A novel visceral pain model of uterine cervix inflammation in rat.

Treatment of visceral pain originating from the uterine cervix is a substantial clinical problem. The underlying mechanisms of such visceral pain remain unclear mainly due to a lack of reliable model. This study aimed to develop and evaluate the performance of a rat model of pain induced by uterine cervix inflammation. Rats were randomized to six groups according to the solution injected into the uterine cervix: normal saline, vehicle, capsaicin (0.3 mg, 0.6 mg, 0.9 mg), capsaicin 0.9 mg + morphine (n = 15 in each group). Spontaneous behaviors after cervical injection were recorded by a computerized video system and analyzed offline. An equation for calculating a novel pain score was derived from particular behaviors, based on Pearson's correlation analysis and regression analysis. c-Fos expression in the spinal cord was detected. The pain score and c-fos expression in the spinal cord were highest in the 0.9 mg capsaicin group and lowest in the normal saline and vehicle groups (P < 0.05). Intrathecal morphine significantly decreased the pain score (P < 0.05) and c-fos expression in the spinal cord (P < 0.05). Injection of capsaicin into the uterine cervix in rats could be a practical model of inflammatory cervical pain, which can be evaluated using our novel pain score. This model will provide further insight into the mechanism underlying visceral pain originating from the uterine cervix.

Ainsliaea fragrans champ. Extract prevents cervicitis in BALB/c mice and regulates MyD88-NF-κB signaling pathway in MALP-2-stimulated RAW264.7 cells.

Ethnopharmacological relevance Ainsliaea fragrans Champ. (A. fragrans) is used to treat infection of the lower genital tract in gynecology, such as cervicitis and pelvic inflammatory disease. This study analyzed the therapeutic efficiency of A. fragrans on cervicitis and the inhibition mechanism of AF-p2 in MALP-2-stimulated RAW264.7 cells. Materials and methods The anti- Ureaplasma urealyticum (Uu) activity of A. fragrans and AF-p2 were determined by antimicrobial susceptibility testing. The activity of A. fragrans extracts (AFext) was evaluated in female BALB/c mice with cervicitis induced by Uu. Furthermore, the therapeutic mechanism of AFext and AF-p2 on myeloid differentiation factor 88 (MyD88) pathway were studied in macrophage activating lipopeptide-2 (MALP-2) irritated RAW264.7 cells. Results AFext could suppress the proliferation of Uu in vitro, including the azithromycin resistant strains. Meanwhile, AFext prevented cervicitis caused by Uu infection in BALB/c mice. Moreover, both AFext and AF-p2 could significantly suppress the nitric oxide (NO) production as well as other proinflammatory cytokines (IL-1ß,IL-6,TNF-α) in MALP-2 stimulated RAW264.7 cells. Moreover, AF-p2 also down-regulated iNOS, p65, Iκ-Bα, MyD88 and cyclooxygenase-2 (COX-2) levels in RAW264.7 cells. Conclusion This study indicated that AFext had a therapeutic effect in cervicitis induced by Uu infection. Furthermore, the lead compound AF-p2 showed an anti-infectious effect in MALP-2 irritated RAW264.7 cells through downregulating MyD88-NF-κB signaling pathway.

An integrative investigation of the therapeutic mechanism of Ainsliaea fragrans Champ. in cervicitis using liquid chromatography tandem mass spectrometry based on a rat plasma metabolomics strategy.

Cervicitis is an extremely common gynecological disease and can be induced by diverse factors such as Neisseria gonorrhoeae, Chlamydia trachomatis, and Mycoplasma genitalium infections. Long-term unhealed cervicitis may lead to a series of diseases including endometritis, salpingitis, pelvic inflammatory disease, and chorioamnionitis. However, the pathogenesis of cervicitis remains unknown. Ainsliaea fragrans Champ. (AFC) has been widely used in clinical treatment of cervicitis. In the present study, we performed an integrative investigation involving histopathology analysis and non-target plasma metabolomics analysis in a cervicitis rat model induced by phenol mucilage, using ultra-performance liquid chromatography coupled with a tandem quadrupole time-of-flight mass spectrometry approach. Based on the integrative investigation, marked metabolomic differences were identified between the cervicitis and control groups using multivariate analysis. As a result, 32 potential biomarkers were identified in the response to cervicitis, and were involved in arachidonic acid metabolism, linoleic acid metabolism, primary bile acid biosynthesis, taurine and hypotaurine metabolism, pantothenate and CoA biosynthesis, and glycerophospholipid metabolism. After treatment, a total of 27 potential biomarkers exhibited altered levels in the AFC group compared to the model group, and 12 metabolites including 1-stearoylglycerophosphoinositol, bolasterone, lysoPC(16:0), lysoPC(20:4), lysoPC(P-16:0), lysoPC(P-18:0), lysoPC(P-18:1), stearoylcarnitine, taurine, lysoPC(17:0), 20-hydroxyeicosatetraenoic acid, and 1-arachidonoylglycerophosphoinositol returned to their normal levels. This study suggested that the therapeutic mechanism of AFC is related to those altered endogenous metabolites.

