Identification of a vaginal pacemaker: An immunohistochemical and morphometric study.

Vaginal electric waves spread caudally in the vagina. We investigated the hypothesis that electric waves originate from a centre of interstitial cells of Cajal (ICCs) in the proximal vagina. Specimens (0.75 x 0.75 cm) were obtained from the vaginal walls of 23 cadavers (age 38.2 +/- 10.2 years). Sections were prepared for immunohistochemical investigations using the specific ICC marker, C-kit. Morphometric studies for image analysis using a Leica imaging system were performed. C-kit positive cells were detected in vaginal smooth muscle. Results from image analyser revealed that mean area percent of positive immunoreactivity for C-kit in the upper part of posterior vaginal wall was significantly higher (p < 0.0001) than of areas in other vaginal walls, and also significantly higher (p < 0.05) in circular than in longitudinal muscle layer. Studies have shown that the greatest collection of ICCs occurred in the upper part of the posterior vaginal wall. The vaginal electric waves are suggested to originate from this 'centre' and spread caudally.     

The electrovaginogram: study of the vaginal electric activity and its role in the sexual act and disorders

Objectives We investigated the hypothesis that the vagina generates electric waves which effect vaginal contraction during penile thrusting.Methods In 24 healthy female volunteers, the electric waves of the vagina were recorded by two electrodes applied to its wall. The vaginal pressure was registered by a manometric tube. The electric waves and vaginal pressure were recorded at rest and on vaginal distension by condom in increments of 10 ml of carbon dioxide. The test was repeated after vaginal anesthetization proximally and distally to the electrodes.Results Slow waves (SWs) were recorded from the two electrodes with regular rhythm and similar frequency, amplitude and conduction velocity. They were randomly followed or superimposed by action potentials (APs). Vaginal pressure increase was coupled with APs. Large-volume condom distension significantly increased the electric waves variables and pressure. Upon vaginal anesthetization, the electric waves were recorded proximal but not distal to the anesthetized area.Conclusions Electric waves could be recorded from the vagina. They spread caudad. A pacemaker was postulated to exist at the upper vagina evoking these waves. The electric waves seem to be responsible for the vaginal contractile activity. Large-volume vaginal distension effected an increase in the vaginal electric waves and pressure which probably denotes increased vaginal muscle contraction. It appears that penile thrusting during coitus stimulates the vaginal pacemaker which effects an increase in vaginal electric activity and muscle contractility and thus leading to an increase in sexual arousal. The vaginal pacemaker seems to represent the G spot, which is claimed to be a small area of erotic sensitivity in the vagina. The electrovaginogram may act as a diagnostic tool in the investigation of sexual disorders.     

Study of interstitial cells in the penis: human study

IntroductionSpecialized pacemaker cells, similar to the interstitial cells of Cajal (ICC) of the gut, have been detected in the urinary organs and are thought to pace their motile activity.AimWe investigated the hypothesis that such cells could also exist in the corpora cavernosa (CC) of the human penis.MethodsDuring the treatment of Peyronie’s disease in 11 subjects (age 42.6 ± 3.2 SD years), 3 × 3 mm strips were excised from each of the two CC and subjected to C-kit immunohistochemistry. Controls for the specificity of the antisera consisted of incubation of the tissue with normal rabbit serum substituted for the primary antiserum.Main Outcome MeasuresInterstitial cells similar to ICC could exist in the CC based on C-kit immunohistochemistry.ResultsC-kit positive branched interstitial cells were detected in the CC. They were clearly distinguishable from the smooth muscle cells that were C-kit negative and non-branched. Although the mast cells were C-kit positive, they had a smooth body surface.ConclusionsInterstitial cells have been identified in the CC. They are similar to the ICC and may be responsible for initiating the slow waves recorded from the smooth muscle cells and for controlling their activity. We assume that a deficiency or absence of these cells may affect the erectile function of the patient. Further studies are needed to explore the role of these cells in erection. Shafik A. Study of interstitial cells in the penis: Human study. J Sex Med 2007;4:66–71.     

