Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium ...Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium were obtained from 5 normally cycling women of reproductive age. Half of each tissue was cultured in the DMEM medium containing diethylstilbesterol (25 μg/mL) plus medroxyprogesterone acetate (25 μg/mL) (E + P culture). As a control, the other half was cultured in the medium alone. After 100 h incubation, the tissues were assessed for the formation of NCS with transmission electron microscope.Results NCS was observed in the endometrial epithelium treated with E + P or the medium alone. Moreover, giant mitochondria and glycogen accumulation were both seen in epithelia derived from both types of cultures.Conclusion Progesterone would be not indispensable for the formation of NCS in human endometrium. Transition of proliferative endometrium to the secretory stage in vitro could occur even in the absence of both estrogen and progesterone.展开更多
Objective To investigate the expression of Calbindin-d28k （CaBP-d28k） in human endometrium. Methods Thirty-three samples of human normal endometrial tissues were divided into 6 groups： early proliferative stage （n...Objective To investigate the expression of Calbindin-d28k （CaBP-d28k） in human endometrium. Methods Thirty-three samples of human normal endometrial tissues were divided into 6 groups： early proliferative stage （n =6）, mid proliferative stage （n =5）, late proliferative stage （n=5）, early secretory stage （n=7）, mid secretory stage （n=5） and late secretory stage （n=5）. The expression and change of CaBP-d28k protein and gene were determined by immunohistochemistry and reverse transcription polymerase chain reaction methods. Results In endometrial samples, the expression of CaBP-d28k protein was mainly observed in the cytoplasm of luminal and glandular epithelium. In the menstrual cycle, the level of CaBP-d28k protein in the epithelium was the lowest during the early and mid proliferative stages, and was the highest during the mid secretory stage, then decreased in the late secretory stage （P〈0.05）. In the stroma, the expressed type of CaBP-d28k protein was the same as in the epithelium, but was lower than that in the epithelium（P〈0.05）. The CaBP-d28k mRNA was at the lowest level in the early proliferative stage（P〈0.05）, and significantly increased in the late proliferative, and early, mid secretory stages （P〈0. 05）. Conclusion Both CaBP-d28k protein and gene were expressed in human endometrium, and their expression had cyclic changes.展开更多
The functional significance of the endometrial hCG/ LH receptors has been related to a rapid release of prostaglandins. However, as compared to gonads and myometrium, in-endometrium mechanisms of transmembrane signall...The functional significance of the endometrial hCG/ LH receptors has been related to a rapid release of prostaglandins. However, as compared to gonads and myometrium, in-endometrium mechanisms of transmembrane signalling of the hCG/LH receptors are probably not conventional and remain unclear. Here we investigated, in vivo, the potential of hCG to interact with, and stimulate the membrane effector enzyme, adenylyl cyclase (AC), in human endometrium. Hormonal and nonhormonal activation of AC was tested in membrane fractions prepared from endometrial biopsies obtained from patients undergoing evaluation cycles for hormone replacement therapy (HRT) and controlled ovarian hyperstimulation (COH). AC activity was determined by the direct conversion of the substrate ATP into cAMP under unstimulated conditions and in the presence of the non-hormonal activators guanyl nucleotide and forskolin. Also AC activity was tested in the presence of hCG under conditions allowing maximal enzyme stimulation. Isoproterenol and prostaglandin E2 (PGE2) were included for comparison. Immunoblot analyses demonstrated the presence of hCG/LH receptors and Gsα protein and other members of the G protein family in the membrane fractions. Endometrial membranes also exhibited high levels of AC activity compared to luteal membranes used as control. Stimulation by GMP-P(NH)P alone was 196 ± 63 (n = 8) (pmol/mg/ min ± SD). Neither hCG nor isoproterenol showed stimulation of endometrial AC (210 ± 65, and 197 ± 53, respectively;n = 66 assays). But PGE2 stimulated the enzyme system significantly (264 ± 63, p < 0.05;n = 66 assays). These data show that membrane fractions from human endometrium express all the AC system components, namely, hCG/LH receptors, Gsα protein and AC;however, hCG does not stimulate the endometrial AC system. Our data indicate that, in great contrast to gonadal receptors, endometrial hCG/ LH receptors are not coupled to the transmembrane AC effector. The well known release of eicosanoids in response to hCG suggests that these receptors are functional in human endometrium but throughout a signalling system different from AC. This enzyme is certainly coupled to and directly activated by eicosanoids and other embryonic signals.展开更多
Objective: The presence of nerve fibers in human endometrium remains unsettled but recent immunocytochemical studies have shown that there was increased innervation in the endometrium from women with endometriosis and...Objective: The presence of nerve fibers in human endometrium remains unsettled but recent immunocytochemical studies have shown that there was increased innervation in the endometrium from women with endometriosis and some nerve fibers in the normally cycling human endometrium. In the current study, we used uterine tissue cryosections from normal cycling women, which previously provided better immunocytochemical staining for lymphatic vessels than in paraffin sections. Materials and Methods: A total of 16 cases from normally cycling women were included representing menstrual, early proliferative, early to late secretary phase. Neurofilament and CD 56 were used as immunocytochemical markers for nerve fibers with cryosections. Results: There were consistent presence of nerve fibers in myometrium and basalis. Few small nerve fibers were identified in early proliferative endometrium and more nerve fibers were present in lower-half functionalis from mid-secretary phase. Late-secretary functionalis showed less nerve fibers in the upper-half than the lower-half functionalis, implying growing nerve fibers from lower functionalis to upper functionalis in late-secretary phase. Conclusion: Nerve fibers appeared to cyclically grow from basalis to lower functionalis and then from lower functionalis to upper functionalis concomitantly with blood vessels in normally cycling human endometrium. These cycling endometrial nerve fibers consisted mostly of nonmyelinated small nerve fibers, which may transmit pelvic pain in the normally cycling women.展开更多
