Objective To explore the construction of α-1,3-galactosyltransferase (GGTA1) gene-knockout (GTKO) Diannan miniature pigs and the kidney xenotransplantation from pigs to rhesus macaques, and to assess the effectiveness of GTKO pigs. Methods The GTKO Diannan miniature pigs were constructed using the CRISPR/Cas9 gene-editing system and somatic cell cloning technology. The phenotype of GTKO pigs was verified through polymerase chain reaction, Sanger sequencing and immunofluorescence staining. Flow cytometry was used to detect antigen-antibody (IgM) binding and complement-dependent cytotoxicity. Kidney xenotransplantation was performed from GTKO pigs to rhesus macaques. The humoral immunity, cellular immunity, coagulation and physiological indicators of the recipient monkeys were monitored. The function and pathological changes of the transplanted kidneys were analyzed using ultrasonography, hematoxylin-eosin staining, immunohistochemical staining and immunofluorescence staining. Results Single-guide RNA (sgRNA) targeting exon 4 of the GGTA1 gene in Diannan miniature pigs was designed. The pGL3-GGTA1-sgRNA1-GFP vector was transfected into fetal fibroblasts of Diannan miniature pigs. After puromycin selection, two cell clones, C59# and C89#, were identified as GGTA1 gene-knockout clones. These clones were expanded to form cell lines, which were used as donor cells for somatic cell nuclear transfer. The reconstructed embryos were transferred into the oviducts of trihybrid surrogate sows, resulting in 13 fetal pigs. Among them, fetuses F04 and F11 exhibited biallelic mutations in the GGTA1 gene, and F04 had a normal karyotype. Using this GTKO fetal pig for recloning and transferring the reconstructed embryos into the oviducts of trihybrid surrogate sows, seven surviving piglets were obtained, all of which did not express α-Gal epitope. The binding of IgM from the serum of rhesus monkey 20# to GTKO pig PBMC was reduced, and the survival rate of GTKO pig PBMC in the complement-dependent cytotoxicity assay was higher than that of wild-type pig. GTKO pig kidneys were harvested and perfused until completely white. After the left kidney of the recipient monkey was removed, the pig kidney was heterotopically transplanted. Following vascular anastomosis and blood flow restoration, the pig kidney rapidly turned pink without hyperacute rejection (HAR). Urine appeared in the ureter 6 minutes later, indicating successful kidney transplantation. The right kidney of the recipient was then removed. Seven days after transplantation, the transplanted kidney had good blood flow, the recipient monkey's serum creatinine level was stable, and serum potassium and cystatin C levels were effectively controlled, although they increased 10 days after transplantation. Seven days after transplantation, the levels of white blood cells, lymphocytes, monocytes and eosinophils in the recipient monkey increased, while platelet count and fibrinogen levels decreased. The activated partial thromboplastin time, thrombin time and prothrombin time remained relatively stable but later showed an upward trend. The recipient monkey survived for 10 days. At autopsy, the transplanted kidney was found to be congested, swollen and necrotic, with a small amount of IgG deposition in the renal tissue, and a large amount of IgM, complement C3c and C4d deposition, as well as CD68⁺ macrophage infiltration. Conclusions The kidneys of GTKO Diannan miniature pigs may maintain normal renal function for a certain period in rhesus macaques and effectively overcome HAR, confirming the effectiveness of GTKO pigs for xenotransplantation.