Epithelioid vascular leiomyoma of the uterus mimicking glomangiomyoma

Case report: An unusual case of epithelioid vascular leiomyoma mimicking glomangiomyoma arising in the uterine corpus of a 55-year-old Japanese woman is presented. The surgically resected uterine mass, measuring 4.0 × 3.5 cm², demonstrated a dark red well-circumscribed tumor. Histologically, the rounded epithelioid cells around the dilated vessels showed gradual transition to spindled smooth muscle cells. Immunohistochemistry was positive for smooth muscle actin, but negative for collagen IV. Conclusion: The experience in this case emphasizes that glomangiomyoma-like feature is interesting and might indeed be a new histological variant of uterine leiomyomas.     

Uterine effect of cervical buffeting: role of the cervicouterine reflex in coitus

OBJECTIVE: To investigate the hypothesis that cervical buffeting during coitus effects reflex relaxation and dilatation of the corpus uteri (CorU) smooth muscle; that mechanism acts to suck the ejected semen into the CorU. STUDY DESIGN: The CorU response to electrical and mechanical stimulation of the CU was recorded in 22 healthy women (mean age, 38.2 +/- 7.6 SD years). Cervical electrical stimulation was effected by an electromyographic needle electrode and mechanical stimulation by pencil electrode. The test was repeated after individual anesthetization of the CU and CorU and after normal saline injection. RESULTS: Slow waves, superimposed or followed by action potentials, were recorded at rest from the CorU electrode. Electrical or mechanical stimulation of the CU effected a significant reduction of the electric wave parameters and pressure in the CorU; this response was reproducible. Separate stimulation of the anesthetized CU or CorU did not evoke the response, while saline infiltration did. CONCLUSION: CU stimulation resulted in diminished CorU electrical activity and pressure; that presumably indicates uterine wall relaxation. Repeated CU buffeting apparently leads to an intermittent uterine pressure decline, with resulting semen siphonage from the vagina to the uterus. Diminished CorU electrical activity and pressure on cervical stimulation or buffeting might be mediated through the cervicouterine inhibitory reflex. This reflex seems to be evoked by penile thrusting during coitus and may prove of diagnostic significance in sexual disorders.     

Study of the effect of cervix uteri buffeting on the vaginal musculature with identification of the cervico-vaginal inhibitory reflex

Objectives During penile thrusting, the penis repeatedly buffets the cervix uteri (CU). We investigated the hypothesis that CU stimulation induces a change in the motor activity of the vaginal wall musculature.Materials and methods The response of the vaginal wall musculature to electrical and mechanical CU stimulation was recorded in 27 healthy women (age 34.8±7.3 years). The test was repeated after separate anesthetization of CU and vagina and after normal saline injection.Results Resting electric activity in the form of slow waves (SWs), followed or superimposed by action potentials (APs), was recorded from two electrodes applied to the vagina. The waves displayed the same frequency, amplitude and conduction velocity from the two electrodes. Electrical or mechanical stimulation of the CU effected disappearance of the electric waves; this response was reproducible. Individual stimulation of the anesthetized CU or vagina did not evoke the response, but saline infiltration did.Conclusions CU stimulation resulted in disappearance of the vaginal electric activity which presumably denotes vaginal tone disappearance. Meanwhile, stimulation of the anesthetized CU or vagina did not evoke that response. The data suggested the presence of a reproducible reflex relationship between the CU and vaginal musculature that seems to be mediated through a reflex which we call "cervico-vaginal inhibitory reflex". Disappearance of the vaginal tone probably enhances vaginal penetration and penile thrusting during the sexual act and penile buffeting of the CU. The cervico-vaginal reflex may be incorporated as an investigative tool in sexual act disorders.     

