Hair follicle stem cells: In vitro and in vivo neural differentiation

Hair follicle stem cells(HFSCs) normally give rise to keratinocytes, sebocytes, and transient amplifying progenitor cells. Along with the capacity to proliferate rapidly, HFSCs provide the basis for establishing a putative source of stem cells for cell therapy. HFSCs are multipotent stem cells originating from the bulge area. The importance of these cells arises from two important characteristics, distinguishing them from all other adult stem cells. First, they are accessible and proliferate for long periods. Second, they are multipotent, possessing the ability to differentiate into mesodermal and ectodermal cell types. In addition to a developmental capacity in vitro, HFSCs display an ability to form differentiated cells in vivo. During the last two decades, numerous studies have led to the development of an appropriate culture condition for producing various cell lineages from HFSCs. Therefore, these stem cells are considered as a novel source for cell therapy of a broad spectrum of neurodegenerative disorders. This review presents the current status of human, rat, and mouse HFSCs from both the cellular and molecular biology and cell therapy perspectives. The first section of this review highlights the importance of HFSCs and in vitro differentiation, while the final section emphasizes the significance of cell differentiation in vivo.

Human hair follicle-derived mesenchymal stem cells:Isolation,expansion,and differentiation

Hair follicles are easily accessible skin appendages that protect against cold and potential injuries.Hair follicles contain various pools of stem cells,such as epithelial,melanocyte,and mesenchymal stem cells(MSCs)that continuously self-renew,differentiate,regulate hair growth,and maintain skin homeostasis.Recently,MSCs derived from the dermal papilla or dermal sheath of the human hair follicle have received attention because of their accessibility and broad differentiation potential.In this review,we describe the applications of human hair follicle-derived MSCs(hHF-MSCs)in tissue engineering and regenerative medicine.We have described protocols for isolating hHF-MSCs from human hair follicles and their culture condition in detail.We also summarize strategies for maintaining hHF-MSCs in a highly proliferative but undifferentiated state after repeated in vitro passages,including supplementation of growth factors,3D suspension culture technology,and 3D aggregates of MSCs.In addition,we report the potential of hHF-MSCs in obtaining induced smooth muscle cells and tissue-engineered blood vessels,regenerated hair follicles,induced red blood cells,and induced pluripotent stem cells.In summary,the abundance,convenient accessibility,and broad differentiation potential make hHF-MSCs an ideal seed cell source of regenerative medical and cell therapy.

HSP25 Affects the Proliferation and Differentiation of Rat Dental Follicle Cells

Aim To detect the expression of HSP25 in rat dental follicles both in vivo and vitro, and explore the underlying mechanism of HSP25 on the proliferation and differentiation of rat dental follicle cells （DFCs）. Methodology Immunohistochemistry was performed to detect the expression of HSP25 in mandibles of postnatal rats on days 1, 3, 5, 7, 9 and 11 in vivo. In vitro, the expression of HSP25 in DFCs was detected by an indirect immunofluorescence assay. Thiazolyl blue tetrazolium bromide （MTT） assay, flow cytometry and alkaline phosphatase （ALP） assay were used to identify the time-course effect mediated by different concentrations of recombinant murine HSP25 of 0, 1, 10, 50 and 100 ng/mL on rat DFCs. Results Expression of HSP25 was not detected in dental follicles of the rats until day 5 after birth, but became up-regulated in a time-dependent manner till day 11. HSP25 was detected in the cytoplasm of cultured rat DFCs. No significant difference could be observed in the proliferation of DFCs after stimulation with different concentrations of HSP25 on days 1, 2 and 3 （P〉0.05）. HSP25 at concentrations of 50 ng/mL and 100 ng/mL up-regulated the ALP activity of DFCs on day 9 （P〈0.05）. Conclusion HSP25-immunoreactivity increased chronologically during the development of dental follicles. The protein had no significant effect on cell proliferation but may play a role in cementoblast/osteoblast differentiation of DFCs.

