Intrauterine transplantation of autologous bone marrow derived mesenchymal stem cells followed by conception in a patient of severe intrauterine adhesions

On a woman with severe intrauterine adhesions, hysteroscopy followed by cyclical hormone replacement therapy was tried for 5 months, for development of the endometrium. When this failed, autologous stem cells were tried as an alternative therapy. Adult autologous bone marrow mesenchymal stem cells isolated from patient’s own bone marrow and were cultured and placed in the endometrial cavity under ultrasound guidance after curettage. Patient was then given cyclical hormonal therapy. Endometrium was assessed intermittently using ultrasound. Three months later, endometrium partly recovered with improved ultrasonic echo. This resulted in spontaneous pregnancy followed by confirmation of gestational sac, yolk sac, and primitive heart tube pulse on ultrasound. Autologous bone marrow derived mesenchymal stem cells could regenerate injured endometrium not responding to conventional treatment and can be used as an alternative in females with severe Asherman’s syndrome.

Intrauterine Adhesions (IUAs) or Asherman’s Syndrome (AS) and the Stem Cells Treatment: A Systemic Review and Meta-Analysis

Background: Intrauterine Adhesions (IUAs) or Asherman’s Syndrome (AS) usually contains symptoms such as decreased menstrual flow or even amenorrhea, chronic pelvic pain, recurrent abortion and infertility. The current treatment of IUAs includes hysteroscopic adhesiolysis, oral hormone and biological barriers, but each of them has limitation. Stem cell therapy may be an expanding field seeking for therapy in IUAs. Objective: We will discuss current advances in stem cell therapy as a treatment for endometrial pathophysiology. Materials and Methods: We search on PubMed, Embase and Cochrane library and select several keywords on researches, then review the cell biology theories and animal experiments, finally do meta-analysis in human clinical trials. Results: 77 articles on PubMed, 71 articles on Embase and 17 articles on Cochrane Library, as a result, 37 articles are included under the criteria, which are intrauterine adhesions (IUAs), Asherman’s Syndrome (AS), cell therapy, stem cells, bone marrow stem cells, clinical trials, recent 10 years and human or animal experiments. The included criteria: original articles, cohort study, case control study, animal experiments, human clinical trials, high quality, 10 years recent. The excluded articles are case reports, meeting reports, low quality or more than 10 years ago. Conclusion: Stem cell may be a new therapeutic schedule for IUAs in the future clinical treatment, but it is necessary to compare it with traditional therapy such as oral hormone, also the development of random clinical tests should proceed. For clinical treatment on IUAs, stem cells could be a new choice.

Diagnostic Accuracy of Hysterosalpingography in the Identification of Intrauterine Adhesions in Infertile Women in a Referral Centre, Southern Nigeria

Background: Examination of the endometrial cavity is crucial in the management of infertile women. This evaluation is done by Hysterosalpingography (HSG) in the University of Port Harcourt Teaching Hospital. The diagnostic value and accuracy of hysterosalpingography varies in various centres and has not been assessed in the University of Port Harcourt Teaching Hospital (UPTH), hence this study. Methodology: A prospective cross sectional study done among 101 infertile women at the gynaecology clinic of the UPTH from December 2018 to July 2019. Socio-demographic data of women who met the eligibility criteria were collected. Their hysterosalpingographic and outpatient hysteroscopic findings were statistically analyzed using IBM SPSS for windows version 20.0. The validity and reliability statistics of hysterosalpingography for the identification of endometrial adhesions were determined and significance was reported at p-value < 0.05. The analyzed data was presented in tables and figure. Result: The respondents were all married, 66.3% of them were nulliparous, while 54.5% were above 35 years of age. Most, 93.1% of the studied group had at least secondary education and had duration of infertility of more than 2 years. There was a moderate strength of agreement in the diagnostic accuracy of HSG with hysteroscopy in the diagnosis of intrauterine adhesions;showing sensitivity and specificity of 59.6% and 90.9% respectively. The accuracy of HSG in the identification of intrauterine adhesions in this study was 73.3%. The agreement in the diagnostic accuracy of HSG using Hysteroscopy as a gold standard for intrauterine adhesions was therefore moderately significant (k = 0.482). Conclusion: HSG had an overall fair strength of agreement with office hysteroscopy in the identification of intrauterine adhesions. It is a specific but not a sensitive predictor of intrauterine adhesion.

