Combined detection and subclass characteristics analysis of CTCs and CTECs by SE-iFISH in ovarian cancer

Objective:Hematogenous metastasis is essential for the progression of ovarian cancer(OC),and circulating tumor cells(CTCs)are part of the metastatic cascade.However,the detection rate of CTC is low due to the use of less sensitive detection methods.Therefore,this study aimed to detect CTCs and circulating tumorigenic endothelial cells(CTECs)in patients with OC using subtraction enrichment and immunostaining and fluorescence in situ hybridization(SE-iFISH).Methods:We enrolled a total of 56 subjects,including 20 OC patients and 36 ovarian benign tumor patients.CTCs and CTECs were captured by subtraction enrichment(SE)and counted and classified according to immunofluorescence staining of tumor markers(TMs)carbohydrate antigen 125(CA125)and human epididymis protein 4(HE4)combined with fluorescence in situ hybridization(iFISH)of chromosome 8(Chr8)aneuploidy.The diagnostic value and subtype characteristics of CTCs and CTECs were investigated.Results:The detection rate of CTCs by SE-iFISH was high.Compared with CA125 and HE4,Chr8 aneuploidy was the major identification feature of CTC.CTC counts in OC were statistically higher than those in benign groups.CTC and CTEC with≥pentaploidy were detected in both groups,illustrating the poor diagnostic value of CTC or CTEC.Distributions of triploid and tetraploid CTC subtypes were significantly different,and combined detection of triploid and tetraploid CTCs showed the best diagnostic value.In contrast,the distribution of CTECs in the OC and benign groups had no statistically significant difference.Small CTCs accounted for over 1/3 of the total CTC count.We also found that small CTCs and CTECs primarily comprised triploid cells,while large CTCs and CTECs mainly comprised pentaploidy and beyond.Conclusions:The application of SE-iFISH offered a more comprehensive understanding of heterogeneous CTCs and CTECs in OC.Analysis of subclass characteristics of the CTCs and CTECs according to Chr8 aneuploidy and cell size may broaden their potential clinical utility and deepen mechanistic studies in OC.

Precise capture of circulating endometrial cells in endometriosis

Background:Endometriosis(EM)is a complex benign gynecological disease,but it has malignant biological behavior and can invade any part of the body.Clinical manifestations include pelvic pain,dysmenorrhea,infertility,pelvic nodules,and masses.Our previous study successfully detected circulating endometrial cells(CECs)in the peripheral blood of patients with EM.The purpose of this study is to overcome the limitation of cell size in the previous microfluidic chip method,to further accurately capture CECs,understand the characteristics of these cells,and explore the relationship between CECs and the clinical course characteristics of patients with EM.Methods:Human peripheral venous blood used to detect CECs and circulating vascular endothelial cells(CVECs)was taken from EM patients(n=34)hospitalized in the Peking University People’s Hospital.We used the subtraction enrichment and immunostaining fluorescence in situ hybridization(SE-iFISH)method to exclude the interference of red blood cells,white blood cells,and CVECs,so as to accurately capture the CECs in the peripheral blood of patients with EM.Then we clarified the size and ploidy number of chromosome 8 of CECs,and a second grouping of patients was performed based on clinical characteristics to determine the relationship between CECs and clinical course characteristics.Results:The peripheral blood of 34 EM patients and 12 non-EM patients was evaluated by SE-iFISH.Overall,34 eligible EM patients were enrolled.The results showed that the detection rates of CECs were 58.8%in EM patients and 16.7%in the control group.However,after classification according to clinical characteristics,more CECs could be detected in the peripheral blood of patients with rapidly progressive EM,with a detection rate of 94.4%(17/18).In total,63.5%(40/63)of these cells were small cells with diameters below 5μm,and 44.4%(28/63)were aneuploid cells.No significant association was found between the number of CECs and EM stage.Conclusion:The number and characteristics of CECs are related to the clinical course characteristics of patients with EM,such as pain and changes in lesion size,and may be used as biomarkers for personalized treatment and management of EM in the future.

