Objective To evaluate the inhibitory effects of prophylactic administration of α-zearalanol(α-ZAL)on bone microarchitecture and bone resorption activity in ovariectomized osteoporotic rats,and to investigate its regulatory effects on the osteogenic and adipogenic differentiation of bone marrow mesenchymal stem cells(BMSCs).Methods A total of 606-month-old unmated female Sprague-Dawley(SD)rats weighing(300±20)g were randomly divided into the sham surgery group(Sham group),ovariectomy group(OVX group),solvent group(Oil group),estradiol benzoate treatment group(Post-E2 group),α-ZAL prevention group(Pre-ZAL group),and α-ZAL treatment group(Post-ZAL group),with 10 rats in each group.An osteoporosis rat model was established using the ovariectomy method.Rats in the Sham group underwent the same surgical procedures except for ovarian removal.Seventy-two hours after ovarian removal,the Oil group received intramuscular injections of 0.5 mL of oil solvent,and the Pre-ZAL group received intramuscular injections of α-ZAL(1.5 mg·kg-1),administered every 3 days for 120 consecutive days.The Post-E2 group and Post-ZAL group began intramuscular injections of estradiol benzoate(1.5 mg·kg-1)and α-ZAL(1.5 mg·kg-1),respectively,90 days after ovariectomy,administered every 3 days for 120 consecutive days.After drug administration,bone density and bone tissue microstructure morphology were analyzed using a micro-CT small animal in vivo imaging system and staining methods.Osteoclasts were isolated and their activity was detected.Femoral BMSCs were obtained to assess their osteoblast and adipocyte differentiation capabilities,and uterine tissue morphological changes were observed via histological sections.Results Compared with the OVX group,BMD in the Sham group,Post-E2 group,Pre-ZAL group,and Post-ZAL group increased by 133.12%,75.97%,69.64%,and 24.69%,respectively(all P<0.01).BMD in the Pre-ZAL group was 36.09%higher than in the Post-ZAL group(P<0.01),and there was no significant difference in BMD between the Post-E2 and Pre-ZAL groups(P>0.05).Tb.N in the Sham group,Post-E2 group,Pre-ZAL group,and Post-ZAL group increased by 160.08%,118.14%,94.76%,and 46.76%,respectively,compared with the OVX group(all P<0.01).Tb.Ar increased by 324.21%,203.83%,177.99%,and 82.71%,respectively(all P<0.01).Tb.N in the Pre-ZAL group increased by 32.71%compared to the Post-ZAL group(P<0.05),while Tb.Ar increased by 52.15%(P<0.01).Tb.Sp in the Sham,Post-E2,and Pre-ZAL groups decreased by 58.53%,42.18%,and 35.61%,respectively,compared with the OVX group(all P<0.01).The MAR of the upper tibial cancellous bone in the Sham,Post-E2,and Pre-ZAL groups increased by 257.81%,156.72%,and 142.63%,respectively,compared with the OVX group(all P<0.01),BFR increased by 192.19%,137.23%,and 88.13%,respectively(all P<0.01).MAR and BFR in the Pre-ZAL group increased by 58.10%and 43.63%,respectively,compared with the Post-ZAL group(both P<0.01).There were no significant differences in MAR and BFR between the Post-E2 group and the Pre-ZAL group(P>0.05).MMP-9,TRAP,and CK mRNA expression was significantly downregulated in both the Post-E2 group and the Pre-ZAL group(P<0.01).The osteoblast differentiation capacity of BMSCs in the Post-E2 group and all Post-ZAL groups was enhanced,with a significant increase in the number of mineralized nodules,and the expression levels of OCN,COL1,and OPN mRNA were significantly increased(P<0.01),while the ability to differentiate into adipocytes was weakened.The number of intracellular lipid droplets in BMSCs was significantly reduced,the lipid droplet volume was smaller,and the expression levels of PPAR-γ2 and aP2 mRNA were decreased(P<0.05).There were no significant differences between the Post-E2 group and the Pre-ZAL group(P>0.05).There was no significant increase in body weight in the Post-E2,Pre-ZAL,and Post-ZAL groups,but uterine weight significantly increased in the Post-E2 group(P<0.05),with marked uterine epithelial hyperplasia.Uterine weight in the Pre-ZAL and Post-ZAL groups showed no significant difference compared to the OVX group(P>0.05),and no significant changes were observed in uterine epithelium.Conclusion α-ZAL can effectively protect bone mass,improve bone microstructure,and reduce estrogen-related uterine adverse reactions by regulating the osteogenic/adipogenic differentiation balance of BMSCs,providing a potential new therapeutic strategy for the prevention and treatment of postmenopausal osteoporosis.

