Macrophage polarization in adenomyosis: A review.

Adenomyosis (AM) is a common gynecological disorder characterized by the presence of endometrial glands and stroma within the uterine myometrium. It is associated with abnormal uterine bleeding (AUB), dysmenorrhea, and infertility. Although several mechanisms have been proposed to elucidate AM, the exact cause and development of the condition remain unclear. Recent studies have highlighted the significance of macrophage polarization in the microenvironment, which plays a crucial role in AM initiation and progression. However, a comprehensive review regarding the role and regulatory mechanism of macrophage polarization in AM is currently lacking. Therefore, this review aims to summarize the phenotype and function of macrophage polarization and the phenomenon of the polarization of adenomyosis-associated macrophages (AAMs). It also elaborates on the role and regulatory mechanism of AAM polarization in invasion/migration, fibrosis, angiogenesis, dysmenorrhea, and infertility. Furthermore, this review explores the underlying molecular mechanisms of AAM polarization and suggests future research directions. In conclusion, this review provides a new perspective on understanding the pathogenesis of AM and provides a theoretical foundation for developing targeted drugs through the regulation of AAM polarization.

Intraoperative evaluation of local cerebral hemodynamic change by laser speckle contrast imaging for predicting postoperative cerebral hyperperfusion during STA-MCA bypass in adult patients with moyamoya disease.

Cerebral hyperperfusion (CHP) occurred frequently after direct superficial temporal artery-middle cerebral artery (STA-MCA) bypass surgery for moyamoya disease (MMD). We analyzed cortical microvascular density (CMD) and the change of cerebral blood flow (LΔCBF) using intraoperative laser speckle contrast imaging (LSCI) on 130 hemispheres of 95 consecutive adult patients with MMD. The demographic characteristics, cortical hemodynamic sources, bypass methods, intraoperative blood flow data, and relative CBF changes on single-photon emission computed tomography (SPECT) examination (SΔrCBF) were compared between the groups with and without CHP. The median values for CMD, LΔCBF, and SΔrCBF were significantly higher in the CHP group than in the non-CHP group (CMD 0.240 vs 0.206, P = 0.004; LΔCBF 2.285 vs 1.870, P < 0.001; SΔCBF 1.535 vs 1.260, P < 0.001). Multivariate analysis revealed that hemodynamic sources of recipient parasylvian cortical arteries from MCA (M-PSCAs), end-to-side (E-S) bypass method, CMD ≥ 0.217, and LΔCBF ≥ 1.985 were the risk factors for CHP. Intraoperative LSCI was useful for evaluating hemodynamics and predicting CHP in patients with MMD.

Ginsenoside Rh1, a novel casein kinase II subunit alpha (CK2α) inhibitor, retards metastasis via disrupting HHEX/CCL20 signaling cascade involved in tumor cell extravasation across endothelial barrier.

Tumor cell extravasation across endothelial barrier has been recognized as a pivotal event in orchestrating metastasis formation. This event is initiated by the interactions of extravasating tumor cells with endothelial cells (ECs). Therefore, targeting the crosstalk between tumor cells and ECs might be a promising therapeutic strategy to prevent metastasis. In this study, we demonstrated that Rh1, one of the main ingredients of ginseng, hindered the invasion of breast cancer (BC) cells as well as diminished the permeability of ECs both in vitro and in vivo, which was responsible for the attenuated tumor cell extravasation across endothelium. Noteworthily, we showed that ECs were capable of inducing the epithelial-mesenchymal transition (EMT) and invadopodia of BC cells that are essential for tumor cell migration and invasion through limiting the nuclear translocation of hematopoietically expressed homeobox (HHEX). The decreased nuclear HHEX paved the way for initiating the CCL20/CCR6 signaling axis, which in turn contributed to damaged endothelial junctions, uncovering a new crosstalk mode between tumor cells and ECs. Intriguingly, Rh1 inhibited the kinase activity of casein kinase II subunit alpha (CK2α) and further promoted the nuclear translocation of HHEX in the BC cells, which resulted in the disrupted crosstalk between chemokine (C-C motif) ligand 20 (CCL20) in the BC cells and chemokine (C-C motif) receptor 6 (CCR6) in the ECs. The prohibited CCL20-CCR6 axis by Rh1 enhanced vascular integrity and diminished tumor cell motility. Taken together, our data suggest that Rh1 serves as an effective natural CK2α inhibitor that can be further optimized to be a therapeutic agent for reducing tumor cell extravasation.

