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 of electroacupuncture at"Zusanli"(ST36)on type 3 innate lymphoid cells(ILC3s)secreting interleukin(IL)-22 and intestinal cholinergic neurons in the colon of mice with inflammatory bowel disease(IBD),and to explore the mechanisms underlying electroacupuncture-mediated amelioration of IBD.Methods Twenty-four male C57BL/6J mice were divided into a normal group(n=8)and a modeling group(n=16)using the random number table method.The modeling group received 4％dextran sulfate sodium solution ad libitum for 7 days to establish the IBD model.On the fourth day after modeling,the modeling group was divided into a model group and an electroacupuncture group using the random number table method,with eight mice per group.Electroacupuncture intervention was applied bilaterally at"Zusanli"(ST36)in mice of the electroacupuncture group,once daily for 20 min each time,for seven consecutive days.After intervention,stool characteristics and occult blood were assessed.The disease activity index(DAI)of mice in each group was scored,and colon length was measured.The morphology of the colon tissue in mice was observed using hematoxylin-eosin staining,and the histopathological score was determined.A cytometric bead array was used to detect the contents of IL-1β,tumor necrosis factor-α(TNF-α),and IL-6 in colon tissue.Flow cytometry was used to determine the IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue.The content of IL-22 in colon tissue was detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the number of choline acetyltransferase(ChAT)+/cellular Fos(c-Fos)+co-labeled neurons in colon tissue.Results Compared with the normal group,mice in the model group exhibited different degrees of loose stools,strongly positive fecal occult blood test,elevated DAI scores,shortened colon length,increased colon histopathological score;increased contents of IL-1β,TNF-α,and IL-6 in colon tissue;and a decreased IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue(P＜0.05).Compared with the model group,mice in the electroacupuncture group exhibited improved fecal traits,weakly positive fecal occult blood test,decreased DAI score,increased colon length,reduced colon histopathological score;decreased contents of IL-1β,TNF-α,and IL-6 in colon tissues;an increased IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue;increased IL-22 content in colon tissue;and increased ChAT+/c-Fos+co-labeled neurons in colon tissue(P＜0.05).Conclusion Electroacupuncture at"Zusanli"(ST36)can significantly ameliorate colonic damage and reduce inflammatory responses in IBD mice,and its mechanism may be related to the activation of intestinal cholinergic neurons and the enhancement of colonic ILC3s function.

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.

Objective:To explore the effect of levonorgestrel-releasing intrauterine system (LNG-IUS) placed in the uterine cavity after transcervical preparation for the treatment of adenomyosis (AM).Methods:A retrospective study analyzed 219 cases of AM patients treated at Department of Obstetrics and Gynaecology, the Second Hospital of Hebei Medical University from March 2022 to February 2023. Among them, 112 cases were treated with LNG-IUS after hysteroscopy due to abnormal uterine bleeding or abnormal uterine echo suggested by gynecological ultrasound, and were recorded as the hysteroscopy+LNG-IUS group; 107 cases without the above manifestations were treated with LNG-IUS alone, and were recorded as the LNG-IUS group. The two groups were observed for dysmenorrhea severity, menstrual volume, endometrial thickness, anemia, serum CA125 levels, uterine volume, and adverse reactions such as abnormal vaginal bleeding and intrauterine contraceptive device displacement or shedding before and 1, 3, 6, and 12 months after treatment.Results:1) Postoperative pathology of hysteroscopy+LNG-IUS group after hysteroscopy included endometrial polyps [39.3% (44/112)], proliferative endometrium [16.1% (18/112)], uterine leiomyoma [11.6% (13/112)], secretory endometrium [8.9% (10/112)], irregular endometrial hyperplasia [6.2% (7/112)], simple hyperplasia [5.4% (6/112)], AM lesions [4.5% (5/112)], and others [8.0% (9/112)]. 2) The degree of dysmenorrhea and menstrual volume, as well as other indicators, improved after treatment in both groups (all P<0.001). 3) When comparing the two groups, the degree of dysmenorrhea, menstrual volume, endometrial thickness, anemia, and serum CA125 at 1, 3, 6, and 12 months of treatment, the hysteroscopy+LNG-IUS group was significantly better than those in the LNG-IUS group (all P<0.05), the difference of uterine volume at 12 months of treatment between the hysteroscopy+LNG-IUS group [(109.76±32.90) cm 3] and the LNG-IUS group [(120.84±31.30) cm 3] was statistically significant ( P=0.011). 4) The cumulative incidence of adverse reactions of the two groups was statistically significant ( P=0.001) between the hysteroscopy+LNG-IUS group [24.1% (27/112)] and the LNG-IUS group [52.3% (56/107), P=0.001]. The hysteroscopy+LNG-IUS group had lower rates of vaginal irregular bleeding [8.0% (9/112)] and ring displacement or shedding [3.6% (4/112)] than those in the LNG-IUS group [23.4% (25/107), P=0.002; 11.2% (12/107), P=0.030]. Conclusion:After treatment with hysteroscopy, the placement of LNG-IUS and the simple intrauterine placement of LNG-IUS both significantly improve the severity of dysmenorrhea, endometrial thickness, anemia, menstrual volume, CA125, and uterine volume. The treatment effect of the LNG-IUS group after hysteroscopy is better and adverse reactions are milder, and it is expected to become the preferred option for the long-term management of conservative treatment of AM.

