Objective To investigate the alleviating effect of jujuboside B (JuB) on acute epilepsy in mice and its protective effect on hippocampal neurons. Methods An in vitro epilepsy model was established by stimulating primary hippocampal neurons with cyclothiazide (CTZ). Spontaneous epileptiform discharge was recorded using patch clamp technique. An acute epilepsy model was induced in adult male C57BL/6 mice by intraperitoneal injecting pentylenetetrazol (PTZ) following pretreatment with JuB, and severity of epilepsy was recorded. Mice were euthanized and brain tissues were collected 24 hours after model establishment. The expression levels of mitoptosis related proteins in the hippocampus were detected by Western blotting. Neuronal damage in the hippocampal CA1 and CA3 regions was observed using pathological staining. Results JuB reduced the frequency of CTZ-induced epileptiform discharges (P＜0.001). Pretreatment with 30 mg/kg and 50 mg/kg JuB decreased the maximal behavioral seizure score and prolonged the latency to Racine stage Ⅲ seizures in PTZ-induced epileptic mice (P＜0.001). Compared with the PTZ group, JuB treatment downregulated Bax protein level (P＜0.01) and upregulated Bcl-2 protein level (P＜0.05) in acute epileptic mice. Furthermore, JuB protected neuronal viability in the hippocampal CA1 (P＜0.05) and CA3 (P＜0.01) regions, and ameliorated pathological morphological changes including cellular disarray, unclear boundaries, pyknosis, fragmentation, and dissolution. Conclusions JuB exhibits antiepileptic effects both in vivo and in vitro. It exerts potential antiepileptic effects by inhibiting mitoptosis and attenuating neuronal damage in the hippocampus in an acute epilepsy model.

Objective: To investigate the biological effects of carvacrol on rats with stomach-heat and stomach-cold and its regulation on transient receptor potential(TRP) channels in rats with stomach-heat, and to study the cold and heat properties of carvacrol and its possible mechanism. Methods: According to the random number method, 100 SD rats were divided into stomach-heat blank group, stomach-heat model group, Coptidis Rhizoma group, stomach-heat low-dose and high-dose carvacrol group, stomach-cold blank group, stomach-cold model group, Baked ginger group, stomach-cold low-dose group and high-dose carvacrol group, 10 rats in each group. The rat model of stomach-heat was established by intragastric administration of pepper aqueous solution (0.80 g/kg) and anhydrous ethanol, and the rat model of stomach-cold was established by intragastric administration of water extract of Anemarrhena asphodeloides and sodium hydroxide (10.40 g/kg). On the day of modeling, the rats in the Baked ginger group were given Baked ginger decoction (0.78 g/kg), and the rats in the Coptidis Rhizoma group were given Coptidis Rhizoma decoction (0.43 g/kg).The stomach-cold and stomach-heat low-dose group of carvacrol was given carvacrol emulsion (40 mg/kg), high-dose group was given carvacrol emulsion (80 mg/kg).All rats of the blank and model groups were given the equal volume of emulsion prepared by 5% dimethyl sulfoxide, 1% Tween 80, 1% polyethylene glycol 400, and 93% normal saline, once a day, for 7 days. The general condition of rats was observed and the body mass was recorded. The pathological morphology of gastric tissue was observed by hematoxylin-eosin staining. The changes of material and energy metabolism, cyclic nucleotide (cAMP), thyroid hormone and gastrointestinal hormone in each group were determined by enzyme-linked immunosorbent assay. The expression levels of transient receptor potential vanilloid subtype 1 (TRPV1), transient receptor potential channel M8 (TRPM8) and uncoupling protein-1 (UCP1) in rats with gastric fever were detected by Western blotting. Results: Compared with the stomach-heat blank group, the body mass of rats in the stomach-heat model group decreased at the fifth and seventh day (P<0.05). The contents (or ratio) of hepatic glycogen (HGlyc), total cholesterol (TC), triglyceride (TG), and vasoactive intestinal peptide (VIP) were decreased (P<0.05), and Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, cytochrome C oxidase (COX), NADH dehydrogenase (ND), cyclic adenosine phosphate (cAMP), cAMP/cyclic guanosine phosphate (cGMP), triiodothyronine (T3), thyroxine (T4), gastrin (GAS), motilin (MTL), and α-amylase (α-AMS) all increased (P<0.05). Compared with the stomach-heat model group, the body mass of rats in the Coptidis Rhizoma group decreased at the third, fifth, and seventh day, the contents (or ratio) of HGlyc, TC, TG, VIP and α-AMS were increased, and Na+ -K+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, and GAS all decreased (P<0.05). The body mass of rats in the stomach-heat low-dose carvacrol group decreased at the seventh day. The contents (or ratio) of HGlyc, TC, and VIP were increased, Na+ -K+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, and MTL all decreased, the expression of TRPV1 and UCP1 in gastric tissue decreased, while TRPM8 increased (P<0.05) in rats of the stomach-heat low-dose and high-dose carvacrol groups. Compared with the stomach-cold blank group, the body mass of rats in the stomach-cold model group decreased at the third, fifth, and seventh day, the contents (or ratio) of HGlyc, TC, TG, α-AMS, and VIP all increased, while Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, GAS, and MTL all decreased (P<0.05). Compared with the stomach-cold model group, the body mass of rats in the Baked ginger group was increased at the seventh day, and the contents (or ratio) of HGlyc, VIP, and α-AMS all decreased, while Na+ -K+ -ATPase, COX, ND, cAMP/cGMP, T3, T4, GAS, and MTL all increased (P<0.05). The contents of HGlyc, cAMP, α-AMS, and VIP of rats in the stomach-cold low and high-dose carvacrol group all decreased (P<0.05). TG in the stomach-cold low-dose carvacrol group was increased. TC, Ca2+ -Mg2+ -ATPase, and cGMP all increased, while cAMP/cGMP decreased (P<0.05) in the high-dose carvacrol group. Conclusion In this study, the rat model of stomach-cold and stomach-heat were successfully established by using cold and heat factors. The result showed that carvacrol had a certain inhibitory effect on body mass, material energy metabolism, cyclic nucleotide level, thyroid hormone and gastrointestinal function in rats with stomach-heat, indicating that the drug was cold. Carvacrol′s cold medicinal property could be biologically explained by TRPV1 activation, UCP1 induction, and TRPM8 suppression.

Objective To investigate the activation of microglia and astrocytes, the secretion of pro-inflammatory factors, and the survival of neurons in the hippocampus of mice with acute seizures induced by pentylenetetrazol (PTZ) 24 hours after the onset of seizures. Methods Adult male C57BL/6 mice were randomly assigned to the control group and the PTZ-induced acute epileptic seizure group using random numbers, with 28 mice in each group. The activation status of microglia and astrocytes in the CA1 region of the hippocampus was evaluated by immunofluorescence 24 hours after the onset of seizures. RNA was extracted from the hippocampal tissue to detect the expression level of inflammatory factor mRNA, and HE staining was used to assess the survival of neurons in the hippocampus. Results Twenty-four hours after PTZ-induced acute seizures in mice, the numbers of activated Iba1⁺ microglia (55.72±4.29 vs 35.71±9.66, P＜0.001) and GFAP⁺ astrocytes (51.61±8.21 vs 37.64±5.27, P＜0.01) in the CA1 region were significantly increased compared with the control group; the proportion of M1 microglia was significantly increased (0.58±0.02 vs 0.35±0.08, P＜0.0001), while the proportion of M2 microglia was significantly decreased (0.25±0.08 vs 0.44±0.09，P＜0.01); the mRNA level of pro-inflammatory factor IL-1β in the hippocampus was significantly higher than that in the control group (2.49±1.61 vs 1.02±0.68, P＜0.05); the numbers of neurons in the CA1 (284.91±73.32 vs 498.88±176.79, P＜0.05) and CA3 (247.42±50.20 vs 457.78±150.08, P＜0.05) regions during the acute phase was significantly reduced compared with the control group. Conclusions Twenty-four hours after PTZ-induced acute epileptic seizures in mice, microglia and astrocytes in the CA1 region of the hippocampus are activated, with microglia mainly presenting the M1 phenotype to exert pro-inflammatory effects, secretions of pro-inflammatory factors increasing, and surviving hippocampal neurons decreasing.