Expression of hypoxia-inducing factor-1α and matrix metalloproteinase-9 in the recipient parasylvian cortical arteries with different hemodynamic sources in adult moyamoya disease.

Objective: In our latest research, we have demonstrated that the recipient parasylvian cortical arteries (PSCAs) with hemodynamic sources from the middle cerebral artery (M-PSCAs) has a higher risk of postoperative cerebral hyperperfusion (CHP) syndrome than those from non-M-PSCAs in adult moyamoya disease (MMD) patient. However, whether there are differences between M-PSCAs and non-M-PSCAs in vascular specimens characteristics has not been studied. In this study, we further investigate the vascular specimen of recipient PSCAs by histological and immunohistochemical methods. Methods: 50 vascular specimens of recipient PSCAs were obtained from 50 adult MMD patients during the combined bypass surgeries in our departments of Zhongnan hospital. 4 recipient PSCAs samples were also obtained in the same way from the middle cerebral artery occlusion patients. The samples were received the pathological sectioning, hematoxylin and eosin staining, and immunohistochemistry, then the vascular wall thickness, matrix metalloproteinase-9 (MMP-9) and hypoxia-inducing factor-1α (HIF-1α) were analyzed. Results: M-PSCAs adult MMD patients had a thinner intima than non-M-PSCAs in the recipient PSCAs specimens. In recipient non-M-PSCAs vascular specimens, the immunoreactivity indicating HIF-1α and matrix metalloproteinase-9 (MMP-9) was significantly higher than M-PSCAs groups. The logistic regression analyses showed that the M-PSCAs was an independent risk factor of postoperative cerebral hyperperfusion (CHP) syndrome (OR 6.235, 95% CI1.018-38.170, P = 0.048) in MMD. Conclusion: Our results indicate that M-PSCAs adult MMD patients had thinner intima than non-MCAs adult MMD patients in the PSCAs. More importantly, HIF-1α and MMP-9 were overexpressed in non-M-PSCAs vascular specimens.

Transcriptome analysis of eutopic endometrial stromal cells in women with adenomyosis by RNA-sequencing.

This study aimed to identify differentially expressed genes (DEGs) and molecular pathways in eutopic endometrial stromal cells (EuESCs) from adenomyosis (AM) patients and to provide a new insight into the disease mechanisms. The gene expression profiles in adenomyotic EuESCs (A-EuESCs) and normal ESCs (N-ESCs) were analyzed by RNA-sequencing (RNA-Seq) and validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways enrichment analyses were performed to obtain insights into the functions of DEGs. The protein-protein interaction (PPI) network was constructed using the STRING database and visualized by Cytoscape software, and their hub genes were identified. A total of 458 up-/363 down-regulated genes were identified in A-EuESCs versus N-ESCs. The GO enrichment analysis showed that these genes were significantly enriched in calcium-dependent cell-cell adhesion. The most significant term of the KEGG pathway analysis was cytokine-cytokine receptor interaction. There were 145 nodes in the PPI network of the 157 DEGs, which were identified in significant enrichment pathway by the KEGG pathway analysis in N-ESCs and A-EuESCs. The PPI network revealed that IL-6 was a central hub gene. Besides, IL-6 was found as a central hub gene in the pro-inflammatory/chemotactic subnetwork, and EGF was noted as a central hub gene in the angiogenesis subnetwork. Our study indicated the alterations of transcriptomic profiles in A-EuESCs and provided new insights into the pathogenesis of AM. The A-EuESCs in women with AM have fundamental abnormalities that may predispose to pro-invasion/migration and angiogenesis.

TCF3 is epigenetically silenced by EZH2 and DNMT3B and functions as a tumor suppressor in endometrial cancer.

