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Cryopreservation of bovine sperm causes single‑strand DNA breaks that are localized in the toroidal regions of chromatin

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Journal of Animal Science and Biotechnology 2025年第1期16卷 92-106页

作者： Jordi Ribas‑Maynou Rodrigo Muiño Carolina Tamargo Marc Yeste Biotechnology of Animal and human Reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaS17003 GironaSpain Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaS17003 GironaSpain Present Address:Unit of Cell Biology and Medical Genetics Department of Cell BiologyPhysiology and ImmunologyAutonomous University of BarcelonaS08193 BellaterraSpain department of Animal pathology Faculty of Veterinary MedicineUniversity of Santiago de CompostelaS15705 LugoSpain department of Animal Selection and Reproduction The Regional Agri-Food Research and Development Service of Asturias(SERIDA)S33394 GijónSpain Catalan Institution for Research and Advanced Studies(ICREA) S08010 BarcelonaSpain

Background Sperm cryopreservation is widely used in the cattle industry,as it allows for disassociating the localiza-tion of sires and the collection of semen from the timing of artificial *** freeze-thawing is known to impair sperm DNA integrity,whether the damage induced consists of single-(SSB)or double-strand breaks(DSB)has not been *** addition,no previous study has addressed if DNA breaks preferentially reside in specific genome regions such as those forming the toroid linker regions,or are rather spread throughout the regions linked to *** main aim of the present work,therefore,was to elucidate the type and localization of the DNA damage generated by cryopreservation and to evaluate its impact on artificial insemination outcomes in *** The incidence of SSB and DSB was evaluated in 12 ejaculates before and after cryopreservation with the Comet assay,and the localization of the DNA breaks was assessed using pulsed-field gel electrophoresis(PFGE).Before cryopreservation,the incidence of SSB was 10.99%±4.62%and involved 20.56%±3.04%of sperm cells,whereas these figures significantly(P0.990)before and after cryopreservation(before:incidence of 13.91%±1.75%of sperm DNA affecting 56.04%±12.49%of sperm cells;after:incidence of 13.55%±1.55%of sperm DNA involving 53.36%±11.00%of sperm cells).Moreover,PFGE revealed that the percentage of sperm DNA fragments whose length was shorter than a toroid(<31.5 kb)was greater(P<0.0001)after(27.00%±4.26%)than before freeze-thawing(15.57%±4.53%).These differences indicated that the DNA breaks induced by cryopreservation affect the regions condensed in protamines,which are structured in *** the other hand,in vivo fertility rates were associated to the incidence of SSB and DSB in frozen-thawed sperm(P=0.032 and P=0.005),but not w

关键词： Cattle DNA damage Fertility Sperm Toroid linker regions

High Expression of INF2 Predicts Poor Prognosis and Promotes Hepatocellular Carcinoma Progression

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生物化学与生物物理进展 2025年第1期52卷 194-208页

作者： WANG Hai-Biao LIN Man YE Fu-Sang SHI Jia-Xin LI Hong YE Meng WANG Jie department of Hepatobiliary and Pancreatic Surgery Ningbo Medical Center of LiHuiLi HospitalNingbo 315040China department of Radiotherapy and Chemotherapy The First Affiliated Hospital of Ningbo UniversityNingbo 315010China department of Biochemistry and Molecular Biology Zhejiang Key Laboratory of PathophysiologyScience Health Center of Ningbo UniversityNingbo 315211China department of pathology Ningbo Clinical Pathology Diagnosis CenterNingbo 315021China

Objective INF2 is a member of the formins *** expression and regulation of INF2 have been associated with the progression of various tumors,but the expression and role of INF2 in hepatocellular carcinoma(HCC)remain *** is a highly lethal malignant *** the limitations of traditional treatments,this study explored the expression level,clinical value and potential mechanism of INF2 in HCC in order to seek new therapeutic *** In this study,we used public databases to analyze the expression of INF2 in pan-cancer and HCC,as well as the impact of INF2 expression levels on HCC *** real time polymerase chain reaction(RT-qPCR),Western blot,and immunohistochemistry were used to detect the expression level of INF2 in liver cancer cells and human HCC *** correlation between INF2 expression and clinical pathological features was analyzed using public databases and clinical data of human HCC ***,the effects of INF2 expression on the biological function and Drp1 phosphorylation of liver cancer cells were elucidated through in vitro and in vivo ***,the predictive value and potential mechanism of INF2 in HCC were further analyzed through database and immunohistochemical *** INF2 is aberrantly high expression in HCC samples and the high expression of INF2 is correlated with overall survival,liver cirrhosis and pathological differentiation of HCC *** expression level of INF2 has certain diagnostic value in predicting the prognosis and pathological differentiation of *** vivo and in vitro HCC models,upregulated expression of INF2 triggers the proliferation and migration of the HCC cell,while knockdown of INF2 could counteract this ***2 in liver cancer cells may affect mitochondrial division by inducing Drp1 phosphorylation and mediate immune escape by up-regulating PD-L1 expression,thus promoting tumor *** INF2 is highly expressed in H

