Nonobstructive azoospermia (NOA), one of the most severe types of male infertility, etiology often remains unclear in most cases. Therefore, this study aimed to detect four biallelic detrimental variants (0.5%) in the minichromosome maintenance domain containing 2 ( MCMDC2 ) genes in 768 NOA patients by whole-exome sequencing (WES). Hematoxylin and eosin (H&E) demonstrated that MCMDC2 deleterious variants caused meiotic arrest in three patients (c.1360G>T, c.1956G>T, and c.685C>T) and hypospermatogenesis in one patient (c.94G>T), as further confirmed through immunofluorescence (IF) staining. The single-cell RNA sequencing data indicated that MCMDC2 was substantially expressed during spermatogenesis. The variants were confirmed as deleterious and responsible for patient infertility through bioinformatics and in vitro experimental analyses. The results revealed four MCMDC2 variants related to NOA, which contributes to the current perception of the function of MCMDC2 in male fertility and presents new perspectives on the genetic etiology of NOA.
Humans ; Male ; Azoospermia/genetics* ; Meiosis/genetics* ; Spermatogenesis/genetics* ; Adult ; Exome Sequencing ; Microtubule-Associated Proteins/genetics* ; Alleles ; Infertility, Male/genetics*

OBJECTIVE: To explore and verify the effect and potential mechanism of Brucea javanica Seed Oil Emulsion Injection (YDZI) and Shengmai Injection (SMI) on peripheral microcirculation dysfunction in treatment of gastric cancer (GC). METHODS: The potential mechanisms of YDZI and SMI were explored through network pharmacology and verified by cellular and clinical experiments. Human microvascular endothelial cells (HMECs) were cultured for quantitative real-time polymerase chain reaction, Western blot analysis, and human umbilical vein endothelial cells (HUVECs) were cultured for tube formation assay. Twenty healthy volunteers and 97 patients with GC were enrolled. Patients were divided into surgical resection, surgical resection with chemotherapy, and surgical resection with chemotherapy combining YDZI and SMI groups. Forearm skin blood perfusion was measured and recorded by laser speckle contrast imaging coupled with post-occlusive reactive hyperemia. Cutaneous vascular conductance and microvascular reactivity parameters were calculated and compared across the groups. RESULTS: After network pharmacology analysis, 4 ingredients, 82 active compounds, and 92 related genes in YDZI and SMI were screened out. β-Sitosterol, an active ingredient and intersection compound of YDZI and SMI, upregulated the expression of vascular endothelial growth factor A (VEGFA) and prostaglandin-endoperoxide synthase 2 (PTGS2, P<0.01), downregulated the expression of caspase 9 (CASP9) and estrogen receptor 1 (ESR1, P<0.01) in HMECs under oxaliplatin stimulation, and promoted tube formation through VEGFA. Chemotherapy significantly impaired the microvascular reactivity in GC patients, whereas YDZI and SMI ameliorated this injury (P<0.05 or P<0.01). CONCLUSIONS YDZI and SMI ameliorated peripheral microvascular reactivity in GC patients. β-Sitosterol may improve peripheral microcirculation by regulating VEGFA, PTGS2, ESR1, and CASP9.
Humans ; Microcirculation/drug effects* ; Drugs, Chinese Herbal/administration & dosage* ; Stomach Neoplasms/physiopathology* ; Emulsions ; Male ; Plant Oils/administration & dosage* ; Brucea/chemistry* ; Middle Aged ; Female ; Drug Combinations ; Human Umbilical Vein Endothelial Cells/metabolism* ; Seeds/chemistry* ; Injections ; Vascular Endothelial Growth Factor A/metabolism* ; Aged ; Network Pharmacology

In recent years,significant breakthroughs have been achieved in the immunotherapy for Alzheimer's disease.In line with global advancements,two anti-amyloid-β monoclonal antibodies have been approved and successfully launched in China for clinical use.Lecanemab and Donanemab were officially used in June 2024 and April 2025 in China,respectively.In order to standardize the rational and safe application of anti-amyloid-β monoclonal antibodies for Alzheimer's disease in China,this article integrates recom-mendations from the clinical trials and real-world experience from the author's team and domestic peers to further update the recom-mendations for the clinical use of anti-amyloid-β monoclonal antibody based on the 2024 version.It includes indications for therapy,pre-treatment evaluation and preparation,administration protocols and safety measures during treatment,and post-treatment monitor-ing strategies.

