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 evaluate the clinical efficacy of endoclip papillaplasty (ECPP) for preventing recurrent choledocholithiasis after endoscopic retrograde cholangiopancreatography (ERCP).Methods:A retrospective analysis was conducted on 1 941 patients who underwent ERCP for choledocholithiasis in Henan Provincial People's Hospital from January 2019 to December 2023. A total of 250 patients who received ECPP were assigned to the ECPP group, while 251 matched controls were selected via 1∶1 year-stratified sampling into the control group. After follow-up, 209 ECPP cases and 190 controls were ultimately included in the analysis. Stone removal success rate, incidence of perioperative complications, and postoperative choledocholithiasis recurrence were compared between the two groups. Univariate and multivariate logistic regression were used to determine the risk factors for choledocholithiasis recurrence after ERCP.Results:Both groups achieved 100.0% stone removal success rate. There was no significant difference in the incidence of intraoperative perforation [0.5% (1/209) VS 1.1% (2/190), χ2=0.01, P=0.934], postoperative hyperamylasemia [21.5% (45/209) VS 17.4% (33/190), χ2=1.10, P=0.295] or post-ERCP pancreatitis [3.8% (8/209) VS 8.1% (9/190), χ2=0.20, P=0.653] between the ECPP group and the control group. The ECPP group showed significantly lower bleeding rate [5.1% (11/209) VS 12.3% (23/190), χ2=5.98, P=0.014] and choledocholithiasis recurrence rate [10.5% (22/209) VS 18.9% (36/190), χ2=5.68, P=0.017] compared with the control group. The multivariate logistic regression identified dilated common bile duct diameter ( OR=1.881, 95% CI: 1.101-3.213, P=0.021) as an independent risk factor for choledocholithiasis recurrence, while being female ( OR=0.482, 95% CI: 0.266-0.875, P=0.016) and ECPP ( OR=0.497, 95% CI:0.278-0.887, P=0.018) were protective factors. Conclusion:ECPP effectively reduces choledocholithiasis recurrence rate and bleeding risk after ERCP. ECPP and being female serve as protective factors for choledocholithiasis recurrence, while dilated bile duct diameter is an independent risk factor.

Objective:To evaluate the clinical efficacy of endoclip papillaplasty (ECPP) for preventing recurrent choledocholithiasis after endoscopic retrograde cholangiopancreatography (ERCP).Methods:A retrospective analysis was conducted on 1 941 patients who underwent ERCP for choledocholithiasis in Henan Provincial People's Hospital from January 2019 to December 2023. A total of 250 patients who received ECPP were assigned to the ECPP group, while 251 matched controls were selected via 1∶1 year-stratified sampling into the control group. After follow-up, 209 ECPP cases and 190 controls were ultimately included in the analysis. Stone removal success rate, incidence of perioperative complications, and postoperative choledocholithiasis recurrence were compared between the two groups. Univariate and multivariate logistic regression were used to determine the risk factors for choledocholithiasis recurrence after ERCP.Results:Both groups achieved 100.0% stone removal success rate. There was no significant difference in the incidence of intraoperative perforation [0.5% (1/209) VS 1.1% (2/190), χ2=0.01, P=0.934], postoperative hyperamylasemia [21.5% (45/209) VS 17.4% (33/190), χ2=1.10, P=0.295] or post-ERCP pancreatitis [3.8% (8/209) VS 8.1% (9/190), χ2=0.20, P=0.653] between the ECPP group and the control group. The ECPP group showed significantly lower bleeding rate [5.1% (11/209) VS 12.3% (23/190), χ2=5.98, P=0.014] and choledocholithiasis recurrence rate [10.5% (22/209) VS 18.9% (36/190), χ2=5.68, P=0.017] compared with the control group. The multivariate logistic regression identified dilated common bile duct diameter ( OR=1.881, 95% CI: 1.101-3.213, P=0.021) as an independent risk factor for choledocholithiasis recurrence, while being female ( OR=0.482, 95% CI: 0.266-0.875, P=0.016) and ECPP ( OR=0.497, 95% CI:0.278-0.887, P=0.018) were protective factors. Conclusion:ECPP effectively reduces choledocholithiasis recurrence rate and bleeding risk after ERCP. ECPP and being female serve as protective factors for choledocholithiasis recurrence, while dilated bile duct diameter is an independent risk factor.