关键词： HCC INF2 expression prognosis Drp1

Clinical significance of programmed cell death-ligand expression in small bowel adenocarcinoma is determined by the tumor microenvironment

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World Journal of Gastroenterology 2023年第40期29卷 5566-5581页

作者： Aitoshi Hoshimoto Atsushi Tatsuguchi Ryohei Hamakubo Takayoshi Nishimoto Jun Omori Naohiko Akimoto Shu Tanaka Shunji Fujimori Tsutomu Hatori Akira Shimizu Katsuhiko Iwakiri department of Gastroenterology Nippon Medical SchoolTokyo 113-8603Japan department of analytic human pathology Nippon Medical SchoolTokyo 113-8603Japan department of pathology Nippon Medical SchoolChiba Hokusoh HospitalChiba 270-1694Japan

BACKGROUND Comprehensive genomic analysis has shown that small bowel adenocarcinoma(SBA)has different genomic profiles from gastric and colorectal ***,it is essential to establish chemotherapeutic regimens based on SBA *** expression of programmed cell death-ligand 1(PD-L1)and programmed cell death-ligand 2(PD-L2)in SBA is not fully ***-PD-L1/PD-1 therapy uses tumor-infiltrating lymphocytes(TILs);therefore,the status of TILs in the tumor microenvironment(TME)may influence their *** ratio of FoxP3+to CD8+T cells has been reported to be useful in predicting the prognosis of digestive system *** To investigate the clinicopathological significance of PD-L1/2 expression according to the status of TILs in SBA *** We performed immunohistochemical analysis for PD-L1,PD-L2,CD8,FoxP3,and DNA mismatch repair(MMR)proteins using formalin-fixed,paraffin-embedded tissues from 50 patients diagnosed with primary *** immunoreactivities of PD-L1 and PD-L2 were determined separately in tumor cells and tumor-infiltrating immune cells throughout the tumor center and invasive margins,and finally evaluated using the combined positive score(CPS).We assessed CD8+and FoxP3+T cells in the intratumoral and tumor-surrounding ***,we calculated and summed the ratio of FoxP3 to CD8+T cell ***-related cell densities were graded as low or *** results were compared with clinicopathological factors and patient *** distribution of cancer-specific survival(CSS)was estimated using the Kaplan–Meier method,and the log-rank test was used to test for significant differences in CSS.A Cox proportional hazard model was also used to assess the effect of tumor variables on *** PD-L1 expression was positive in 34%in tumor cells(T-PD-L1)and 54%in tumor-infiltrating immune cells(I-PDL1)of the cases examined.T-PD-L2 was positive in 34%and I-PD-L2 was positive in 42%of the ***-L1

关键词： Small bowel adenocarcinoma Programmed cell death-ligand 1 Programmed cell death-ligand 2 Tumor microenvironment Tumor-infiltrating lymphocytes Regulatory T-cells

A FURTHER STUDY ON CAPILLARY-LIKE BILE DUCTULES PROLIFERATION IN CHRONIC ACTIVE HEPATITIS

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中华医学杂志(英文版) 1990年第8期103卷 623-628页

作者：梁英锐程瑞雪沈明郭亚先苏先狮 department of pathology Human Medical University Changsha.

64 cases of chronic active hepatitis (CAH) and 84 cases of non-CAH liver diseases were studied with keratin stain. The capillary-like bile ductules (CLBD) were proliferating amd their morphology was identical to that with Type V collagen stain reported *** proliferation were more marked in CAH than in other liver diseases, and it was considered to he one of the characteristics of CAH and could he used for differential diagnosis. The ultrastructure of CLBD was specific in morphology. The HBV-DNA in CLBD shown by the technique of in situ hybridization suggested that HBV light infect the cells of CLBD.

