Objective Orosomucoid1 （ORM1） is a novel target in the quest for anti-fatigue pharmacotherapy. Preliminary investigations have illuminated oleuropein （OLE） as a promising candidate molecule, poised to enhance ORM1 expression. To elucidate the influence of OLE on ORM1 protein expression and assess its ramifications on muscle endurance. Methods The impact of OLE on ORM1 protein expressions within hepatocytes and liver tissue was meticulously quantified through Western blotting; the effects of OLE on muscle endurance were evaluated via the rotarod and forced swimming tests; glycogen content within liver and muscle tissues was determined utilizing a specialized kit; and PAS staining was employed to visualize glycogen deposition in the gastrocnemius muscle. Results OLE demonstrated a capacity to elevate ORM1 protein expression in hepatocytes in a time- and dose-dependent manner, concurrently prolonging the duration of swimming and rotarod performance in mice, also in a time- and dose-dependent manner. Furthermore, OLE augmented ORM1 expression in liver tissue, elevated serum ORM1 levels, and enhanced glycogen reserves within the liver and muscle. Conclusion OLE may serve to amplify muscle endurance by elevating ORM1 levels in vivo and augmenting glycogen stores within skeletal muscle.

ObjectiveTo investigate the characteristics of mitochondrial translational initiation factor 2 （MTIF2） gene methylation and its association with the development and progression of hepatocellular carcinoma （HCC）. MethodsMethSurv and EWAS Data Hub were used to perform the standardized analysis and the cluster analysis of MTIF2 methylation samples， including survival curve analysis， methylation signature analysis， the association of tumor signaling pathways， and a comparative analysis based on pan-cancer database. The independent-samples t test was used for comparison between two groups； a one-way analysis of variance was used for comparison between multiple groups， and the least significant difference t-test was used for further comparison between two groups. The Cox proportional hazards model was used to perform the univariate and multivariate survival analyses of methylation level at the CpG site. The Kaplan-Meier method was used to investigate the survival differences between the patients with low methylation level and those with high methylation level， and the Log-likelihood ratio method was used for survival difference analysis. ResultsGlobal clustering of MTIF2 methylation showed that there was no significant difference in MTIF2 gene methylation level between different races， ethnicities， BMI levels， and ages. The Kaplan-Meier survival curve analysis showed that the patients with N-Shore hypermethylation of the MTIF2 gene had a significantly better prognosis than those with hypomethylation （hazard ratio ［HR］=0.492， P<0.001）， while there was no significant difference in survival rate between the patients with different CpG island and S-Shore methylation levels （P>0.05）. The methylation profile of the MTIF2 gene based on different ages， sexes， BMI levels， races， ethnicities， and clinical stages showed that the N-Shore and CpG island methylation levels of the MTIF2 gene decreased with the increase in age， and the Caucasian population had significantly lower N-Shore methylation levels of the MTIF2 gene than the Asian population （P<0.05）； the patients with clinical stage Ⅳ had significantly lower N-Shore and CpG island methylation levels of the MTIF2 gene than those with stage Ⅰ/Ⅱ （P<0.05）. Clinical validation showed that the patients with stage Ⅲ/Ⅳ HCC had a significantly lower methylation level of the MTIF2 gene than those with stage Ⅰ/Ⅱ HCC and the normal population （P<0.05）. ConclusionN-Shore hypomethylation of the MTIF2 gene is a risk factor for the development and progression of HCC.