Objective:To evaluate the effect of heavy smoking on the dose-effect relationship of ciprofol for painless gastroscopy when combined with alfentanil.Methods:This was a prospective study. American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ male patients, aged 18-64 yr, with body mass index of 18-28 kg/m 2, undergoing elective painless gastroscopy from October 2023 to February 2024 in Baoding First Central Hospital, were divided into non-smoking group and heavy smoking group (smoking index>400) according to the status of smoking. Alfentanil 5 μg/kg and ciprofol were intravenously injected, and gastroscopy was performed after the patient′s eyelash reflex disappeared and the jaw was relaxed. The study was performed by the Dixon′s up-and-down method, and the initial dose of ciprofol was 0.4 mg/kg. The dose of ciprofol increased or decreased by 0.04 mg/kg each time based on the positive or negative response of the previous patient. A positive response was defined as the Modified Observer′s Assessment of Alertness/Sedation Scale score being greater than 1 point at 3 min after ciprofol injection or the occurrence of coughing, swallowing, body movement, or other responses that affected the operation during the insertion of the endoscope. The median effective dose (ED 50) and 95% confidence interval of propofol for painless gastroscopy were determined by the probit analysis. Results:Twenty-five patients were finally included in non-smoking group and 23 patients in heavy smoking group. The ED 50 (95% confidence interval) of ciprofol when combined with alfentanil was 0.205 (0.159, 0.244) mg/kg in non-smoking group and 0.252 (0.184, 0.295) mg/kg in heavy smoking group. The ED 50 was significantly higher in heavy smoking group than in non-smoking group ( u=390, P=0.009). Conclusions:Heavy smoking can weaken the sedative potency of propofol for sedation when used for painless gastroscopy when combined with alfentanil.

Objective:To evaluate the effect of extracorporeal shock wave on the phosphoproteomics of the spinal cord in rats with diabetic neuralgia.Methods:Thirty-six healthy male SPF-grade Sprague-Dawley rats, aged 2 months, weighing 200-250 g, were divided into 3 groups ( n=12 each) using the random number table method: control group (group C), diabetic neuralgia group (group D), and extracorporeal shock wave + diabetic neuralgia group (group E). The rats were continuously fed a common diet in group C, while the rats were fed a high-sugar and high-fat diet for 8 weeks in D and E groups. Streptozotocin 35 mg/kg was intraperitoneally injected, and the successful induction of diabetic neuralgia was defined as the blood glucose >14.6 mmol/L and the mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) ≤85% of baseline values. Group E received extracorporeal shock wave treatment after developing the model, with 1, 000 shocks per session at a frequency of 10 Hz and an energy of 1.0 bar, once per week for a total of 4 sessions. The MWT and TWL were measured before developing the model (T 0) and at 1, 2, 3 and 4 weeks after developing the model (T 1-T 4). After the last extracorporeal shock wave treatment, the rats were anesthetized and sacrificed, and lumbar spinal cord tissues were obtained for proteomic analysis and for detection of the expression of glial fibrillary acidic protein (GFAP), interleukin-1beta (IL-1β), and tumor necrosis factor-alpha (TNF-α) (by immunohistochemistry). Results:Compared with group C, the MWT and TWL were significantly decreased at T 1-T 4 in D and E groups ( P<0.05). Compared with group D, the MWT and TWL were significantly increased at T 1-T 4 in group E ( P<0.05). The results of phosphoproteomics screening revealed 284 differentially phosphorylated proteins in D and C groups, 282 in E and C groups, and 303 in E and D groups ( P<0.05). The results of immunohistochemistry showed that the expression of GFAP, IL-1β and TNF-α was significantly up-regulated in group D compared with group C ( P<0.05); the expression of GFAP, IL-1β and TNF-α was significantly down-regulated in group E compared with group D ( P<0.05). Conclusions:The mechanism by which extracorporeal shock wave alleviates diabetic neuralgia is related to inhibition of astrocyte activation and excessive phosphorylation of mGluR5 in rats.

