Laparoscopic pectopexy with native tissue repair for pelvic organ prolapse.

PURPOSE: The use of mesh for vaginal repair is currently problematic; consequently, there is increased interest in native tissue repair. Combining native tissue repair with sufficient mesh-applied apical repair might provide effective treatment. We describe the study focusing on the combination of pectopexy and native tissue repair. METHODS: Between April 2020 and November 2021, 49 patients with symptomatic stage III or IV were treated with laparoscopic pectopexy combined with native tissue repair. The mesh was solely used for apical repair. All other clinically relevant defects were treated with native tissue repair. The perioperative parameters including surgical time, blood loss, hospital stay, and complications were recorded. The anatomical cure rate was evaluated according to the Pelvic Organ Prolapse Questionnaire (POP-Q) assessment. Validated questionnaires of the Pelvic Floor Distress Inventory (PFDI-20) and the Pelvic Floor Impact Questionnaire (PFIQ-7) were recorded to evaluate the symptom severity and quality of life. RESULTS: The mean duration of follow-up was 15 months. All domains of POP-Q, PFDI-20, and PFIQ-7 scores improved significantly after surgery. No major complications, mesh exposure, or mesh complication occurred during the follow-up period. CONCLUSION: The overall repair concept of laparoscopic pectopexy as the core, assisted by vaginal natural tissue repair for severe pelvic organ prolapse can achieve satisfactory clinical results and improve patient satisfaction.

FOXA1 can be modulated by HDAC3 in the progression of epithelial ovarian carcinoma.

FOXA1 is associated with malignant tumors, but the function of FOXA1 in EOC is unclear. HDAC3 can influence the proliferation, migration and invasion ability of EOC. In this study, we wanted to explore the function of FOXA1 in ovarian cancer and the relationship between HDAC3 and FOXA1.The expression of HDAC3 and FOXA1 was detected by immunohistochemical staining of primary lesions from 127 epithelial ovarian carcinoma patients. A proliferation assay, a Transwell assay, an apoptosis assay and animal experiments were used to assess the proliferation, invasion and apoptosis abilities of ovarian cancer cells before and after transfection with FOXA1. The relevance of the in vitro findings was confirmed in xenografts. The H-scores for FOXA1 and HDAC3 staining in FIGO stage III-IV were noticeably higher and predicted adverse clinical outcomes in patients with ovarian cancer. The expression level of HDAC3 was significantly correlated with the expression level of FOXA1. Invasion, proliferation and apoptosis capacity and tumor formation were decreased in the FOXA1-knockdown cells. Experiments in xenografts confirmed that HDAC3 mediated tumor formation. In conclusion, FOXA1 can be modulated by HDAC3 through the Wnt/ß-catenin signaling pathway, and FOXA1 plays essential roles in the proliferation, apoptosis and invasion of EOC cell lines and xenograft experiments.

Glutathionylation Decreases Methyltransferase Activity of PRMT5 and Inhibits Cell Proliferation.

Glutathionylation is an important posttranslational modification that protects proteins from further oxidative damage as well as influencing protein structure and activity. In the present study, we demonstrate that the cysteine-42 residue in protein arginine N-methyltransferase 5 (PRMT5) is glutathionylated in aged mice or in cells that have been exposed to oxidative stress. Deglutathionylation of this protein is catalyzed by glutaredoxin-1 (Grx1). Using mutagenesis and subsequent biochemical analyses, we show that glutathionylation decreased the binding affinity of PRMT5 with methylosome protein-50 (MEP50) and reduced the methyltransferase activity of PRMT5. Furthermore, overexpression of PRMT5-C42A mutant caused a significant increase in histone methylation in HEK293T and A549 cells and promoted cell growth, whereas overexpression of the PRMT5-C42D mutant, a mimic of glutathionylated PRMT5, inhibited cell proliferation. Taken together, our results demonstrate a new mechanism of regulation of PRMT5 methyltransferases activity and suggest that PRMT5 glutathionylation is partly responsible for reactive oxygen species-mediated cell growth inhibition.

Long term follow-up of inguinal endometriosis.