Objective: The presence of lymphatic vessels in endometrium has been controversial and recent immunocytochemical studies with routinely paraffin embedded sections revealed lymphatic vessels in basalis and occasionally...Objective: The presence of lymphatic vessels in endometrium has been controversial and recent immunocytochemical studies with routinely paraffin embedded sections revealed lymphatic vessels in basalis and occasionally in functionalis. We aimed to investigate endometrial lymphatic vessels by immunocytochemical staining using cryosections, which provided better and consistent immunostaining for lymphatic vessels with a lymphatic marker, D2-40. We aimed further to explore the structure-function relationship of lymphatic vessels in the menstrual cycle. Materials and Methods: Sixteen cases of endometrium from menstrual, early-proliferative to latesecretary phase were immunostained for D2-40 and lymphatic vessels were morphometrically analyzed for functionalis, basalis and myometrium, respectively. Results: Lymphatic vessels were consistently most numerous in myometrium, followed by basalis in all phases whereas menstrual endometrium showed small, fragmented aggregates of lymphatic vessels in thin basalis. Earlyto mid-secretary endometrium revealed many lymphatic vessels in basalis and lower-functionalis with few lymphatic vessels in upper-functionalis. Late-secretary endometrium revealed more lymphatic vessels in upper-functionalis with dilated walls, which then burst at the surface of functionalis. Conclusions: These degenerating lymphatic vessels with markedly dilated lumen in upper-functionalis may contribute to lymphatic leakage in late-secretary phase. These immunostained lymphatic vessels in functionalis support proliferating and degenerating lymphatic vessel cycle synchronized with the menstrual cycle of endometrial arteries to maintain adequate fluid leakage.展开更多
Context: Cyclic changes of endometrial arteries are well established but possible cyclic changes of lymphatic and venous vessels have not been fully documented. There are no published morphological reports to support ...Context: Cyclic changes of endometrial arteries are well established but possible cyclic changes of lymphatic and venous vessels have not been fully documented. There are no published morphological reports to support cyclic changes of endometrial lymphatic and venous vessels. Objective: Using cryosections of human endometrium, this study aimed to unveil possible cyclic changes of lymphatic and venous vessels. We previously reported cyclic changes of lymphatic vessels in human endometrium using D2-40. Design: A total of 16 cases representing menstrual, proliferative and mid and late secretary phase were studied. For Immunocytochemical staining, lymphatic vessel endothelial hyaluronan receptor 1 and von Willebr and factor were used for lymphatic and venous vessels, respectively. We used polyclonal LYVE-1 in this study, which revealed more lymphatic vessels than using D2-40. Results: Residual lymphatic and venous vessels were present in menstrual basalis. In Day 5 - 9 endometrium, there were sparse lymphatic vessels but were numerous growing venous vessels in thin proliferating functionalis. In Day 14 - 22 endometrium, there were scattered lymphatic vessels and numerous venous vessels in functionalis. In Day 25 - 26 endometrium, there were many dilated lymphatic vessels and numerous dilated, disintegrating venous vessels in upper functionalis than lower functionalis. Conclusion: The above findings support that lymphatic vessels are sparse but venous vessels are numerous in early proliferative functionalis. Lymphatic vessels grow from basalis to thin functionalis. In premenstrual phase, lymphatic vessels proliferate from lower to upper functionalis, and both lymphatic and venous vessels disintegrate for shedding by this immunocytochemical study using lymphatic and venous markers. Thus, all lymphatic, venous and arterial vessels undergo menstrual cyclic changes and shed for menstruation.展开更多
基金This study was supported by the National Science Fund of P.R.China (No.39970765)
文摘Objective To determine whether formation of the nucleolar channel system (NCS) in human endometrium depends on the presence of progesteronal steroids. Materials & Methods Tissues of late proliferative endometrium were obtained from 5 normally cycling women of reproductive age. Half of each tissue was cultured in the DMEM medium containing diethylstilbesterol (25 μg/mL) plus medroxyprogesterone acetate (25 μg/mL) (E + P culture). As a control, the other half was cultured in the medium alone. After 100 h incubation, the tissues were assessed for the formation of NCS with transmission electron microscope.Results NCS was observed in the endometrial epithelium treated with E + P or the medium alone. Moreover, giant mitochondria and glycogen accumulation were both seen in epithelia derived from both types of cultures.Conclusion Progesterone would be not indispensable for the formation of NCS in human endometrium. Transition of proliferative endometrium to the secretory stage in vitro could occur even in the absence of both estrogen and progesterone.