Electrophysiologic activity of the tunica albuginea and corpora cavernosa: possible role of tunica albuginea in the erectile mechanism

IntroductionIt is claimed that the tunica albuginea (TA) shares in the erectile mechanism by compressing the emissary veins passing through it. However, the TA does not contain smooth muscle fibers.AimWe investigated the hypothesis that TA lacks a contractile activity on the emissary veins passing through it.MethodsFourteen healthy male volunteers (mean age 35.2 ± 4.3 years) were studied. The electromyographic (EMG) activity of the TA and corpora cavernosa (CC) was individually recorded in the flaccid and erectile phases by EMG needle electrodes. Recording was performed in the upper, middle, and lower third of the TA and CC on one and then on the contralateral side.Main Outcome MeasuresThe TA lacks a contractile activity on the emissary veins passing through it.ResultsThe EMG of the CC in the flaccid phase recorded regular slow waves and random action potentials. The wave variables in the erectile phase exhibited a significant decrease (P < 0.01) compared with the variables in the flaccid phase of the same subject. The TA EMG showed no electric waves in the flaccid or erectile phases. These recordings were similar from the upper-, middle-, and lower-third of the penis, and were reproducible from the contralateral CC.ConclusionsElectric waves were recorded from the CC in the flaccid phase; wave variables decreased at erection. In contrast, the TA showed no electric waves in the flaccid or erectile phases. It appears that the TA acts as a CC covering sheet which expands passively at erection, and shares in compressing the subtunical venular plexus between it and the tumescent CC. Shafik A, Shafik AI, El Sibai O, and Shafik AA. Electrophysiologic activity of the tunica albuginea and corpora cavernosa: Possible role of tunica albuginea in the erectile mechanism.     

Adaptations of the rat vagina in pregnancy to accommodate delivery

OBJECTIVE: To characterize ultrastructural changes in the rat vagina in pregnancy, delivery, and postpartum, focusing on collagen architecture and smooth muscle cell morphology. METHODS: The vagina of four virgin, four midpregnant, four late pregnant, four immediate, and four late post-vaginal-delivery rats were examined by transmission electron microscopy. Images were classified into one of four categories based on collagen fibril area fraction, with group 1 containing the highest number of collagen fibers per unit area and group 4 containing the lowest. Smooth muscle cells were characterized into three cell types ("synthetic," "intermediate," and "contractile") based on the volume fraction of cytoplasm occupied by organelles compared with myofibrils. RESULTS: Quantitative analysis demonstrated that 76% of collagen fibers in virgin rats were categorized as group 1 or 2 compared with 49% in midpregnant, 40% in late pregnant, and 23% in immediate postpartum animals (P=0.006). Late postpartum tissue seemed similar to virgin tissue (77%). Midpregnant (37%), late-pregnant (34%) and immediate postpartum animals (43%) contained a higher proportion of synthetic smooth muscle cells compared with virgins (20%) and late postpartum animals (21%) (P=.02). Contractile smooth muscle cells predominated in virgin (64%) and late postpartum animals (70%) compared with midpregnant (42%), late pregnant (50%) and immediate postpartum (50%, P=.05). CONCLUSION: In pregnancy, collagen fiber area decreased while smooth muscle cells transformed from a contractile to a synthetic phenotype. The late postpartum period returned to prepregnant levels for both collagen and smooth muscle cell morphologies. It is likely that these changes represent adaptations to minimize trauma to the vagina during passage of the fetus.     

Spontaneous Ca2+ Waves in Rabbit Corpus Cavernosum: Modulation by Nitric Oxide and cGMP

