentrations (5, 10, and 20 microg/ mL), the expressions of HPV16 E6, E7, and mutant p53 protein decreased gradually (P < 0.05), while the Rb protein expression increased gradually (P < 0.05). CONCLUSIONS: BJOE showed obvious in vitro inhibitory effects on HPV type 16 infected cells. Its underlying mechanisms might be possibly associated with down-regulating expressions of HPV16 E6 and E7 oncogenes.

Inhibitory effects of methotrexate on spontaneous motility and Cajal-like type of tubal interstitial cells in rabbit oviduct.

OBJECTIVE: To study the adverse biomechanical effects of methotrexate (MTX) on spontaneous tubal motility and on a widely distributed Cajal-like type of tubal interstitial cells (t-ICC) in rabbits. In our previous study, MTX was confirmed to cause acute endosalpingitis, and ultrastructural and steroid receptor damage in rat's endosalpinx in a dose-dependent manner. DESIGN: Differences in spontaneous tubal contractions and cellular distribution of t-ICC in isthmus were evaluated in response to MTX. SETTING: Medical school research laboratory. ANIMAL(S): Twenty nonpregnant female New Zealand albino rabbits in estrus stage were divided equally into four groups. INTERVENTION(S): Rabbits received IM MTX (1, 5, 10 mg/kg body weight) and controls received physiological saline. MAIN OUTCOME MEASURE(S): On day 7, in vitro motility studies measuring spontaneous tubal contractions were performed, and cellular distribution of t-ICC was determined by immunohistochemistry. RESULT(S): Methotrexate produced a concentration-dependent inhibition of spontaneous isthmus contractions (frequency in 5, 10 mg/kg groups, and amplitude in 1, 5, 10 mg/kg MTX groups). It decreased significantly compared with the control group. Meanwhile, MTX at 5, 10 mg/kg decreased the population of c-kit immunoreactive t-ICC significantly. CONCLUSION(S): The decreased t-ICC may contribute to the diminished tubal smooth muscle contractility caused by MTX as observed. Tubal interstitial cells might be new potential targets for a variety of dysfunctional tubal motility diseases.