Objective:To explore the safety and efficacy of intraosseous regional administration (IORA) of vancomycin for preventing infection in primary total knee arthroplasty (TKA).Methods:A total of 124 patients with knee osteoarthritis undergoing TKA between February 2024 and May 2024 at nine hospitals were enrolled. Preoperative infection prophylaxis involved either IORA (0.5 g vancomycin administered via intraosseous regional infusion before incision) or intravenous infusion (1 g vancomycin via peripheral vein). The IORA group included 15 males and 47 females with a median age of 66.5 years (range, 60.0-70.0 years), while the intravenous group included 14 males and 48 females with a median age of 66.0 years (range, 61.8-70.3 years) years. Intraoperative samples were collected including fat and synovium tissues after incision, before prosthesis placement, and after tourniquet release; distal femoral cancellous bone during femoral osteotomy; proximal tibial cancellous bone during tibial osteotomy; proximal intercondylar cancellous bone before prosthesis placement; and peripheral blood from non-infused arms at surgery initiation and after tourniquet release. Vancomycin concentrations were measured using liquid chromatography-tandem mass spectrometry. Vital sign changes were recorded from admission to 5~10 minutes post-IORA (IORA group) or post-incision (intravenous group). Follow-ups were conducted on postoperative day 1 and 3, and at 1 and 3 months, to document complications including IORA-related adverse events, periprosthetic joint infections, surgical site infections, red man syndrome, acute kidney injury, deep vein thrombosis and so on.Results:Vancomycin concentrations in bone, fat, and synovial tissue samples were significantly higher in the IORA group than in the intravenous group ( P<0.05), while vancomycin concentrations in blood samples were significantly lower in the IORA group than in the intravenous group ( P<0.05). Only 7.3%(41/558) of tissue samples in the IORA group had vancomycin concentrations below 2.0 μg/g (the minimum inhibitory concentration of vancomycin against coagulase-negative staphylococcus), compared to 59.3%(331/558) in the intravenous group (χ 2=11.285, P<0.001). In the intravenous group, 16.9%(21/124) of blood samples had vancomycin concentrations exceeding 15.0 mg/L (the threshold associated with a significantly increased risk of nephrotoxicity), while all concentrations in the IORA group were below this threshold, the difference was statistically significant (χ 2=22.943, P<0.001). There were no statistically significant difference ( P>0.05) in vital signs changes before and after vancomycin administration between the two groups. Two patients in the intravenous group experienced incision exudate, while no other related complications occurred in either group. Conclusions:Compared to the traditional intravenous infusion of 1 g vancomycin, intraosseous injection of a low dose (0.5 g) of vancomycin achieves higher local tissue concentrations in the knee joint with a lower incidence of adverse reactions and is safe for infection prophylaxis. Despite guidelines not recommending the routine use of vancomycin for preventing infection after primary TKA, intraosseous injection of 0.5 g vancomycin may be considered intraoperatively for primary TKA in the following scenarios: patients in medical institutions with a high prevalence of methicillin-resistant staphylococcus aureus (MRSA) infections, patients with potential preoperative MRSA colonization, or patients with cephalosporin allergy.

This study was aimed at understanding the antimicrobial resistance patterns,virulence characteristics,and phyloge-netic relationships of foodborne Listeria monocytogenes in Weifang.A total of 67 strains of Listeria monocytogenes were isolated from livestock,poultry meat,and clinical samples in Weifang between 2020 and 2023.The susceptibility of these isolates was determined through broth microdilution.Whole-genome sequencing and genetic characterization of these isolates were conducted.The 67 strains were divided into 12 STs,among which ST121,ST8,ST9,and ST87 predominated(76.12%).Eight groups of closely related strains were identified through cgMLST typing.Three Listeria pathogenicity islands and two genomic islands were identified in all strains:100%of the strains carried LIPI-1,5.97%carried LIPI-3,14.93%carried LIPI-4,2.99%carried LGI-2,and 4.48%of the strains carried LGI-3.No antibiotic resistance genes were found in any strains.All isolates were susceptible to ampicillin,penicillin,merope-nem,trimethoprim-sulfamethoxazole,and vancomycin.Five isolates were resistant to tetracycline,and three strains of ST87,one strain of ST8,one strain of ST224,and two strains of ST87 were simultaneously resistant to erythromycin.The tet(M)tetracycline re-sistance genes and msr(D)and mef(A)erythromycin resistance genes from three strains of ST87 and one strain of ST8 were carried by a phage similar to phi1605 in Erysipelothrix,with>95%identity.The tet(M)gene from the ST224 isolates was carried by a transposon similar to Tn5801_B15 in Enterococcus faecalis,with>95%identity.Drug-resistant strains of Listeria monocytogenes were found in livestock and poultry meat on sale in Weifang,particularly strains of type ST87 and ST224 simultaneously carrying highly pathogenic virulence islands,thus posing a threat to food safety and public health.These findings therefore warrant attention from relevant depart-ments and strengthened monitoring efforts.