Anti-Müllerian hormone and antral follicle count predict ovarian response in women less than 45 years following GnRH antagonist multiple-dose protocol

Objective:To speculate which of the following parameters:antral follicle count(AFC),anti-Müllerian hormone(AMH),follicle-stimulating hormone(FSH)and age can be used as a predictor of ovarian response to gonadotropin-releasing hormone(GnRH)antagonist stimulation multiple-dose protocol in women under 45 years,and to determine the cutoff value of these parameters and their correlations for predicting low and high ovarian response.Methods:This prospective study included 462 women with the mean age of(29.3±6.5)years.All women were subjected to the GnRH antagonist stimulation multiple-dose protocol.On the second day of the menstrual cycle,ultrasonography was conducted to determine AFC in both ovaries.Peripheral blood samples were collected to evaluate the level of estradiol,FSH,luteinizing hormone,prolactin,thyroid-stimulating hormone,and AMH.The women were divided into three groups:low response(AHH<1 ng/mL,n=173),normal response(AMH=1.0-3.5 ng/mL,n=175),and high response(AMH>3.5 ng/mL,n=114).Results:A significant decrease was found in the age and FSH level in the high response group compared to other groups(P<0.001).Conversely,a significant increase was shown in AMH,estradiol on human chorionic gonadotropin(hCG)day,AFC,mature oocytes,fertilized oocytes,and embryos transferred in the high response group compared to the other two groups(P<0.001).The receiver operating characteristic(ROC)curves demonstrated that AFC and AMH had the highest accuracy,followed by basal FSH level and age in the prediction of low ovarian reserves(P<0.001)with cutoff values of≤4.50 and≤0.95 for AFC and AMH,respectively.Moreover,the ROC analysis showed that AFC had the highest accuracy,followed by AMH level and age in the prediction of high ovarian reserves with a cutoff value of≥14.50,≥3.63,and≤27.50 years,respectively(P<0.01).A significant decrease was observed in women's age,estradiol level,and oocyte fertilization rate in pregnant women compared to non-pregnant women(P<0.001).Additionally,significant negative correlations were found between the AFC,the number of mature oocytes,fertilized oocytes,embryos transferred,and the age of pregnant women(P<0.001).Conclusions:AFC and AMH predict low and high ovarian response to GnRH antagonist stimulation multiple-dose protocol in women under 45 years.

Ovarian Follicle Disaggregation to Assess Granulosa Cell Viability

Background: Mammalian ovaries contain follicles containing an oocyte enclosed by layers of granulosa cells (GC). Follicle growth and oocyte maturation are largely dependent on GC numbers and viability, but there is no established, reliable method for assessing the number of viable GC within an isolated follicle. Methods: Centrifugation conditions and the Trypan Blue (TB) Exclusion assay were optimised for low cell densities compatible with the numbers of GC in follicles. Mouse ovarian follicles were disaggregated to produce a single cell suspension of GC which were examined by TB (n = 4), but also by crystal violet assay in a 96-well plate format after 24 h in vitro (n = 3). GC viability in vitro was characterised further by using enzyme-linked immunoassays to quantify GC production of anti-Mullerian hormone (AMH) and estrogen. Results: The centrifugation and low cell density TB protocol could accurately measure the viability of 78 GC in 10 &mu;L, with an intra-assay coefficient of variation (CoV) 22%, and inter-assay CoV 7%. The best follicle disaggregation method (30 min 37°C exposure to 2 mg/mL collagenase prior to 30 min exposure to 0.025% hyaluronidase) yielded (656 &plusmn;87) GC per antral follicle of which 82% &plusmn;5% were viable. Culturing 312 - 20,000 GC per well for 24 hours and assessing viability by crystal violet assay generated a linear correlation between OD value and viable GC number (R2 = 0.98) and estrogen concentration per well (R2 = 0.92). 20,000 GC per well produced 143 &plusmn;16 pg/mL estrogen during 24 hours in vitro, but no detectable AMH. Conclusion: This is the first report describing the isolation of viable, estrogen-producing GC from murine follicles, and their subsequent culture. These procedures are transferrable to other species including humans and can be applied to screening the reproductive toxicity of pharmaceutical agents.