This study was aimed at investigating differentially expressed genes(DEGs)in Clonorchis sinensis(C.sinensis)meta-cercariae and newly excysted juveniles(NEJs)cultured in vitro for 1 hour or 3 hours,through transcriptomic analysis.Our objective was to explore the mechanisms underlying host invasion.Metacercariae were digested and isolated from Pseudorasbora parva infected with C.sinensis.The metacercariae excysted and developed into NEJs in vitro.Subsequently,the mRNA of metacercariae and NEJs cultured in vitro for 1 hour or 3 hours was extracted for transcriptomic sequencing analysis to screen for DEGs,and to conduct GO and KEGG analyses.A protein-protein interaction network(PPI)was constructed to identify hub genes.A total of 1 218 DEGs were de-tected.The main enriched GO terms of DEGs included transcription regulator activity and gated channel activity(primarily K+).The KEGG pathways significantly enriched in DEGs included cholesterol metabolism,lysosome,synthesis,secretion,and action of para-thyroid hormone.ZFAND4-2,BIRC6,and other genes were screened and identified as hub genes through PPI network analysis.Addi-tionally,abundant differential expression of cathepsin-related genes,including Cathepsin L and Cathepsin F,were observed before and after excystment in C.sinensis.Therefore,significant transcriptional level changes occurred in the metacercariae of C.sinensis be-fore and after excystation,and enrichment was observed primarily in signaling pathways,such as activation of growth and material me-tabolism,that regulate parasite growth and development.Meanwhile,biological events conducive to parasite invasion,migration,and adhesion were triggered.

This study was aimed at investigating differentially expressed genes(DEGs)in Clonorchis sinensis(C.sinensis)meta-cercariae and newly excysted juveniles(NEJs)cultured in vitro for 1 hour or 3 hours,through transcriptomic analysis.Our objective was to explore the mechanisms underlying host invasion.Metacercariae were digested and isolated from Pseudorasbora parva infected with C.sinensis.The metacercariae excysted and developed into NEJs in vitro.Subsequently,the mRNA of metacercariae and NEJs cultured in vitro for 1 hour or 3 hours was extracted for transcriptomic sequencing analysis to screen for DEGs,and to conduct GO and KEGG analyses.A protein-protein interaction network(PPI)was constructed to identify hub genes.A total of 1 218 DEGs were de-tected.The main enriched GO terms of DEGs included transcription regulator activity and gated channel activity(primarily K+).The KEGG pathways significantly enriched in DEGs included cholesterol metabolism,lysosome,synthesis,secretion,and action of para-thyroid hormone.ZFAND4-2,BIRC6,and other genes were screened and identified as hub genes through PPI network analysis.Addi-tionally,abundant differential expression of cathepsin-related genes,including Cathepsin L and Cathepsin F,were observed before and after excystment in C.sinensis.Therefore,significant transcriptional level changes occurred in the metacercariae of C.sinensis be-fore and after excystation,and enrichment was observed primarily in signaling pathways,such as activation of growth and material me-tabolism,that regulate parasite growth and development.Meanwhile,biological events conducive to parasite invasion,migration,and adhesion were triggered.