Correlation of FoxF2 expression with MMPs, ADAMs and TGF-β1 pathway in intrauterine adhesions

Objective: To study the correlation of forkhead box F2 (FoxF2) expression with matrix metalloproteinases (MMPs), a disintegrin and metalloproteases (ADAMs), transforming growth factor-β1 (TGF-β1) pathway in intrauterine adhesions. Methods: The patients who underwent hysteroscopic surgery and were diagnosed with intrauterine adhesions according to postoperative pathology results in our hospital between March 2015 and January 2018 were selected as the IUA group, and the patients who underwent surgical resection and were diagnosed with uterine fibroid according to postoperative pathology results in our hospital during the same period were selected as the control group. The intrauterine adhesion tissues of the IUA group and the normal endometrial tissues of the control group were collected to measure the expression levels of FoxF2, MMPs and ADAMs-related molecules as well as TGF-β1 pathway molecules. Results: FoxF2, ADAM15, ADAM17, TIMP1, TGF-β1, Smad2, Smad3 and CTGF mRNA expression in the intrauterine adhesion tissues of IUA group were significantly higher than those of control group whereas MMP9 mRNA expression was significantly lower than that of control group;FoxF2 mRNA expression in the intrauterine adhesion tissues of IUA group was negatively correlated with MMP9 mRNA expression, and positively correlated with ADAM15, ADAM17, TIMP1, TGF-β1, Smad2, Smad3 and CTGF mRNA expression. Conclusion: The high expression of FoxF2 in intrauterine adhesions can inhibit protease activity and enhance TGF-β1 pathway function.

Safety of menstrual blood-derived stromal cell transplantation in treatment of intrauterine adhesion

BACKGROUND Intrauterine adhesion(IUA)can cause serious damage to women’s reproductive health,yet current treatment methods are difficult to achieve satisfactory results.In our previous studies,we demonstrated that menstrual-derived stromal stem cells(MenSCs),with high proliferative capacity and self-renewal ability,have a powerful therapeutic effect in patients with severe IUA.However,safety assessment of MenSCs transplantation is essential for its further application.AIM To evaluate the short-,medium-,and long-term biosafety of MenSCs via intrauterine transplantation in a rat model of IUA,with a focus on toxicity and tumorigenicity.METHODS MenSCs were injected into the sub-serosal layer of the uterus in an IUA rat model,for 3 d,3 mo,and 6 mo separately,to monitor the corresponding acute,sub-chronic,and chronic effects.Healthy rats of the same age served as negative controls.Toxicity effects were evaluated by body weight,organ weight,histopathology,hematology,and biochemistry tests.Tumorigenicity of MenSCs was investigated in Balb/c-nu mice in vivo and by colony formation assays in vitro.RESULTS Compared with the same week-old control group,all of the IUA rats receiving MenSC transplantation demonstrated no obvious changes in body weight,mainorgan weight,or blood cell composition during the acute,sub-chronic,and chronic observation periods.At the same time,serum biochemical tests showed no adverse effects on metabolism or liver and kidney function.After 4 wk of subcutaneous injection of Men SCs in Balb/c-nu nude mice,no tumor formation or cell metastasis was observed.Moreover,there was no tumor colony formation of Men SCs during soft agar culture in vitro.CONCLUSION There is no acute,sub-chronic,or chronic poisoning,infection,tumorigenesis,or endometriosis in rats with IUA after Men SC transplantation.The above results suggest that intrauterine transplantation of Men SCs is safe for endometrial treatment.

Clinical and investigative correlates of etiologic risk factors on treatment outcome of intrauterine adhesion in women with infertility: A descriptive study