OBJECTIVE To explore the differences in the clinical laboratory test indexes between the patients with two different TCM syndrome types of eczema(dampness-heat infiltration type and non-dampness-heat infiltration type)and observe the distribution and drug resistance rate of pathogens isolated from the skin lesions so as to pro-vide bases for syndrome differentiation and reasonable use of antibiotics.METHODS A total of 180 patients with eczema who were positive for bacterial culture of skin secretions and were treated in Beijing Hospital of Traditional Chinese Medicine from Jan.2021 to Dec.2023 were enrolled in the study and were divided into the dampness-heat infiltration group with 134 cases and the non-dampness-heat infiltration group with 46 cases.The data of clini-cal laboratory test indexes were collected from the two groups of patients,the secretion specimens were sampled from the skin lesion sites,the isolated pathogens were identified by VITEK 2 Compact automatic microorganism analysis system,and the drug susceptibility testing was performed.RESULTS The direct bilirubin level of the dampness-heat infiltration group was(3.99±1.62)umol/L,higher than(3.46±1.12)umol/L of the non-damp-ness-heat infiltration group(P＜0.05),but both were in the normal range;there were no significant differences in other test indexes between the two groups.Totally 180 strains of pathogens were isolated from the 180 patients with eczema,156(86.67％)of which were gram-positive bacteria;Staphylococcus aureus(98 strains),Staphy-lococcus epidermidis(29 strains)and Staphylococcus haemolyticus(15 strains)were the predominant species of the gram-positive bacteria.The gram-negative bacteria accounted for 12.22％(22 strains).The drug resistance rate of the S.aureus strains to penicillin was up to 82.65％,and the isolation rate of methicillin-resistant Staphy-lococcus aureus(MRSA)was 12.24％(12/98);the drug resistance rates of the S.epidermidis strains to penicil-lin and erythromycin were 75.86％,and the drug resistance rate of the S.haemolyticus strains to erythromycin was 100.00％.CONCLUSIONS The gram-positive bacteria(dominated by the S.aureus)are dominant among the pathogens isolated from the skin lesion specimens of the eczema patients and are highly resistant to penicillin and erythromycin.The isolation rate of MRSA is relatuvely low.There is limited clinical significant difference in the direct bilirubin between the two groups.It is necessary to further explore more reliable indexes for syndrome dif-ferentiation.

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.

Objective To investigate the effects and potential mechanism of tuina on pain and depressive behaviors in rats with neuropathic pain(NP).Methods A total of 102 SD rats were randomly divided into blank group,sham-operation group,model group,tuina group,inhibitor group and inhibitor+tuina group,with 17 rats in each group.The NP model was established by chronic constriction injury of the sciatic nerve.Starting from the 8th day post-operation,the rats underwent a 14-day tuina intervention and stereotactic injection of the SIRT1 inhibitor EX-527(20 μg/μL,0.5 μL)into the hippocampal CA1 region.Pain behaviors were assessed using the mechanical withdrawal threshold test one day before operation and on days 7,14 and 21 post-operation.Depressive behaviors were evaluated using the forced swimming test and sucrose preference test.Nissl staining was employed to observe neuronal morphology and quantity in the hippocampal tissue,while Golgi staining was used to examine dendritic spine density,hippocampal expression of SIRT1/BDNF/TrkB signaling pathway related protein and mRNA were analyzed using immunofluorescence,Western blot and RT-qPCR.Results Compared with the sham-operation group,the model group showed a significant decrease in mechanical withdrawal threshold(P<0.001),prolonged immobility time in the forced swimming test and reduced sucrose preference(P<0.001)on days 7,14 and 21 post-operation;the morphology of hippocampal CA1 neurons was abnormal,with a significant decrease in the number of Nissl positive cells(P<0.001)and a significant decrease in dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus were significantly reduced(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly reduced(P<0.001).Compared with the model group,the tuina group showed a significant increase in mechanical withdrawal threshold(P<0.01,P<0.001)on days 14 and 21 post-operation,shortened immobility time in the forced swimming test(P<0.01,P<0.001)and increased sucrose preference(P<0.001);the hippocampal CA1 neuronal morphology was improved,with significantly increased Nissl positive cells(P<0.001)and dendritic spine density(P<0.001);the expressions of SIRT1,BDNF and TrkB in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.001),and the protein and mRNA expressions of SIRT1,BDNF and TrkB were significantly increased(P<0.001).The beneficial effects of tuina were significantly inhibited when the SIRT1 inhibitor EX-527 was used.Conclusion Tuina may alleviate pain and depressive behaviors in NP rats by activating the SIRT1/BDNF/TrkB signaling pathway and improving hippocampal neuronal structural plasticity.