This study was aimed at understanding the antimicrobial resistance patterns,virulence characteristics,and phyloge-netic relationships of foodborne Listeria monocytogenes in Weifang.A total of 67 strains of Listeria monocytogenes were isolated from livestock,poultry meat,and clinical samples in Weifang between 2020 and 2023.The susceptibility of these isolates was determined through broth microdilution.Whole-genome sequencing and genetic characterization of these isolates were conducted.The 67 strains were divided into 12 STs,among which ST121,ST8,ST9,and ST87 predominated(76.12%).Eight groups of closely related strains were identified through cgMLST typing.Three Listeria pathogenicity islands and two genomic islands were identified in all strains:100%of the strains carried LIPI-1,5.97%carried LIPI-3,14.93%carried LIPI-4,2.99%carried LGI-2,and 4.48%of the strains carried LGI-3.No antibiotic resistance genes were found in any strains.All isolates were susceptible to ampicillin,penicillin,merope-nem,trimethoprim-sulfamethoxazole,and vancomycin.Five isolates were resistant to tetracycline,and three strains of ST87,one strain of ST8,one strain of ST224,and two strains of ST87 were simultaneously resistant to erythromycin.The tet(M)tetracycline re-sistance genes and msr(D)and mef(A)erythromycin resistance genes from three strains of ST87 and one strain of ST8 were carried by a phage similar to phi1605 in Erysipelothrix,with>95%identity.The tet(M)gene from the ST224 isolates was carried by a transposon similar to Tn5801_B15 in Enterococcus faecalis,with>95%identity.Drug-resistant strains of Listeria monocytogenes were found in livestock and poultry meat on sale in Weifang,particularly strains of type ST87 and ST224 simultaneously carrying highly pathogenic virulence islands,thus posing a threat to food safety and public health.These findings therefore warrant attention from relevant depart-ments and strengthened monitoring efforts.

Objective:To explore the safety and efficacy of intraosseous regional administration (IORA) of vancomycin for preventing infection in primary total knee arthroplasty (TKA).Methods:A total of 124 patients with knee osteoarthritis undergoing TKA between February 2024 and May 2024 at nine hospitals were enrolled. Preoperative infection prophylaxis involved either IORA (0.5 g vancomycin administered via intraosseous regional infusion before incision) or intravenous infusion (1 g vancomycin via peripheral vein). The IORA group included 15 males and 47 females with a median age of 66.5 years (range, 60.0-70.0 years), while the intravenous group included 14 males and 48 females with a median age of 66.0 years (range, 61.8-70.3 years) years. Intraoperative samples were collected including fat and synovium tissues after incision, before prosthesis placement, and after tourniquet release; distal femoral cancellous bone during femoral osteotomy; proximal tibial cancellous bone during tibial osteotomy; proximal intercondylar cancellous bone before prosthesis placement; and peripheral blood from non-infused arms at surgery initiation and after tourniquet release. Vancomycin concentrations were measured using liquid chromatography-tandem mass spectrometry. Vital sign changes were recorded from admission to 5~10 minutes post-IORA (IORA group) or post-incision (intravenous group). Follow-ups were conducted on postoperative day 1 and 3, and at 1 and 3 months, to document complications including IORA-related adverse events, periprosthetic joint infections, surgical site infections, red man syndrome, acute kidney injury, deep vein thrombosis and so on.Results:Vancomycin concentrations in bone, fat, and synovial tissue samples were significantly higher in the IORA group than in the intravenous group ( P<0.05), while vancomycin concentrations in blood samples were significantly lower in the IORA group than in the intravenous group ( P<0.05). Only 7.3%(41/558) of tissue samples in the IORA group had vancomycin concentrations below 2.0 μg/g (the minimum inhibitory concentration of vancomycin against coagulase-negative staphylococcus), compared to 59.3%(331/558) in the intravenous group (χ 2=11.285, P<0.001). In the intravenous group, 16.9%(21/124) of blood samples had vancomycin concentrations exceeding 15.0 mg/L (the threshold associated with a significantly increased risk of nephrotoxicity), while all concentrations in the IORA group were below this threshold, the difference was statistically significant (χ 2=22.943, P<0.001). There were no statistically significant difference ( P>0.05) in vital signs changes before and after vancomycin administration between the two groups. Two patients in the intravenous group experienced incision exudate, while no other related complications occurred in either group. Conclusions:Compared to the traditional intravenous infusion of 1 g vancomycin, intraosseous injection of a low dose (0.5 g) of vancomycin achieves higher local tissue concentrations in the knee joint with a lower incidence of adverse reactions and is safe for infection prophylaxis. Despite guidelines not recommending the routine use of vancomycin for preventing infection after primary TKA, intraosseous injection of 0.5 g vancomycin may be considered intraoperatively for primary TKA in the following scenarios: patients in medical institutions with a high prevalence of methicillin-resistant staphylococcus aureus (MRSA) infections, patients with potential preoperative MRSA colonization, or patients with cephalosporin allergy.