Evaluation of glucose metabolism in women with multiple ovarian follicles

Objective ：To investigate glucose metabolism in women with multiple ovarian follicles （MOF） and explore the relationship between glucose metabolism, insulin resistance and body weight. Methods：We evaluated 46 women with MFO and 30 normal women as controls. All the subjects were given 75g of glucose orally in order to perform the oral glucose tolerance test （OGTT） and insulin releasing test （IRT）, and they were also evaluated for insulin resistance using the insulin resistance index with homeostatic model assessment （HOMA）. Results：The occurrence of impaired glucose tolerance in women with MOF was 10.87%, which was significantly higher than that in the control group （3.33% ,P 〈 0.05）. The rate of insulin resistance was 30.43% in the study group as compared to 10.00% in the control group. The results showed that there was significant difference between the two groups（P 〈 0.05）. The levels of FSH,LH,PRL,E2,T and P between the two groups had no significant difference （P 〉 0.05）. BMI in women with impaired glucose tolerance was correlated positively to insulin resistance （r = 0.567, P 〈 0.05）. Conclusion：Abnormal glucose metabolism was observed in women with unitary multiple ovarian follicles, and this could be attributed to obesity and insulin resistance. Women with MOF and associated obesity should be subjected to OGTT so that their glucose levels can be monitored as a preventive measure.

Effects of Antisense Oligodeoxynucleotide to Follicle-stimulating Hormone Receptor on the Cell Proliferation and Apoptosis in Cells Derived from Human Ovarian Mucinous Cystadenocarcinoma in Vitro

The human ovarian mucinous cystadenocarcinoma （hOMC） cells were co-cultured with antisense oligodeoxynucleotide （antisense ODN）, nonsense ODN, and follicle-stimulating hormone （FSH） at different concentrations for the purpose of observing the effects of antisense ODN to FSH receptor （FSHR） on the proliferation and apoptosis of cultured hOMC cells in vitro. The inhibitory rates of growth were measured by using MTT method on the 2nd, 4th, 6th, 8th and 10th days after the interference of antisense ODN, nonsense ODN, and FSH, respectively. The apoptotic rates and the cell cycles were determined by means of flow cytometry, the apoptosis indexes were detected by using TUNEL, and the expression of caspase-3 was measured by using SP immunohistochemistry. Compared with that in the control group, the proliferative activity of hOMC cells was increased obviously in FSH groups （P〈0.05 or P〈0.01）, decreased distinctly in antisense ODN groups （P〈0.05 or P〈0.01）, and unchanged in nonsense ODN groups, respectively. Meanwhile, antisense ODN could significantly antagonize the FSH-promoted cell proliferative activity （P〈0.01）. Compared with those in the control group, the apoptotic rates and the expression of caspase-3 were dramatically increased in the mid- and high-dose antisense ODN groups （P〈0.05 or P〈0.01）, while the number of cells in G1/G0 phase was significantly decreased and that in S phase distinctly increased （P〈0.01）, There was no change in nonsense ODN groups （P〉0.05）, It was suggested that FSH may improve the development of hOMC cells, However, antisense ODN could inhibit proliferative activity and the FSH-promoted proliferative activity in hOMC cells, at the same time, antisense ODN could inhibit hOMC cell growth by inducing apoptosis.