IntroductionDetumescent tone and subsequent relaxation by nitric oxide (NO) are essential processes that determine the erectile state of the penis. Despite this, the mechanisms involved are incompletely understood. It is often assumed that the tone is associated with a sustained high cytosolic Ca²⁺ level in the corpus cavernosum smooth muscle cells, however, an alternative possibility is that oscillatory Ca²⁺ signals regulate tone, and erection occurs as a result of inhibition of Ca²⁺ oscillations by NO.AimsThe aim of this study is to determine if smooth muscle cells displayed spontaneous Ca²⁺ oscillations and, if so, whether these were regulated by NO.MethodsMale New Zealand white rabbits were euthanized and smooth muscle cells were isolated by enzymatic dispersal for confocal imaging of intracellular Ca²⁺ (using fluo-4AM) and patch clamp recording of spontaneous membrane currents. Thin tissue slices were also loaded with fluo-4AM for live imaging of Ca²⁺.Main Outcome MeasureCytosolic Ca²⁺ was measured in isolated smooth muscle cells and tissue slices.ResultsIsolated rabbit corpus cavernosum smooth muscle cells developed spontaneous Ca²⁺ waves that spread at a mean velocity of 65 µm/s. Dual voltage clamp/confocal recordings revealed that each of the Ca²⁺ waves was associated with an inward current typical of the Ca²⁺-activated Cl^- currents developed by these cells. The waves depended on an intact sarcoplasmic reticulum Ca²⁺ store, as they were blocked by cyclopiazonic acid (Calbiochem, San Diego, CA, USA) and agents that interfere with ryanodine receptors and IP₃-mediated Ca²⁺ release. The waves were also inhibited by an NO donor (diethylamine NO; Tocris Bioscience, Bristol, Avon, UK), 3-(5-hydroxymethyl-2-furyl)-1-benzyl indazole (YC-1) (Alexis Biochemicals, Bingham, Notts, UK), 8-bromo-cyclic guanosine mono-phosphate (Tocris), and sildenafil (Viagra, Pfizer, Sandwich, Kent, UK). Regular Ca²⁺ oscillations were also observed in whole tissue slices where they were clearly seen to precede contraction. This activity was also markedly inhibited by sildenafil, suggesting that it was under NO regulation.ConclusionsThese results provide a new basis for understanding detumescent tone in the corpus cavernosum and its inhibition by NO. Sergeant GP, Craven M, Hollywood MA, McHale NG, and Thornbury KD. Spontaneous Ca²⁺ waves in rabbit corpus cavernosum: Modulation by nitric oxide and cGMP. J Sex Med **;**:**–**.     

Morphological and structural changes of umbilical veins and clinical significance in preeclampsia

Objective: Preeclampsia is a common complication of pregnancy that is characterized by high blood pressure. Preeclampsia can have both negative maternal and fetal effects, including impaired fetal development from reduced placental perfusion. Because umbilical vessels provide fetal blood supply, morphological changes in these vessels can critically affect fetal function. This study investigated morphological and structural changes in umbilical veins from 17 controls and 19 preeclampsia cases. Methods: 36 umbilical cords were collected, and take umbilical cord of placental, middle, and fetal segment to H&E and orange G staining. Microscope images were obtained by digital microscope, and wall thickness, lumen diameter, wall-luminal ratio and tunica media thickness, Area of smooth muscle, area of nuclei, nuclei density, and nuclear area density of smooth muscle in tunica media were tested by Image-pro Plus. Results: Lumen diameter of umbilical veins gradually decreased, while wall thickness, tunica media thickness, and wall-luminal ratio gradually increased, from placental to fetal segments of umbilical veins. Area of smooth muscle, area of nuclei, nuclei density, and nuclear area density did not significantly differ from placental to fetal segments of umbilical veins. Compared to controls, (1) Preeclampsia cases had a smaller lumen diameter and greater wall thickness, tunica media thickness, wall-luminal ratio; (2) Preeclampsia cases had higher nuclei density and nuclear area density, and had no difference in area of smooth muscle and area of nuclei. In fetal segments of umbilical veins from preeclampsia cases, correlation analysis showed that wall-luminal ratio negatively correlated with gestation age, birth weight, and Apgar score. Conclusions: These results indicate that umbilical vein of preeclampsia cases has a thickened tunica media from smooth muscle proliferation, which may be an adapted reaction to hypertension. Altogether, this study demonstrates that morphological changes in umbilical veins adversely affect fetal development in preeclampsia cases.     