Primary ciliary dyskinesia (PCD) is a genetically heterogeneous disorder characterized by impaired motility of cilia and flagella. Mutations in cilia- and flagella-associated protein 300 ( CFAP300 ) are associated with human PCD and male infertility; however, the underlying pathogenic mechanisms remain poorly understood. In a consanguineous Chinese family, we identified a homozygous CFAP300 loss-of-function variant (c.304delC) in a proband presenting with classical PCD symptoms and severe sperm abnormalities, including dynein arm deficiency and acrosomal malformation, as confirmed by transmission electron microscopy (TEM). Histological analysis revealed multiple morphological abnormalities of the sperm flagella in CFAP300 -mutant individual, whereas immunofluorescence demonstrated markedly reduced CFAP300 expression in the spermatozoa of the proband. Furthermore, tandem mass tag (TMT)-based quantitative proteomics showed that the CFAP300 mutation reduced key spermatogenesis proteins (e.g., sperm flagellar 2 [SPEF2], solute carrier family 25 member 31 [SLC25A31], and A-kinase anchoring protein 3 [AKAP3]) and mitochondrial ATP synthesis factors (e.g., SLC25A31, cation channel sperm-associated 3 [CATSPER3]). It also triggered abnormal increases in autophagy-related proteins and signaling mediator phosphorylation. These molecular alterations are likely to contribute to progressive deterioration of sperm ultrastructure and function. Notably, successful pregnancy was achieved via intracytoplasmic sperm injection (ICSI) using the proband's sperm. Overall, this study expands the known CFAP300 mutational spectrum and offers novel mechanistic insights into its role in spermatogenesis.
Humans ; Male ; Infertility, Male/pathology* ; Acrosome/pathology* ; Sperm Tail/pathology* ; Pedigree ; Spermatozoa ; Adult ; Loss of Function Mutation ; Ciliary Motility Disorders/genetics* ; Spermatogenesis/genetics* ; Female

Objective To investigate the protective effect of high-altitude hypoxia acclimatization against hepatic ischemia-reperfusion injury(HIRI)in rats,as well as the mechanism of action of high-altitude hypoxia acclimatization in activating autophagy.Methods A total of 56 male Sprague-Dawley rats were randomly divided into plain sham-operation group(P-S group),plain model group(P-M group),acute high-altitude hypoxia sham-operation group(AHH-S group),acute high-altitude hypoxia model group(AHH-M group),high-altitude hypoxia acclimatization sham-operation group(HHA-S group),high-altitude hypoxia acclimatization model group(HHA-M group),and high-altitude hypoxia acclimatization model group with the adenosine monophosphate-activated protein kinase(AMPK)inhibitor compound C(HHA-M-CC group),with 8 rats in each group.The rats in the acute high-altitude hypoxia groups and the high-altitude hypoxia acclimatization groups were placed in a low-pressure oxygen chamber at an altitude of 5 000 meters for 1 week and 12 weeks,respectively;the rats in the sham-operation groups were given laparotomy to expose the portal vein without vascular clamping;the rats in the HHA-M-CC group were given abdominal injection of 20 mg/kg CC at 1 hour before surgery,while those in the other groups were given injection of an equal volume of normal saline.An automatic biochemical analyzer was used to measure the levels of liver function parameters including alanine aminotransferase(ALT),aspartate aminotransferase(AST),and total bilirubin(TBil);HE staining was used to observe liver histopathological changes;transmission electron microscopy was used to observe the formation of autophagosomes in liver tissue;RT-qPCR was used to measure the mRNA expression levels of AMPK and Unc-51 like autophagy activating kinase 1(ULK1)in liver tissue;Western Blot was used to measure the protein expression levels of phosphorylated AMPK(p-AMPK),phosphorylated ULK1(p-ULK1),Beclin-1,and microtubule-associated protein 1 light chain 3 Ⅱ(LC3Ⅱ).An analysis of variance was used for comparison of continuous data between multiple groups,and the least significant difference t-test was sued for comparison between two groups.Results Compared with the AHH-M and HHA-M-CC groups,the HHA-M group had significantly reductions in the levels of ALT,AST,and TBil(all P<0.05),alleviation of liver histopathological injury,a significant reduction in Suzuki score(all P<0.05),a reduction in the degree of abnormal morphological structure of hepatocytes under transmission electron microscopy,and significant increases in the number of autophagosomes,the mRNA expression levels of AMPK and ULK1(all P<0.05),and the protein expression levels of p-AMPK,p-ULK1,Beclin-1,and LC3Ⅱ(all P<0.05).Conclusion High-altitude hypoxia acclimatization can alleviate HIRI in SD rats by activating the AMPK/ULK1 signaling pathway and enhancing autophagy in hepatocytes.

