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.

BACKGROUND:Bibliometrics and visual analyses based on thematic literature are particularly important for understanding the foundation and frontiers of postmenopausal osteoporosis research. OBJECTIVE:To perform bibliometric,citation,and visualization analyses of highly cited SCI papers in postmenopausal osteoporosis research over the last 20 years. METHODS:The top 100 highly cited papers on postmenopausal osteoporosis published between 2003 and 2022 included in SCI-EXPANDED catalog of the Web of Science database were obtained for bibliometric measure and visual analysis using CiteSpace software. RESULTS AND CONCLUSION:The top 100 highly cited papers have a total of 67 377 citations in the Web of Science Core Collection,with an annual average of 49.17 citations per paper.Postmenopausal osteoporosis research primarily involves medical,engineering,biological,and multidisciplinary fields.The subcategories are dominated by endocrinology and metabolism,and medicine:internal medicine.Stable and close cooperative network relationships have been formed globally.United States,University of California System,Cummings,and Steven R are the country,research institution,and author,respectively,with the most highly-cited publications.The frontiers of postmenopausal osteoporosis research mainly include calcium and vitamin D supplementation and fracture risk,clinical studies of bisphosphonates in the treatment of postmenopausal osteoporosis,atypical femur fracture,clinical studies of new drugs and sequential treatment of postmenopausal osteoporosis,predictors of fracture risk,mid-and long-term follow-up of osteoporotic vertebral compression fractures,genetic polymorphisms and hereditary factors,formulation and updating of clinical practice guidelines for postmenopausal osteoporosis.Large cohort studies,high-quality randomized controlled trials,systematic reviews,meta-analyses,and clinical practice guidelines are the great engines that drive the development of clinical research in postmenopausal osteoporosis.We should make efforts in the above areas to improve China's international influence in the field of osteoporosis.

Objective To evaluate the efficacy of endovascular treatment for ruptured descending thoracic aortic aneurysm(rDTAA),and to summarize the treatment experience.Methods The clinical data of 8 patients with rDTAA,who were admitted to the Department of Vascular Interventional Surgery,Zhangjiajie Municipal People's Hospital of China to receive treatment between January 2016 and June 2023,were retrospectively analyzed.The patients included 4 males and 4 females,with a median age of 70.5 years(61-78 years).Preoperative computed tomography angiography(CTA)was performed in all the 8 patients,and the diagnosis of rDTAA complicated by hematoma in the thoracic cavity and/or mediastinum was confirmed.Emergency thoracic endovascular aortic repair(TEVAR)was carried out in all the 8 patients.Results Complete isolation of the thoracic descending aortic aneurysm was achieved in 6 patients,and 2 of them received reconstruction of left subclavian artery by using chimney stent technique.The remaining two patients had a small amount of type Ⅰa endoleak,which disappeared after implantation of a cuff stent at the proximal site.During hospitalization,one patient died of septic shock caused by thoracic aortic-esophageal fistula,and 3 patients developed respiratory failure after operation,which was recovered after thoracic drainage,anti-infection medication and ventilator-assisted breathing therapy.Two patients developed postoperative ischemic stroke,one of them presented as grade Ⅲ motor muscle weakness on the affected side accompanied by speech impairment,and the another patient presented as grade Ⅳ motor muscle weakness on the affected side accompanied by speech impairment.During the follow-up period,one patient died of sudden chest pain 7 months after surgery,and another patient who had not received chimney stent implantation developed type Ⅰa endoleak 18 months after operation,which disappeared after implantation of a cuff stent at the proximal site.The remaining 5 patients showed good clinical condition during the follow-up period.Conclusion For the treatment of rDTAA,TEVAR is clinically safe and effective.

Objective To evaluate the effect of preoperative visits using virtual reality(VR)tech-nology on alleviating perioperative anxiety in children undergoing adenotonsillectomy for obstructive sleep ap-nea syndrome(OSAS).Methods Sixty children with OSAS scheduled for elective adenotonsillectomy were selected,including 32 males and 28 females,aged 6 to 12 years,ASA physical status Ⅰ or Ⅱ.The chil-dren were randomly divided into two groups using random number table:control group and VR group,30 children in each group.The control group received routine education and preoperative visits,while the VR group used VR smart glasses to play videos of three-dimensional scenes of the operating room in addition to routine visits.The modified Yale preoperative anxiety scale-short form(m-YPAS-SF)was used to assess perioperative anxiety levels.The heart rates(HR)of the children on the day before surgery,upon entering the preparation room,and immediately after anesthesia induction,and the induction compliance checklist(ICC)was used to evaluate anesthesia induction compliance,as well as the duration of stay in the recovery room and the length of hospital stay were recorded.Results Compared with control group,the m-YPAS-SF scores in VR group were significantly lower(P＜0.05),the HR was significantly lower upon entering the preparation room and immediately after anesthesia induction(P＜0.05),and the ICC scores were signifi-cantly lower(P＜0.05),recovery room stay duration and length of hospital stay were significantly short-ened(P＜0.05).Conclusion Preoperative visits using VR can effectively alleviate perioperative anxiety in children with OSAS undergoing adenotonsillectomy which can improve anesthesia compliance and reduce the length of hospital stay.