Background: Intrauterine adhesion is a clinical entity that may present with subfertility. However, outcome of management depends on severity of intrauterine adhesion and type of treatment intervention employed. Objective: To evaluate clinical and investigative correlation of etiologic risk factors of intrauterine adhesion on fertility and pregnancy outcome following treatment. Patients and Methods: A 5 years prospective observational study. Treatment employed includes transvaginal blind intrauterine adhesiolysis, insertion of inert intrauterine device or inflated Foley’s catheter balloon and oestrogen therapy. Inclusion criteria are intrauterine adhesion as the only identifiable cause of infertility and post treatment follows up for at least a year. Results: Over a period of 5 years, 63 patients with mean age of 31.6 years and age range of 21 to 42 years were managed. Etiologic risk factors were dilatation and curettage 33 (52.4%), vacuum aspiration 13 (20.6%), myomectomy 11 (17.5%) and caesarean section in 6 (9.5%) patients. Of the 63 patients, 28 conceived giving a pregnancy rate of 44.4%. Nine out of the 28 patients that conceived had miscarriage, giving a miscarriage rate of 32.1%. The highest pregnancy rate and lowest miscarriage rate were recorded in the subgroup that had vacuum aspiration as their etiologic risk factor. Late pregnancy complications encountered are preterm contractions 5 (26.3%), placenta praevia 4 (21.1%), morbidly adherent placenta 5 (26.3%) and preterm delivery in 2 (10.5%) patients. Route of delivery was per vaginum in 13 (68.4%) patients and by caesarean section in 6 (31.6%) patients. Live birth rate was 89.5% (17/19 deliveries). There was no maternal mortality recorded. Conclusion: Compared to other risk factors, uterine vacuum aspiration was associated with higher fertility rate and better pregnancy outcome.

The Clinical Value of Ultrasound Image Texture Analysis in the Diagnosis of Uterine Adhesions

Purpose: This review examines the diagnostic value of transvaginal 3D ultrasound image texture analysis for the diagnosis of uterine adhesions. Materials and Methods: The total clinical data of 53 patients with uterine adhesions diagnosed by hysteroscopy and the imaging data of transvaginal three-dimensional ultrasound from the Second Affiliated Hospital of Chongqing Medical University from June 2022 to August 2023 were retrospectively analysed. Based on hysteroscopic surgical records, patients were divided into two independent groups: normal endometrium and uterine adhesion sites. The samples were divided into a training set and a test set, and the transvaginal 3D ultrasound was used to outline the region of interest (ROI) and extract texture features for normal endometrium and uterine adhesions based on hysteroscopic surgical recordings, the training set data were feature screened and modelled using lasso regression and cross-validation, and the diagnostic efficacy of the model was assessed by applying the subjects’ operating characteristic (ROC) curves. Results: For each group, 290 texture feature parameters were extracted and three higher values were screened out, and the area under the curve of the constructed ultrasonographic scoring model was 0.658 and 0.720 in the training and test sets, respectively. Conclusion Relative clinical value of transvaginal three-dimensional ultrasound image texture analysis for the diagnosis of uterine adhesions.

Expression of miR-195 in intrauterine adhesion and its relationship with TGF-β1/Smads and FGF2/FGFR1/ERK pathways

Objective:To investigate the expression of miR-195 in intrauterine adhesion(IUA)and its relationship with TGF-β1/Smads and FGF2/FGFR1/ERK pathways.Methods:118 cases of IUA patients who underwent hysteroscopic treatment in our hospital between September 2017 and February 2019 were regarded as IUA group,80 cases of dysfunctional uterine bleeding patients who underwent hysteroscopic curettage in our hospital during the same period were regarded as control group.Differences in the expression levels of miR-195 as well as TGF-β1/Smads and FGF2/FGFR1/ERK signaling pathway-related molecules in the focal tissues obtained by hysteroscopy were compared between the two groups.Pearson test was used to evaluate the correlation of miR-195 expression in the intrauterine adhesion tissues with TGF-β1/Smads and FGF2/FGFR1/ERK pathways in IUA patients.Results:miR-195 expression in intrauterine adhesion tissues of IUA group was higher than that in endometrial tissues of control group(P<0.05).TGF-β1,Smad2 and Smad3 mRNA expression in intrauterine adhesion tissues of IUA group were higher than those in endometrial tissues of control group;FGF2,FGFR1 and ERK mRNA expression in intrauterine adhesion tissues were higher than those in endometrial tissues of control group(P<0.05).Pearson test showed that the miR-195 expression in intrauterine adhesion tissues of IUA group was positively correlated with the TGF-β1/Smads pathway-related molecules TGF-β1,Smad2 and Smad3 mRNA expression,and positively correlated with the FGF2/FGFR1/ERK pathway-related molecules FGF2,FGFR1 and ERK mRNA expression(P<0.05).Conclusion:miR-195 is highly expressed in IUA lesion tissues and may promote disease progression by activating TGF-β1/Smads and FGF2/FGFR1/ERK signaling pathways.