Effect of inflammatory mediators on the physiology of the human Fallopian tube

The physiological basis for the use of non-steroidal anti-inflammatory drugs (NSAIDs) in the treatment of acute pelvic pain in women was examined in terms of: (i) the electrophysiological responses of epithelial cells in the human Fallopian tube; and (ii) the longitudinal and circular contractions of the myosalpinx. Epithelial cells were grown as a polarized layer in primary culture, and transepithelial potential difference (p.d.) and short-circuit current (I_scc) were recorded using a modified Ussing chamber. The inflammatory mediators histamine (0.1–100.0 µmol l^-1) and platelet activating factor (PAF) (1.9–1900.0 nmol l^-1) increased p.d. and I_scc in a dose-dependent manner. Pre-incubation with the NSAID diclofenac sodium (100 µmol l^-1) inhibited the histamine- and PAF-induced stimulation of p.d. and I_scc. Aspirin (100 µmol l^-1), ibuprofen (100 µmol l^-1), indomethacin (100 µmol l^-1) or naproxen (100 µmol l^-1) were only partially effective. Histamine (0.1–1000.0 µmol l^-1) increased the frequency of contractions of longitudinal and circular smooth muscle in segments of Fallopian tube in vitro, in a dose-dependent manner. Pre-incubation with diclofenac significantly reduced the histamine-induced stimulation of tubal smooth muscle contraction at the higher doses of histamine. The other NSAIDs had no effect. These data provide evidence that diclofenac downregulates acute inflammation in the human Fallopian tube and may be of use as an anti-inflammatory agent in the treatment of pelvic inflammatory disease.     

Placental tumor necrosis factor-α protein expression during normal human gestation

Objective: Placental tumor necrosis factor-α (TNF-α) is a cell signaling protein. During pregnancy, TNF-α induces synthesis of matrix metalloproteinases (MMPs) which allows cytotrophoblasts to reach the spiral arteries deeper within the uterine decidua. TNF-α also augments apoptosis of vascular smooth muscle cells surrounding these arteries. In this study, chorionic villi TNF-α protein expression throughout normal human gestation were investigated.

Methods: Placental chorionic villi tissues obtained from elective surgical terminations of pregnancy and from uncomplicated term births were assayed using EIA kits (Cayman Chemicals, Ann Arbor, MI, Item # 589201).

Results: The median, 25th percentile and 75th percentile values in the first (N?=?99), second (N?=?58) and third trimester (N?=?42) were: 36.46, 27.25, 45.90?pg/100?mg tissue; 55.43, 40.09, 110.88?pg/100?mg tissue; and 16.63, 9.32, 31.92?pg/100?mg tissue, respectively.

Conclusions: Variations in placental TNF-α protein expression noted at different trimesters may suggest gestational age specific roles for the cytokine. The increase in TNF-α protein expression observed in the second trimester may be involved in upregulating synthesis of MMP and in augmenting apoptosis of vascular smooth muscle cells of the spiral arteries. A failure in this second trimester increase in TNF-α protein could contribute to gestational compromise.     

Socio-cultural representations of the vagina

Although the vagina is a rare topic in the social science literature, numerous socio-cultural representations of the vagina can be found throughout Western societies. Such representations offer a range of cultural resources for making sense of the vagina and its functions, and have implications for women's health and well-being. In this paper, we identify and overview seven persistent negative representations of the vagina: the vagina as inferior to the penis; the vagina as absence; the vagina as (passive) receptacle for the penis; the vagina as sexually inadequate; the vagina as disgusting; the vagina as vulnerable and abused; and the vagina as dangerous. In the last sections, we argue that in order to promote women's sexual and reproductive health, it is necessary to challenge such negative representations, and we offer some alternative - and much more positive - representations of the vagina.     

Streptococcus agalactiae in pregnancies complicated by preterm prelabor rupture of membranes

Objective: The main aim of this study was to evaluate the presence of Streptococcus agalactiae (S. agalactiae) in the vagina and the amniotic fluid in pregnancies complicated by preterm prelabor rupture of membranes (PPROM). The next aim was to evaluate the incidence of S. agalactiae early onset sepsis in newborns from PPROM pregnancies, with respect to the presence of S. agalactiae in the vagina and the amniotic fluid.

Methods: Singleton gestations with PPROM between 24?+?0 and 36?+?6 were included. A vaginal swab was obtained, and amniocentesis was performed at admission. The presence of S. agalactiae in the vagina and in the amniotic fluid was assessed by culture and by real-time polymerase chain reaction, respectively.