BACKGROUND: Inguinal endometriosis (IEM) is a rare extra pelvic endometriosis. Here, we study the clinical characteristics, management strategies, and long-term gynecological outcomes of IEM patients at Beijing Chaoyang Hospital. CASE PRESENTATION: Three patients presented with a total of four lesions (one on the left side, one on the right side, and one bilaterally). The diameters of the four lesions were 2 cm, 2 cm, 3.5 cm and 1.5 cm, respectively. Two patients were admitted with inguinal hernias. Two patients were admitted with endometrioses-one with ovarian endometriosis and one with pelvic endometriosis. The hernia sac was repaired concomitantly via excision of the round ligament in two patients. One patient underwent a concomitant laparoscopy for gynecologic evaluations, including an ablation to the peritoneal endometriosis, and resection of the left uterosacral ligament endometriosis and pelvic adhesiolysis. All lesions were located on the extraperitoneal portion of the round ligament and were diagnosed histologically. No recurrence was observed in the inguinal region. All patients diagnosed with adenomyosis were treated with medication alone without any complaints. CONCLUSIONS: Inguinal endometriosis can occur simultaneously with pelvic endometriosis. In most cases, a concomitant hernia sac appears together with groin endometriosis. Clinical management should be individualized and performed in tandem with general practitioners and obstetrics & gynecology experts. Pelvic disease, in particular, should be followed-up by a gynecologist.

Clinicopathological characteristics and prognosis of uterine serous carcinoma: A SEER program analysis of 1016 cases.

AIM: To identify the clinicopathological features and survival outcomes of uterine serous carcinoma (USC) and the prognostic factors influencing survival. METHODS: The retrospective, population-based cohort study enrolled patients with USC diagnosed between 2001 and 2015 for the Surveillance, Epidemiology, and End Results (SEER) program. Kaplan-Meier analysis was performed to identify survival outcomes, multivariable Cox regression models were used to determine the risk factors influencing the disease-specific survival (DSS) and overall survival (OS). RESULTS: A total of 1016 patients with USC from the SEER database were enrolled. The median age at diagnosis was 65 years. The 5-year OS was 48.5%, and the 5-year DSS rates were 58.0%, respectively. In the univariate analyses, AJCC stage, SEER summary stage, number of lymph nodes resected, and adjuvant therapy were significant predictors for OS and DSS, while grade, was significant only for OS. Multivariate Cox regression models demonstrated that poor grade, stage III/IV, distant disease, the number of lymph nodes resected being <4 and no adjuvant treatment were independent risk factors for poor OS, while stage III/IV, regional or distant disease, the number of lymph nodes <4 and no adjuvant treatment were independent risk factors for poor DSS. Multivariate Cox regression models also identified that chemotherapy and combination therapies were the independent risk factors for improved OS and DSS of early-stage USC. CONCLUSION: USC had a relatively poor prognosis compared with endometriod carcinoma. Moreover, advanced stage and fewer lymph nodes resected were independent negative prognostic factors for survival, while adjuvant therapy was significant for improved survival.

The Relationship between HDAC3 and Malignant Tumors: A Mini Review.

Histone deacetylases 3 (HDAC3) is a member of the histone deacetylases family. This family is associated with cellular physiological function, such as signal transduction, cell cycle, proliferation, apoptosis, and cardiac development. HDAC3 plays an important role in the progression of malignant tumors, especially in terms of proliferation, apoptosis, metastasis, angiogenesis, and anticancer drug resistance. This review discusses the basic elements of HDAC3 and the relationship between HDAC3 and malignant tumors.

Does Circular RNA Exert Significant Effects in Ovarian Cancer?