Purpose To investigate the clinical,imaging,pathological,and genetic characteristics of neuroepithelial tumors with EWSR1 translocation.Methods The clinicopatho-logical data of 6 patients with EWSR1 translocation in neuroepi-thelial tumors were collected,routine HE and immunohistochem-ical staining were performed,the information of high-throughput sequencing was summarized,and the relevant literature was re-viewed.Results The median age of the 6 patients was 11.5 years(ranging from 1.9 to 17 years),including 1 male and 5 females.The tumors located in temporal lobe,frontal lobe,pari-etal lobe,suprasellar region,or lateral ventricle.The clinical manifestations mainly started with seizures.Brain MRI showed abnormal signal focus in the cerebral hemisphere near the cortex in 4 cases,and ventricle/periventricular regions in 2 cases,with an almost clear boundary in 5 cases.Microscopically,the histo-logical changes were diverse,including low-grade gliomas/gli-oneuronal tumors in 3 cases,high-grade gliomas in 2 cases,and glioneuronal tumor with high-grade feature in 1 case.Immuno-histochemically,tumor cells expressed GFAP,S-100,Syn,and Olig2 partially.2 cases exhibited slightly positive of NeuN and 1 case exhibited little dot-like staining of EMA.Next generation sequencing revealed EWSR1 rearrangement in all 6 cases,with chaperone genes including PATZ1 in 5 cases,and PLAGL1 in 1 case.3 cases were treated with chemotherapy after surgery,and no recurrence or progression was found during follow-up.Con-clusion The neuroepithelial tumors with the fusion of EWSR1 and non-ETS commonly occur in the cerebral hemisphere of teenagers and children.Most of the boundaries lesion are still clear,the histomorphological spectrum is diverse,and the bio-logical behavior is presented as a low to moderate malignancy,which provides the possibility for expanding the molecular classi-fication of CNS neuroepithelial tumor.

BACKGROUND: Ischemic acute kidney injury (AKI) is a common syndrome associated with considerable mortality and healthcare costs. Up to now, the underlying pathogenesis of ischemic AKI remains incompletely understood, and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking. Here, this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing (scRNA-seq) analysis in kidneys. METHODS: In this study, scRNA-seq technology was applied to kidneys from two ischemic AKI patients, and three human public scRNA-seq datasets were collected as controls. Differentially expressed genes (DEGs) and cell clusters of kidneys were determined. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis, as well as the ligand-receptor interaction between cells, were performed. We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion (I/R) injury induced AKI mice through immunohistochemistry staining. RESULTS: 15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control. The injured proximal tubules (PT) displayed a proapoptotic and proinflammatory phenotype. PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47 , RASSF4 , EBAG9 , IER3 , SASH1 , SEPTIN7 , and NUB1 , which have not been reported in ischemic AKI previously. Several hub genes were validated in kidneys from human AKI and renal I/R injury mice, respectively. Furthermore, PT highly expressed DEGs enriched in endoplasmic reticulum stress, autophagy, and retinoic acid-inducible gene I (RIG-I) signaling. DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain (NOD)-like receptor signaling, estrogen signaling, interleukin (IL)-12 signaling, and IL-17 signaling. Overexpressed genes in kidney-resident immune cells including macrophages, natural killer T (NKT) cells, monocytes, and dendritic cells were associated with leukocyte activation, chemotaxis, cell adhesion, and complement activation. In addition, the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI. CONCLUSION Together, this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients, altered signaling pathways, and potential cell-cell crosstalk in the development of AKI. These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.
Humans ; Mice ; Animals ; Transcriptome/genetics* ; Ligands ; Kidney/metabolism* ; Acute Kidney Injury/metabolism* ; Ischemia/metabolism* ; Reperfusion Injury/metabolism* ; Sequence Analysis, RNA ; Adaptor Proteins, Signal Transducing/metabolism* ; Tumor Suppressor Proteins/metabolism*