Results: In total, 336 women were included. The presence of S. agalactiae in the vaginal and amniotic fluid was found in 9% (31/336) and 1% (3/336) of women. One woman had S. agalactiae in the amniotic fluid but was negative for the presence of S. agalactiae in the vaginal fluid. Early onset neonatal sepsis developed in one newborn from pregnancies complicated by the presence of S. agalactiae in the amniotic fluid.

Conclusion: The presence of S. agalactiae in the vagina and amniotic fluid complicated approximately each 10th and each 100th PPROM pregnancy. Cultivation-negative findings of S. agalactiae in the vagina did not exclude the positivity of the amniotic fluid for S. agalactiae and the development of early onset sepsis in newborns.     

Vascular smooth muscle oxygen consumption is reversibly stimulated by sera from women with preeclampsia

Objective: Preeclampsia is a complication of pregnancy that causes maternal vasoconstriction and hypertension. The disease may progress to eclampsia, which is thought to be related to cerebral vasospasm. Although there is evidence for more than one circulating factor that causes endothelial cell dysfunction in preeclampsia, little work has focused on the possibility that vascular smooth muscle function might be directly stimulated by a circulating factor. The aim of this study was to determine whether such a factor or factors could be detected by the vessels. Study Design: Excessive vascular smooth muscle oxygen consumption was used as a screen for metabolic stimulation because pathologic arterial constriction would require oxidative metabolism to generate adenosine triphosphate. De-endothelialized porcine carotid artery (a well-validated model of human arterial contractile function) was exposed to sera from patients with preeclampsia (1:30 dilution) in a sealed chamber with an oxygen electrode, and the rate of oxygen consumption by the tissue was measured. Comparisons with the effects of sera from matched normal pregnant patients and from nonpregnant women were made. Results: Exposure of vascular smooth muscle to sera from women with preeclampsia for 90 minutes resulted in greater oxygen consumption by the tissue (0.66 ± 0.16 μmol O₂ /min per gram of dry weight) than did exposure to sera of matched pregnant and nonpregnant control subjects (0.34 ± 0.08 μmol O₂ /min per gram of dry weight, P < .001, and 0.29 ± 0.03 μmol O₂ /min per gram of dry weight, P < .001, respectively). This stimulation was completely reversed by rinsing. Conclusions: There is a factor in the circulation of women with preeclampsia that has the reversible effect on vascular smooth muscle of accelerating oxygen consumption. We discuss the implications of this observation in terms of known aspects of vascular smooth muscle contractile function. (Am J Obstet Gynecol 1998;179:1534-8.)     

Phosphodiesterase type 1, calcitonin gene-related peptide and vasoactive intestinal polypeptide are involved in the control of human vaginal arterial vessels

Objective

The vagina makes a major contribution to the normal female sexual response cycle. An increase in vaginal blood flow is considered a key event in the mechanism of sexual arousal. Recent research has focused mainly on the cyclic GMP pathway and phosphodiesterase type 5 (PDE5, cyclic GMP specific PDE) in the control of vaginal vascular smooth muscle, whereas only little is known on the role of other key proteins and mediators of cyclic nucleotide mediated signaling in this process. The aim of the present study was to evaluate in the human vagina, by means of immunohistochemistry, the expression and distribution of phosphodiesterase type 1 (PDE1, known to hydrolize both cyclic AMP and cyclic GMP) in relation to calcitonin gene-related peptide (CGRP), vasoactive intestinal polypeptide (VIP) and protein gene product 9.5 (PGP 9.5).

Study design

Sections of human vagina (full wall specimens) were incubated with antibodies directed against PDE1, CGRP, VIP, PGP 9.5 and alpha-actin, followed by exposure to fluorochrome-labelled secondary antibodies. Visualization was commenced by means of laser fluorescence microscopy.