Endometrial cancer (EC) is the most common gynecological malignancy worldwide. However, the molecular mechanisms underlying EC progression are still largely unknown, and chemotherapeutic options for EC patients are currently very limited. In this study, we found that histone methyltransferase EZH2 and DNA methyltransferase DNMT3B were upregulated in EC samples from patients, and promoted EC cell proliferation as evidenced by assays of cell viability, cell cycle, colony formation. Mechanistically, we found that EZH2 promoted EC cell proliferation by epigenetically repressing TCF3, a direct transcriptional activator of CCKN1A (p21WAF1/Cip1), in vitro and in vivo. In addition, we found that DNMT3B specifically methylated the TCF3 promoter, repressing TCF3 expression and accelerating EC cell proliferation independently of EZH2. Importantly, elevated expression of EZH2 or DNMT3B in EC patients inversely correlated with expression of TCF3 and p21, and was associated with shorter overall survival. We show that combined treatment with GSK126 and 5-Aza-2d treatment wit synergistically inhibited methyltransferase activity of EZH2 and DNMT3B, resulting in a profound block of EC cell proliferation as well as EC tumor progression in cell line-derived xenograft (CDX) and patient-derived xenograft (PDX) mouse models. These findings reveal that TCF3 functions as a tumor suppressor epigenetically silenced by EZH2 and DNMT3B in EC, and support the notion that targeting the EZH2/DNMT3B/TCF3/p21 axis may be a novel and effective therapeutic strategy for treatment of EC.

Endometriotic Peritoneal Fluid Promotes Myofibroblast Differentiation of Endometrial Mesenchymal Stem Cells.

During the development of endometriosis, the presence of fibrotic tissues in and surrounding endometriotic lesions may lead to subsequent adhesion, anatomic distortion, and chronic pain. Therefore, studies aimed at clarifying the underlying mechanisms of fibrogenesis in endometriosis could potentially provide a novel strategy for effective treatment. Mesenchymal stem cells (MSCs) play a key role in fibrotic diseases by differentiating into myofibroblasts in appropriate microenvironment. In this study, we collected endometrial and endometriotic tissues from patients with endometriosis (n = 32) and control patients without endometriosis (n = 20) to compare the expression of fibrotic proteins and investigate the effect of endometriotic peritoneal fluid (PF) on myofibroblast differentiation of endometrial MSCs. We found that the expression of fibrotic proteins, including alpha-smooth muscle actin (α-SMA), type I collagen (collagen I), connective tissue growth factor (CTGF), and fibronectin, and the extent of fibrosis extremely enhanced in ectopic endometria compared with eutopic endometria from the same patients with endometriosis and normal endometria from patients without endometriosis. We next isolated and identified endometrial MSCs and found that treatment with endometriotic PF strongly induced endometrial MSCs to differentiate into myofibroblasts concomitant with the activation of Smad2/3. Moreover, ectopic endometrial MSCs expressed elevated collagen I, α-SMA, fibronectin, and CTGF. Sushi domain containing-2 (SUSD2), a marker of endometrial MSCs, and α-SMA, a well-recognized marker for myofibroblasts, colocalized extensively in ectopic endometria while seldom in normal and eutopic endometria. These findings suggest that ectopic endometrial MSCs are probably more susceptible to myofibroblast differentiation because of the long-term influence of endometriotic PF. All together, we report for the first time that endometriotic PF promotes myofibroblast differentiation of endometrial MSCs. This understanding will greatly improve our understanding of the pathophysiology of endometriosis and help design better therapeutics.

Activin a promotes myofibroblast differentiation of endometrial mesenchymal stem cells via STAT3-dependent Smad/CTGF pathway.

