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 identify influencing factors associated with the prognosis of sepsis patients receiving blood component transfusion, and to provide a more rational and scientific transfusion strategy for clinical management. Methods: Clinical data of 232 patients with sepsis treated at the Second Xiangya Hospital of Central South University between January 2022 and December 2023 were retrospectively analyzed. These patients were categorized into the transfusion group (n=64) and the non-transfusion group (n=168) based on whether they received transfusions, and the patients in the transfusion group were further divided into non-survivor group (n=26) and survivor group (n=38) based on their survival outcome. Baseline characteristics and clinical characteristics were compared between two groups. Factors impacting the prognosis of sepsis patients undergoing blood component transfusion were identified using logistic regression. Results: Compared to the non-transfusion group, the transfusion group showed significantly higher levels of coagulation indicators (prothrombin time, activated partial thromboplastin time, international normalized ratio, D-dimer) and inflammatory markers (C-reactive protein, procalcitonin, interleukin-6), while the level of hemoglobin, platelet, lymphocyte, fibrinogen, albumin, blood glucose, and oxygen saturation were significantly lower (P<0.05). The [M(P , P )] for C-reactive protein (mg/L), hemoglobin (g/L), and platelet count (×10 /L) in the transfusion vs non-transfusion groups were 178.0(156.1-178) vs 102.7(74.0-119.6), 88.5(72.3-113.0) vs 110.5(101-121.8), and 63.0(26.5-156.5) vs 202.5(108.3-286.8), respectively (all P<0.05). Logistic regression analysis revealed that hemoglobin level, platelet count, lactate concentration, and the storage duration of transfused red blood cells were independent risk factors affecting the survival outcomes of sepsis patients receiving transfusions (P<0.05). In septic transfusion patients, the [M(P , P )] lactate concentration (mmol/L) and RBC storage time (d) in the non-survivor vs survivor groups were 3.5(1.9-7.7) vs 2.1(1.3-3.5), 18.0 (13.0-18.0) vs 12.0(9.0-14.0), respectively (both P<0.05). Conclusion: Compared to non-transfused sepsis patients, those receiving transfusions exhibited poorer baseline conditions, more severe infections, and worse survival outcomes. More importantly, the study found that the timing of transfusion decisions and the quality control of blood products (such as storage duration) may directly impact patient prognosis, providing critical evidence for optimizing transfusion strategies in septicemia patients.

Objective: To identify influencing factors associated with the prognosis of sepsis patients receiving blood component transfusion, and to provide a more rational and scientific transfusion strategy for clinical management. Methods: Clinical data of 232 patients with sepsis treated at the Second Xiangya Hospital of Central South University between January 2022 and December 2023 were retrospectively analyzed. These patients were categorized into the transfusion group (n=64) and the non-transfusion group (n=168) based on whether they received transfusions, and the patients in the transfusion group were further divided into non-survivor group (n=26) and survivor group (n=38) based on their survival outcome. Baseline characteristics and clinical characteristics were compared between two groups. Factors impacting the prognosis of sepsis patients undergoing blood component transfusion were identified using logistic regression. Results: Compared to the non-transfusion group, the transfusion group showed significantly higher levels of coagulation indicators (prothrombin time, activated partial thromboplastin time, international normalized ratio, D-dimer) and inflammatory markers (C-reactive protein, procalcitonin, interleukin-6), while the level of hemoglobin, platelet, lymphocyte, fibrinogen, albumin, blood glucose, and oxygen saturation were significantly lower (P<0.05). The [M(P , P )] for C-reactive protein (mg/L), hemoglobin (g/L), and platelet count (×10 /L) in the transfusion vs non-transfusion groups were 178.0(156.1-178) vs 102.7(74.0-119.6), 88.5(72.3-113.0) vs 110.5(101-121.8), and 63.0(26.5-156.5) vs 202.5(108.3-286.8), respectively (all P<0.05). Logistic regression analysis revealed that hemoglobin level, platelet count, lactate concentration, and the storage duration of transfused red blood cells were independent risk factors affecting the survival outcomes of sepsis patients receiving transfusions (P<0.05). In septic transfusion patients, the [M(P , P )] lactate concentration (mmol/L) and RBC storage time (d) in the non-survivor vs survivor groups were 3.5(1.9-7.7) vs 2.1(1.3-3.5), 18.0 (13.0-18.0) vs 12.0(9.0-14.0), respectively (both P<0.05). Conclusion: Compared to non-transfused sepsis patients, those receiving transfusions exhibited poorer baseline conditions, more severe infections, and worse survival outcomes. More importantly, the study found that the timing of transfusion decisions and the quality control of blood products (such as storage duration) may directly impact patient prognosis, providing critical evidence for optimizing transfusion strategies in septicemia patients.