Objective:To evaluate the effect of age on the sedative potency of remimazolam.Methods:This was a prospective study. Patients of either sex, aged 40-80 yr, with body mass index of 18-28 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, scheduled for elective surgery with general anesthesia, were divided into middle age group (40-64 yr) and elderly group (65-80 yr) based on the age of the patients. The test was performed by the Dixon′s up-and-down method. The initial dose of remimazolam was 0.3 mg/kg. The Modified Observer′s Assessment of Alertness/Sedation Scale score ≤1 and bispectral index value≤60 within 3 min after administration were considered as an effective sedation. The dose of remimazolam was increased/decreased by 0.03 mg/kg based on the sedative efficacy in the previous patient. The 50% effective dose (ED 50) and 95% confidence interval ( CI) of remimazolam were calculated by the probit method. The time to the loss of consciousness was recorded. Results:The ED 50 (95% CI) of remimazolam was 0.345 (0.306-0.384) mg/kg in middle age group, while the ED 50 (95% CI) of remimazolam was 0.322 (0.303-0.339) mg/kg in elderly group. The ED 50 was significantly higher in middle age group than in elderly group ( u=417.00, P=0.022). The time to loss of consciousness was significantly longer in elderly group than in middle age group ( t=-2.96, P=0.008). Conclusions:For middle-aged and elderly patients aged 40-80 yr, the sedative potency of remimazolam increases with age.

Objective:To compare the anesthetic potency of ciprofol combined with alfentanil in patients with different body mass indexes (BMIs) during gastroscopy.Methods:American Society of Anesthesiologists Physical Status classification I or Ⅱ patients, aged 18-64 yr, with a BMI of 18-36 kg/m 2, undergoing elective painless gastroscopy at Baoding First Central Hospital from October to December 2023, were divided into 3 groups: normal group (18 kg/m 2≤BMI<24 kg/m 2), overweight group (24 kg/m 2≤BMI<28 kg/m 2) and obesity group (BMI≥28 kg/m 2). The 95% effective dose (ED 95) of ciprofol during painless gastroscopy was determined by the biased coin design. Alfentanyl 5 μg/kg and ciprofol were intravenously injected for anesthesia induction. Gastroscopy was performed after the patient′s eyelash reflex disappeared, the jaw muscle was relaxed and the Modified Observer′s Assessment of Alertness/Sedation Scale score ≤ 1. The initial dose of ciprofol was 0.2 mg/kg with a dose gradient of 0.04 mg/kg. The positive response were defined as the Modified Observer′s Assessment of Alertness/Sedation Scale score still> 1 after 3 min of ciprofol injection or any other responses affecting the procedure, such as body movement, bucking, or swallowing during insertion of the endoscope. If the response was positive, the next patient received a higher dose of ciprofol, or conversely if negative, the next patient received the same dose (95% probability) or a lower dose (5% probability). Each group was completed with 40 cases. Results:The ED 95 and 95% confidence intervals of ciprofol in normal, overweight and obese groups were 0.266 (0.246, 0.285) mg/kg, 0.218 (0.189, 0.244) mg/kg, 0.191 (0.156, 0.220) mg/kg, respectively. The ED 95 of ciprofol was significantly lower in overweight and obese groups than in normal group ( P<0.05). Conclusions:Overweight and obesity can increase the anesthetic potency of ciprofol in patients undergoing gastroscopy.

By combining the ability of SMOTE data augmentation method to process complex audio data with the accurate prediction ability of GA-SVM,a GA-SVM model based on SMOTE data augmentation algorithm is proposed and applied to the diagnosis of thyroid diseases.The results demonstrate the excellent performance of the proposed method in the classification and diagnosis of thyroid diseases.Compared with other methods,the proposed method has better diagnostic efficacy and higher applicability in the classification and diagnosis of thyroid diseases.