BACKGROUND: Endometriosis, characterized by the presence of functional endometrial tissues outside the uterus, is one of the most common gynecological disorders. Endometrial mesenchymal stem cells (MSCs) are crucial for the occurrence and development of endometriosis. Ectopic endometrial MSCs exist in the peritoneal cavity. Thus, the bioactive factors in endometriotic peritoneal fluid may regulate the biological behaviors of endometrial MSCs. METHODS: In this study, after assessing the concentration of Activin A in peritoneal fluid using ELISA, we isolated and cultured endometrial MSCs and investigated whether Activin A stimulated endometrial MSCs to differentiate into myofibroblasts and clarified the underlying mechanisms by quantitative real-time PCR, Western blot analysis, immunofluorescent staining, RNA interference and Chromatin immunoprecipitation. We also employed the inhibitors of Activin A to explore the possibility of suppressing the development of fibrosis in endometriosis using primary endometrial MSCs cultures and a mouse model of endometriosis. RESULTS: Here, we revealed that Activin A significantly elevated in endometriotic peritoneal fluid and activin receptor-like kinase (ALK4), the specific receptor for Activin A, obviously enhanced in ectopic endometrial MSCs compared with eutopic endometrial MSCs from women with or without endometriosis. Next, we found that Activin A drived myofibroblast differentiation of endometrial MSCs, with extremely enhanced expression of connective tissue growth factor (CTGF). CTGF was shown to be required for Activin A-induced expression of ACTA2, COL1A1 and FN1 in endometrial MSCs. CTGF induction by Activin A in endometrial MSCs involved the activation of Smad2/3, as evidenced by the phosphorylation and nuclear translocation of Smad2/3 as well as the binding of Smad2/3 to CTGF promoter. Furthermore, Smad/CTGF pathway in endometrial MSCs required activation of STAT3 while independent of PI3K, JNK and p-38 pathways. In addition, we also demonstrated that inhibition of Activin A pathway impeded myofibroblast differentiation of endometrial MSCs and ameliorated fibrosis in endometriosis mice. CONCLUSIONS: Activin A promotes myofibroblast differentiation of endometrial mesenchymal stem cells via STAT3-dependent Smad/CTGF pathway. The results provided the first evidence that STAT3 acted as a crucial Activin A downstream mediator to regulate CTGF production. Our data may supplement the stem cell theory of endometriosis and provide the experimental basis to treat endometriosis-associated fibrosis by manipulating Activin A signaling.

Correlation between plasma ferritin level and gestational diabetes mellitus and its impact on fetal macrosomia.

AIMS/INTRODUCTION: To explore the relationship between plasma iron levels and gestational diabetes mellitus, as well as its impact on macrosomia. MATERIALS AND METHODS: We retrospectively compared ferritin level and other characteristics between pregnant women with gestational diabetes mellitus (GDM) and pregnant women without GDM. The correlation between the levels of plasma ferritin, glucose and hemoglobin was explored. Meanwhile, we assessed the risk factors of macrosomia. Furthermore, we explored the relationship between ferritin level and the incidence of macrosomia. RESULTS: A total of 793 pregnant women were enrolled in the present study, of which 92 pregnant women had GDM and 701 pregnant women were healthy. Meanwhile, 51 pregnant women gave birth to infants with macrosomia and another 742 women had normal infants. Compared with non-GDM women, pregnant women with GDM were older, with higher pre-pregnancy body mass index, plasma ferritin, fasting plasma glucose, 1-h postprandial glucose, 2-h plasma glucose and hemoglobin. In addition, our results showed a significant positive correlation between the levels of ferritin and fasting plasma glucose when ferritin levels were >70 ng/mL. Our results also showed that pre-pregnancy overweight or obesity, a high concentration of ferritin, as well as abnormal levels of fasting plasma glucose, 1-h plasma glucose and 2 h plasma glucose were risk factors for macrosomia. Furthermore, as the level of ferritin increased, so did the incidence of macrosomia. CONCLUSIONS: The current study provides evidence that pregnant women with high levels of ferritin might be prone to GDM. In addition, a high level of ferritin might be an independent risk factor for macrosomia. Therefore, the negative effect of iron supplementation in non-anemic pregnant women might be noteworthy.