ObjectiveTo investigate the anatomical features of three-dimensional （3D） reconstruction of the hepatic pedicle based on the Laennec’s capsule， as well as its application value in the development of extra-sheath dissection/occlusion clamp and precise hepatectomy. MethodsA retrospective analysis was performed for the abdominal contrast-enhanced CT data of 100 patients without anatomical abnormalities of the hepatic pedicle in The General Hospital of Western Theater Command from January 2021 to June 2024. The Hisense CAS system combined with the 3D U-net deep learning algorithm was used for 3D reconstruction of the hepatic pedicle at the level of Laennec’s capsule， and the hepatic pedicle was measured in terms of the length， outer diameter， and angle of the main trunk and branches. An extra-sheath hepatic pedicle dissection/occlusion clamp was developed based on the above measurements， and a total of 30 patients scheduled for right hemihepatectomy were enrolled and randomly divided into device group and control group， with 15 patients in each group. The two groups were compared in terms of hepatic pedicle handling time， time of operation， intraoperative blood loss， and the incidence rate of bile duct injury. The independent-samples t test was used for comparison of continuous data between two groups， and the Fisher’s exact test was used for comparison of categorical data between two groups. ResultsThe results of 3D reconstruction revealed four variants in the main trunk branches of the hepatic pedicle， with type Ⅰ （left-right branching） accounting for 88% （88/100）， type Ⅱ （trifurcation type） accounting for 5% （5/100）， type Ⅲ （right anterior branching） accounting for 5% （5/100）， and type Ⅳ （special type） accounting for 2% （2/100）. The outer diameter of the main hepatic pedicle was 24.10±6.16 mm， the length of the left main branch was 20.59±6.38 mm， and the length of the right main branch was 21.99±7.98 mm. Compared with the control group， the device group had significantly shorter hepatic pedicle handling time （14.10±1.30 minutes vs 17.50±2.00 minutes， t=-5.620， P=0.001） and time of operation （217.00±28.28 minutes vs 241.87±19.49 minutes， t=-2.804， P=0.009）. The device group had a significantly lower incidence rate of bile duct injury than the control group （0 vs 20%， P=0.031）. Conclusion3D reconstruction based on the Laennec’s capsule can accurately display the anatomical variations of the hepatic pedicle. The extra-sheath hepatic pedicle dissection/occlusion clamp developed based on such data can optimize the process of hepatic pedicle management and improve surgical safety， and therefore， it holds promise for clinical application.

Objective: To screen and identify the key active molecules, signaling pathways, and therapeutic targets of Shuxuening (SXN) injection for treating liver cirrhosis (LC) and to evaluate its therapeutic potential using a mouse model. Methods: Target genes of SXN and LC were retrieved from public databases, and enrichment analysis was performed. A protein–protein interaction (PPI) network was constructed using the Search Tool for the Retrieval of Interacting Genes/Proteins (STRING), and hub genes were identified using Molecular Complex Detection (MCODE). LC was induced in rats and mice via intraperitoneal injections of diethylnitrosamine and carbon tetrachloride (CCl4) for 12 weeks. Starting at week 7, SXN was administered intraperitoneally to the mice in the treatment group. Serum and liver tissues of the mice were collected for the detection of indicators, pathological staining, and expression analysis of hub targets using quantitative real-time polymerase chain reaction (qRT-PCR). Results: We identified 368 overlapping genes (OLGs) between SXN and LC targets. These OLGs were subsequently used to build a PPI network and to screen for hub genes. Enrichment analysis showed that these genes were associated with cancer-related pathways, including phosphoinositide-3-kinase/Akt and mitogen-activated protein kinase signaling and various cellular processes, such as responses to chemicals and metabolic regulation. In vivo experiments demonstrated that SXN treatment significantly improved liver function and pathology in CCl4-induced LC mice by reducing inflammation and collagen deposition. Furthermore, qRT-PCR demonstrated that SXN regulated the expression of MAPK8, AR and CASP3 in the livers of LC mice. Conclusion This study highlighted the therapeutic effects of SXN in alleviating LC using both bioinformatics and experimental methods. The observed effect was associated with modulation of hub gene expression, particularly MAPK8, and CASP3.