Follicle and melanocyte stem cells, and their application in neuroscience A Web of Science-based literature analysis

OBJECTIVE： To identify global research trends of follicle and melanocyte stem cells, and their application in neuroscience. DATA RETRIEVAL： We performed a bibliometric analysis of studies from 2002 to 2011 on follicle and melanocyte stem cells, and their application in neuroscience, which were retrieved from the Web of Science, using the key words follicle stem cell or melanocyte stem cell, and neural, neuro or nerve. SELECTION CRITERIA： Inclusion criteria： （a） peer-reviewed published articles on follicle and melanocyte stem cells, and their application in neuroscience, which were indexed in the Web of Science; （b） original research articles, reviews, meeting abstracts, proceedings papers, book chapters, editorial material, and news items. Exclusion criteria： （a） articles that required manual searching or telephone access; （b） documents that were not published in the public domain; and （c） a number of corrected papers from the total number of articles. MAIN OUTCOME MEASURES： （1） Distribution of publications on follicle and melanocyte stem cells by years, journals, countries, institutions, institutions in China, and most cited papers. （2） Distribution of publications on the application of follicle and melanocyte stem cells in neuroscience by years, journals, countries, institutions, and most cited papers. RESULTS： Of the 348 publications from 2002 to 2011 on follicle and melanocyte stem cells, which were retrieved from the Web of Science, more than half were from American authors and institutes. The most prolific institutions in China for publication of papers on follicle and melanocyte stem cells were the Fourth Military Medical University and Third Military Medical University. The most prolific journals for publication of papers on follicle and melanocyte stem cells were the Journal of Investigative Dermatology, Pigment Cell ＆ Melanoma Research. Of the 63 publications from 2002 to 2011 on the application of follicle and melanocyte stem cells in neuroscience, which were retrieved from the Web of Science, more than half were from American authors and institutes, and no papers were from Chinese authors and institutes. The most prolific journals for publication of papers on the application of follicle and melanocyte stem cells in neuroscience were the Journal of Investigative Dermatology, Pigment Cell ＆ Melanoma Research. CONCLUSION： Based on our analysis of the literature and research trends, we found that follicle stem cells might offer further benefits in neural regenerative medicine.

Three Dimensional <i>In Vitro</i>Culture of Murine Secondary Follicles in a Defined Synthetic Matrix

Ovarian follicle growth in three dimensional (3D) matrices in vitro has limitations: a) matrices don’t expand as follicles grow, b) requirements for enzyme-mediated retrieval, and c) animal-derived components prevent clinical application. Therefore, we evaluated N-Isopropylacrylamide (SFX-1), a novel synthetic 3D culture matrix, for follicle culture. Groups of three murine secondary follicles were encapsulated in 50 μL of DMEM/F12-1%ITS-10%FCS (DMEM/F12) or SFX-1 (3:2 v/v DMEM/F12) or Matrigel (1:1 DMEM/F12) and cultured for 48 h. Matrigel contains growth factors but SFX-1 has no animal-derived factors. Each culture condition was examined in 6 wells containing 18 follicles, in four replicate experiments (n = 4). Photomicrographs were used to determine follicle diameters and morphological integrity. Follicles were Live-Dead (LD) stained or disaggregated to generate cells for viability assessment using Trypan Blue (TB). Estradiol, progesterone and anti-mullerian hormone (AMH) in conditioned media were measured using Enzyme-linked Immunoassay. All culture conditions supported similar increases in follicle diameter. DMEM/F12 did not maintain morphological integrity which prevented follicle retrieval after 48 h;25% were retrieved from DMEM/F12, but 44% and 41% follicles were retrieved from SFX-1 and Matrigel respectively. Follicles retrieved from Matrigel could not be disaggregated, which prevented TB viability assessment. LD estimations of viable cells/follicle were lower than TB, but culture conditions had no effect on viability;SFX-1 64% ± 8% and DMEM/F12 69% ± 9%. SFX-1 and Matrigel supported similar levels of progesterone synthesis, only Matrigel supported estrogen synthesis, but none of the culture conditions supported AMH production. SFX-1 was not cytotoxic and was comparable to Matrigel. Further development of SFX-1 for use with human follicles is supported.