Wenshen Xiaozheng Tang induces apoptosis and inhibits migration of ectopic endometriotic stromal cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Wenshen Xiaozheng Tang (WXT), a traditional Chinese medicine prescription, exerted a good therapeutic effect on endometriosis. However, the underlying mechanism is unclear. In the present study, we sought to evaluate the effect of WXT on the proliferation and migration of ectopic endometriotic stromal cells and explore the potential molecular mechanism. MATERIALS AND METHODS: Primary stromal cells derived from ectopic endometriotic lesions of patients with endometriosis were isolated and cultured. The inhibition effect of WXT on cell proliferation was determined by MTT. Apoptosis of ectopic endometriotic cells treated with WXT was analyzed with Annexin V-FITC/7-AAD staining. The activation of caspases was detected by western blot analysis. The influence of WXT on migration of ectopic endometriotic cells was measured by scratch wound healing assay and Transwell assay. The DNA binding activity of NF-κB and the expression of nuclear p65 protein were determined by electrophoretic mobility shift assay and western blot analysis, respectively. The impact of WXT on the expression of NF-κB regulated gene products involved in apoptosis and migration was determined by western blot analysis. RESULTS: WXT inhibited the proliferation of ectopic endometriotic cells in a time- and dose-dependent manner. In addition, WXT treatment resulted in significant induction of apoptosis through the activation of caspases and inhibition of migration in ectopic endometriotic cells. WXT notably suppressed constitutive NF-κB-DNA-binding activity as well as TNF-α induced nuclear translocation of NF-κB p65 subunit in ectopic endometriotic cells. Moreover, WXT diminished the expression of NF-κB regulated gene products involved in apoptosis and migration, including c-IAP1, c-IAP2, XIAP, survivin, Mcl-1, COX-2 and MMP-9. CONCLUSIONS: Our results indicate that WXT induces apoptosis and inhibits migration of ectopic endometriotic stromal cells.

Unruptured pregnancy in a noncommunicating rudimentary horn at 37 weeks with a live fetus: a case report.

We report an extremely rare case of an unruptured non-communicating rudimentary horn full-term pregnancy. A woman who had a uterine malformation was misdiagnosed as uterus didelphys and gave birth to a live, healthy fetus. The correct diagnosis was not made until cesarean section at 37 4/7 weeks. The case suggests that women who are pregnant in a rudimentary horn could obtain a full-term delivery and give birth to a live and healthy baby.

[Adjunctive treatment of GnRHa combined wenshen xiaozheng decoction in treating endometriosis after laparoscopy: a clinical observation].

OBJECTIVE: To observe the therapeutic efficacy and safety of gonadotropin-releasing hormone agonist (GnRHa) combined Wenshen Xiaozheng Decoction (WXD) in auxiliary treating endometriosis after laparoscopy. METHODS: One hundred and thirty-four endometriosis patients with confirmative pathological diagnosis were assigned to three groups depending on whether they would receive adjuvant therapy or Chinese medicine treatment, i.e., the control group, the observation 1 group, and the observation 2 group. The 22 patients in the control group received no adjuvant therapy after laparoscopy. The 42 patients in the observation 1 group were treated with GnRHa 3.6 mg by subcutaneous injection starting from the 1st day to the 5th day of menstruation, once per 28 days. The 70 patients in the observation 2 group were treated with GnRHa 3.6 mg by subcutaneous injection in combination with WXD starting from the 1st day to the 5th day of menstruation, once per 28 days. They also took WXD for 7 doses, one cycle per every 28 days. The treatment lasted for three to six months. Serum levels of estradiol (E2), follicle-stimulating hormone (FSH), luteinizing hormone (LH), and cancer antigen 125 (CA125), as well as clinical efficacy, and adverse drug reactions were observed before and after treatment. RESULTS: There was statistical difference in serum levels of E2, FSH, or LH between the control group and the observation 1 and 2 groups (P < 0.05). There was no statistical difference in serum levels of E2, FSH, or LH between the observation 1 group and the observation 2 group (P > 0.05). There was statistical difference in the clinical efficiency among the 3 groups (P < 0.05). There was statistical difference in the pre-post difference of CA125 levels among the three groups (P < 0.01). Compared with the control group, there was no statistical difference in the pre-post difference of CA125 levels between the observation 1 group and the observation 2 group (P > 0.05). No obvious adverse reaction occurred during the treatment. CONCLUSIONS: GnRHa combined WXD showed confirmative clinical efficacy in treating endometriosis after laparoscopy. It also could lower serum levels of E2, FSH, and LH levels. So it was an ideal solution for treatment of endometriosis.