Circular RNA, also called circRNA, is a type of widespread RNA that is mainly derived from the spliceosome, from which DNA is transcribed into messenger RNA (mRNA). CircRNAs are characterized by the formation of a covalently closed continuous loop, and they play a variety of gene regulatory roles. CircRNAs were initially thought to represent splicing errors, but an increasing amount of evidence indicates that they may play significant roles in a variety of human diseases, including cancers, cardiovascular disease, neurological disorders, and pre-eclampsia. Studies have suggested that circRNAs regulate miRNA activity that arises from miRNA-mediated sponge effects. It has been shown that CDR1as acts as a sponge for miR-7 and that a testis-specific circRNA within the sex-determining region of Y (SRY) acts as a sponge for miR-138. Because miR-7 and miR-138 modulate several oncogenes and tumor suppressor genes, the interactions between ciRS-7 and miR-7 and between SRY and miR-138 may play important roles in cancer-related pathways. Hence, currently available data suggest that circRNAs may exert significant effects on diagnoses, prognoses, and therapies in ovarian cancer.

HDAC3 positively regulates HE4 expression to promote ovarian carcinoma progression.

OBJECTIVE: To identify the relationship between Histone deacetylase 3 (HDAC3) and Human epididymis protein 4 (HE4) and to explore the mechanisms underlying their effects on the malignant behaviors of ovarian carcinoma cells. METHODS: The expression levels of HDAC3 in ovarian carcinoma tissues were identified by immunohistochemistry, Western blot and real-time PCR. A wound healing assay, a Transwell assay and a CCK8 proliferation assay were used to assess the proliferation, invasion and metastatic capacities of ovarian carcinoma cells before and after transfection and HDAC3 protein treatment. HDAC3 and HE4 protein expression level in epithelial ovarian tissues were detected by immunohistochemistry, and the relationship between them was examined. RESULTS: HE4 was identified as an HDAC3-interacting protein. HDAC3 promotes ovarian carcinoma cell proliferation, invasion and migration by increasing the expression of HE4. HE4 and HDAC3 expression levels were significantly higher in malignant epithelial ovarian tissues than they were in benign and normal epithelial ovarian tissues. HDAC3 gene interference downregulated the expression of the PI3K/AKT signaling pathway-associated molecules P-PI3K/PI3K and P-AKT/AKT. CONCLUSION: HDAC3 expression is higher in ovarian carcinoma and promotes ovarian carcinoma cell proliferation, invasion and migration. HDAC3 and HE4 binding activates the PI3K/AKT signaling pathway, enhances ovarian carcinoma and promotes ovarian carcinoma cell proliferation, invasion and migration. Therefore, inhibiting the relationship between HDAC3 and HE4 may therefore have potential therapeutic value in patients with ovarian carcinoma.

MicroRNA regulation of transthyretin in trophoblast biofunction and preeclampsia.

Preeclampsia (PE) is the major cause of maternal, fetal and neonatal mortality affecting approximately 2-7% of pregnancies. Transthyretin (TTR) is down-regulated in PE pregnancies serum and placenta. Our bioinformatic analysis showed that TTR is a predicted target for miR-200a-3p and miR-141-3p. The aim of this study was to determine whether miR-200a-3p and miR-141-3p are involved in preeclampsia through its targeting of TTR in human placental trophoblasts. In human PE placenta, TTR transcript and protein levels were significantly lower associated with high expression of miR-141-3p and 200a-3p. We found that miR-200a-3p and miR-141-3p inhibited TTR expression by directly binding to the 3'UTR of TTR, which is reversed by mutation in the microRNA binding site. In preeclamptic plasm, TTR levels were significantly downregulated. TTR was validated as a direct target of miR-200a-3p and miR-141-3p using dual luciferase assays in JEG3 cells. Transwell insert invasion assays showed that TTR mediated the invasion-inhibitory effect of miR-200a-3p and miR-141-3p in JEG3 cells. These data provides new insight into physiological role of miR-141-3p and miR-200a-3p in regulating TTR during trophoblast dysfunction and PE development.

A novel MtHSP70-FPR1 fusion protein enhances cytotoxic T lymphocyte responses to cervical cancer cells by activating human monocyte-derived dendritic cells via the p38 MAPK signaling pathway.