Aim To construct an mBSA/IL-1 β-induced inflam-matory monoarthritis model,and to explore the differences in the inflammatory monoarthritis mouse model induced by recombinant human IL-1 β and recombinant mouse IL-1 β combined with mB-SA,providing experimental evidence for the study of the patho-genesis and drug treatment of arthritis.Methods Twenty C57BL/6 mice were randomly divided into two groups,with the left knee joints as the model joints.One group was the mBSA+recombinant human IL-1β group(n=10),and the other was the mBSA+recombinant mouse IL-1β group(n=10).The de-gree of knee joint swelling and body weight changes in mice were observed.HE staining was used to detect the pathological chan-ges in the knee joints,and safranin O staining for the cartilage damage in the knee joints,and immunohistochemistry for the ex-pression of macrophages in the synovial tissue of the knee joints.Results Compared with the control joints(right knee joints),joint swelling occurred in the left knee joints of both groups of mice.Compared with the recombinant mouse IL-1β group,body weight of the mice in the recombinant human IL-1 β group de-creased more significantly,the pathology and cartilage damage of the knee joints were significantly higher,and the number of macrophages in the synovial tissue of the left knee joints in-creased significantly.Conclusion Recombinant human IL-1 βand recombinant mouse IL-1 β combined with mBSA induce in-flammatory monoarthritis in C57BL/6 mice.Compared with re-combinant mouse IL-1β,the arthritis induced by recombinant human IL-1β is more severe,suggesting that recombinant human IL-1β combined with mBSA is more conducive to the establish-ment of the inflammatory monoarthritis model.

Aim To investigate the role of tryptophan 2,3-dioxygenase(TDO2)in cisplatin-acute kidney in-jury(Cis-AKI)and to explore the mechanism of TDO2 in relation to apoptosis in tubular epithelial cells(TECs)to investigate the mechanism of TDO2 associ-ated with apoptosis.Methods An AKI model was es-tablished by intraperitoneal injection of cisplatin(Cis).Colorimetric assay was used to detect CRE and BUN levels,and PAS staining was employed to observe renal injury in mice.Immunohistochemistry was used to detect TDO2 protein expression and distribution and macrophage(F4/80+)infiltration;immunofluores-cence was used to detect the co-localization of TDO2 with the tubular marker LTL;TUNEL staining was used to detect apoptosis in mouse kidney;flow cytome-try was used to detect overexpression of human renal cortical proximal tubular epithelial cells(HK2)and apoptosis after administration of the TDO2 inhibitor 680C91;Western blot was used to detect TDO2 and NF-κB pathway protein levels in HK2 cells after over-expression and inhibition of TDO2.Results In the o-verall animal experiments,Cis-AKI mice showed signif-icantly higher levels of CRE and BUN and obvious tu-bular damage compared with the control group;at the same time,the renal tissues of Cis-AKI mice showed increased expression of F4/80,and the proportion of apoptotic cells in kidney cells was increased.Immuno-histochemistry and immunofluorescence showed that the expression of TDO2 increased,mainly localized in TECs.In cellular experiments,HK2 cells overexpress-ing TDO2 increased the proportion of apoptosis,and the expression of TDO2,p-IKBα,and p-p65 proteins was elevated,and p-IKBα/IκBα and p-p65/p65 were ele-vated;furthermore,the proportion of apoptosis was re-duced by the administration of 680C91,and the expres-sion of p-IκBα,and p-p65 proteins decreased,and the expression of p-IKBα/IKBα,and p-p65/p65 de-creased.Conclusions Elevated TDO2 in TECs is in-volved in the pathological mechanism of Cis-AKI,which may be related to its induction of apoptosis in TECs and activation of the NF-κB signaling pathway and consequently renal injury.

Prostate cancer is one of the most common male urological malignancies,in which bone metastasis of desmo-plasia-resistant prostate cancer is an important stage in the progression of the disease,which seriously affects the quality of life and survival of patients.With the development of nuclide therapy technology in recent years,223-Ra,as a new type of alpha-targeted therapy,has shown good efficacy in the treatment of desmoplasia-resistant prostate cancer bone metastasis.The purpose of this pa-per is to review the characteristics,mechanism of action,treatment,and the main research results of its treatment of desmoplasia-resistant prostate cancer bone metastasis,and provide a comprehensive review of the clinical application of 223-Ra in the treatment of desmoplasia-resistant prostate cancer bone metastasis for the clinical application of 223-Ra in prostate cancer bone metastasis.