The disturbance of TH17-Treg cell balance in adenomyosis.

OBJECTIVE: To investigate the balance between regulatory T cells (Treg) and T-helper 17 cells (T(H)17) in peripheral blood and uteri of women with adenomyosis (AM), and to evaluate their potential correlation with dysmenorrhea and CA-125 levels. DESIGN: Laboratory study using human peripheral blood and tissues. SETTING: Academic hospital. PATIENT(S): Forty-five patients with AM (study group) and 25 women without AM (control group). INTERVENTION(S): The peripheral blood and tissues harvested from all groups were subjected to flow cytometry, ELISA, quantitative real-time polymerase chain reaction, and immunohistochemistry. The severity of dysmenorrhea was distinguished by visual analog scale (VAS). MAIN OUTCOME MEASURE(S): T(H)17 and Treg cell frequency, mRNA and protein levels of transcription factors and cytokines in all groups, and their correlation between the T(H)17-Treg ratio and dysmenorrhea severity or CA-125 level. RESULT(S): The disturbance of T(H)17-Treg balance was demonstrated in peripheral circulation and uteri of patients with both diffuse and focal AM, and it correlated positively with dysmenorrhea severity and CA-125. CONCLUSION(S): The findings suggest that T(H)17-Treg imbalance may play a crucial role in the immunopathogenesis of AM, and may be thus a potential target of AM therapy. CLINICAL TRIAL REGISTRATION NUMBER: ChiCTR-CCC-13003500.

[Treatment of adenomyosis patients by bushen huoxue sanyu decoction: a clinical study].

OBJECTIVE: To observe the clinical efficacy of Bushen Huoxue Sanyu Decoction (BHSD) in treatment of adenomyosis (AM) patients. METHODS: Seventy AM patients of Shen deficiency blood stasis syndrome (SDBSS) were randomly assigned to two groups, the CM treatment group (50 cases) and the Mirena group (20 cases). Patients in the CM treatment group were treated with BHSD, one dose per day. Levonorgestrel intrauterine system (Mirena) was placed in the uterine cavity of those in the Mirena group. The therapeutic course for all was 3 months. Changes of dysmenorrhea, menstrual quantity, SDBSS, CM syndrome, uterine volume, and serum CA125 levels were observed before and after treatment. RESULTS: Compared with before treatment in the same group, scores for dysmenorrhea integral, scores for menstrual quantity, scores for SDBSS, and scores for CM syndrome all decreased in the two groups after treatment (P < 0.01). Compared with before treatment in the same group, the uterine volume was reduced after treatment in the two groups (P < 0.05) and serum carbohydrate antigen CA125 levels decreased between the two groups (P < 0.05, P < 0.01). Compared with the Mirena group, scores for dysmenorrhea integral increased and scores for SDBSS decreased in the CM treatment group (P < 0.01, P < 0.05). There was no statistical difference in the uterine volume or serum carbohydrate antigen CA125 levels (P > 0.05). CONCLUSIONS: BHSD could effectively alleviate main symptoms of AM patients of QSBSS such as dysmenorrhea, profuse menstrual blood volume, and increased uterine volume, and lower scores for QSBSS and the total score for CM syndrome.

Serum microRNA-205 as a novel biomarker for cervical cancer patients.