文摘Objective To investigate the expression of Calbindin-d28k （CaBP-d28k） in human endometrium. Methods Thirty-three samples of human normal endometrial tissues were divided into 6 groups： early proliferative stage （n =6）, mid proliferative stage （n =5）, late proliferative stage （n=5）, early secretory stage （n=7）, mid secretory stage （n=5） and late secretory stage （n=5）. The expression and change of CaBP-d28k protein and gene were determined by immunohistochemistry and reverse transcription polymerase chain reaction methods. Results In endometrial samples, the expression of CaBP-d28k protein was mainly observed in the cytoplasm of luminal and glandular epithelium. In the menstrual cycle, the level of CaBP-d28k protein in the epithelium was the lowest during the early and mid proliferative stages, and was the highest during the mid secretory stage, then decreased in the late secretory stage （P〈0.05）. In the stroma, the expressed type of CaBP-d28k protein was the same as in the epithelium, but was lower than that in the epithelium（P〈0.05）. The CaBP-d28k mRNA was at the lowest level in the early proliferative stage（P〈0.05）, and significantly increased in the late proliferative, and early, mid secretory stages （P〈0. 05）. Conclusion Both CaBP-d28k protein and gene were expressed in human endometrium, and their expression had cyclic changes.
文摘The functional significance of the endometrial hCG/ LH receptors has been related to a rapid release of prostaglandins. However, as compared to gonads and myometrium, in-endometrium mechanisms of transmembrane signalling of the hCG/LH receptors are probably not conventional and remain unclear. Here we investigated, in vivo, the potential of hCG to interact with, and stimulate the membrane effector enzyme, adenylyl cyclase (AC), in human endometrium. Hormonal and nonhormonal activation of AC was tested in membrane fractions prepared from endometrial biopsies obtained from patients undergoing evaluation cycles for hormone replacement therapy (HRT) and controlled ovarian hyperstimulation (COH). AC activity was determined by the direct conversion of the substrate ATP into cAMP under unstimulated conditions and in the presence of the non-hormonal activators guanyl nucleotide and forskolin. Also AC activity was tested in the presence of hCG under conditions allowing maximal enzyme stimulation. Isoproterenol and prostaglandin E2 (PGE2) were included for comparison. Immunoblot analyses demonstrated the presence of hCG/LH receptors and Gsα protein and other members of the G protein family in the membrane fractions. Endometrial membranes also exhibited high levels of AC activity compared to luteal membranes used as control. Stimulation by GMP-P(NH)P alone was 196 ± 63 (n = 8) (pmol/mg/ min ± SD). Neither hCG nor isoproterenol showed stimulation of endometrial AC (210 ± 65, and 197 ± 53, respectively;n = 66 assays). But PGE2 stimulated the enzyme system significantly (264 ± 63, p < 0.05;n = 66 assays). These data show that membrane fractions from human endometrium express all the AC system components, namely, hCG/LH receptors, Gsα protein and AC;however, hCG does not stimulate the endometrial AC system. Our data indicate that, in great contrast to gonadal receptors, endometrial hCG/ LH receptors are not coupled to the transmembrane AC effector. The well known release of eicosanoids in response to hCG suggests that these receptors are functional in human endometrium but throughout a signalling system different from AC. This enzyme is certainly coupled to and directly activated by eicosanoids and other embryonic signals.