Host Defense Peptide Expression in Human Cervical Cells and Regulation by 1,25-Dihydroxyvitamin D3 in the Presence of Cytokines and Bacterial Endotoxin.

Host defense peptides (HDPs) in the pregnant female reproductive tract provide protection against infection. The relationship between HDPs and infection/inflammation is poorly understood. Therefore, we investigated the regulation of HDPs by 1α, 25-dihydroxyvitamin D3 (1,25-(OH)2) in the presence/absence of infectious/inflammatory agents. Endocervical epithelial cells (END1/E6E7, n = 6) were exposed to 1,25-(OH)2, calcipotriol, interleukin 1ß (IL-1ß), granulate-macrophage colony-stimulating factor (GM-GSF), and lipopolysaccharide (LPS). Elafin, human beta defensin (hBD2), cathelicidin, secretory leucocyte protease inhibitor, interleukin 8, 1,25-(OH)2 receptor, and toll-like receptor 4 (TLR4) expression was determined using quantitative polymerase chain reaction and/or enzyme-linked immunosorbent assay. Host defense peptide gene and protein expression was assessed in cervicovaginal cells/fluid, respectively, from first trimester pregnant women (n = 8-12). Interleukin 1ß induced elafin and hBD2. The 1,25-(OH)2 induced cathelicidin expression in the presence of IL-1ß and LPS. The 1,25-(OH)2 also attenuated IL-1ß-induced IL-8 expression and LPS enhancement of TLR4. Host defense peptides and TLR4 profiles in cervicovaginal cells and fluid samples from pregnant women were similar to END1/E6E7 cells. In conclusion, HDPs are differentially regulated in END1/E6E7 cells. The 1,25-(OH)2 induction of cathelicidin and suppression of IL-8 highlights a mechanism by which 1,25-(OH)2 supplementation could enhance the pregnant innate immune defenses.

[THE ROLE OF NO AND MEXIDOL IN CONTRACTILITY OF OVARIAN AND CERVICAL PARTS OF UTERUS UNDER THE CONDITION OF IMMUNE-MEDIATED INJURY IN MICE].

Contractility of ovarian (OP) and cervical parts (CP) of uterus under the condition of immune-mediated injury which was induced by immunization with bovine serum albumin (BSA) was investigated. It was shown that under the activation of energy-synthesizing function of mitochondria with Mexidol the frequency of reductions in both uterine parts decreased, the amplitude and contractility index in the OP and CP as well as the duration of the active state in CP increased. Mexidol under the condition of immunization with BSA leads to the decrease in amplitude in 2,6 time and contractility index in 2,2 time in OP and to the increase of them in CP. It was shown that contractility features of ovarian and cervical parts of uterine under the condition of BSA- induced immunization were caused by changes of mitochondria functional state and were associated with nitric oxide.

Prevalence of visible disruption of cervical epithelium and cervical ectopy in African women using Depo-Provera.