Clinical Study of Changmaile II Combined with Estrogen and Progesterone in Preventing and Treating Intrauterine Adhesion

Purpose: To explore the clinical efficacy of Changmaile II combined with cyclic estrogen/progesterone therapy in treating hypomenorrhea caused by intrauterine adhesion (IUA) and its effect on the endometrium after the separation procedure for intrauterine adhesion. Methods: Sixty patients with IUA confirmed by hysteroscopy in the Second People’s Hospital of Fujian University of Traditional Chinese Medicine from March 2020 to December 2020 were selected and randomly divided into the control group and the observation group, with 30 cases in each group. After the separation procedure for intrauterine adhesion, the observation group underwent the cyclic estrogen/progesterone therapy alone, while the observation group was administered the in-hospital preparation of Changmaile II in addition to the therapy of the observation group. Menstrual recovery, endometrial thickness, and the diagnostic grading scores of IUA after 3 menstrual cycles were measured in both groups. Results: After the treatment for 3 menstrual cycles, the total effective rate was higher in the observation group (73.33%) than in the control group (53.33%) (P < 0.05). The period flow was heavier in the observation group than in the control group (P < 0.05). The endometrial thickness of both groups was thicker than before treatment (P < 0.05), nonetheless the difference between the groups was not statistically significant (P > 0.05). After treatment, the diagnostic grading scores of IUA were higher in the observation group than in the control group (P < 0.05). Conclusions: Changmaile II combined with estrogen and progesterone artificial cycle therapy could increase the menstrual volume and endometrial thickness of patients with intrauterine adhesion, while promoting the endometrial repair, with better efficacy than artificial cycle therapy alone.

Correlation of the changes of TLR4/NF-κB pathway function in intrauterine adhesion tissue with the characteristics of cytokine secretion and collagen metabolism

Objective:To study the correlation of the changes of Toll-like receptor 4 (TLR4) / nuclear factorκB (NF-κB) pathway function in intrauterine adhesion (IUAs) tissue with the characteristics of cytokine secretion and collagen metabolism.Methods:The patients with IUAs who were treated in our hospital between February 2015 and March 2018 were selected as the IUAs group, and the patients who underwent hysteroscopy due to infertility and were pathologically confirmed to have normal endometrium during the same period were selected as the control group. The expression levels of TLR4/NF-κB pathway molecules and collagen metabolism genes as well as the contents of cytokines and collagen metabolism markers in the adhesion tissues of IUAs group and the normal endometrial tissue of control group were measured.Results: TLR4, NF-κB, a disintegrin and metalloproteinase 15 (ADAM15), ADAM17, matrix metalloproteinase 9 (MMP9) and plasminogen activator inhibitor 1 (PAI-1) mRNA expression as well as transforming growth factor-β1 (TGF-β1), Smad2/3, insulin-like growth factor 1 (IGF-1), IGF-1 receptor (IGF-1R), basic fibroblast growth factor (bFGF), periostin/osteoblast-specific factor 2 (Postn), type I collagen (Col-I) and actin-α (α-SMA) contents in the adhesion tissues of IUAs group were significantly higher than those of control group while urokinase-type plasminogen activator (uPA) mRNA expression was significantly lower than that of control group;TLR4 and NF-κB mRNA expression were positively correlated with TGF-β1, Smad2/3, IGF-1, IGF-1R, bFGF, Postn, Col-I,α-SMA, ADAM15, ADAM17, MMP9 and PAI-1, and negatively correlated with uPA.Conclusion:The excessive activation of TLR4/NF-κB pathway in IUAs is associated with the cytokine secretion and collagen metabolism abnormalities.