OBJECTIVE: To evaluate the potential effects of recombinant mycobacterium tuberculosis heat shock protein 70-formyl peptide receptor 1 (MtHSP70-FPR1) fusion protein on human monocyte-derived dendritic cell (moDC) maturation; cytotoxic T lymphocyte (CTL) responses to cervical cancer (CC) cells; and the roles of the p38 MAPK, ERK, and JNK pathways in its transition. METHODS: Monocytes were positively selected with a MACS column with antiCD14 antibody-conjugated microbeads from umbilical cord blood. MoDCs were stimulated with MtHSP70-FPR1, MtHSP70, a mix of MtHSP70 and FPR1, FPR1, or phosphate buffer solution (PBS) as control. Flow cytometry was used to analyze the surface molecule expression of moDCs and IFN-γ-producing CD8+ T cells. T cell proliferation was assessed using [3][H]-thymidine assays. The cytotoxicity of moDC-activated T cells against CC cells was evaluated by MTT assays. Cytokine production was determined by enzyme-linked immunosorbent assay. Western blotting was used to investigate protein expression. RESULTS: Compared with MtHSP70, MtHSP70 + FPR1, FPR1, or PBS-mediated moDCs, MtHSP70-FPR1-pulsed moDCs expressed higher levels of CD80, CD86, CD83, HLA-DR, and CCR7; secreted more IL-12p70, TNF-ɑ and IL-1ß; and elicited stronger CTL priming and proliferation, resulting in an effective, HLA-I-dependent killing effect on CC cells. The p38 MAPK, ERK, and JNK pathways were all activated in MtHSP70-FPR1-mediated moDC maturation, but the p38 MAPK pathway played a vital role. CONCLUSIONS: The excellent capability of MtHSP70-FPR1 fusion protein to induce phenotypical and functional maturation of moDCs and CC-specific CTL responses partly illustrates the potential clinical benefits of DC-based immunotherapy for CC.

Supplementation of folic acid in pregnancy and the risk of preeclampsia and gestational hypertension: a meta-analysis.

OBJECTIVES: We aimed to systematically assess the relationship between folic acid supplementation in pregnancy and risk of preeclampsia and gestational hypertension. METHODS: The relevant studies were included by retrieving the Embase, PubMed and Cochrane library databases. Data extraction was conducted by two investigators independently. The risk ratio (RR) and 95% confidence interval (CI) were used as effect indexes to evaluate the relationship between folic acid supplementation and risk of gestational hypertension or preeclampsia. A subgroup analysis was performed according to the supplementation patterns of folic acid. The homogeneity of the effect size was tested across the studies, and publication biases were examined. RESULTS: In total, 13 cohort studies and 1 randomized controlled trial study was included, containing 160,562 and 149,320 women with and without folic acid supplementation during pregnancy. Pooled results showed that risk of gestational hypertension was not associated with the supplementation of folic acid. However, folic acid supplementation during pregnancy could significantly reduce the risk of preeclampsia. Moreover, the results of subgroup analysis showed that the decreased preeclampsia risk was associated with supplementation of multivitamins containing folic acid rather than folic acid alone. CONCLUSIONS: Our findings indicate that the supplementation of multivitamins containing folic acid during pregnancy could significantly lower preeclampsia risk.

N-myc Downstream-Regulated Gene 1 and Endometriosis: A Minireview.

NDRG1 (N-myc downstream-regulated gene 1) was previously considered to be a differentiation-related gene. However, many other functions of NDRG1 have since been identified, including proliferation, migration, invasion, and vascularization of tumor cells. Currently regarded as a tumor suppressor in most studies, NDRG1 is abundant in prostate, brain, kidney, placental, and intestinal tissues. It is expressed in normal endometrium, with higher expression occurring in the secretory phase. NDRG1 was first identified as an inhibitor of signaling pathways associated with the pathology of endometriosis. The NDRG1 protein regulation of endometriosis is assumed to be associated with several important pathways. This review summarizes the relationship between NDRG1 and endometriosis, focusing on the potential function of NDRG1 in endometriosis through signaling pathways and discusses the additional research that is required for future studies.

MtHsp70-CLIC1-pulsed dendritic cells enhance the immune response against ovarian cancer.