Lethal（2）giant larvae is required in the follicle cells for formation of the initial AP asymmetry and the oocyte polarity during Drosophila oogenesis

The intricately regulated differentiation of the somatic follicle cell lineages into distinct subpopulations with specific functions plays an essential role in Drosophila egg development. At early oogenesis, induction of the stalk cells generates the first anteroposterior （AP） asymmetry in the egg chamber by inducing the posterior localization of the oocyte. Later, the properly specified posterior follicle cells signal to polarize the oocyte along the AP and dorsoventral （DV） axes at mid-oogenesis. Here, we show that lethal（2）giant larvae （lgl）, a Drosophila tumor suppressor gene, is required in the follicle cells for the differentiation of both stalk cells and posterior follicle cells. Loss-of-function mutations in lgl cause oocyte mispositioning in the younger one of the fused chambers, due to lack of the stalk. Removal oflgl function from the posterior follicle cells using the FLP/FRT system results in loss of the oocyte polarity that is elicited by the failure of those posterior cells to differentiate normally. Thus, we provide the first demonstration that lgl is implicated in the formation of the initial AP asymmetry and the patterning of the AP and DV axes in the oocyte by acting in the specification of a subset of somatic follicle cells.

An active RUNX1-ID1/ID3 axis governs differentiation and chemoresistance of cancer stem cell population in epithelial ovarian cancer cells

Progression,relapse,and therapy resistance are the most challenging features of cancer therapy that have been postulated to be driven by Cancer Stem Cell(CSC)population.This enigmatic subpopulation of cancer cells has therefore emerged as promising therapeutic candidate.We earlier reported enrichment of CSC-like side population(SP)with increasing resistance towards Cisplatin and Paclitaxel either alone or in combination in epithelial ovarian cancer(EOC)cells.This SP population is a small proportion of the total population of cancer cells characterised with high expression of drug transporters,a unique feature of stem cells and thereby can be isolated through their efflux properties of DNA binding dyes.While the bulk non-SP(NSP)population of the cancer cells lack overexpression of the drug transporters and thus can be identified as the dye containing population.In this study,we show that increased expression of Runt related transcription factor 1(RUNX1)maintains undifferentiated state of CSC-like SP cells through upregulation of inhibitors of DNA binding/differentiation genes(ID1 and ID3)in late cisplatinpaclitaxel resistant cells.Higher RUNX1 expression was found to correlate with decreased median overall survival and disease-free survival in The Cancer Genome Atlas(TCGA)data set of high grade serous ovarian cancer(HGSOC)patients.The protein-protein interaction network analysis of 397 upregulated genes in RUNX1-high samples of TCGA data show significant enrichment of pathways known to negatively regulate CSC differentiation.Intriguingly RUNX1 inhibition not only induces CSC differentiation but also downregulates anti-apoptotic protein BCL2 in both SP and NSP cells and potentiates cytotoxic effects of Cisplatin-Paclitaxel in chemoresistant EOC cells.Inhibition of BCL2 through Venetoclax treatment,a small molecule BH3 mimic,sensitized these cells to platinum taxol treatment.Altogether,our data reveal new regulatory roles by RUNX1 to modulate CSC differentiation via ID1 and ID3 and to promote chemoresistance through BCL2 upregulation.

IDENTIFICATION OF DIFFERENTIAL GENES IN OVARIAN CANCER USING REPRESENTATIONAL DIFFERENCE ANALYSIS OF cDNA

Objoctive To identify differential genes between normal ovarian epithelium tissue and ovarian epithelial cancer using representational difference analysis of cDNA （cDNA-RDA）. Methods cDNA-RDA was performed to identify the differentially expressed sequences between cDNAs from cancer tissue and cDNAs from normal ovarian tissue in the same patient who was in the early stage of ovarian serous cystadenocarcinoma. These differentially expressed fragments were cloned and analyzed, then sequenced and compared with known genes. Results Three differentially cxpressed cDNA fragments were isolated using cDNA from normal ovarian tissue as tester and cDNA from cancer tissue as driver amplicon by cDNA-RDA. DP Ⅲ- 1 and DP Ⅲ-2 cDNA clone showed significant homology to the cDNA of alpha actin gene; DPⅢ-3 cDNA clone showed significant homology to the cDNA oftransgelin gene. Conclusion cDNA-RDA can bc used to sensitively identify the differentially expressed genes in ovarian serous cystadenocarcinoma. Ovarian serous cystadenocarcinoma involves alteration of multiple genes.