羊膜细胞外基质与膀胱细胞外基质材料修复大鼠子宫内膜损伤

背景:大量研究已证实,羊膜细胞外基质与膀胱细胞外基质材料均可作为干细胞载体用于子宫内膜损伤的治疗,但是有关两种材料的比较研究相对少见。目的:对比羊膜细胞外基质与膀胱细胞外基质材料作为干细胞载体治疗子宫内膜损伤的差异。方法:采用全骨髓贴壁法分离纯化SD大鼠骨髓间充质干细胞。分别制备SD大鼠羊膜细胞外基质与膀胱细胞外基质材料,然后将骨髓间充质干细胞分别接种于两种材料表面,检测细胞增殖与黏附情况。将40只SD大鼠随机分为4组(n=10),除假手术组外,子宫内膜损伤组、羊膜细胞外基质组、膀胱细胞外基质组均通过机械干预的方式建立子宫内膜损伤模型,分别将羊膜细胞外基质/骨髓间充质干细胞复合物、膀胱基质细胞外基质/骨髓间充质干细胞复合物移植至羊膜细胞外基质组、膀胱细胞外基质组大鼠损伤内膜部位,移植后14,28 d取材检测,苏木精-伊红染色观察大鼠子宫内膜组织形态,酶联免疫吸附法分析子宫内膜组织中碱性成纤维细胞生长因子、胰岛素样生长因子1与血管内皮生长因子水平,免疫组化染色分析子宫内膜组织中波形蛋白与CD34表达。结果与结论:(1)两种细胞外基质材料均有利骨髓间充质干细胞的增殖,相较于膀胱细胞外基质材料,羊膜细胞外基质材料可促进骨髓间充质干细胞的黏附;(2)相较于假手术组,子宫内膜损伤组碱性成纤维细胞生长因子、胰岛素样生长因子1与血管内皮生长因子水平降低(P <0.01),子宫内膜组织形态发育不良,内膜厚度与腺体数量减少,波形蛋白与CD34阳性表达减少(P <0.01);相较于子宫内膜损伤组,羊膜细胞外基质组、膀胱细胞外基质组碱性成纤维细胞生长因子、胰岛素样生长因子1与血管内皮生长因子水平均升高(P <0.05或P <0.01),子宫内膜组织形态明显改善,内膜厚度与腺体数量增加,波形蛋白与CD34阳性表达增加(P <0.05或P <0.01),并且羊膜细胞外基质组的改善作用优于膀胱细胞外基质组(P <0.05);(3)结果表明,相较于膀胱细胞外基质材料,羊膜细胞外基质材料作为骨髓间充质干细胞的载体可进一步促进损伤子宫内膜的修复。

聚己内酯-透明质酸静电纺丝膜联合间充质干细胞修复子宫内膜损伤

背景:人子宫内膜间充质干细胞能够直接修复受损的子宫内膜,促进血管生成、恢复子宫形态结构,然而将干细胞直接注入受损子宫内膜后的细胞存活率低、滞留时间短,修复效果有限。目的:观察聚己内酯-透明质酸静电纺丝膜复合人子宫内膜间充质干细胞修复大鼠子宫内膜损伤的效果。方法:①细胞实验:采用胶原酶消化法提取人子宫内膜间充质干细胞,静电纺丝技术制备聚己内酯-透明质酸静电纺丝膜。将人子宫内膜间充质干细胞分别接种于聚苯乙烯培养板与聚己内酯-透明质酸静电纺丝膜上,通过DNA定量分析、WST-1细胞活性实验、鬼笔环肽染色、扫描电镜观察细胞的增殖与黏附能力,qRT-PCR检测静电纺丝膜上细胞CD90、Meflin的mRNA表达。②动物实验:取27只处于动情期的雌性SD大鼠,通过机械搔刮法建立宫腔粘连模型后随机分为3组,每组9只:空白对照组不进行任何治疗,对照组将聚己内酯-透明质酸静电纺丝膜植入宫腔损伤部位,实验组将聚己内酯-透明质酸静电纺丝膜/人子宫内膜间充质干细胞补片植入宫腔损伤部位。术后第3,7,14天取材,采用苏木精-伊红染色观察子宫形态结构及腺体数量,qRT-PCR和免疫荧光染色观察子宫组织CD31、血管内皮生长因子的表达。结果与结论:①细胞实验:与聚苯乙烯培养板相比,聚己内酯-透明质酸静电纺丝膜可促进人子宫内膜间充质干细胞的增殖与黏附,并且聚己内酯-透明质酸静电纺丝膜支持人子宫内膜间充质干细胞基因CD90和Meflin的表达;②动物实验:苏木精-伊红染色显示,聚己内酯-透明质酸静电纺丝膜/人子宫内膜间充质干细胞补片可促进子宫内膜损伤后形态结构的恢复,术后第14天的内膜厚度与腺体数量均多于空白对照组、对照组(P<0.05);qRT-PCR和免疫荧光染色检测显示,实验组术后第7,14天的CD31、血管内皮生长因子mRNA与蛋白表达均高于空白对照组、对照组(P<0.05);③结果表明:聚己内酯-透明质酸静电纺丝膜可以提高干细胞的存活率、延长干细胞与受损组织的接触时间,二者复合移植可更好地修复受损子宫内膜组织。