文摘Objective: The presence of nerve fibers in human endometrium remains unsettled but recent immunocytochemical studies have shown that there was increased innervation in the endometrium from women with endometriosis and some nerve fibers in the normally cycling human endometrium. In the current study, we used uterine tissue cryosections from normal cycling women, which previously provided better immunocytochemical staining for lymphatic vessels than in paraffin sections. Materials and Methods: A total of 16 cases from normally cycling women were included representing menstrual, early proliferative, early to late secretary phase. Neurofilament and CD 56 were used as immunocytochemical markers for nerve fibers with cryosections. Results: There were consistent presence of nerve fibers in myometrium and basalis. Few small nerve fibers were identified in early proliferative endometrium and more nerve fibers were present in lower-half functionalis from mid-secretary phase. Late-secretary functionalis showed less nerve fibers in the upper-half than the lower-half functionalis, implying growing nerve fibers from lower functionalis to upper functionalis in late-secretary phase. Conclusion: Nerve fibers appeared to cyclically grow from basalis to lower functionalis and then from lower functionalis to upper functionalis concomitantly with blood vessels in normally cycling human endometrium. These cycling endometrial nerve fibers consisted mostly of nonmyelinated small nerve fibers, which may transmit pelvic pain in the normally cycling women.
文摘Objective: The presence of lymphatic vessels in endometrium has been controversial and recent immunocytochemical studies with routinely paraffin embedded sections revealed lymphatic vessels in basalis and occasionally in functionalis. We aimed to investigate endometrial lymphatic vessels by immunocytochemical staining using cryosections, which provided better and consistent immunostaining for lymphatic vessels with a lymphatic marker, D2-40. We aimed further to explore the structure-function relationship of lymphatic vessels in the menstrual cycle. Materials and Methods: Sixteen cases of endometrium from menstrual, early-proliferative to latesecretary phase were immunostained for D2-40 and lymphatic vessels were morphometrically analyzed for functionalis, basalis and myometrium, respectively. Results: Lymphatic vessels were consistently most numerous in myometrium, followed by basalis in all phases whereas menstrual endometrium showed small, fragmented aggregates of lymphatic vessels in thin basalis. Earlyto mid-secretary endometrium revealed many lymphatic vessels in basalis and lower-functionalis with few lymphatic vessels in upper-functionalis. Late-secretary endometrium revealed more lymphatic vessels in upper-functionalis with dilated walls, which then burst at the surface of functionalis. Conclusions: These degenerating lymphatic vessels with markedly dilated lumen in upper-functionalis may contribute to lymphatic leakage in late-secretary phase. These immunostained lymphatic vessels in functionalis support proliferating and degenerating lymphatic vessel cycle synchronized with the menstrual cycle of endometrial arteries to maintain adequate fluid leakage.
文摘Context: Cyclic changes of endometrial arteries are well established but possible cyclic changes of lymphatic and venous vessels have not been fully documented. There are no published morphological reports to support cyclic changes of endometrial lymphatic and venous vessels. Objective: Using cryosections of human endometrium, this study aimed to unveil possible cyclic changes of lymphatic and venous vessels. We previously reported cyclic changes of lymphatic vessels in human endometrium using D2-40. Design: A total of 16 cases representing menstrual, proliferative and mid and late secretary phase were studied. For Immunocytochemical staining, lymphatic vessel endothelial hyaluronan receptor 1 and von Willebr and factor were used for lymphatic and venous vessels, respectively. We used polyclonal LYVE-1 in this study, which revealed more lymphatic vessels than using D2-40. Results: Residual lymphatic and venous vessels were present in menstrual basalis. In Day 5 - 9 endometrium, there were sparse lymphatic vessels but were numerous growing venous vessels in thin proliferating functionalis. In Day 14 - 22 endometrium, there were scattered lymphatic vessels and numerous venous vessels in functionalis. In Day 25 - 26 endometrium, there were many dilated lymphatic vessels and numerous dilated, disintegrating venous vessels in upper functionalis than lower functionalis. Conclusion: The above findings support that lymphatic vessels are sparse but venous vessels are numerous in early proliferative functionalis. Lymphatic vessels grow from basalis to thin functionalis. In premenstrual phase, lymphatic vessels proliferate from lower to upper functionalis, and both lymphatic and venous vessels disintegrate for shedding by this immunocytochemical study using lymphatic and venous markers. Thus, all lymphatic, venous and arterial vessels undergo menstrual cyclic changes and shed for menstruation.