ObjectiveXenotransplantation is an effective way to address the shortage of human organ donors, but it faces serious immune rejection reactions, including hyperacute rejection caused by blood type differences. Establishing a stable, convenient, and reliable method for pig blood type identification can quickly screen suitable donor pigs for xenograft research.MethodsBanna miniature inbred pigs, Diannan small eared pigs, and Bama Xiang pigs were selected as the research objects. DNA was extracted from the blood, oral buccal mucosa, and fetal fibroblasts of the three strains of pigs using DNA extraction kits. The target fragment of the ABO homologous gene EAA intron 7 in pigs was amplified using PCR method. Blood agglutination reaction was used to detect hemolysis in pig anterior vena cava whole blood after adding anti A and B antibodies. Immunohistochemical method was used to detect the expression level of A antigen in pig heart, liver, spleen, lung, and kidney tissues. Immunofluorescence method was used to detect the expression level of A antigen in pig oral mucosa. By comparing the results of different methods for determining pig blood types, the stability and reliability of the PCR method were verified, and a convenient PCR based pigblood type identification method was established.Results Firstly, the blood PCR results of 69 inbred strains of Banna miniature pigs, 7 Diannan small eared pigs, and 34 Bama Xiang pigs showed 20 AO blood types, 66 AA blood types, and 24 O blood types. The PCR results of fetal fibroblasts from 47 Diannan small eared pigs showed that all 47 fetuses were O blood type. Among them, the oral mucosal PCR results of 8 gene edited cloned pigs were consistent with those of donor fetal fibroblasts, all of which were O blood type. The oral mucosal PCR results of 8 wild-type pigs (2 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs) were consistent with the blood PCR identification results. Then, 11 inbred lines of Banna miniature pigs, 4 Diannan small eared pigs, and 2 Bama Xiang pigs were randomly selected for blood agglutination reaction validation, and the results were consistent with the PCR identification results of both blood samples and oral mucosa samples. Moreover, immuno-histochemical analysis was performed on the heart, liver, lung, kidney, and spleen tissues of one Banna miniature pig inbred line and two Bama Xiang pigs, and the results were consistent with blood PCR identification and blood agglutination reaction results. Finally, oral mucosal samples were collected from 2 inbred strains of Banna miniature pigs and 1 Bama Xiang pig for immunofluorescence detection, and the results were consistent with the blood PCR identification results.ConclusionBy collecting fetal cells and oral mucosal samples from live pigs for PCR detection, the blood type of pigs can be accurately and efficiently identified, providing a convenient method for blood type screening of xenograft donor pigs.

Ferroptosis is defined as an iron-dependent, non-apoptotic cell death pathway, with specific morphological phenotypes and biochemical changes. There is a growing realization that ferroptosis has significant implications for several neurodegenerative diseases. Even though ferroptosis is different from other forms of programmed death such as apoptosis and autophagic death, they involve a number of common protein molecules. This review focuses on current research on ferroptosis and summarizes the cross-talk among ferroptosis, apoptosis, and autophagy that are implicated in neurodegenerative diseases. We hope that this information provides new ideas for understanding the mechanisms and searching for potential therapeutic approaches and prevention of neurodegenerative diseases.
Apoptosis ; Autophagy ; Cell Death ; Ferroptosis ; Humans ; Neurodegenerative Diseases

To evaluate the effects of resuscitation with normal saline and sodium potassium magnesium calcium and glucose injection on renal structure and function in septic rats. Rat model of sepsis was established by ligation and perforation of cecum. Male SD rats were divided into four groups: sham operation group, sepsis group, saline resuscitation group, sodium potassium magnesium calcium and glucose injection resuscitation group. Blood gas analysis was performed at the end of resuscitation. The rats were sacrificed 72 hours after resuscitation. Blood samples were taken to measure the plasma levels of blood urea nitrogen (BUN), creatinine, interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factorα (TNFα). Caspase-3 expression was detected by immunohistochemistry in kidney sections. The degree of renal injury was evaluated by regular HE staining and electron microscope. Compared with normal saline resuscitation, sodium potassium calcium magnesium glucose injection resuscitation could decrease the levels of BUN, serum creatinine, IL-1β, IL-6 and TNFα ( P<0.05) , reduce the expression of caspase-3 ( P<0.05) , and improve the renal injury score ( P<0.05) . Sodium potassium calcium magnesium glucose injection resuscitation can significantly improve the renal function of sepsis rats with less pathological damage of the kidney.