Effect of autologous bone marrow stem cells-scaffold transplantation on the ongoing pregnancy rate in intrauterine adhesion women:a randomized,controlled trial

Intrauterine adhesion is a major cause of female reproductive disorders.Although we and others uncontrolled pilot studies showed that treatment with autologous bone marrow stem cells made a few patients with severe intrauterine adhesion obtain live birth,no large sample randomized controlled studies on this therapeutic strategy in such patients have been reported so far.To verify if the therapy of autologous bone marrow stem cells-scaffold is superior to traditional treatment in moderate to severe intrauterine adhesion patients in increasing their ongoing pregnancy rate,we conducted this randomized controlled clinical trial.Totally 195 participants with moderate to severe intrauterine adhesion were screened and 152 of them were randomly assigned in a 1:1 ratio to either group with autologous bone marrow stem cells-scaffold plus Foley balloon catheter or group with only Foley balloon catheter(control group)from February 2016 to January 2020.The per-protocol analysis included 140 participants:72 in bone marrow stem cells-scaffold group and 68 in control group.The ongoing pregnancy occurred in 45/72(62.5%)participants in the bone marrow stem cells-scaffold group which was significantly higher than that in the control group(28/68,41.2%)(RR=1.52,95%CI 1.08–2.12,P=0.012).The situation was similar in live birth rate(bone marrow stem cells-scaffold group 56.9%(41/72)vs.control group 38.2%(26/68),RR=1.49,95%CI 1.04–2.14,P=0.027).Compared with control group,participants in bone marrow stem cells-scaffold group showed more menstrual blood volume in the 3rd and 6th cycles and maximal endometrial thickness in the 6th cycle after hysteroscopic adhesiolysis.The incidence of mild placenta accrete was increased in bone marrow stem cells-scaffold group and no severe adverse effects were observed.In conclusion,transplantation of bone marrow stem cells-scaffold into uterine cavities of the participants with moderate to severe intrauterine adhesion increased their ongoing pregnancy and live birth rates,and this therapy was relatively safe.

The study on the safety and efficacy of amnion graft for preventing the recurrence of moderate to severe intrauterine adhesions

Transcervical resection of adhesion(TCRA)is the standard treatment for the intrauterine adhesions,but the recurrence of adhesions is a tough problem for the gynecologist.In addition,the therapeutic strategy after TCRA about prevention of recurrence remains controversial especially for the patients with moderate to severe intrauterine adhesions(IUAs).Hence,we designed this study to explore the safety and efficacy of fresh amnion grafts for preventing the recurrence after TCRA for patients with moderate to severe IUAs.One hundred patients with moderate to severe IUAs who presented with a history of hypomenorrhea,amenorrhea and infertility were included in the study from January 2015 to December 2017.Patients were divided into amnion group(52 patients)and chitosan group(48 patients).Fresh amnion grafts or intrauterine injections of chitosan were administered after TCRA.Transvaginal ultrasonography(TVUS)and hysteroscopy were performed at the first and third month after the operation.The surgical procedures for all patients were completed successfully without relevant complications.In amnion group,8 patients exhibited relapse in the first month and 2 patients in three months after surgery;in chitosan group,23 women exhibited relapse in the first month and 18 patients in three months after surgery.Statistical analysis revealed that the recurrence rate of adhesion in amnion group was significantly lower than those of chitosan group in the first and three months after surgery(P1=0.000,P2=0.000).After TCRA,fresh amnion graft plays a significant role in preventing further adhesions than injections of chitosan.