Objective To develop a preoperative question prompt list(QPL)for older patients with benign prostatic hyperplasia(BPH)and evaluate its effectiveness in application.Methods This trial adopted a randomized controlled design.The QPL was developed by literature review,expert discussions,and Delphi consultation.Convenience sampling was used to subject 76 older BPH inpatients treated in our department,and then they were randomly divided into control(routine communication,n=38)and intervention(QPL-assisted communication,n=38)groups.Number of the questions patient asking,communication duration,information recall,and communication quality were compared between the 2 groups.Results In the 2 rounds of expert consultation,the response rate of questionnaire was 94.44%and 100%,the authority coefficient was 0.89 and 0.93,the coefficient of variation was 0.05～0.22 and 0～0.11,and Kendall's coefficients was 0.645(Chi-square=87.782,P<0.001)and 0.733(Chi-square=74.789,P<0.001),respectively.The final QPL included 3 themes and 7 questions.The intervention group asked more questions(4.03±1.89 vs 2.11±1.27,P<0.05)but spent similar time for communication(8.18±2.11 vs 7.67±1.72 min,P>0.05).At 1 d before discharge,better information recall(8.74±1.12 vs 6.49±1.68,P<0.001)and communication quality(60.06±6.25 vs 54.86±7.98,P<0.05)were observed in the intervention group when compared with the control group.Conclusion Our developed preoperative communication QPL is of scientificalness and effectiveness for elderly BPH patients.This tool can not only encourage question-asking behavior,but also improve information recall and communication quality in the patients.

Objective To analyze the differences in the epidemiological characteristics,susceptible popula-tions,and laboratory index of patients with common acute upper respiratory tract infections(novel coronavir-us infection,influenza A and influenza B)between plateau and plain areas.Methods Clinical data of 408 008 patients with symptoms of fever or upper respiratory tract infection in the fever clinic of the Second Affiliated Hospital of Zhejiang University and General Hospital of Xizang Military Command from January 2023 to Au-gust 2024 were collected.The epidemic characteristics,susceptible populations,and peripheral blood test data were compared and analyzed.Results The total positive rate of novel coronavirus infection,influenza A virus and influenza B virus infection in plain group(17.86％)was higher than that in plateau group(14.49％),and the difference was statistically significant(P＜0.05).The positive rates(17.98％and 17.76％)of male and female upper respiratory tract infection pathogens(novel coronavirus,influenza A virus and influenza B virus)in plain group were higher than those in plateau group(13.94％and 16.00％),and the differences were statis-tically significant(P＜0.05).The total positive rates of three kinds of upper respiratory tract infection patho-gens were 16.21％,18.27％and 14.63％in the plain group,and 14.62％,14.06％and 21.26％in the plateau group,respectively.According to the results of pathogen analysis of susceptible populations,whether it was plateau or plain,the positive rate of influenza A virus was higher in the minor group,the positive rate of influ-enza B virus was highest in the adult group,and the positive rate of novel coronavirus was highest in the elder-ly group.In terms of epidemic season,plateau and plain areas were different,and the epidemic occurred earlier in the plain area.In terms of peripheral blood test indicators,there were statistically significant differences in lymphocyte count,monocyte count,neutrophil to lymphocyte ratio and other indicators between plateau group and plain group(P＜0.05),while there were no statistically significant differences in white blood cell count between plateau group and plain group(P＞0.05).Conclusion The epidemiological characteristics of acute upper respiratory tract infection in plateau area are obviously different from those in plain area,which may be related to the natural environment and human geography environment.