Objective Through the safety evaluation of Chinese patent medicines containing cinnabar,the correlation between the evaluation score and the content of drug instructions was analyzed.And the methods and suggestions for improving the label quality of Chinese patent medicines were proposed.Methods Establish a database of Chinese patent medicines and sim-ple preparations containing cinnabar that meet the screening criteria in the 2020 edition of the Chinese Pharmacopoeia,and quan-titatively score such Chinese patent medicines and simple preparations from the two first level dimensions of safety and toxicity characteristics of Chinese patent medicines and simple preparations,and at the same time,grade the level of safety evidence.Results From the perspective of safety evaluation dimension,Chinese patent medicines and simple preparations with relatively high safety score(safety score ≥ 5 points)has a certain correlation with the content of adverse drug reactions clearly reported in the instructions of this kind of medicine,and 54.55％of the evidence risk of this kind of Chinese patent medicines and simple preparations is controllable,with good safety.From the perspective of the evaluation dimension of toxicity characteristics of Chi-nese patent medicines and simple preparations,the types of toxic ingredients in the compatibility of prescriptions and the scores of daily maximum dosage have certain guiding significance for clinical medication precautions.Conclusion For improving the ad-verse reaction information in the instructions of Chinese patent medicines and simple preparations containing cinnabar,and remin-ding the special population of drug use restrictions,it not only has an important impact on drug safety evaluation and grading,but also provides warning and reference for clinical and patient safety in drug use.

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.

Objective To investigate the effects of electroacupuncture at"Zusanli"(ST36)on type 3 innate lymphoid cells(ILC3s)secreting interleukin(IL)-22 and intestinal cholinergic neurons in the colon of mice with inflammatory bowel disease(IBD),and to explore the mechanisms underlying electroacupuncture-mediated amelioration of IBD.Methods Twenty-four male C57BL/6J mice were divided into a normal group(n=8)and a modeling group(n=16)using the random number table method.The modeling group received 4％dextran sulfate sodium solution ad libitum for 7 days to establish the IBD model.On the fourth day after modeling,the modeling group was divided into a model group and an electroacupuncture group using the random number table method,with eight mice per group.Electroacupuncture intervention was applied bilaterally at"Zusanli"(ST36)in mice of the electroacupuncture group,once daily for 20 min each time,for seven consecutive days.After intervention,stool characteristics and occult blood were assessed.The disease activity index(DAI)of mice in each group was scored,and colon length was measured.The morphology of the colon tissue in mice was observed using hematoxylin-eosin staining,and the histopathological score was determined.A cytometric bead array was used to detect the contents of IL-1β,tumor necrosis factor-α(TNF-α),and IL-6 in colon tissue.Flow cytometry was used to determine the IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue.The content of IL-22 in colon tissue was detected using an enzyme-linked immunosorbent assay.Immunofluorescence was used to detect the number of choline acetyltransferase(ChAT)+/cellular Fos(c-Fos)+co-labeled neurons in colon tissue.Results Compared with the normal group,mice in the model group exhibited different degrees of loose stools,strongly positive fecal occult blood test,elevated DAI scores,shortened colon length,increased colon histopathological score;increased contents of IL-1β,TNF-α,and IL-6 in colon tissue;and a decreased IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue(P＜0.05).Compared with the model group,mice in the electroacupuncture group exhibited improved fecal traits,weakly positive fecal occult blood test,decreased DAI score,increased colon length,reduced colon histopathological score;decreased contents of IL-1β,TNF-α,and IL-6 in colon tissues;an increased IL-22+ILC3s proportion in lamina propria lymphocytes of colon tissue;increased IL-22 content in colon tissue;and increased ChAT+/c-Fos+co-labeled neurons in colon tissue(P＜0.05).Conclusion Electroacupuncture at"Zusanli"(ST36)can significantly ameliorate colonic damage and reduce inflammatory responses in IBD mice,and its mechanism may be related to the activation of intestinal cholinergic neurons and the enhancement of colonic ILC3s function.