Isoflavones which mainly distributed in leguminous plants have plenty of health benefits.Isoflavone synthase(IFS)is a membrane-associated cytochrome P450 enzyme(CYP450)which carries out the unique aryl-ring migration and hydroxylation.So far,few crystal structures of plant P450s have been obtained.We determined the crystal structure of IFS from Medicago truncatula at 1.9 ? by MAD method using a selenomethionine substituted crystal and conducted molecular docking and mutagenesis study.The structure of IFS complexed with imidazole exhibits the helix Ⅰa-loop-helix Ⅰβ motif which cor-responds to helix Ⅰ of other P450s.Compared with structures of common P450s,IFS/imidazole structure contains an extra domain,i.e.,the γ-domain.The structure reveals a homodimer in which the γ-domain of one molecule interacts with the β-domain of another.The plane of heme group makes an angle of ap-proximately 40° with the helix Ⅰa-loop-helix Ⅰβ motif.Molecular docking combined with mutagenesis study suggested that Trp-128 and Asp-300 might play important roles in substrate binding and recogni-tion.Phe-301,Ser-303 and Gly-305 from the helix Ⅰa-loop-helix Ⅰβ motif may play important roles in the aryl-ring migration.These novel structural features reveal insights into the unique reaction mechanism of IFS and provide a basis for engineering IFS in leguminous crops for health purpose.

Several imaging examination means for the military training-related injuries at the knee joint were introduced in terms of the research progress,advantages and limitations,including X-ray examination,multi-slice spiral CT,ultrasound examination and MRI.It's pointed out the progress of imaging devices and image post-processing techniques and the involvement of AI diagnosis contributed to the development of the imaging diagnoses of the military training-related injuries at the knee joint.[Chinese Medical Equipment Journal,2025,46(1):101-107]

Although no cross protection was observed between different serotypes of foot-and-mouth disease virus(FMDV),there were cross-reactivity between different serotypes of antibodies produced after vaccination,the aim of this paper was to prepare the neutralizing monoclonal anti-body against Foot-and-mouth disease virus(FMDV)serotype O,and to develop the method to dis-tinguish antibody against FMDV serotype O and A based on mAb.The inactivated FMDV serotype O was used as antigen in mAb production,a series of GST fusion overlapping peptides and trun-cated peptides expressed in Escherichia coli were used to identify antigenic epitope recognized by monoclonal antibodies.In order to verify feasibility of the screened monoclonal antibodies in diag-nosis,20 positive serum of FMDV serotype O and A,20 negative serum with known background were detected by blocking ELISA.Results were as follows:five monoclonal antibodies were suc-cessfully screened.The five monoclonal antibodies showed good reactivity with FMDV serotype O,but did not react with FMDV serotype A by Western blot and IFA,these mAbs showed neutrali-zing ability to FMDV/O/MY98/GZBY/2013 by VNT.The same epitope was identified by five monoclonal antibodies,the minimum epitope was145 RGDLQVLA152,Arg145 and Gln149 were key a-mino acids of the epitope.Sequence alignment analysis revealed that the identified epitopes were conserved among most of O type FMDV strains,but Gln149 was mutated among all A,Asia 1 and SAT1-3 type FMDV strains.The mAb-8C5D3 distinguished between antibody of FMDV serotype O and FMDV serotype A by blocking ELISA.The results provided materials for development of O type FMDV antibody detection kit and evaluation of vaccine immune effect.

In recent years,the intensification of dairy farming has significantly improved production efficiency but has also led to a rise in the incidence of metabolic diseases.Conditions such as keto-sis,fatty liver,and hypocalcemia pose serious threats to dairy cattle health and productivity.These diseases not only profoundly affect individual physiological function but also impose considerable economic pressures and challenges on farm management.As research advances,exosomes have e-merged as a novel intercellular signaling molecule,showing unique potential in regulating dairy cattle's nutritional metabolism.Studies suggest that exosomes hold promise as biomarkers for dis-ease and can even serve as carriers for disease detection and prevention.Acting as a crucial mediator of intercellular communication,exosomes play an important role in modulating the metabolic processes of dairy cattle.This review aims to systematically explore the role of exosomes in bovine nutritional metabolism and to provide new perspectives and theoretical support for their potential as tools for diagnosing,treating,and preventing metabolic diseases in dairy cattle,thus advancing research and practice in this field.