Influence of toxic substances on dynamical behavior of a delayed diffusive predator-prey model

In this paper,we propose and investigate a delayed diffusive predator-prey model affected by toxic substances.We first study the boundedness and persistence property of the model.By analyzing the associated characteristic equation,we obtain the conditions for the existence of steady state bifurcation,Hopf bifurcation and Turing bifurcation.Furthermore,we also study the Hopf bifurcation induced by the delay.Finally,our theoretical results are verified by numerical simulation.The numerical observation results are in good agreement with the theoretically predicted results.Theoretical and numerical simulations indicate that toxic substances have a great impact on the dynamics of the system.

Relationship between the change of ATP level controlled by ectonucleotidases and endometriosisassociated infertility and pain

Background Thedysfunction ofthe immune system is one of the pathogeneses of endometriosis.Immune cells can not onlyaffect the microenvironmentof the endometrium by secreting cytokines and defensins but also promote angiogenesis,growth and invasion of endometrial stromal cells.Result ATP is a key mediator in the immune mechanism of endometriosis and plays a crucial role in endometriosis.While ATP acts as a purinergic signaling molecule,it has a close relationship with the pain of endometriosis via activating ATP receptors,including P2X3,P2X4,P2X7 and P2Y receptors,after being activated by the immune system.Besides,ATP levels reflect the impairment of mitochondrial function in granulosa cells,which could lead to infertility.The modulation of ATP expression levels is controlled by ectonucleotidases.The content of ectonucleotidases is altered in endometriosis which may be emerging non-invasive biomarkers.Conclusion In the present review,we briefly introduce the relationship between the change of ATP level controlled by ectonucleotidases and endometriosis-associated infertility and pain,and illustrate our prospects for future research.

Characterization of candidate factors associated with the metastasis and progression of high-grade serous ovarian cancer

Background:High-grade serous ovarian cancer(HGSOC)is the biggest cause of gynecological cancer-related mortality because of its extremely metastatic nature.This study aimed to explore and evaluate the characteristics of candidate factors associated with the metastasis and progression of HGSOC.Methods:Transcriptomic data of HGSOC patients’samples collected from primary tumors and matched omental metastatic tumors were obtained from three independent studies in the National Center for Biotechnology Information(NCBI)Gene Expression Omnibus(GEO)database.Differentially expressed genes(DEGs)were selected to evaluate the effects on the prognosis and progression of ovarian cancer using data from The Cancer Genome Atlas(TCGA)database.Hub genes’immune landscapes were estimated by the Tumor Immune Estimation Resource(TIMER)database.Finally,using 25 HGSOC patients'cancer tissues and 10 normal fallopian tube tissues,immunohistochemistry(IHC)was performed to quantify the expression levels of hub genes associated with International Federation of Gynecology and Obstetrics(FIGO)stages.Results:Fourteen DEGs,ADIPOQ,ALPK2,BARX1,CD37,CNR2,COL5A3,FABP4,FAP,GPR68,ITGBL1,MOXD1,PODNL1,SFRP2,and TRAF3IP3,were upregulated in metastatic tumors in every database while CADPS,GATA4,STAR,and TSPAN8 were downregulated.ALPK2,FAP,SFRP2,GATA4,STAR,and TSPAN8 were selected as hub genes significantly associated with survival and recurrence.All hub genes were correlated with tumor microenvironment infiltration,especially cancer-associated fibroblasts and natural killer(NK)cells.Furthermore,the expression of FAP and SFRP2 was positively correlated with the International Federation of Gynecology and Obstetrics(FIGO)stage,and their increased protein expression levels in metastatic samples compared with primary tumor samples and normal tissues were confirmed by IHC(P=0.0002 and P=0.0001,respectively).Conclusions:This study describes screening for DEGs in HGSOC primary tumors and matched metastasis tumors using integrated bioinformatics analyses.We identified six hub genes that were correlated with the progression of HGSOC,particularly FAP and SFRP2,which might provide effective targets to predict prognosis and provide novel insights into individual therapeutic strategies for HGSOC.

求解无约束优化问题的分式模型信赖域算法

本文提出一个求解无约束优化问题的分式模型信赖域拟Newton算法.在新算法中,分式模型信赖域子问题是用简单折线法求解的.在合理假设条件下,算法的全局收敛性获得了证明.数值实验结果表明新算法是可行、有效的.