Objective To explore the real experience during treatment of patients with multiple myeloma, so as to provide evidences for clinical nursing staff to implement targeted nursing measures and to carry out research on nursing staff of high evidence level. Methods Using phenomenological methods, nine cases of patients with multiple myeloma conducted semi-structured and in-depth interviews and then using Colaizzi 7-step for analysis. Results Three major topics were extracted: poor physiological experience (feeling abnormal, numbness, pain, urinary incontinence), positive response (family support, Increased medical standards, self-regulation), self-perceived burden (economic stress, short cycle of relapse, psychological burden). Conclusion Treatment of multiple myeloma have many adverse reactions, and patients bear too much psychological burden. Medical staff need to be evaluated in a timely manner, strengthen symptom management, provide comprehensive information support, analysis of patient self-perception burden causes, propose targeted interventions, and build continuation care system to improve patient treatment experience.

Objective To study the effect and mechanism of down-regulating Silt2/Robo 1 signaling pathway on rabbit iliac artery after angioplasty restenosis. Methods The 30 male New Zealand white rabbits were divided randomly into 3 groups , namely the blank group , the control group , and the experimental group , 10 rabbits in each group. Hign-fat feeding , the rabbits were produced endothelial denudation of iliac artery stenosis model. Another 4 weeks of feeding , percutaneous balloon angioplasty was performed. Then R5 antibody was injected into the abdominal cavity. After 4 weeks of feeding ,angiography again. The results of angiography was analysied by image workstation. The concentrations of Slit2 and Robo1 was detected by ELISA. The iliac artery tissue examined by HE staining. Results The rabbit iliac artery after angioplasty restenosis animal model was set up successfully. Compared with the control group and the experimental group , the serum concentration of Slit2 and Robo1 were significantly higher (P < 0.01) than the blank group. But in the experimental group, the Slit2 and Robo1 serum concentrations were significantly lower than those in the control group (P < 0.05) after R5 antibody intervention. The area ratio stenosis and diameter stenosis rate of iliac artery were reduced that confirmed by angiography. Conclusion The expression of Slit2/Robo1 was significantly higher in the rabbit model of vascular restenosis. R5 antibody can effectively inhibit the expression of Slit2/Robo1. Down regulation of Slit2/Robo1 signaling pathway in the treatment of restenosis after angioplasty in rabbits.

OBJECTIVETo assess the feasibility and safety of using the modified active fixation pacing leads model to pace the right ventricular outflow tract septum.

METHODSA total of 136 patients undergoing artificial heart pacemaker implantation with active fixation pacing leads were randomized into two groups to receive conventional right ventricular outflow tract pacing (CRVOTP) or modified right ventricular outflow tract pacing (MRVOTP). The electrode lead wire core was modeled in a double-curved three-dimensional shape in CRVOTP group and in a J-shaped bend in MRVOTP group before fixation at the right ventricular outflow tract septum.

RESULTSRight ventricular outflow tract septum pacing was achieved successfully in all the patients. None of patients experienced serious complications. No significant differences were found between the two groups in the number of times of electrode fixation, pacing thresholds, impedance, R wave height or QRS wave width during the operation, but MRVOTP was associated with a reduced time of X -ray exposure and operation (P<0.05) due to the convenience in electrode modeling and in passing the leads through the tricuspid annulus and the direct access to the right ventricular outflow tract septum. Postoperative follow-up of the patients showed no incidence of active fixation pacing lead dislocation and comparable pacing thresholds of the ventricular electrodes, impedance, R wave height and QRS wave width between the two groups.

CONCLUTIONSUsing the modified active fixation pacing leads model to pace the right ventricular outflow tract septum can reduce the time of X -ray exposure and operation with a low probability of lead damage.

Objective This retrospective study is to analysis the special medical conditions that most Chinese secondary hospitals are facing with,and to review the safeguards and pitfalls for arterial switch operations,in order to probe intothe feasibility of this procedure for Chinese secondary hospitals and provide our experiences to help other surgeons to avoid pitfalls on complex procedures.Methods Between January 2006 and December 2011,totally 21 newborns and infants with TGA/VSD and TGA/IVS underwent arterial switch operation.There were 15 males and 6 females.In the TGA/VSD group,there were 16 cases,ranging from 30 days to 1 year at surgeries,and weight from 3.4-8.5 kg with average of (5.33 ± 1.42) kg.In the TGA/IVS group,there were 5 cases,ranging from 13 days to 1 month at surgeries,and weight from 3.1-5.5 kg with average of (3.75 ± 1.17)kg.All patients underwent one stage of arterial switch operation.Routine follow-up checking points are set at discharging,3 months,half year and every year after operation.Results The early death rate is 9.5％ (2/21),and the reexploration rate is 9.5％ (2/21).In the TGA/VSD group,average cardiopulmonary bypass time is (151 ± 33) minuntes with the aortic crossclamp time is (1 19 ± 26) minutes.Ventilator support time is 24-159 hours,and the length of ICU stay is 3-17 days.1 case has residual VSD with the diameter less than 2 mm.The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s and 2.2 m/s,and another 2 cases increase severely to 3.1 m/s and 3.7 m/s.The aortic flow velocity in 3 cases increase to 2.0m/s.ECG instructs no case has myoinfarction signs.In the TGA/IVS group,average cardiopulmonary bypass time is (170 ± 52) minuntes with the aortic crossclamp time is (137 ± 48) minutes.Ventilator support time is 51-144 hours,and the length of ICU stay is 4-14 days;The pulmonary flow velocity in 2 cases increase mildly to 2.0 m/s.The aortic flow velocity in 1 cases increase to 2.0 m/s.ECG instructs no case has myoinfarction signs.Conclusion Unbalanced medical resources distribution causes significant differences between heart centers and the secondary hospitals in China,especially on the complex congenital heart diseases procedures.However,with relatively solid background on correction of the simple congenital heart diseases,the Chinese secondary hospitals can still perform arterial switch operation with satisfactory mortality and morbidity,and provide more prompt medical services for more population.