Associations between Depo-Provera (injectable, progesterone-only contraceptive) use and visible disruption of cervical epithelium and cervical ectopy were investigated using data collected as part of a cervical cancer screening study in periurban Cape Town, South Africa. Women were interviewed about their contraceptive use, and underwent a gynecologic examination that included two 35-mm photographs of the cervix after application of 5% acetic acid. Photographs of 723 subjects were reviewed (blind to clinical information and using systematic criteria developed before review) for evidence of atrophy and epithelial disruption, including inflammation and ulceration. The percentage of the cervix covered with columnar epithelium (ectopy) was also estimated from the photographs. A random sample of 85 photographs was reviewed again for reliability. A total of 121 current users of Depo-Provera were no more likely to have evidence of epithelial disruption (38%) than 574 nonusers (39%), odds ratio (OR) = 1.37, 95% CI: 0.89-2.11 adjusting for age and parity. The prevalence of significant ectopy (columnar epithelium covering > 10% of the cervix) was also no different among current Depo-Provera users (OR = 1.22, 95% CI: 0.80-1.86 adjusting for age and parity). Reliability of visual scoring of epithelial disruption and ectopy was excellent (kappa = 0.8). Although the underlying prevalence of visible disruption of cervical epithelium was very high, current use of Depo-Provera was not associated with increased prevalence of visible disruption of the cervical epithelium or with ectopy in this sample of African women.

PIP: The relationship between cervical epithelium disorders and cervical ectopy and Depo-Provera use was investigated among African women. The women, aged 35-65 years, had gynecologic examinations, which included various cervical cancer screening tests including two 35-mm colored photographs of the cervix. Cervical photographs from 723 participants were reexamined for the signs of disruption. Results showed that among 121 current Depo-Provera users, 38% showed no steady increase in epithelial disruption compared with 34% of past users and 49% of nonusers. There was no difference in the odds ratio (OR = 1.37; 95% CI: 0.89-2.11). The percentage of women with significant ectopy was higher in current and past users of Depo-Provera, but no trends remain after age adjustment and parity (OR = 1.22; 95% CI: 0.80-1.86). The credibility of the diagnosis of cervical disruption combined (either ulceration, inflammation, or athropy) was excellent (K = 0.8). However, the results were doubtful in light of the concept that simple epithelial thinning may account for putative effects of Depo-Provera. The validity of this method needs further investigations.

Effects of multiple applications of benzalkonium chloride and nonoxynol 9 on the vaginal epithelium in the pigtailed macaque (Macaca nemestrina).

OBJECTIVE: Safe and effective vaginally applied microbicides could help to control the continuing spread of sexually transmitted diseases. STUDY DESIGN: This study used nonhuman primates to test the effects of multiple applications of nonoxynol 9, benzalkonium chloride, or a combination on vaginal flora and lower reproductive tract tissues. Fourteen monkeys (Macaca nemestrina) received daily vaginal applications of nonoxynol 9, benzalkonium chloride, or both for 3 to 4 days. Vaginal microflora and colposcopic observations were made at baseline and during and after completion of treatments. Cervical biopsy specimens were collected from a subset of animals. RESULTS: Cervical erythema and vaginal erythema were observed in all 3 treatment groups. Cervical papillae and epithelial disruption were present in both the nonoxynol 9 and the nonoxynol 9 plus benzalkonium chloride groups. Vaginal epithelial disruption was noted in both the benzalkonium chloride and the nonoxynol 9 plus benzalkonium chloride groups. Cervical biopsy specimens from each group revealed acute inflammatory infiltrates with occasional plasma cells and lymphoid follicles. Detection of most microorganisms, including viridans streptococci, decreased in the benzalkonium chloride and the nonoxynol 9 plus benzalkonium chloride groups. Detection of Lactobacillus species decreased in the benzalkonium chloride group. All microflora levels recovered after several days without microbicide use. CONCLUSIONS: Although nonoxynol 9 is currently the only microbicide approved for use as a spermicide in the United States, its repeated use may be detrimental to the epithelial tissues of the female reproductive tract. Benzalkonium chloride, currently approved for use in other countries, not only may damage epithelial tissues but also appears to reduce the population of potentially protective Lactobacillus species in the vagina.

[Chromo-colposcopic images during oral contraception]. / Immagini cromocolposcopiche in corso di contraccezione orale.

The Authors report chromocolposcopic pictures with AZEA (Azure 2 + alcohol-soluble eosine + ethanol) and related histologic pictures in six women under oral contraception. Columnar epithelium develops violet or blue-violet areas histologically corresponding to endocervicitis. Aceto-white epithelium develops reddish color in case of medium-mature metaplasia, brown-violet in case of mature metaplasia, lilac in case of superficial koilocytosis on metaplasia. Native epithelium appears dark brown, or yellow-brown in case of koilocytosis.