Results

Microscopic examination revealed a dense meshwork of PGP 9.5-positive nerve fibers innervating the sections of vaginal wall. Small vessels interspersing the tissue presented dense staining for PDE1 in their smooth musculature. Blood vessels were seen surrounded by PDE1-immunoreactive longitudinal smooth muscle fibers. The vessels were also found innervated by PGP-positive varicose nerve fibers characterized by the expression of CGRP. Some fibers presented immunosignals specific for VIP.

Conclusion

Key mediators of the cyclic AMP and cyclic GMP pathways are co-localized in nerves seen in close proximity to vascular smooth muscle expressing PDE1. These findings suggest that both signaling cascades are involved in the control of vaginal blood flow.     

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.     

Smooth Muscle Cells in Pelvic Washings at Time of Benign Hysterectomy

Study ObjectiveTo evaluate if smooth muscle cells can be detected in pelvic washings at the time of intact hysterectomy.DesignA multicentered pilot cohort study (Canadian Task Force classification II-2).SettingTwo academically affiliated tertiary referral centers.PatientsPatients undergoing total hysterectomy for benign indications without morcellation by minimally invasive gynecologic surgeons were enrolled from January 2018 to July 2018.InterventionsPelvic washings were collected at 2 times during surgery: after abdominal entry and after vaginal cuff closure. Cell blocks were generated, and slides were stained using hematoxylin and eosin, smooth muscle actin, and desmin and interpreted by 1 expert pathologist at each institution.Measurements and Main ResultsThirty-eight subjects were recruited; 3 subjects were excluded because of unplanned morcellation. Smooth muscle uterine cells were detected in 1 prewash specimen and 2 postwash specimens. The group with positive washings was noted to have longer procedure times (136 vs 114 minutes), lower blood loss (25 vs 86 mL), and higher uterine weight (242 vs 234 g) compared with negative washings group.ConclusionTissue dissemination of uterine cells may be possible at the time of hysterectomy. Larger prospective studies are needed to better describe the incidence of and risk factors for tissue dissemination.     

Up-regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells

IntroductionFunctional failure of smooth muscle cells and endothelial cells in corpus cavernosum contributes to erectile dysfunction (ED) in aging men. Given that vascular endothelial growth factor (VEGF) may improve the function of smooth muscle cells and endothelial cells through different mechanisms, it is thus expected that increasing the expression of VEGF may have beneficial effects on erectile function.AimThe aim of this article is to explore the possibility that VEGF can be induced by ribonucleic acid activation (RNAa) technology, and VEGF induction by RNAa has the potential of treating ED.MethodsPrimary human corpus cavernosum smooth muscle cells (CCSMCs) were isolated and cultured in vitro. The expression of α‐smooth muscle actin was detected by immunohistochemistry to identify CCSMCs. A previously identified VEGF promoter‐targeted small activator RNA (saRNA, double‐stranded [ds]VEGF‐706) and a negative control dsRNA were chemically synthesized. Cultured human CCSMCs were transfected with the saRNAs. The expression of VEGF messenger RNA (mRNA) and protein in transfected CCSMCs was evaluated by real‐time polymerase chain reaction (RT‐PCR) and Western blotting assay, respectively. Immunofluorescent staining was also used to confirm VEGF protein expression in cultured CCSMCs.Main Outcome MeasureThe expression of VEGF was assessed by RT quantitative PCR, Western blotting, and immunofluorescence assays.ResultsAfter transfection, RT quantitative PCR analysis showed that the expression of VEGF mRNA was significantly induced in dsVEGF‐706 transfected cells compared with cells receiving control treatments (P < 0.05). Consistent with mRNA induction, Western blotting and immunofluorescence analysis showed that VEGF protein expression was also induced by dsVEGF‐706.ConclusionVEGF expression can be activated by RNAa in primary human CCSMCs, suggesting a potential application of RNAa‐mediated VEGF activation for the treatment of ED. Chen R, Wang T, Rao K, Yang J, Zhang S, Wang S, Liu J, and Ye Z. Up‐regulation of VEGF by small activator RNA in human corpus cavernosum smooth muscle cells. J Sex Med 2011;8:2773–2780.