OBJECTIVE: Serum microRNAs (miRNAs) are a novel class of diagnostic and prognostic biomarkers for numerous cancers. However, the level and clinical relevance of circulating miR-205 transcripts in human serum of cervical cancer patients are unclear. The purpose of this study was to determine serum miR-205 levels in cervical cancer patients and explore its association with clinicopathological factors and prognosis. METHODS: Serum miR-205 expression was investigated in 60 cervical cancer patients and 60 healthy normal controls by using real-time PCR. Correlations between miR-205 expression and the clinicopathological features and prognosis of cervical cancer patients were then evaluated. Receiver operating characteristic curves were used to evaluate the sensitivity and specificity of serum miR-205. RESULTS: Serum miR-205 was significantly upregulated in cervical cancer patients compared with healthy donors (p < 0.01), and a high level of miR-205 expression was correlated with poor tumor differentiation (p = 0.009), lymph node metastasis (p = 0.015) and increased tumor stage (p = 0.001). The serum miR-205 level was capable of separating advanced stage from early stage metastatic cervical cancer from non-metastatic samples and poorly differentiated tumors from differentiated tumors with an area under the curve values of 0.74, 0.694 and 0.717, respectively. The expression of miR-205 was also higher in the cervical cancer tissues compared with the para-carcinoma tissues. In addition, Kaplan-Meier survival analysis showed that cervical cancer patients with high miR-205 expression tended to have shorter overall survival. In multivariate Cox regression analysis, miR-205 was identified as an independent prognostic marker. CONCLUSIONS: Serum miR-205, which is upregulated in cervical cancer, represents a predictive biomarker for the prognosis of cervical cancer patients.

Papillary serous carcinoma of the cervix mixed with squamous cells: A report of the first case.

OBJECTIVE: Primary papillary serous carcinoma (PPSC) of the cervix is rarely recognized, with the aggressive and unpredictable course. Here we report a case of primary adenosquamous papillary serous carcinoma of the cervix in a woman who underwent comprehensive treatment. CASE: A 53-year-old woman presented with irregular vaginal bleeding in hospital. The patient with a diagnosis of PPSC by an intracolposcopic biopsy received radical hysterectomy with bilateral salpingo-oophorectomy, right pelvic lymphadenectomy, left pelvic lymph node dissection, and postoperative concurrent chemoradiotherapy. Postoperative immunohistochemistry showed that CK5/6, CK7, P16, CEA, CA12-5 and P53 were positive. During 17 months after operation, the patient demonstrated distant metastases of lymph nodes and finally died of brain metastasis. CONCLUSIONS: Papillary serous adenocarcinoma of the cervix mixed with squamous cell carcinoma has not been reported since now, and here, this is the first documented case. Despite surgery and concurrent chemoradiotherapy, which were reported as effective therapeutic strategies for papillary serous adenocarcinoma of the cervix, the patient showed a poorer prognosis. Taken together, papillary serous adenosquamous carcinoma of the cervix could be more malignant than pure papillary serous adenocarcinoma.

[Effects of miR-135a on HOXA10 expression, proliferation and apoptosis of ovarian cancer cells].

OBJECTIVE: To investigate the effects of miR-135a on HOXA10 expression, proliferation and apoptosis of SKOV3 cells. METHODS: (1) Through computer-aided algorithms,the predicted target gene of miR-135a (HOXA10)were determined. (2) miR-135a mimics, miR-135a inhibitor and negative control were transfected into SKOV3 cells, respectively.Reverse transcription (RT)-PCR, western blot analysis were used to examine the expression levels of HOXA10 at different times (24, 48 and 72 hours). (3) A luciferase reporter assay was used to confirm the direct regulation between miR-135a and HOXA10. (4) SKOV3 cells proliferation at different times (24,48 and 72 hours) was detected by methyl thiazolyl tetrazolium (MTT) assay [quantified by absorbance(A)]. Western blot was used to examine the expression of apoptosis-associated protein bcl-2, bax and caspase-3 in SKOV3 cells after 48 hours transfection. RESULTS: (1) HOXA10 was predicted to be the target gene of miR-135a by computer-aided algorithms. (2) RT-PCR shown that HOXA10 mRNA levels were decreased over time (24, 48 and 72 hours) after miR-135a mimics transfection in SKOV3 cells (0.94 ± 0.04 vs 0.78 ± 0.03 vs 0.70 ± 0.03, P < 0.05). While, the expression of HOXA10 mRNA was increased over time after miR-135a inhibitor transfection (1.14 ± 0.05 vs 1.16 ± 0.03 vs 2.60 ± 0.08,P < 0.05). After transfected with miR-135a mimics or miR-135a inhibitor over 48 and 72 hours, the HOXA10 expression levels in SKOV3 cells were significantly lower or higher than each control group, respectively (all P < 0.01). Western blot analysis of HOXA10 expression in SKOV3 cells confirmed the results of RT-PCR detected. (3) After cotransfection of miR-135a plasmid and pMIR-REPORT luciferase plasmid containing HOXA10, luciferase reporter assays showed that the luciferase activity reduced by 67.8% (P < 0.01). (4) MTT showed that SKOV3 cells growth after miR-135a mimics transfection for 48 and 72 hours were significantly lower than those in control group (0.38 ± 0.03 vs 0.52 ± 0.05, 0.67 ± 0.05 vs 0.75 ± 0.06;respectively,all P < 0.05).While, SKOV3 cells transfected with miR-135a inhibitor for 72 hours grew significantly faster than that in control group (0.95 ± 0.05 vs 0.75 ± 0.06, P < 0.01). After miR-135a mimics transfection, the level of bcl-2 protein was significantly lower than that in control group (0.28 ± 0.06 vs 0.76 ± 0.09,P < 0.01). The activity of caspase-3 was significantly higher than that in control group (115.0 ± 2.4 vs 95.4 ± 2.1, P < 0.01). While, there was no statistical difference of bax expression (P = 0.142). However, after miR-135a inhibitor transfection, the expression level of bcl-2 protein was significantly higher than that in control group (0.92 ± 0.03 vs 0.76 ± 0.09, P = 0.037) and the activity of caspase-3 was significantly lower than that in control group (59.5 ± 4.1 vs 95.4 ± 2.1, P < 0.01). There was also no statistical difference of bax expression (P = 0.066). CONCLUSION: miR-135a may play an important role in cell proliferation and apoptosis of ovarian cancer cells by regulating HOXA10 and its downstream pathways.