Objective:To construct a eukaryotic expression vector of leptin gene,and to transfect it into human placental mesenchy-mal stem cells(HPMSCs)and appraise its expression.Methods:Primers were designed and the leptin gene was obtained by RT-PCR from human adipose tissue.The aimed segments were inserted into the eukaryotic expression vector pIRES2-EGFP,plasmid pIRES2-EGFP-LEP was constructed and identified by restricted enzymatic resection,and then transfected it into HPMSCs by liposome.The ex-pression of leptin in the transfected cells was detected by RT-PCR and Western blotting,the multi-differentiation potential of the cells was indentified.Results:The length of specific fragment was 500 bp,the recombinant plasmid pIRES2-EGFP-LEP presented 5.3 kb and 500 bp bands by restriction enzyme digestion.Leptin gene was expressed in transfected HPMSCs and the transfected HPMSCs maintained multi-directional differentiation potential.Conclusion:The eukaryotic expression vector of leptin can be transfected and ex-pressed in HPMSCs.

Objective To assess the feasibility and safety of using the modified active fixation pacing leads model to pace the right ventricular outflow tract septum. Methods A total of 136 patients undergoing artificial heart pacemaker implantation with active fixation pacing leads were randomized into two groups to receive conventional right ventricular outflow tract pacing (CRVOTP) or modified right ventricular outflow tract pacing (MRVOTP). The electrode lead wire core was modeled in a double-curved three-dimensional shape in CRVOTP group and in a J-shaped bend in MRVOTP group before fixation at the right ventricular outflow tract septum. Results Right ventricular outflow tract septum pacing was achieved successfully in all the patients. None of patients experienced serious complications. No significant differences were found between the two groups in the number of times of electrode fixation, pacing thresholds, impedance, R wave height or QRS wave width during the operation, but MRVOTP was associated with a reduced time of X-ray exposure and operation (P<0.05) due to the convenience in electrode modeling and in passing the leads through the tricuspid annulus and the direct access to the right ventricular outflow tract septum. Postoperative follow-up of the patients showed no incidence of active fixation pacing lead dislocation and comparable pacing thresholds of the ventricular electrodes, impedance, R wave height and QRS wave width between the two groups. Conclusion Using the modified active fixation pacing leads model to pace the right ventricular outflow tract septum can reduce the time of X-ray exposure and operation with a low probability of lead damage.

Objective To assess the feasibility and safety of using the modified active fixation pacing leads model to pace the right ventricular outflow tract septum. Methods A total of 136 patients undergoing artificial heart pacemaker implantation with active fixation pacing leads were randomized into two groups to receive conventional right ventricular outflow tract pacing (CRVOTP) or modified right ventricular outflow tract pacing (MRVOTP). The electrode lead wire core was modeled in a double-curved three-dimensional shape in CRVOTP group and in a J-shaped bend in MRVOTP group before fixation at the right ventricular outflow tract septum. Results Right ventricular outflow tract septum pacing was achieved successfully in all the patients. None of patients experienced serious complications. No significant differences were found between the two groups in the number of times of electrode fixation, pacing thresholds, impedance, R wave height or QRS wave width during the operation, but MRVOTP was associated with a reduced time of X-ray exposure and operation (P<0.05) due to the convenience in electrode modeling and in passing the leads through the tricuspid annulus and the direct access to the right ventricular outflow tract septum. Postoperative follow-up of the patients showed no incidence of active fixation pacing lead dislocation and comparable pacing thresholds of the ventricular electrodes, impedance, R wave height and QRS wave width between the two groups. Conclusion Using the modified active fixation pacing leads model to pace the right ventricular outflow tract septum can reduce the time of X-ray exposure and operation with a low probability of lead damage.