OBJECTIVES: Long-term elevated blood pressure may lead to kidney damage, yet the pathogenesis of hypertensive kidney damage is still unclear. This study aims to explore the role and significance of leucine-rich alpha-2-glycoprotein-1 (LRG-1) in hypertensive renal damage through detecting the levels of LRG-1 in the serum and kidney of mice with hypertensive renal damage and its relationship with related indexes. METHODS: C57BL/6 mice were used in this study and randomly divided into a control group, an angiotensin II (Ang II) group, and an Ang II+irbesartan group. The control group was gavaged with physiological saline. The Ang II group was pumped subcutaneously at a rate of 1.5 mg/(kg·d) for 28 days to establish the hypertensive renal damage model in mice, and then gavaged with equivalent physiological saline. The Ang II+irbesartan group used the same method to establish the hypertensive renal damage model, and then was gavaged with irbesartan. Immunohistochemistry and Western blotting were used to detect the expression of LRG-1 and fibrosis-related indicators (collagen I and fibronectin) in renal tissues. ELISA was used to evaluate the level of serum LRG-1 and inflammatory cytokines in mice. The urinary protein-creatinine ratio and renal function were determined, and correlation analysis was conducted. RESULTS: Compared with the control group, the levels of serum LRG-1, the expression of LRG-1 protein, collagen I, and fibronectin in kidney in the Ang II group were increased (all P<0.01). After treating with irbesartan, renal damage of hypertensive mice was alleviated, while the levels of LRG-1 in serum and kidney were decreased, and the expression of collagen I and fibronectin was down-regulated (all P<0.01). Correlation analysis showed that the level of serum LRG-1 was positively correlated with urinary protein-creatinine ratio, blood urea nitrogen, and blood creatinine level in hypertensive kidney damage mice. Serum level of LRG-1 was also positively correlated with serum inflammatory factors including TNF-α, IL-1β, and IL-6. CONCLUSIONS Hypertensive renal damage mice display elevated expression of LRG-1 in serum and kidney, and irbesartan can reduce the expression of LRG-1 while alleviating renal damage. The level of serum LRG-1 is positively correlated with the degree of hypertensive renal damage, suggesting that it may participate in the occurrence and development of hypertensive renal damage.
Animals ; Mice ; Mice, Inbred C57BL ; Fibronectins ; Irbesartan ; Creatinine ; Kidney/physiology* ; Hypertension/complications* ; Angiotensin II ; Collagen Type I

Detailed characterizations of genomic alterations have not identified subtype-specific vulnerabilities in adult gliomas. Mapping gliomas into developmental programs may uncover new vulnerabilities that are not strictly related to genomic alterations. After identifying conserved gene modules co-expressed with EGFR or PDGFRA (EM or PM), we recently proposed an EM/PM classification scheme for adult gliomas in a histological subtype- and grade-independent manner. By using cohorts of bulk samples, paired primary and recurrent samples, multi-region samples from the same glioma, single-cell RNA-seq samples, and clinical samples, we here demonstrate the temporal and spatial stability of the EM and PM subtypes. The EM and PM subtypes, which progress in a subtype-specific mode, are robustly maintained in paired longitudinal samples. Elevated activities of cell proliferation, genomic instability and microenvironment, rather than subtype switching, mark recurrent gliomas. Within individual gliomas, the EM/PM subtype was preserved across regions and single cells. Malignant cells in the EM and PM gliomas were correlated to neural stem cell and oligodendrocyte progenitor cell compartment, respectively. Thus, while genetic makeup may change during progression and/or within different tumor areas, adult gliomas evolve within a neurodevelopmental framework of the EM and PM molecular subtypes. The dysregulated developmental pathways embedded in these molecular subtypes may contain subtype-specific vulnerabilities.
Humans ; Brain Neoplasms/pathology* ; Neoplasm Recurrence, Local/metabolism* ; Glioma/pathology* ; Neural Stem Cells/pathology* ; Oligodendrocyte Precursor Cells/pathology* ; Tumor Microenvironment

We applied literature review and the Delphi method to develop technical operation of manual lactation technique. The databases and websites were systematically searched, such as UpToDate, the Cochrane Library, CNKI, VIP, Wanfang, and SinoMed. The evidence related to the technique was evaluated by Appraisal of Guidelines for Research and Evaluation (AGREE Ⅱ), and A Measure Tool to Assess Systematic Reviews 2 (AMSTAR 2). the included evidence was classified and summarized to form the preliminary draft of the technical operation specification. A total of 26 experts were selected to evaluate the draft, and some recommendations were deleted and modified according to the screening criteria and expert opinions, resulting in the final technical specifications for manual lactation technique applicable to health care professionals. The study included 4 guidelines, 5 expert consensus articles, 2 clinical decisions, and 3 systematic reviews, and the overall quality of the evidence was fair. A total of 26 questionnaires were collected in each of 2 rounds of expert consultation, with a 100% return rate and an overall authority coefficient of 0.91. The technical practice specification was formed in 11 aspects, including assessment content, indications, contraindications, operation methods, adverse events, and treatment methods, with a total of 50 recommendations. The technical operation specification is comprehensive, and the recommendations are clearly expressed, which is in line with the real clinical situation and can provide effective reference for the clinical practice of this technique.