In situ delivery of apoptotic bodies derived from mesenchymal stem cells via a hyaluronic acid hydrogel:A therapy for intrauterine adhesions

Stem cell-based and stem cell-derived exosome-based therapies have shown promising potential for endometrial regeneration and the clinical treatment of intrauterine adhesions(IUAs).Evidence shows that apoptosis occurs in a majority of grafted stem cells,and apoptotic bodies(ABs)play a critical role in compensatory tissue regeneration.However,the therapeutic potential of AB-based therapy and its mechanism have not been explored in detail.Here,a cell-free therapeutic strategy was developed by incorporating mesenchymal stem cell-derived ABs into a hyaluronic acid(HA)hydrogel to achieve endometrial regeneration and fertility restoration.Specifically,we found that the ABs could induce macrophage immunomodulation,cell proliferation,and angiogenesis in vitro.The HA hydrogel promoted the retention of ABs and facilitated their continuous release.In a murine model of acute endometrial damage and a rat model of IUAs,in situ injection of the AB-laden HA hydrogel could efficiently reduce fibrosis and promote endometrial regeneration,resulting in the fertility restoration.Consequently,ABs show good potential as therapeutic vesicles,and the AB-laden HA hydrogel appears to be a clinically feasible and cell-free alternative for endometrial regeneration and IUA treatment.

Creation of a rabbit model for intrauterine adhesions using electrothermal injury

The pathogenesis and therapeutic treatment of intrauterine adhesions （IUAs） remain unsolved, highlighting the need for stable and effective experimental animal models. In this study, uterine electrocoagulaUon of twenty-one female New Zealand White rabbits was carried out to establish an IUA model. As rabbits have two completely separate uterine horns, each rabbit had its own internal control： one uterine horn was given an electrothermal injury （Group A, n=21）, and the contralateral uterine horn received no treatment and served as the control （Group B, n=21）. The en- dometrial morphology, number of endometrial glands, area of endometrial fibrosis, and number of implanted fetuses were compared between the two groups. In Group A, the numbers of endometrial glands on Days 7 and 14 and the number of implanted fetuses were significantly lower than those in Group B（P〈0.05, P〈0.05, and P〈0.01, respectively）, while the ratio of the area with endometrial stromal fibrosis to the total endometrial area was significantly increased （P〈0.01）. These results suggest that this method of electrothermal injury is effective for the establishment of a rabbit lUA model between 7 and 14 d after surgery.

An Impact of Suspected Peritubal Adhesions by Hysterosalpingography on Outcomes of Intrauterine Insemination

Objective To assess the fertility prospect of women with suspected peritubal adhesions diagnosed by hysterosalpingography （HSG） treated with intrauterine insemination （IUI）. Methods The study group consisted of 93 patients diagnosed as bilateral suspected peritubal adhesions by HSG, and 175 patients with no tubal pathology were classified as control group. A total of 496 cycles of lUI were finished in 268 infertile women. Results There were no differences in basic clinical parameters between the two groups. Cumulative pregnancy rates after two cycles of lUI were 19.4%for the study group, and 34.3%for the control. Cumulative pregnancy rate in the women with abnormal contrast media loculation was significantly lower than that in women of the control （7.4% vs 34.3%, P=0.037）. The cumulative pregnancy rates were similar between the women with abnormal tubal contour and the normal ones （36. 7% vs 34.3%, P=0.800）. Conclusion Suspected peritubal adhesions in HSG were worthy more attention, since the outcome of lUI in women with loculation of contrast material, combined with or without abnormal tubal contour resulted in a significantly lower pregnancy rate than the normal ones. IUI should be delayed before tubal patency was confirmed during these patients.

中重度宫腔粘连应用中医临床决策系统效果分析

[目的]探讨中医临床决策系统在治疗中重度宫腔粘连(intrauterine adhesion,IUA)中的应用及效果。[方法]采用随机对照试验进行前瞻性研究,选择2022年1月至12月在杭州市某三级甲等中医院行宫腔镜下子宫粘连松解术的中重度IUA患者,根据是否通过中医优势病种临床决策系统进行辨证论治分为对照组(51例)、决策系统组(53例)、临床医师组(54例),比较3组患者治疗前后月经量改善、美国生育协会(American Fertility Society,AFS)评分、子宫内膜厚度、子宫内膜血流参数、中医伴随症候疗效。[结果]治疗后3组患者月经量比较,决策系统组、临床医师组较对照组均有显著改善,差异有统计学意义(P<0.05),决策系统组与临床医师组差异无统计学意义(P>0.05)。治疗后3组患者宫腔AFS评分比较,决策系统组、临床医师组较对照组显著下降(P<0.01),决策系统组与临床医师组差异无统计学意义(P>0.05)。治疗后3组患者子宫内膜厚度显著增加,子宫内膜血流参数显著下降,差异有统计学意义(P<0.01),但治疗前后组间差异均无统计学意义(P>0.05)。治疗后3组患者中医伴随症候疗效比较,临床医师组较对照组、决策系统组明显改善,差异有统计学意义(P<0.05),对照组与决策系统组差异无统计学意义(P>0.05)。[结论]中医临床决策系统用于中重度IUA的治疗可显著改善患者月经量,降低术后AFS评分,防止粘连再发生。