AIM:To investigate the effect of cryptotanshinone(CPT)on myocardial hypertrophy induced by isoprenaline(ISO)in rats and explore its potential mechanism.METHODS:The experimental design consisted of two parts.The first aimed to investigate the effects of CPT on cardiac function,pathological manifestations,and the Janus ki-nase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in rats with myocardial hyper-trophy.The rats were divided into six groups,namely the control,CPT control,and model groups and low-(15 mg·kg-1·d-1),medium-(30 mg·kg-1·d-1),and high-dose(60 mg·kg-1·d-1)CPT treatment groups,with six rats per group.The sec-ond part aimed to validate the role of the JAK2/STAT3 signaling pathway in the CPT-mediated myocardial hypertrophy treatment.Rats were divided into four groups,namely the control,model,high-dose CPT treatment,and coumermycin A1(CA1,a JAK2/STAT3 agonist)intervention(rats received ISO injection followed by 60 mg·kg-1·d-1 of high-dose CPT and 1 mg·kg-1·d-1 of CA1 for 15 d)groups,with five rats per group.Myocardial hypertrophy was induced in rats via intra-peritoneal injection of ISO(5 mg/kg),and CPT intervention lasted for 15 days.Cardiac function-related parameters were assessed using echocardiography,and pathological changes were evaluated through hematoxylin-eosin,Masson,and wheat germ agglutinin staining.Protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and JAK2/STAT3 signaling pathway-related proteins were detected by Western blot analysis.RESULTS:Compared with the model group,CPT administration improved cardiac dysfunction-related ul-trasound markers and significantly reduced ISO-induced cardiomyocyte hypertrophy and myocardial fibrosis in rats with hy-pertrophy in a dose-dependent manner(P<0.05).Additionally,CPT decreased the levels of ANP,BNP,and β-MHC in-duced by ISO modeling(P<0.05),and inhibiting the phosphorylation of JAK2 and STAT3(P<0.05).Furthermore,a partial reversal of the therapeutic effect on myocardial hypertrophy induced by ISO modeling was observed when CA1 was administered(P<0.05).CONCLUSION:The CPT exhibits potential as a therapeutic agent for cardiac hypertrophy by effectively attenuating ISO-induced cardiac hypertrophy in rats through JAK2/STAT3 signaling inhibition.