Therapeutic potential of Wenshen Xiaozheng tang, a traditional Chinese medicine prescription, for treating endometriosis.

The objective of this study is to investigate the effect of Wenshen Xiaozheng Tang (WXT) on the development of endometriosis in a rat model. Sprague-Dawley rats in which endometriotic implants were induced were divided randomly into 3 groups. The rats in the low-dose and high-dose WXT groups were administered WXT 8.57 and 17.14 g/kg/d, respectively. The rats in the control groups received an equal volume of dissolvent, as did the sham-operated rats. After treatment for 4 weeks, WXT significantly decreased the mean lesion size as well as the peritoneal fluid and serum levels of tumor necrosis factor α and interleukin 1ß. Cyclooxygenase-2, matrix metalloproteinase 9, plasminogen activator inhibitor 1, and intercellular adhesion molecule 1 messenger RNA (mRNA) levels were downregulated, and the mRNA expression of tissue inhibitor of metalloproteinase 1 was upregulated in the endometriotic lesions of WXT versus control group. Our data suggested that WXT may suppress the development of endometriosis by inhibiting the production of proinflammatory cytokines and regulating the expression of invasion-related genes in the endometriotic lesions.

Immunological regulation of Chinese herb Guizhi Fuling Capsule on rat endometriosis model.

AIM OF THE STUDY: To investigate the immunological regulation of Guizhi Fuling Capsule (GZFLC) on rat endometriosis. MATERIALS AND METHODS: Twenty-seven rats, in which endometriotic implants were induced by transplanting autologous uterine tissue to the peritoneum, were randomly divided into three groups equally: (1) the GZFLC group of low dose (480 mg/kg/day); (2) the GZFLC group of high dose (1,920 mg/kg/day); and (3) the model group(saline solution). Another 10 rats were treated as sham operation group. After rats were treated for four weeks, we examined the alterations of implants volume, the percentage of CD4(+) T lympholeukocyte, the activity of NK cell and the expression of cytokines (MCP-1 and ICAM-1) on each group. RESULTS: Statistical analysis showed that posttreatment volumes were significantly reduced compared with pretreatment in GZFLC groups, whereas there was no significant change in the model group. The percentage of CD4(+) T lympholeukocyte and the activity of NK cell in GZFLC groups significantly increased to the level of the sham group compared with the model. RT-PCR and immunohistochemistry showed that the endometria of the sham operation and treatment groups were similar on expression level of MCP-1 and ICAM-1. CONCLUSIONS: GZFLC plays an important role in the regression of endometriotic implants by immunological regulation in the rat model.