Objective:To investigate the association between the polymorphism of the hepatic lipase gene(LIPC)and the sensitivity to exercise intervention in the Han Chinese population with prediabetes mellitus(PDM)in the cold northern region of China,and to identify molecular markers related to exercise sensitivity in glucose me-tabolism disorders.Method:From January to September 2022,32 PDM patients were recruited in Harbin sports university,and 40％—59％VO2max aerobic exercise(3 times/week,30-60min/time)combined with 50％—70％1RM elastic band training(2 times/week,15-30 min/time)was performed for 12 weeks.FPG,HbA1c,FINs,TC,TG,HDL-c and LDL-c were measured before and after the intervention,SNP microarray chip was used to analyze the LIPC-514C/T locus in saliva,and SPSS26.0 was used to compare the differences in efficacy indicators among PDM patients with different LIPC-514C/T genotypes.Result:①The CC type of LIPC-514C/T locus accounted for 37.5％,and CT/TT accounted for 62.5％in PDM patients.The gene frequency distribution was consistent with Hardy-Weinberg equilibrium(P>0.05).②The HDL-c level of LIPC-514 CC type in PDM patients was lower than those in CT/TT type,with a signifi-cant difference(P<0.05),while there was no significant difference in other indicators(P>0.05).③After 12-week exercise intervention,the levels of FPG,HbA1c,TC,TG,LDL-c and HOMA-IR were significantly lower(P<0.05),and HDL-c,FINs and HOMA-IS were significantly higher than those before intervention(P<0.05).④The change in FPG,HOMA-IR,TG and HDL-c in CC type PDM patients was significantly higher than those in CT/TT genotype after intervention(P<0.05).Conclusion:A 12-week moderate-intensity aerobic exercise combined elastic band resistance strength training can improve the glucose and lipid metabolism in PDM in cold region.The effect of exercise intervention in re-ducing blood compared to the CT/TT genotype of the LIPC-514C/T locus,which can be used as a molecular marker to evaluate the efficacy of exercise intervention in the Han Chinese population with glucose metabolism disorders in cold region of northern China.

Lung cancer poses a serious threat to global public health security.Chemotherapy,as the main strategy for cancer treatment,faces challenges such as high toxicity and drug resistance.Anticancer peptides have the potential of being developed into new anticancer drugs due to their advantages of broad-spectrum anticancer activity,rapid action,and difficulty in generating drug resistance,but they also face shortcomings such as weak activity and strong toxic side effects.The weakly acidic microenvironment of tumors(pH 6.5-6.8)provides a good idea for the design of anticancer peptides of high-efficiency and low-toxicity.Previously,we designed the acid-sensitive antibacterial peptide pHly-1 using the wolf spider(Lycosa singoriensis)toxin Lycosin-Ⅰ as a template.In this study,we found that pHly-1 also had acid-sensitive anticancer activity.Further alanine scanning analysis of pHly-1 was carried out,and we ob-tained a mutant pHTP-2 with better acid sensitivity,whose IC50(half maximal inhibitory concentration)against A549 cells was 15.68 μmol/L at pH 6.6 and was greater than 100 μmol/L at pH 7.4.At pH 6.6,pHTP-2 could act on various lung cancer cell lines and induce the death of A549 cells by rapid ly-sis;at pH 7.4,500 μmol/L pHTP-2 had weak toxicity to red blood cells(the hemolysis rate was ap-proximately 38％)and primary myocardial cells(the inhibition rate was 49.7％,with P＜0.05).Analy-sis of its charge,particle size,morphology,and secondary structure showed that at pH 6.6,the histidine in the sequence of pHTP-2 was protonated,increasing the positive charge(P＜0.01),decreasing the hy-drated particle size(P＜0.05)and forming an α-helical structure to induce membrane lysis of A549 cells.At pH 7.4,it was deprotonated,the positive charge decreases,a β-sheet structure was formed and self-aggregation occurred,limiting its effect on the A549 cell membrane and showing weak activity.In summary,pHTP-2 could respond to the weakly acidic microenvironment of tumors to exert selective cyto-toxic activity,effectively overcoming the shortcomings of anticancer peptides such as low efficiency and high toxicity.Our findings suggest that it is a high-quality lead molecule for anticancer drugs.