A clinicopathological study of 39 cases of atypical polypoid adenomyoma of the uterus

Objective:To clarify the clinical and pathological characteristics of atypical polypoid adenomyoma(APA)in order to improve the criteria used to diagnose and treat this disease.Study design:In 39 cases of APA,retrospective analysis was performed of theclinical data,pathological characteristics,and the follow-up findings in patients admitted to the Peking University People's Hospital from 2007 to 2019.Results:The median age was 39.6 years(range 24–60 years).Thirty seven patients were premenopausal(i.e.94.9%)and eighteen patients were nullipara(i.e.46.2%).Abnormal uterine bleeding was the most common presenting symptom.Hysteroscopic transcervical resection(i.e.TCR)identified lesions in 35 cases whereas:(a)hysterectomy identified them in 3 cases;(b)dilatation and curettage(D&C)identified them in 1 case.Immunohistochemical analysis was performed on 24 samples.In the glandular component,hormone receptors were positively expressed.In all cases,Ki67 expression was detectable in approximately 50%of the cases.In those samples,its presence was definitive in eighteen of the nineteen cases(94.7%,18/19),p53 positive expression was present in most cases(68.8%,11/16),and PTEN positive expression was detected in 40%(4/10).Stroma immunophenotype expression was exhibited as follows:(a)CD10-(12/12),Desminþ(6/7);(b)Vimentinþ(4/4);(c)α-SMAþ(3/3)and;d p16 positve staining in of 80%(4/5).The concurrent amount of atypical endometrial hyperplasia with atypical polypoid adenomyoma was 23.1%(9/39),while its concurrent level of endometrial carcinoma with atypical polypoid adenomyoma was 7.7%(3/39).Fertility preserving treatments were performed in 20 patients with strong childbearing desires.Their pregnancy success was 15%(3/20)and the live birth frequency was 10%(2/10).Follow-up findings were available in 36 cases(92.3%)with a medial follow-up of 48.1 months(range 4–122 months).Its pathological recurrence and frequency of progression into endometrial carcinoma were both 5%(1/20).One case died of another type of malignancy,while the other patients were alive.Conclusions:APA occurs mostly during the years of a women's reproductive period.Its diagnosis is based on the analysis of pathological and immunohistochemical findings.Individuals diagnosed with APA are at risk to coexist with endometrial carcinoma and atypical endometrial hyperplasia.For those individuals who desire retaining fertility,the treatment strategy involves performing TCR completely remove the lesions and close follow-up for surveillance of possible progressive APA recurrence.For those individuals who have no fertility desire,hysterectomy may be a preferred option.

Optimal control of a diffusive eco-epidemiological predator-prey model

In this study,we investigate the optimal control problem for a diffusion eco-epidemiological predator-prey model.We applied two controllers to this model.One is the separation control,which separates the uninfected prey from the infected prey population,and the other is used as a treatment control to decrease the mortality caused by the disease.Then,we propose an optimal problem to minimize the infected prey population at the final time and the cost cause by the controls.To do this,by the operator semigroup theory we prove the existence of the solution to the controlled system.Furthermore,we prove the existence of the optimal controls and obtain the first-order necessary optimality condition for the optimal controls.Finally,some numerical simula-tions are carried out to support the theoretical results.

An experimental study on autologous transplantation of fresh ovarian tissue in sheep

Objective:To investigate current autologous transplantation methods and sites of ovarian tissues in sheep.Methods:Sheep ovaries were resected.Ovarian cortices were sliced and transplanted orthotopically into the ovarian mesangial latum and heterotopically into the greater omentum and under groin skin.The grafts were removed two months after transplantation and examined to evaluate the survival of follicles(hematoxylin-eosin staining)and help determining feasible graft sites and transplantation methods.Results:Graft nodules were found in the transplanted sites.HE staining of the grafts showed that multiple primordial follicles were able to survive in the grafts on both sides of the ovarian mesangial latum,the right side of the greater omentum,and the left inguinal subcutaneous tissue.Secondary or cystic follicles were found in almost all of the grafts.Conclusion:The ovarian mesangial latum,the greater omentum and the inguinal subcutaneous tissue can be used as autologous transplantation sites,where sheep ovarian tissue can survive and the follicles grow and develop in good condition.