Objective To establish a technical system for allogeneic kidney transplantation surgery in pigs using the Banna miniature pig inbred strain,and to evaluate it through routine blood tests,liver and kidney function tests,thus providing reference data for the preparation of allogeneic kidney transplantation models.Methods A total of 4 cases of allogeneic kidney transplantation surgeries were performed,including 1 case of single kidney transplantation in a healthy pig,2 cases of kidney transplantation after unilateral nephrectomy,and 1 case of kidney transplantation after bilateral nephrectomy.Before kidney transplantation,cross-matching and complement-dependent cytotoxicity(CDC)tests were used for matching between donor and recipient pigs.After kidney transplantation,peripheral blood samples were regularly collected from pigs for routine blood tests,liver function tests,and kidney function tests,and color Doppler ultrasound technology was used to detect blood supply to the transplanted kidneys.After reaching the experimental endpoint,both kidneys of pig DR1 and the left kidney of pig DR3 were collected and hematoxylin-eosin(HE)staining was performed to evaluate pathological changes in the transplanted kidneys.Results Recipient pigs DR1 and DR3 died at 17 days and 30 days after surgery respectively,while recipient pigs R and DR2 remained in good condition during the 30-day observation period.The results of liver and kidney function test showed that in pig DR1,alanine aminotransferase(ALT)levels increased on postoperative day 1(＞1 000 U/L),peaked on postoperative day 7(1 300 U/L),and aspartate aminotransferase(AST)levels peaked on postoperative day 1(＞3 000 U/L).On postoperative day 17,ALT and AST levels remained high(ALT,500 U/L;AST,700 U/L).In pigs R,DR2,and DR3,ALT and AST levels returned to normal around day 17.Serum creatinine(Crea)levels in pig R remained stable without postoperative increase.Crea levels in pigs DR1 and DR2 showed transient elevation on postoperative day 1,then gradually returned to normal(＜100 μmol/L).Crea levels in pig DR3 remained below 500 μmol/L from postoperative days 2-10,but increased between days 11-28,reaching up to 1 500 μmol/L,indicating gradual loss of kidney function.Ultrasound results showed that the preoperative resistive index(RI)of recipient pig R was 0.91.On postoperative day 24,renal cortex and medulla showed abundant blood flow signals with RI value of 0.88,which was close to the pre-transplantation RI value.For pig DR2,the RI value on postoperative day 17 was 0.89,with poor renal cortex blood flow and relatively good renal medulla blood flow.In pig DR1 on postoperative day 17,no blood flow signals were detected in the transplanted kidney.HE staining results showed that the non-transplanted healthy right kidney of pig DR1 had normal structure,while the transplanted left kidney showed blurred glomerular structure and nuclear dissolution,indicating that the left kidney had lost function before removal.In the transplanted left kidney of pig DR3,large numbers of red blood cells and lymphocyte infiltration were observed in glomeruli and renal tubules,indicating possible coagulation dysfunction and rejection reactions after kidney transplantation.Conclusion Banna miniature pig inbred strain is used as experimental animals to perform four cases of allogeneic pig-to-pig kidney transplantation.The physiological parameters of the recipient pig and the function of the transplanted kidney are monitored after surgery using routine blood tests,liver and kidney function tests,color Doppler ultrasound,and pathological examinations.The allogeneic pig-to-pig kidney transplantation technical system established in the study can provide a foundation for clinicians to conduct kidney transplantation surgeries.

AIM:To investigate the effect of cryptotanshinone(CPT)on myocardial hypertrophy induced by isoprenaline(ISO)in rats and explore its potential mechanism.METHODS:The experimental design consisted of two parts.The first aimed to investigate the effects of CPT on cardiac function,pathological manifestations,and the Janus ki-nase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in rats with myocardial hyper-trophy.The rats were divided into six groups,namely the control,CPT control,and model groups and low-(15 mg·kg-1·d-1),medium-(30 mg·kg-1·d-1),and high-dose(60 mg·kg-1·d-1)CPT treatment groups,with six rats per group.The sec-ond part aimed to validate the role of the JAK2/STAT3 signaling pathway in the CPT-mediated myocardial hypertrophy treatment.Rats were divided into four groups,namely the control,model,high-dose CPT treatment,and coumermycin A1(CA1,a JAK2/STAT3 agonist)intervention(rats received ISO injection followed by 60 mg·kg-1·d-1 of high-dose CPT and 1 mg·kg-1·d-1 of CA1 for 15 d)groups,with five rats per group.Myocardial hypertrophy was induced in rats via intra-peritoneal injection of ISO(5 mg/kg),and CPT intervention lasted for 15 days.Cardiac function-related parameters were assessed using echocardiography,and pathological changes were evaluated through hematoxylin-eosin,Masson,and wheat germ agglutinin staining.Protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and JAK2/STAT3 signaling pathway-related proteins were detected by Western blot analysis.RESULTS:Compared with the model group,CPT administration improved cardiac dysfunction-related ul-trasound markers and significantly reduced ISO-induced cardiomyocyte hypertrophy and myocardial fibrosis in rats with hy-pertrophy in a dose-dependent manner(P<0.05).Additionally,CPT decreased the levels of ANP,BNP,and β-MHC in-duced by ISO modeling(P<0.05),and inhibiting the phosphorylation of JAK2 and STAT3(P<0.05).Furthermore,a partial reversal of the therapeutic effect on myocardial hypertrophy induced by ISO modeling was observed when CA1 was administered(P<0.05).CONCLUSION:The CPT exhibits potential as a therapeutic agent for cardiac hypertrophy by effectively attenuating ISO-induced cardiac hypertrophy in rats through JAK2/STAT3 signaling inhibition.