[Morphofunctional features of the cervix uteri in women using hormonal contraception]. / Morfofunktsional'nye osobennosti sheiki matki u zhenshchin, primeniaiushchikh gormonal'nuiu kontratseptsiiu.

PIP: The causes of precancerous and cancerous diseases of the cervix are disputed. In women with menstrual disorders usually benign cervical disease is 5 times higher. In the 1960s the theory of hormonal genesis of cervical disease was advanced as similar pathogenetic processes in the cervix, endometrium, myometrium, and breasts resulted in hyperplastic changes in these organs. Ectopia can occur during sexual maturation under the influence of sex hormones. The maximum frequency (65.5%) of ectopia occurred up to age 20. Cervical ectopia can occur under use of oral contraceptives (OCs) for 6-12 months but it vanishes after discontinuation. In a study of 17,942 women aged 18-58 increased risk of preinvasive carcinoma of the cervix was found under longterm use of OCs. Increased frequency of cervical intraepithelial neoplasm from .9/10,000 women/year to 2.2/10,000 women/year was found only under longterm (up to 8 years) of OC use. An epidemiological investigation of 47,000 women using OCs for up to 10 years concluded that there was significant increase of frequency of cervical cancer compared with nonusers. It was 4 times higher in those taking OCs for over 10 years, although longterm use reduced uterine and ovarian cancer. Adenomatous hyperplasia of the endocervix was 14 times more frequent in OC users. In a sample of 128 women, 44% of whom were OC users, 24% had microglandular hyperplasia. Under the use of the 3-phase preparation Trisiston for 6 months-1 year ectopia was diagnosed in 13.6% of women that disappeared after cessation of use. Early cancer and dysplasia disappeared in 1/3 of women taking Enovid for 6-30 months after diagnosis. OCs promote the prophylaxis against genital cancer because women taking OCs undergo gynecological and cytological examinations more often, thus precancerous changes can be diagnosed early. The optimal and the safest method of contraception has to be chosen to minimize the effect on the cervix.^ieng

A phase I trial of beta-all-trans-retinoic acid delivered via a collagen sponge and a cervical cap for mild or moderate intraepithelial cervical neoplasia.

A phase I trial was conducted of the vitamin A derivative beta-all-trans-retinoic acid (vitamin A acid; TRA), delivered via a collagen sponge and cervical cap for mild or moderate intraepithelial cervical neoplasia. On the basis of known skin and mucosal membrane toxicity, a concentration of 0.05% TRA in a cream-based vehicle was selected as the starting dose and was escalated later with the use of a modified Fibonacchi scale. The delivery device and the TRA were changed daily for 4 days, and side effects were assessed on days 1, 2, 3, 4, 8, and 30 by clinical and colposcopic examination. Vaginal, cervical, and systemic toxicity were evaluated in 35 patients. No dose-related systemic effects were found; mild cervical inflammation increased in many patients at higher doses. Unacceptably high vaginal toxicity was reached at a TRA concentration of 0.484%. A concentration of 0.372% TRA is recommended for use in phase II trials in mild and moderate cervical intraepithelial neoplasia.

Complications of pregnancy and labor in former oral contraceptive users.

Complications of pregnancy and delivery, and obstetric interventions, were studied in a sample of Israeli women questioned post-partum about contraceptive use. The 2,953 women who had used the pill were compared with 13,630 controls. There were no significant differences between users and controls in the frequencies of bleeding in pregnancy, premature rupture of membranes, placenta previa, placental abruption, fetal distress or asphyxia, ABO incompatibility, hydramnios, transverse lie, cephalopelvic disproportion, and persistent occipitoposterior or post-partum hemorrhage. New cases of hypertension, varicose veins, thrombophlebitis, urinary tract infection, and cervicitis were reported in primigravidae and multigravidae without past histories of these conditions; there were no differences between oral contraceptive users and controls other than an excess of cervicitis in primigravidae among former users. There was a slight decrease in normal deliveries in former oral contraceptive users due to an increase in inductions of labor. On the other hand, rates of forceps and vacuum deliveries, caesarian sections, and interventions in the third stage did not differ between former oral contraceptive users and controls. These results indicate that former oral contraceptive users can anticipate the same frequency of complications of pregnancy and labor as women who have used other, or no, methods of contraception.