Antibody-drug conjugates (ADCs) represent a promising class of targeted cancer therapeutics that combine the specificity of monoclonal antibodies with the potency of cytotoxic payloads. Despite their therapeutic potential, the use of ADCs faces significant challenges, including off/on-target toxicity and resistance development. This review examines the current landscape of ADC development, focusing on the critical aspects of target selection and antibody engineering. We discuss strategies to increase ADC efficacy and safety, including multitarget approaches, pH-dependent antibodies, and masked peptide technologies. The importance of comprehensive antigen expression profiling in both tumor and normal tissues is emphasized, highlighting the role of advanced technologies, such as single-cell sequencing and artificial intelligence, in optimizing target selection. Furthermore, we explore combination therapies and innovations in linker‒payload chemistry, which may provide approaches for expanding the therapeutic window of ADCs. These advances pave the way for the development of more precise and effective cancer treatments, potentially extending ADC applications beyond oncology.
Humans ; Immunoconjugates/adverse effects* ; Neoplasms/immunology* ; Animals ; Antibodies, Monoclonal/therapeutic use* ; Antineoplastic Agents/therapeutic use*

Acute ischemic stroke (AIS) is a cerebrovascular disease characterized by high morbidity, disability, and mortality, posing a significant threat to human health. Endovascular treatment has now been established as a key method for AIS management, in which stent retrievers that can mechanically remove blood clots play a key role in this technique. In recent years, stent retrievers have evolved in complexity and functionality to improve the ability of clot removing and surgical safety. However, the present instruments still have limitations on treatment efficiency, vascular adaptability, and operational precision, posing an urgent need for innovation in the design of stent retrievers. This paper systematically reviewed the structural features and working principles of AIS stent retrievers from the perspective of efficacy evaluation metrics, historical development, recent advancements in stent retrieval technology, and future prospects.
Humans ; Ischemic Stroke/surgery* ; Stents ; Endovascular Procedures/methods* ; Thrombectomy/methods* ; Device Removal/methods*

Tumor lineage plasticity (LP) is an emerging hallmark of cancer progression. Through pharmacologically probing the function of epigenetic regulators in prostate cancer cells and organoids, we identified bromodomain protein BRD4 as a crucial player. Integrated ChIP-seq and RNA-seq analysis of tumors revealed, for the first time, that BRD4 directly activates hundreds of genes in the LP programs which include neurogenesis, axonogenesis, EMT and stem cells and key drivers such as POU3F2 (BRN2), ASCL1/2, NeuroD1, SOX2/9, RUNX1/2 and DLL3. Interestingly, BRD4 genome occupancy is reprogrammed by anti-AR drugs from facilitating AR function in CRPC cells to activating the LP programs and is facilitated by pioneer factor FOXA1. Significantly, we demonstrated that BRD4 inhibitor AZD5153, currently at clinical development, possesses potent activities in complete blockade of tumor growth of both de novo neuroendocrine prostate cancer (NEPC) and treatment-induced NEPC PDXs and that suppression of tumor expression of LP programs through reduction of local chromatin accessibility is the primary mechanism of action (MOA) by AZD5153. Together, our study revealed that BRD4 plays a fundamental role in direct activation of tumor LP programs and that its inhibitor AZD5153 is highly promising in effective treatment of the lethal forms of the diseases.