Objective To establish a technical system for allogeneic kidney transplantation surgery in pigs using the Banna miniature pig inbred strain,and to evaluate it through routine blood tests,liver and kidney function tests,thus providing reference data for the preparation of allogeneic kidney transplantation models.Methods A total of 4 cases of allogeneic kidney transplantation surgeries were performed,including 1 case of single kidney transplantation in a healthy pig,2 cases of kidney transplantation after unilateral nephrectomy,and 1 case of kidney transplantation after bilateral nephrectomy.Before kidney transplantation,cross-matching and complement-dependent cytotoxicity(CDC)tests were used for matching between donor and recipient pigs.After kidney transplantation,peripheral blood samples were regularly collected from pigs for routine blood tests,liver function tests,and kidney function tests,and color Doppler ultrasound technology was used to detect blood supply to the transplanted kidneys.After reaching the experimental endpoint,both kidneys of pig DR1 and the left kidney of pig DR3 were collected and hematoxylin-eosin(HE)staining was performed to evaluate pathological changes in the transplanted kidneys.Results Recipient pigs DR1 and DR3 died at 17 days and 30 days after surgery respectively,while recipient pigs R and DR2 remained in good condition during the 30-day observation period.The results of liver and kidney function test showed that in pig DR1,alanine aminotransferase(ALT)levels increased on postoperative day 1(＞1 000 U/L),peaked on postoperative day 7(1 300 U/L),and aspartate aminotransferase(AST)levels peaked on postoperative day 1(＞3 000 U/L).On postoperative day 17,ALT and AST levels remained high(ALT,500 U/L;AST,700 U/L).In pigs R,DR2,and DR3,ALT and AST levels returned to normal around day 17.Serum creatinine(Crea)levels in pig R remained stable without postoperative increase.Crea levels in pigs DR1 and DR2 showed transient elevation on postoperative day 1,then gradually returned to normal(＜100 μmol/L).Crea levels in pig DR3 remained below 500 μmol/L from postoperative days 2-10,but increased between days 11-28,reaching up to 1 500 μmol/L,indicating gradual loss of kidney function.Ultrasound results showed that the preoperative resistive index(RI)of recipient pig R was 0.91.On postoperative day 24,renal cortex and medulla showed abundant blood flow signals with RI value of 0.88,which was close to the pre-transplantation RI value.For pig DR2,the RI value on postoperative day 17 was 0.89,with poor renal cortex blood flow and relatively good renal medulla blood flow.In pig DR1 on postoperative day 17,no blood flow signals were detected in the transplanted kidney.HE staining results showed that the non-transplanted healthy right kidney of pig DR1 had normal structure,while the transplanted left kidney showed blurred glomerular structure and nuclear dissolution,indicating that the left kidney had lost function before removal.In the transplanted left kidney of pig DR3,large numbers of red blood cells and lymphocyte infiltration were observed in glomeruli and renal tubules,indicating possible coagulation dysfunction and rejection reactions after kidney transplantation.Conclusion Banna miniature pig inbred strain is used as experimental animals to perform four cases of allogeneic pig-to-pig kidney transplantation.The physiological parameters of the recipient pig and the function of the transplanted kidney are monitored after surgery using routine blood tests,liver and kidney function tests,color Doppler ultrasound,and pathological examinations.The allogeneic pig-to-pig kidney transplantation technical system established in the study can provide a foundation for clinicians to conduct kidney transplantation surgeries.