This article reviewed and summarized the experimental researches on the treatment of gouty arthritis (GA) with traditional Chinese medicine in recent years, which showed that the traditional or modified animal models of disease were widely used, and also concerned about the the model of integrating the disease and symptoms. However, few are about the application of combined disease and syndromes model. The mechanism research result shows that, single Chinese herb, component extract of Chinese herbs and Chinese herbal compound all play an important role in inhibiting inflammation, regulating signal pathway, anti-oxidation, reducing uric acid, increasing feeling of pain and regulating body metabolism.

Objective To detect the pre- and post-treatment levels of serum interleukin (IL-20) in asthmatic patients during the acute exacerbation period and to investigate the role of IL-20 in the pathogenesis of bronchial asthma and relative clinical significance. Methods Forty-five cases of mild to moderate asthma outpatients in the First Affiliated Hospital of Guangxi Medical University from May 2013 to October 2013 were chosen as the asthma group and 32 healthy people who underwent routine physical examination as the control group. All the patients were treated with inhalation salmeterol and fluticasone 50/250 μg/μg (sucking twice a day) together with ventolin if necessary , and even with theophylline sustained-release tablets as the additional treatment in moderate asthma group for 1 week. The serum levels of IL-20 as well as immunoglobulin E (IgE) were detected by ELISA in the asthma group before and after treatment as well as in the control group. Other data including ECP , the number of eosinophil ceils (EOS) and the pulmonary function (FEV1%) were detected as well. The differences of IL-20 levels between the asthma group before and after treatment and the control group were analyzed and the correlation between IL-20 and Ig E, EOS, ECP and FEV1% were analyzed. Results Compared with the healthy control group [(13.58 ± 6.17)pg/L], the levels of IL-20 in the mild and moderate asthma pretreatment group were significantly increased [(23.43 ± 13.60)pg/L and (33.78 ± 22.69)pg/L]. Compared with the mild asthma group, the levels of IL-20 in the moderate asthma pretreatment group were significantly higher. After treatment , the levels of IL-20 [(15.73 ± 8.27)pg/L and (19.64 ± 11.69)pg/L] were decreased respectively in the asthmatic patients. The Pearson correlative analysis showed that there were positive correlations between the levels of 1L-20 and IgE , EOS and ECP respectively and there were negative correlations between IL-20 levels and FEV1%. Conclusion IL-20 may be involved in the pathogenesis of asthma.

Objective To compare the efficacy of uniform multi-drug therapy (UMDT) versus routine multi-drug therapy (RMDT) for the treatment of multi-bacillary (MB) leprosy patients based on bacterial index changes and frequencies of leprosy reaction.Methods This study recruited newly diagnosed leprosy patients after taking informed consent in three districts of Guizhou province as well as in one district of Yunnan province from November 2003 to June 2005.The patients received 6-month UMDT or 2-year RMDT.Clinical follow up and bacterial reexamination were carried out once a year.Changes of bacterial index (BI) and frequencies of leprosy reaction were compared between the patients receiving RMDT and UMDT.Results A total of 166 patients received UMDT and 170 received RMDT in this study.Among the UMDT-treated patients,114 were skin smear positive,and 83 had been followed up for 42 months; of the RMDT-treated patients,149 underwent all the bacterial examinations during a 48-month follow up.The mean bacterial index decreased from 2.84 before treatment to 0.33 at the end of the 42-month follow up in the 83 patients,and from 2.55 to 0.26 at the end of the 48-month follow up in the 149 patients,with no significant difference in the changes of bacterial index between the two groups (t =0.77,P ＞ 0.05).Bacterial index became negative in 73.5％ (61/83) of the UMDT-treated patients and in 77.2％ (115/149) of the RMDT-treated patients (x2 =0.40,P＞ 0.05)at the end of follow up.During the follow up peroid,the incidence of type Ⅰ leprosy reaction was 14.6％ (13/89) in the UMDT group,significantly higher than that in the RMDT group (3.4％ (5/149),x2 =10.08,P＜ 0.01 ).Conclusions There is no significant difference in mean bacterial index changes and bacterial clearance rate during the follow up peroid between UMDT- and RMDT-treated patients.The incidence of type Ⅰ leprosy reaction is higher in the UMDT group than in the RMDT group,and further investigation is needed to clarify the mechanisms underlying the phenomenon.