Approximately 80% of ovarian cancer (OC) is diagnosed at late stages, and most patients die within 5 years of diagnosis due to recurrence or drug resistance. Novel treatments are required to improve patient survival. Immune therapy against cancer is promising; however, therapeutic vaccination has been limited by the inability of tumor antigens to induce effective immune responses. Chloride intracellular channel 1 (CLIC1) was previously identified as a possible tumor marker for OC. In this study, we constructed a recombinant protein by conjugating the extracellular domain of CLIC1 to the carboxyl terminus of Mycobacterium tuberculosis heat shock protein 70 (MtHsp70). Human dendritic cells (DCs) derived from cortical blood were pulsed with the fusion protein, and the antitumor effect of human cytotoxic T lymphocytes (CTLs) stimulated by autologous DCs was assessed in NOG mice. MtHsp70-CLIC1 promoted the phenotypic maturation of human DCs and the secretion of Th1-associated cytokines in vitro. MtHsp70-CLIC1-stimulated CTLs generated a CLIC1-specific immune response both in vitro and in vivo. These results indicate that DCs pulsed with MtHsp70-CLIC1 can enhance antitumor immunity against OC, providing a novel immune therapeutic strategy.

Dual-cutoff of sFlt-1/PlGF ratio in the stratification of preeclampsia: a systematic review and meta-analysis.

PURPOSE: To systematically review the approach of using two independent sFlt-1/PlGF cutoffs that has better sensitivity (cutoff-sen) and specificity (cutoff-spe) separately for risk stratification in the detection of preeclampsia. METHODS: PubMed and Embase databases and reference lists were searched up to June 2016. Inclusion criteria were blood samples for sFlt-1/PlGF with separate cutoffs (cutoff-sen and cutoff-spe) provided. Six relevant studies were identified. Pooling of results was done based on three studies and a systematic review was performed based on all six. RESULTS: The strategy of using a cutoff of ≤33 and ≥85 for early onset preeclampsia, and ≤33 and ≥110 for the late onset preeclampsia was proposed and examined. The pooled sensitivity for cutoff-sen was: 95.3% (90.6-98.1%) and 88.6% (82.9-92.9%) for early and late onset preeclampsia, respectively. The pooled specificity for cutoff-spe was: 97.6% (95.2-98.9%) and 94.2% (91.4-96.3%) for early and late onset preeclampsia respectively. The pooled estimation of the early onset pre-eclamptic pregnancies and control normal pregnancies classified in the equivocal zone was 4.9% (2.0-8.8%) and 32.4% (25.7-39.5%), respectively, and 26.8% (10.3-47.6%) and 8.7% (3.0-17.6%) for late onset patients. CONCLUSION: The new dual-cutoff diagnostic system optimizes the predictive performance of the single cutoff system. Further studies are required to assess the performance of this system and to define the approach and frequency at which subjects in the equivocal zone should be screened.

Transthyretin and Normal Human Pregnancy: Mini Review.

Since transthyretin (TTR) was discovered, it has been regarded as a serum protein carrier of thyroid hormones and retinol. However, many other important functions of TTR have been found recently, and current evidence suggests that it plays a role in human receptivity and normal pregnancy. TTR is abundant in the uterine cavity, uterine secretion, placenta, and serum of pregnant females in the peri-implantation uterus and the first trimester of pregnancy. It may be involved in the delivery of maternal thyroid hormones to the fetus. In addition, it appears to play a key role in the preeclampsia mechanism and may be involved in spiral artery remodeling. This review will summarize what is currently known about TTR and normal pregnancy; it will focus on our findings regarding the role of TTR in the spiral artery remodeling process and the additional research required in the future.

Serum markers of pre-eclampsia identified on proteomics.