Microarray expression analysis of epithelial ovarian cancer with distinct differentiation

To identify gene expression profiling in epithelial ovarian cancer and to explore its correlation with histopathology characterization and prognosis.Gene expression profiles were generated from 10 hu- man ovarian frozen tissue specimens using Agilent Human 1A microarrays.Strikingly.clear differences of gene expression patterns were observed in ovarian cancer as compared to normal tissues.Unique gene pro files were observed in moderately and poorly differentiated epithelial ovarian cancer.It is concluded that different histopathology characterization likely exists extensive molecular heterogeneity.

Determination of the ovine ovarian reserve during the prenatal and neonatal periods

Objective:To determine the ovine ovarian histomorphology and follicular staging at various age periods in Awassi breed.Methods:Ovaries were collected from prenatal fetuses[gestational age(95±5)days],neonatal(day 0),and prepubertal ewe lambs(two and four months of age);each age group included six animals.Ovaries(n=12,each group)were dissected and processed for hematoxylin and eosin staining.Stained sections(n=24,each group)were imaged and utilized for histomorphology assessment,follicle measurement,and classification.Results:Prenatal ovaries were mainly enriched with primordial follicles accompanied by a lower proportion of primary follicles.In addition to primordial and primary follicles,neonatal ovaries demonstrated a proportion of centrally located multilayered and antral follicles.In comparison with neonatal ovaries,the proportion of multilayered and antral follicles was significantly higher in the ovaries of two-month-old lambs;conversely,the proportion of peripherally situated primordial follicles dramatically declined compared to that of earlier age of lamb.Although there was no statistical variation in the sizes of primordial follicles across groups,the mean diameter of the primary follicle in the prenatal ovaries was substantially smaller than in postnatal ovaries.Compared to the neonatal ovaries,the size of the multilayered and antral follicles in the prepubertal ovaries was substantially larger.Conclusions:The earliest follicular developmental stages were established prenatally whereas the advanced growth stages started in the neonatal period and greatly increased in the prepubertal period.

Quercetin modulates ovarian autophagy-related molecules and stereological parameters in a rat model of PCOS

Objective:To examine the effect of quercetin on stereological parameters and autophagy-related genes in ovaries of polycystic ovary syndrome(PCOS)rats.Methods:Fifty female Sprague-Dawley rats were randomly divided into five groups:the control group,the ethanol group,the quercetin group(15 mg/kg/day),the PCOS group,as well as the PCOS+quercetin group.After the induction of PCOS,quercetin was administered orally for 30 days.Histological,stereological and real-time PCR analyses were carried out to evaluate the effect of quercetin on PCOS rats.Results:Stereological analysis revealed that quercetin significantly increased the number of ovarian follicles and the volume of corpus luteum and induced a significant decrease in atretic follicles in comparison to the PCOS group.In addition,quercetin markedly increased mTOR gene expression while decreasing Beclin-1 and LC3 gene expression.Conclusions:Quercetin strongly modulates the expression of ovarian autophagy-related genes and stereological parameters in PCOS rats.Therefore,it can be considered as an ameliorative component for ovarian follicular impairments.