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Reduced mesencephalic astrocyte-derived neurotrophic factor expression by mutant androgen receptor contributes to neurodegeneration in a model of spinal and bulbar muscular atrophy pathology

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Neural Regeneration Research 2025年第9期20卷 2655-2666页

作者： Yiyang Qin Wenzhen Zhu Tingting Guo Yiran Zhang Tingting Xing Peng Yin Shihua Li Xiao-Jiang Li Su Yang Guangdong Key Laboratory of Non-human Primate Research Key Laboratory of CNS Regeneration(Ministry of Education)Guangdong–HongKong–Macao Institute of CNS RegenerationJinan UniversityGuangzhouGuangdong ProvinceChina

Spinal and bulbar muscular atrophy is a neurodegenerative disease caused by extended CAG trinucleotide repeats in the androgen receptor gene,which encodes a ligand-dependent transcription facto *** mutant androgen receptor protein,characterized by polyglutamine expansion,is prone to misfolding and forms aggregates in both the nucleus and cytoplasm in the brain in spinal and bulbar muscular atrophy *** aggregates alter protein-protein interactions and compromise transcriptional *** this study,we reported that in both cultured N2a cells and mouse brain,mutant androgen receptor with polyglutamine expansion causes reduced expression of mesencephalic astrocyte-de rived neurotrophic *** n of mesencephalic astrocyte-derived neurotrophic factor amelio rated the neurotoxicity of mutant androgen receptor through the inhibition of mutant androgen receptor *** down endogenous mesencephalic astrocyte-derived neurotrophic factor in the mouse brain exacerbated neuronal damage and mutant androgen receptor *** findings suggest that inhibition of mesencephalic astrocyte-derived neurotrophic factor expression by mutant androgen receptor is a potential mechanism underlying neurodegeneration in spinal and bulbar muscular atrophy.

关键词： androgen receptor mesencephalic astrocyte-derived neurotrophic factor mouse model neurodegeneration neuronal loss neurotrophic factor polyglutamine disease protein misfolding spinal and bulbar muscular atrophy transcription factor

Optimized management of advanced hepatocellular carcinoma:Four long-lasting responses to sorafenib

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World Journal of Gastroenterology 2011年第19期17卷 2450-2453页

作者： Giovanni Abbadessa Lorenza Rimassa Tiziana Pressiani Cynthia Carrillo-Infante Emanuele Cucchi Armando Santoro department of Medical Oncology and Hematology Istituto Clinico Humanitas department of human pathology and Oncology Program of Genetic OncologyUniversity of Siena department of human pathology and Oncology Program of Molecular PathologyUniversity of Siena department of Radiology and Ultrasound Polidiagnostico CAM

The therapeutic options for hepatocellular carcinoma (HCC) have been so far rather *** has shown an overall survival benefit and has become the new standard of care for advanced ***,in clinical practice,some patients are discontinuing this drug because of side effects,and misinterpretation of radiographic response may contribute to *** highlight the importance of prolonged sorafenib ad-ministration,even at reduced dose,and of qualitative and careful radiographic *** observed two partial and two complete responses,one histologically confirmed,with progression-free survival ranging from 12 to 62 *** of the responses were achieved following substantial dose reductions,and a gradual change in lesion density preceded or paralleled tumor shrinkage,as seen by computed *** report supports the feasibility of dose adjustments to allow prolonged administration of sorafenib,and highlights the need for new imaging criteria for a more appropriate characterization of response in HCC.

关键词： Hepatocellular carcinoma Sorafenib Drug toxicity Response criteria Decision making

Phagocytosis(cannibalism) of apoptotic neutrophils by tumor cells in gastric micropapillary carcinomas

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World Journal of Gastroenterology 2015年第18期21卷 5548-5554页

作者： Valeria Barresi Giovanni Branca Antonio Ieni Luciana Rigoli Giovanni Tuccari Rosario Alberto Caruso department of human pathology University of Messina department of Pediatrics University of Messina

AIM:To identify those with a micropapillary pattern,ascertain relative frequency and document clinicopathological characteristics by reviewing gastric ***:One hundred and fifty-one patients diagnosed with gastric cancer who underwent gastrectomy were retrospectively studied and the presence of a regional invasive micropapillary component was evaluated by light *** available hematoxylin-eosin(HE)-stained slides were histologically reviewed and 5 tumors were selected as putative micropapillary carcinoma when cancer cell clusters without a vascular core within empty lymphatic-like space comprised at least5%of the *** tissues from these 5 invasive gastric carcinomas were immunostained using an antimucin 1(MUC1)antibody(clone MA695)to detect the characteristic inside-out pattern and with D2-40antibody to determine the presence of intratumoral lymph *** of intraepithelial neutrophil apoptosis was evaluated in consecutive histological tissue sections by three independent methods,namely light microscopy with HE staining,the conventional terminal deoxynucleotidyl transferase-mediated d UTP-biotin nick end-labeling(TUNEL)method and immunohistochemistry for activated caspase-3(clone C92-605).RESULTS:Among 151 gastric cancers resected for cure,5(3.3%)were adenocarcinomas with a micropapillary *** of the patients died of disease from 6 to 23 mo and one patient was alive with metastases at 9 *** patients had advanced-stage cancer(≥p T2)and lymph node *** MUC1 immunostaining on the stroma-facing surface(inside-out pattern)of the carcinomatous cluster cells,together with negative immunostaining for D2-40 in the cells limiting lymphatic-like spaces,confirmed the true micropapillary pattern in these gastric *** all five cases,several micropapillae were infiltrated by *** staining,TUNEL assay and immunostaining for caspase-3 demonstrated apoptoticneutrophils within cytoplasmic vacuoles of