AIM: Pre-eclampsia (PE) is a disorder of pregnancy associated with maternal and fetal mortality and morbidity. The aim of the present study was to use proteomics to identify biomarkers of, and elucidate the pathogenesis of, PE. METHODS: Serum samples were analyzed using peptide ligand library beads (PLLB) on liquid chromatography-mass spectrometry/mass spectrometry. Retinol-binding protein 4 (RBP4) was used as the target protein for further validation on enzyme-linked immunosorbent assay, immunohistochemistry and real-time polymerase chain reaction. Transwell invasion assay was used to evaluate whether RBP4 affects the invasive ability of trophoblast tumor cells. RESULTS: Twenty upregulated and 17 downregulated proteins were differentially expressed between severe PE patients and healthy pregnant women. Those proteins were further classified according to molecular function and biological process according to the gene ontology terms. RBP4 concentration was significantly lower in women with severe PE than in those with healthy pregnancy. CONCLUSIONS: RBP4 is able to function as biomarker to distinguish severe PE from normal pregnancy. More importantly, these results may shed light on the role of RPB4 in the pathogenesis in PE. Further studies are required to validate these results, and determine the precise role of RBP4 in the pathogenesis of PE.

A Randomized Controlled Trial on the Efficacy and Safety of a New Crosslinked Hyaluronan Gel in Reducing Adhesions after Gynecologic Laparoscopic Surgeries.

STUDY OBJECTIVE: To evaluate the safety and efficacy of a new crosslinked hyaluronan (NCH) gel in reducing postoperative adhesions. DESIGN: Randomized controlled trial (Canadian Task Force classification I). SETTINGS: Seven departments of obstetrics and gynecology in China. PATIENTS: A total of 216 women scheduled for gynecologic laparoscopic surgery for primary removal of adhesions, myomas, ovarian cysts, or endometriotic cysts. INTERVENTIONS: Patients were randomized to receive either NCH gel or saline with 1:1 allocation. MEASUREMENTS AND MAIN RESULTS: All patients were evaluated using a modified American Fertility Society (mAFS) scoring system for the incidence, extent, and severity of pre-existing and postoperative adhesions at the 10 anatomic sites of ovaries/tubes and at the expanded 23 or 24 anatomic sites throughout the abdominopelvic cavity by laparoscopy. A total of 215 randomized patients were treated with either saline solution (108 of 108) or NCH gel (107 of 108), composing the full analysis set (FAS), and 196 patients (94 of 108 in the saline control group and 102 of 108 in the NCH gel group) completed the entire study, composing the per protocol set (PPS). The postoperative incidence of moderate or severe adhesions evaluated at the 10 sites (the primary endpoint for efficacy) was 27.7% in the control group and 9.8% in the NCH gel group, a difference of 14.4% (95% confidence interval [CI], 2.6%-20.6%) in the PPS, and 37.0% in the control group and 14.0% in the NCH gel group, a difference of 20.0% (95% CI, 8.9%-26.8%) in the FAS. The postoperative incidence of moderate or severe adhesions evaluated at the 24 sites was also significantly lower in the NCH gel group compared with the control group (5.9% vs 14.9%; p = .036) in the PPS. Also in the PPS, the NCH gel group had significantly lower postoperative adhesion scores of severity, extent, and mAFS: 60.0%, 50.8%, and 76.9%, respectively (median scores of the 10 sites; p = .002) and 48.5%, 50.0%, and 72.2% (median scores of the 24 sites; p = .001) lower than those recorded in the control group. No serious adverse events were observed, and the safety profile of NCH gel was comparable to that of saline control. CONCLUSION: This study demonstrates that NCH gel is safe and significantly reduces adnexal adhesion formation and global adhesion formation throughout the abdominopelvic cavity after gynecologic laparoscopic surgery.

Total laparoscopic radical trachelectomy in the treatment of early-stage cervical cancer: review of technique and outcomes.