To investigate the recent prevalence and molecular epidemiological characteristics of por-cine diarrhea viruses in Sichuan Province,this study used fluorescence quantitative PCR to detect porcine diarrhea samples from multiple regions in Sichuan Province from 2021 to 2023.RT-PCR was used to identify the genotypes of PEDV,PoRVA,PDCoV,and PTV,and their genetic variabil-ity,evolutionary characteristics,and recombination events were analyzed.The results showed that PEDV,PoRVA,PDCoV,and PTV were still prevalent in Sichuan region,with overall positive rates of 14.2％(40/281),13.2％(37/281),15.6％(44/281),and 12.5％(35/281),respectively.PEDV mixed infection with other pathogens was the most common.This study obtained a total of six strains of G2b PEDV,three strains of G3 PDCoV,three strains of G9P[13]PoRVA,one strain of G3P[13]PoRVA,three strains of Type 5 PTV,and one strain of Type 9 PTV.Compared to the seven vaccine strains including CV777,DR13,KPEDV-9,Chinju99,KNU-0801,AJ1102,and LW/L,the 6 PEDV strains showed multiple amino acid mutation sites in the COE region and S1D epitope region.Among them,the strains PSCLZ01 and PSCMY04 formed a separate branch in the phylogenetic tree.The three PDCoV strains have a closer genetic evolution distance to the previ-ously prevalent strains in Sichuan,but they also have 6-48 amino acid mutations compared to them.The four PoRVA strains have 104-108 amino acid variations in the VP4 gene compared to the early vaccine strain LLR,and they have 25 common amino acid variations in the VP7 gene.From the phylogenetic tree,the VP7 gene of RSCMY01/G3P[13]belongs to the same branch as the Heilongjiang strain LNCY,but its VP4 gene clusters with the Sichuan strain SCYA-C7,indica-ting that this PoRVA strain may have undergone genetic reassortment during inter-provincial transmission between different genotypes.It is worth noting that in the detected samples of PTV-5 and PTV-9,other diarrheal viruses tested negative,indicating that these two genotypes of PTV may be important pathogens causing porcine diarrhea.Additionally,the S gene of PEDV PSCLZ01 strain and PDCoV PCSCMY02 strain have undergone recombination events,and their parental strains come from different regions,both domestic and international.These findings reveal the main types of porcine diarrheal viruses,as well as their genetic diversity and variations in Sichuan Province in recent years.This study enriches the molecular epidemiological data of porcine diarrhe-al pathogens in the region and provides an important theoretical basis for the prevention,control,and purification of porcine diarrhea in the local area.