Objective To explore the predictive value of serum C-reactive protein(CRP)/albumin(Alb)ratio,cystatin C(CysC),procalcitonin(PCT),and serum amyloid A(SAA)levels for urinary tract infection after percutaneous nephrolithotripsy in patients with upper urinary tract calculi.Methods A retrospective se-lection was made of 127 patients with upper urinary tract calculi who underwent percutaneous nephrolithotrip-sy in this hospital from January 2021 to January 2024 as the research subjects.The patients were divided into the infection group(n=66)and the non-infection group(n=61)according to their postoperative urinary tract infection conditions.Analyze the distribution of pathogenic bacteria in the infection group,as well as the com-parison of clinical data,CRP/Alb ratio,CysC,PCT and SAA levels between the two groups.Multivariate Lo-gistic regression was used to analyze the influencing factors of postoperative urinary tract infection in patients.The receiver operating characteristic(ROC)curve was used to analyze the predictive value of CRP/Alb ratio,CysC,PCT and SAA levels for postoperative urinary tract infection in patients.Results The pathogenic bacte-ria in the infection group were mainly Gram-negative bacteria[44(66.67％)].The proportions of smoking history and operation duration ≥60 minutes in the infection group were higher than those in the non-infection group,and the difference was statistically significant(P＜0.05).The CRP/Alb ratio,CysC,SAA and PCT levels in the infection group were higher than those in the non-infection group,and the difference was statisti-cally significant(P＜0.05).The results of multivariate Logistic regression analysis showed that smoking his-tory,operation duration ≥60 min,upregulation of CRP/Alb ratio,CysC,SAA,and PCT levels were risk fac-tors for postoperative urinary tract infection in patients(P＜0.05).The results of ROC curve analysis showed that the combined prediction of CRP/Alb ratio,CysC,SAA and PCT for postoperative urinary tract infection in patients had a sensitivity of 89.41％,a specificity of 90.22％,and an area under the curve(AUC)of 0.933.The AUC of the combined prediction was higher than that of the individual predictions of CysC,SAA and PCT(Z=3.564,2.846,2.644,P＜0.05).Conclusion The serum CRP/Alb ratio,CysC,PCT,and SAA levels have important predictive value for urinary tract infection after percutaneous nephrolithotripsy in patients with up-per urinary tract calculi.

Objective To explore the effect of Osthole on neuroinflammation in Alzheimer's disease(AD)by regulating the lactylation of pyruvate kinase M2(PKM2).Methods(1)Animal experiments:18 mice were divided into three groups,namely wild-type(WT)group,APP/PS1 group and APP/PS1+Osthole group.Learning-and memory-related biobehavioral indicators were compared among the three groups.Immunohistochemistry was used to detect the positive expression of Iba1 in brain tissue,enzyme-linked immunosorbent assay(ELISA)was employed to detect the levels of interleukin(IL)-6,tumor necrosis factor(TNF)-α,and IL-1β in brain tissue,and Western Blot was used to detect the protein expression levels of Pan lactylation(Pan-kla)and PKM2 lactylation(PKM2-kla)in brain tissue.(2)Cell experiments:an in vitro AD model was constructed by treated in mouse microglia(BV2 cells)with LPS/Aβ1-42,and followed by treatment with Osthole.Cell viability was detected by methyl thiazolyl tetrazolium(MTT),expression of Iba1(a marker of microglial activation)was detected by Western Blot,nitric oxide(NO)production was assessed by Griess reagent,and levels of IL-6,TNF-α and IL-1β were detected by ELISA.BV2 cell-conditioned medium(CM)was co-cultured with neuroblastoma cells(Na2 cells)to assess the protective effect of Osthole on Na2 cells.(3)Molecular docking was performed between Osthole and PKM2,and experimental verification was conducted.Results In animal experiments,deficits of learning and memory in mice were aggravated in APP/PS1 group compared with that in WT group,which were improved upon treatment with Osthole.Furthermore,the APP/PS1 group mice showed an increase in Iba1 positive cells in brain tissue,an increase in the levels of pro-inflammatory factors IL-6,TNF-α and IL-1β,as well as an increase in the levels of Pan-kla and PKM2-kla compared with the WT group,while the above indexes were inhibited by the Osthole treatment.In cell experiments,Osthole had no significant effect on BV2 cell viability at concentrations up to 100 μmol/L.Treatment with LPS/Aβ1-42 upregulated the expression of Iba1,NO production,and levels of pro-inflammatory factors IL-6,TNF-α,and IL-1β in BV2 cells,while Osthole significantly inhibited the expression of these LPS/Aβ1-42-induced indicators.Meanwhile,Osthole attenuated the damage of BV2-CM on Na2 cells.The molecular docking results indicated a good binding affinity between Osthole and PKM2.Treatment with Osthole can down-regulated the levels of lactate,Pan-kla and PKM2-kla in the AD cell model.Conclusion Osthole can improve the condition of AD and reduce neuroinflammation by inhibiting the lactylation of PKM2.