PURPOSE OF REVIEW: Fertility preservation in early-stage cervical cancer by total laparoscopic radical trachelectomy (TLRT) is gaining acceptance as more cases are published in the literature. The objective is to review all the literatures regarding TLRT especially over the last 12 months and to describe the technique, the operative outcomes, the oncologic outcomes and the obstetric outcomes of this procedure. RECENT FINDINGS: As the number of cases reported in the literature increases, the effectiveness of TLRT for treating early-stage cervical cancer continues to gain support. Under the enhanced vision of the laparoscopy, it is easy to preserve the ascending branches of the uterine arteries and to divide the ligaments surrounding the cervix and vagina. Since TLRT was first reported, about 140 cases of TLRT have been reported. The tumour recurrence rate is 2.9%. Fifty-nine out of 140 patients attempted to conceive after TLRT, and forty-six patients succeeded. There were 17 miscarriages, 14 preterm births and 11 term births. SUMMARY: TLRT appears well tolerated and effective when performed in centres with appropriate experience of laparoscopic techniques. Continued research and clinical trials are needed to further elucidate the equivalence or superiority of TLRT to conventional methods in terms of obstetric outcome and patients' quality of life.

Galectin-3 promotes fibrosis in ovarian endometriosis.

Objective: This study aimed to investigate the potential role of galectin-3 (Gal-3) in the pathogenesis of fibrotic alterations in ovarian endometriosis (OVE). Methods: In this study, we collected the ectopic endometrial tissues and eutopic endometrial tissues from 31 OVE patients treated by laparoscopy, and the eutopic endometrial tissues from 23 non-OVE patients with leiomyoma or other benign diseases were used as control. Hematoxylin and eosin (H&E) and Masson's trichrome staining were utilized for histopathological assessment. The primary normal endometrial stromal cells (NESC), ectopic endometrial stromal cells (ECSC), and eutopic endometrial stromal cells (EUSC) were isolated. Gal-3 overexpression plasmids (Gal-OE) and short hairpin RNA targeting Gal-3 (Gal-3-shRNA) were transfected into the immortalized human endometriotic cell line 12Z, respectively. RT-qPCR, Western blot analysis, and immunohistochemistry were used to detect the mRNA and protein expression levels of Gal-3, type I collagen (COL-1), connective tissue growth factor (CTGF) and α-smooth muscle actin (α-SMA), respectively. Results: H&E and Masson staining showed that ovarian ectopic endometrium exhibited glandular hyperplasia, high columnar glandular epithelium, apical plasma secretion, more subnuclear vacuoles, and obvious fibrosis, compared with normal endometrium. The mRNA and protein levels of Gal-3 , CTGF, α-SMA, and COL-1 were all upregulated in the ectopic endometrial tissues of OVE patients compared to the eutopic endometrial tissues from OVE patients and non-OVE patients. Moreover, ECSC expressed higher levels of Gal-3, CTGF, α-SMA, and COL-1 than EUSC and NESC. Follow-up investigations demonstrated that the Gal-3 overexpression substantially increased fibrosis-related markers including CTGF, α-SMA, and COL-1 within the 12Z cell line. Conversely, Gal-3 knockdown showed the opposite effects. Conclusion: Gal-3 promotes fibrosis in OVE, positioning it as a prospective therapeutic target for mitigating fibrosis in endometriosis.

Azacytidine induces necrosis of multiple myeloma cells through oxidative stress.

Azacytidine is an inhibitor of DNA methyltransferase and is known to be an anti-leukemic agent to induce cancer cell apoptosis. In the present study, multiple myeloma cells were treated with azacytidine at clinically relevant concentrations to induce necrosis through oxidative stress. Necrotic myeloma cells exhibit unique characteristics, including enrichment of the cell-bound albumin and overexpression of endoplasmic reticulum (ER)- and mitochondrial-specific chaperones, which were not observed in other necrotic cells, including HUH-7, A2780, A549, and Hoc1a. Proteomic analysis shows that HSP60 is the most abundant up-regulated mitochondrial specific chaperone, and azacytidine-induced overexpression of HSP60 is confirmed by western blot analysis. In contrast, expression levels of cytosolic chaperones such as HSP90 and HSP71 were down-regulated in azacytidine-treated myeloma cells, concomitant with an increase of these chaperones in the cell culture medium, suggesting that mitochondrial chaperones and cytosolic chaperones behave differently in necrotic myeloma cells; ER- and mitochondrial-chaperones being retained, and cytosolic chaperones being released into the cell culture medium through the ruptured cell membrane. Our data suggest that HSP60 is potentially a new target for multiple myeloma chemotherapy.