Hepatic stellate cells(HSCs)are essential drivers of fibrogenesis.Inducing activated-HSC apoptosis is a promising strategy for treating hepatic fibrosis.18beta-glycyrrhetinic acid(18β-GA)is a natural com-pound that exists widely in herbal medicines,such as Glycyrrhiza uralensis Fisch,which is used for treating multiple liver diseases,especially in Asia.In the present study,we demonstrated that 18β-GA decreased hepatic fibrosis by inducing the apoptosis in activated HSCs.18β-GA inhibited the expression of α-smooth muscle actin and collagen type Ⅰ alpha-1.Using a chemoproteomic approach derived from activity-based protein profiling,together with cellular thermal shift assay and surface plasmon reso-nance,we found that 18β-GA covalently targeted peroxiredoxin 1(PRDX1)and peroxiredoxin 2(PRDX2)proteins via binding to active cysteine residues and thereby inhibited their enzymatic activities.18β-GA induced the elevation of reactive oxygen species(ROS),resulting in the apoptosis of activated HSCs.PRDX1 knockdown also led to ROS-mediated apoptosis in activated HSCs.Collectively,our findings revealed the target proteins and molecular mechanisms of 18β-GA in ameliorating hepatic fibrosis,highlighting the future development of 18β-GA as a novel therapeutic drug for hepatic fibrosis.

ObjectiveTo investigate the effect of intraoperative Viatorr stent implantation for shunting of blood flow in the left or right branch of the portal vein on the clinical outcome of patients with cirrhotic portal hypertension undergoing transjugular intrahepatic portosystemic shunt (TIPS). MethodsA retrospective analysis was performed for the clinical data of 120 patients with cirrhotic portal hypertension who underwent TIPS in The General Hospital of Western Theater Command from March 2016 to December 2019, and according to the target position of portal vein puncture determined by intraoperative angiography, the patients were divided into left branch group and right branch group. The two groups were compared in terms of the incidence rates of postoperative recurrence and bleeding, stent dysfunction, and hepatic encephalopathy (HE) and survival. The t-test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The Kaplan-Meier curve was used to calculate rebleeding rate, stent patency rate, incidence rate of HE, and survival rate. ResultsThe surgical success rate was 100% for all 120 patients, with a short-term hemostasis rate of 100%. Among the 120 patients, 52 underwent shunting of the left branch of the portal vein and 68 underwent shunting of the right branch. There was a significant reduction in portal venous pressure after surgery (9.98±2.84 mm Hg vs 24.72±5.11 mm Hg, t=37.76, P＜0.01). The cumulative rebleeding rates at 12 and 24 months after surgery were 3.2% and 11.0%, respectively, and the cumulative incidence rates of HE at 3, 6, 12, and 24 months after surgery were 10.8%, 13.6%, 21.2%, and 24.5%, respectively. Among the 29 patients who experienced HE, 23 had grade Ⅰ-Ⅱ HE and 6 had grade Ⅲ HE. The cumulative incidence rates of stent dysfunction at 12 and 24 months after surgery were 7.1% and 21.4%, respectively. The cumulative survival rates at 12 and 24 months after surgery were 92.0% and 86.5%, respectively. As for comparison of the left branch group and the right branch group, there were no significant differences in postoperative stent patency rate, rebleeding rate, incidence rate of HE, and survival rate(all P＜0.05). ConclusionTIPS is a safe and effective method for the treatment of cirrhotic portal hypertension, and intraoperative Viatorr stent implantation, no matter for establishing the shunt of the left or right branch of the portal vein, will not affect the clinical outcome of patients.