Expression of VEGF165 and VEGF165b during ovarian follicular development

Objective:To investigate the role of vascular endothelial growth factor(VEGF)165a,VEGF165b,and VEGF receptor(VEGFR)in the development of bovine follicles.Methods:We cultured follicular cells that were collected from small,medium,and large sized bovine follicles with estrogen and measured the expression of VEGF,VEGFR2 and VEGF165b by Western blot analysis and immunofluorescence.Results:The expression of VEGF165 increased in all follicle sizes and the expression of VEGF165b was increased in the small and large follicles after culturing in an estrogen containing medium.The expression of VEGFR2 was increased in the medium and large follicles after culturing with estrogen for 96 h.VEGF165 was activated at 100 ng/mL estrogen in the large follicles for 96 h.In addition,VEGFR2 was upregulated in the medium and large follicles after treated with 100 ng/mL estrogen for 96 h.Conclusions:This evidence suggests that the expression of VEGF165 and VEGFR is associated with estrogen stimulation during the development of bovine follicles and in an autocrine or paracrine manner.This reveals an advantage during oocyte maturation in vitro.

Research progress of in vitro activation mechanism and clinical application of female ovarian tissue

The activation and development of primordial follicles is the key to the maturation of female gametes.Premature ovarian insufficiency(POI)patients are unable to complete the primordial follicle activation and development due to follicular dormancy and unbalanced developmental regulation in the body,leading to female infertility.Ovarian tissue in vitro activation(IVA)technology has become a new way to solve the problem of patients who cannot auto-activate primordial follicles to obtain their own mature oocytes.In IVA research,signaling pathways such as PI3K/PTEN/Akt and Hippo have become the focus of current research.This review will describe the relevant research progress and clinical application of the IVA mechanism,and provide a reference for clinical research on ovarian tissue culture and activation in vitro.

Value of ultrasound and magnetic resonance imaging combined with tumor markers in the diagnosis of ovarian tumors

BACKGROUND Compare the diagnostic performance of ultrasound(US),magnetic resonance imaging(MRI),and serum tumor markers alone or in combination for detecting ovarian tumors.AIM To investigate the diagnostic value of US,MRI combined with tumor markers in ovarian tumors.METHODS The data of 110 patients with ovarian tumors,confirmed by surgery and pathology,were collected in our hospital from February 2018 to May 2023.The dataset included 60 cases of benign tumors and 50 cases of malignant tumors.Prior to surgery,all patients underwent preoperative US and MRI examinations,as well as serum tumor marker tests[carbohydrate antigen 125(CA125),human epididymis protein 4(HE4)].The aim of the study was to compare the diagnostic performance of these three methods individually and in combination for ovarian tumors.RESULTS This study found statistically significant differences in the ultrasonic imaging characteristics between benign and malignant tumors.These differences include echo characteristics,presence or absence of a capsule,blood flow resistance index,clear tumor shape,and blood flow signal display rate(P<0.05).The apparent diffusion coefficient values of the solid and cystic parts in benign tumors were found to be higher compared to malignant tumors(P<0.05).Additionally,the time-intensity curve image features of benign and malignant tumors showed significant statistical differences(P<0.05).The levels of serum CA125 and HE4 in benign tumors were lower than those in malignant tumors(P<0.05).The combined use of US,MRI,and tumor markers in the diagnosis of ovarian tumors demonstrates higher accuracy,sensitivity,and specificity compared to using each method individually(P<0.05).CONCLUSION US,MRI,and tumor markers each have their own advantages and disadvantages when it comes to diagnosing ovarian tumors.However,by combining these three methods,we can significantly enhance the accuracy of ovarian tumor diagnosis,enabling early detection and identification of the tumor’s nature,and providing valuable guidance for clinical treatment.