Objective:To explore the changes in number and immune function of mucosal-associated invariant T (MAIT) cells in peripheral blood of patients with newly diagnosed acute myeloid leukemia (AML),and its correlation with the occurrence and development of AML. Methods:Seventy-five clinical samples of patients with newly diagnosed AML and 48 healthy control samples in our hospital from January 2022 to February 2023 were included. Multiparametric flow cytometry was used to detect the number of MAIT cells,membrane surface markers,effector phenotypes and functional indicators in the samples. Results:Compared with healthy controls,the percentage of MAIT cells in CD3+T cells in peripheral blood of newly diagnosed AML patients was significantly reduced (P<0.001). The percentage of MAIT cells in all CD3+T cells in bone marrow of AML patients was similar to that in peripheral blood (P>0.05). Most of MAIT cells in peripheral blood of AML patients were effector memory T cells. Compared with healthy controls,the proportion of effector memory MAIT cells decreased (P<0.05),while the proportion of terminally differentiated effector memory MAIT cells and PD-1+MAIT cells increased significantly (both P<0.05). AML patients' peripheral blood MAIT cells expressed significantly higher levels of granzyme B and perforin than healthy controls (both P<0.05),and secreted significantly lower levels of cytokines such as gamma interferon and tumor necrosis factor α than healthy controls (both P<0.001). Conclusion:Compared with healthy controls,the proportion of MAIT cells in AML patients is reduced and the expression of functional markers is abnormal,suggesting that their function is impaired and may be involved in the occurrence and development of AML.

Objective:To compare the precision and stability of handheld full-course visual navigation and robot-assisted total hip arthroplasty (THA), as well as to evaluate perioperative laboratory indicators and clinical efficacy.Methods:A retrospective analysis was performed on 68 patients with unilateral hip joint disorders who underwent primary THA at the General Hospital of the People's Liberation Army of China from January to June 2024. Patients were divided into two groups based on the assistance method: the Visual Treatment Solution (VTS) group, which included 34 patients (13 males, 21 females; mean age 56.92±8.31 years; body mass index[BMI] 24.20±3.55 kg/m 2), and the MAKO group, also with 34 patients (18 males, 16 females; mean age 57.97±6.54 years; BMI 25.20±3.91 kg/m 2). Among the VTS group, 1 there were 18 cases on the left side and 16 cases on the right side, including 14 cases of femoral head necrosis, 8 cases of congenital hip dysplasia, and 12 cases of hip osteoarthritis. In the MAKO group, there were 19 cases on the left side and 15 cases on the right side, including 13 cases of femoral head necrosis, 10 cases of congenital hip dysplasia, and 11 cases of hip osteoarthritis. Preoperative planning, intraoperative verification deviations, postoperative acetabular prosthesis anteversion and abduction angles, and limb length discrepancies were analyzed. Acetabular angle distribution scatter plots were generated for intraoperative verification and postoperative imaging. The operation time, hospital stay, and laboratory values, including hemoglobin (Hb) loss, fibrinogen degradation products (FDP), D-dimer, high-sensitivity C-reactive protein (hCRP), as well as postoperative pain (measured by the visual analogue scale, VAS), hip Harris score (HHS), and Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores were recorded. Results:No statistically significant differences were observed between postoperative imaging measurements of anteversion and abduction angles and intraoperative verification values ( P>0.05). Postoperative imaging indicated that 91% (31/34) of the VTS group and 100% (34/34) of the MAKO group were within the Lewinnek safety zone. The scatter distribution of angles in the MAKO group was more concentrated than that in the VTS group. Furthermore, the angle deviation between intraoperative verification and preoperative planning was significantly greater in the VTS group for both anteversion (2.00° [-1.00°, 4.00°] vs. 0.00° [-1.00°, 1.00°]) and abduction angles (-1.00° [-5.00°, 0.00°] vs. 0.00° [-1.00°, 1.00°]; P<0.05). The bilateral limb length discrepancies were 1.35(0.67, 2.45) mm in the VTS group and 1.65(0.50, 2.52) mm in the MAKO group, with no significant difference noted ( P>0.05). Both methods achieved effective hip biomechanical reconstruction. Additionally, no significant differences in operation time, Hb loss, FDP, D-dimer, hCRP, or postoperative hospital stay were found ( P>0.05). Follow-up at one and three months postoperatively showed no significant differences in VAS (2.94±0.92 vs. 2.97±0.83), HHS (69.88±4.20 vs. 70.06±3.88), or WOMAC scores (22.44±3.15 vs. 22.76±3.39) between the two groups ( P>0.05). Conclusion:The stability of preoperative planning and design in robot-assisted total hip arthroplasty is superior to that of handheld navigation methods. Nevertheless, both approaches demonstrate comparable postoperative prosthesis stability, laboratory indicators, and clinical outcomes.