关键词： Gastric cancer Micropapillary pattern Mucin 1 Caspase-3 TUNEL assay

Hydrogen promotes the activation of Cu, Zn superoxide dismutase in a rat corneal alkali-burn model

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International Journal of Ophthalmology(English edition) 2020年第8期13卷 1173-1179页

作者： Takeshi Arima Tsutomu Igarashi Masaaki Uchiyama Maika Kobayashi Ikuroh Ohsawa Akira Shimizu Hiroshi Takahashi department of Ophthalmology Nippon Medical SchoolTokyo1138602Japan department of analytic human pathology Nippon Medical SchoolTokyo1138602Japan Biological Process of Aging Tokyo Metropolitan Institute of GerontologyTokyo173-0015Japan

AIM: To investigate the effects of hydrogen(H2) on Cu, Zn superoxide dismutase(SOD1) activation in a rat model of corneal alkali burn. METHODS: In each rat, one cornea was subjected to alkali exposure. Physiological saline(saline group) or H2-dissolved saline(H2 group) was instilled continuously on the cornea for 5 min before and after alkali exposure. Inflammatory cells, neovascularization, and cytoplasmic SOD1 levels were evaluated immunohistochemically in enucleated eyes from both groups. Three-dimensional ultrastructural tissue changes in the eyes were analyzed using low-vacuum scanning electron ***: The numbers of both inflammatory and vascular endothelial cells were significantly reduced in the corneas of the H2 group(P<0.01). Furthermore, H2 treatment increased both cytoplasmic SOD1 levels(P<0.01) and activity in corneal epithelial cells(P<0.01). Notably, the SOD1 activity level in the H2 group was approximately 2.5-fold greater than that in the saline ***: H2 treatment suppresses inflammation and neovascularization in the injured cornea and indirectly suppresses oxidative insult to the cornea by upregulating the SOD1 enzyme protein level and activity.

关键词： hydrogen alkali burn Cu,Zn superoxide dismutase low-vacuum scanning electron microscopy rats

A supramolecular nanoprodrug for prevention of gallstone formation

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Chinese Chemical Letters 2025年第2期36卷 354-359页

作者： Xiangjun Zhang Xiaodi Yang Yan Wang Zhongping Xu Sisi Yi Tao Guo Yue Liao Xiyu Tang Jianxiang Zhang Ruibing Wang College of Pharmacy Chongqing Medical University Chongqing University Fuling Hospital department of Pharmaceutics College of PharmacyThird Military Medical University (Army Medical University) Yu-Yue pathology Scientific Research Center State Key Laboratory of Trauma and Chemical Poisoning Third Military Medical University (Army Medical University) State Key Laboratory of Quality Research in Chinese Medicine Institute of Chinese Medical SciencesUniversity of Macau

Cholelithiasis affects approximately 10%-20% of the adult population globally. And cholesterol accumulation and nucleation of cholesterol crystals are commonly recognized as the primary process in the initiation and progression of gallstones. Hydroxypropyl-β-cyclodextrin(HPCD) is a supramolecular host compound that can solubilize cholesterol, potentially serving as a preventative or therapeutic agent for cholelithiasis. However, we found that the administration of HPCD treatment did not impede the formation of gallstones in mice, mainly attributed to the pre-complexation of its cavity during the transition process. Here we synthesized a prodrug of HPCD and prepared a HPCD nanoparticle(HPCD-NP), which can be transported efficiently to the gallbladder through the hepatobiliary system following an intravenous injection. In the bile, the HPCD-NP degraded into free HPCD, bound to cholesterol crystals and gallstones within the gallbladder and effectively increased cholesterol solubilization, leading to gallstones regression. Given the established safety of both HPCD and cyclodextrin-based nanoparticles in numerous animal and human studies, HPCD-NP shows considerable promise for the prevention and treatment of human cholelithiasis.

关键词： Gallstone Hepatobiliary transport Host-guest interaction Hydroxypropyl-β-cyclodextrin Solubilization