N-乙酰半胱氨酸对冻融人卵巢组织的保护作用

目的以首都医科大学附属北京妇产医院生育力保护中心现有冻存复苏方案为基础,以雌性去势裸鼠为模型,通过人冻融卵巢组织异种移植探讨抗氧化剂N-乙酰半胱氨酸(N-Acetyl-L-Cysteine,NAC)对人卵巢组织的保护作用。方法将4例患者的卵巢组织按照原有冻融方案和改良方案(加入NAC)进行冻融,所有患者取材后留取新鲜卵巢组织进行钙黄绿素AM(Calcein acetoxymethyl ester,Calcein-AM)和苏木精-伊红(hematoxylin-eosin staining,HE)染色评估卵泡活性并计数。将36只雌性去势裸鼠随机分为4组,将复苏后的卵巢组织移植于双侧裸鼠肾被膜下,按照移植卵巢组织的冻融方案分为原有冻融方案组(control group),改良方案组(NAC group),卵巢去势组(ovariectomy group)和正常对照组(normal group,不进行手术)。分别在移植第3天、第7天、第21天处死裸鼠,取血清及卵巢组织移植物。采用酶联免疫吸附试验(enzyme-linked immunosorbent assay,ELISA)法检测血清雌二醇(estradiol,E2)、卵泡刺激素(follicle stimulating hormone,FSH)和抗苗勒管激素(anti-mullerian hormone,AMH)水平,取卵巢组织移植物1片用于HE染色观察卵泡发育情况,1片用于总抗氧化能力(total antioxidant capability,TAC)的检测。结果移植后各时间点两移植组卵泡发育分级差异无统计学意义(P>0.05)。NAC组总抗氧化能力显著高于control组(P<0.05),与新鲜卵巢组织相比差异无统计学意义(P>0.05)。移植第3天control组裸鼠血清中E2水平显著高于卵巢去势组(P<0.05),余各时间点两移植组与卵巢去势组差异无统计学意义(P>0.05)。移植后第3天和第21天两移植组裸鼠血清中FSH水平低于卵巢去势组(P<0.05),与正常对照组差异无统计学意义(P>0.05)。移植后第3天control组AMH显著高于卵巢去势组(P<0.05),与其余各组差异无统计学意义(P>0.05),移植第7天两移植组AMH水平下降,显著低于正常对照组(P<0.05),移植第21天两移植组AMH水平均显著高于卵巢去势组(P<0.05),与正常对照组差异无统计学意义(P>0.05)。结论原有冻存方案及改良冻存方案均能有效恢复卵巢组织的内分泌功能。与原有冻存方案相比,NAC改良方案能够提高移植卵巢组织的抗氧化能力,并减少移植初期原始卵泡的激活,保存更多的原始卵泡数量。

卵巢-附件报告和数据系统超声2022版(O-RADS US v2022)及其联合恶性风险指数4鉴别附件良、恶性肿瘤

目的观察卵巢-附件报告和数据系统超声2022版(O-RADS US v2022)及其联合恶性风险指数4(RMI4)鉴别附件良、恶性肿瘤的价值。方法回顾性分析126例手术病理诊断为附件肿瘤患者,根据O-RADS US v2022将1~3类归为良性病变、4~5类归为恶性病变,以450为RMI4分类的临界值,基于二者进行联合分类。以病理结果为金标准,绘制受试者工作特征(ROC)曲线,计算曲线下面积(AUC),评估单一O-RADS US v2022、RMI4及其联合鉴别附件良、恶性肿瘤的效能。结果126例附件肿瘤中,良性94例、恶性32例。O-RADS US v2022鉴别附件良、恶性肿瘤的敏感度、特异度、准确率及AUC分别为78.13%、80.85%和80.16%、0.795,RMI4分别为71.88%、84.04%和80.95%、0.780;二者联合的特异度及准确率(93.62%、92.06%)均高于单一O-RADS US v2022(χ^(2)=7.322、5.967,P=0.007、0.015)或RMI4(χ^(2)=4.625、5.331,P=0.032、0.021),而敏感度及AUC(87.50%、0.906)差异均无统计学意义(P均>0.05)。结论O-RADS US v2022能有效鉴别附件良、恶性肿瘤,联合RMI4可提高鉴别特异度及准确率。