术前雌激素、孕激素水平与稽留流产清宫术后宫腔粘连发生的关系及其预测价值

目的探讨雌激素、孕激素水平与稽留流产(MA)患者清宫术后发生宫腔粘连(IUA)的关系及其预测价值。方法选取2019年1月至2022年1月在该院妇科就诊行清宫术的128例MA患者作为研究对象,根据术后6个月内宫腔检查情况分为未发生IUA组(MA组,97例)和IUA组(31例),并根据严重程度将IUA组分为轻、中和重度IUA组。比较IUA组和MA组,以及不同严重程度IUA组术前雌二醇(E 2)和孕酮(P)水平,采用多因素Logistic回归分析发生IUA的独立影响因素;分析E 2、P与IUA病情的相关性,采用受试者工作特征(ROC)曲线分析E 2、P水平对发生IUA及其病情严重程度的预测价值。结果IUA组E 2、P水平均明显低于MA组,差异均有统计学意义(P<0.05);多因素Logistic回归分析结果显示,E 2为MA清宫术后发生IUA的独立影响因素(P<0.05);术前E 2和P水平预测MA清宫术后发生IUA的曲线下面积分别为0.714和0.702。重度IUA组E 2、P水平均明显低于轻、中度IUA组,差异均有统计学意义(P<0.05),且E 2、P水平与IUA严重程度均呈负相关(P<0.05);术前E 2、P水平预测IUA轻/中度和重度的曲线下面积分别为0.845和0.923。结论E 2、P水平与MA清宫术后IUA的发生及严重程度均相关,对临床预测MA清宫术后IUA的发生及病情进展具有一定的参考价值。

lncRNA LUCAT1调控AREG的表达促进宫腔粘连的发生

目的探讨lncRNA LUCAT1通过调控双调蛋白(amphiregulin,AREG)对宫腔粘连发生的作用,为宫腔粘连的防治提供新的分子靶点。方法在宫腔粘连组织和经过转化生长因子-β1(transforming growth factor-β1,TGF-β1)处理的子宫内膜基质细胞(endometrial stromal cell,ESC)中采用实时荧光定量聚合酶链反应(real-time quantitative polymerase chain reaction,RT-qPCR)方法检测lncRNA LUCAT1、AREG及纤维化标志物α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)、Ⅰ型胶原蛋白(collagen typeⅠalpha 1 chain protein,COL1A1)的mRNA表达水平,采用Western blot法检测AREG、α-SMA、COL1A1的蛋白表达水平。si-LUCAT1、pcDNA LUCAT1、si-AREG分别转染ESC,RT-qPCR方法进一步检测lncRNA LUCAT1和AREG的mRNA表达水平。然后,si-LUCAT1和pcDNA LUCAT1分别转染入ESC中并经过TGF-β1处理48h,采用Western blot法进一步检测AREG、α-SMA、COL1A1的蛋白表达水平,细胞增殖实验和细胞凋亡实验检测细胞增殖率和细胞凋亡率。结果宫腔粘连组织和经TGF-β1处理的ESC中lncRNA LUCAT1、AREG及纤维化标志物α-SMA、COL1A1的表达水平上调。AREG随lncRNA LUCAT1的变化而变化,而下调AREG后lncRNA LUCAT1的表达无明显变化,lncRNA LUCAT1正向调节AREG。在ESC向成纤维细胞转化过程中,si-LUCAT1显著抑制AREG、α-SMA、COL1A1的蛋白表达水平,抑制细胞增殖促进细胞凋亡,差异有统计学意义(P<0.01);pcDNA LUCAT1显著诱导AREG、α-SMA、COL1A1的蛋白表达,差异有统计学意义(P<0.01)。结论lncRNA LUCAT1通过上调AREG的表达促进宫腔粘连的发生。lncRNA LUCAT1/AREG轴可能为宫腔粘连的防治提供新的分子靶点。