Objective To explore the protective effects of genetically modified porcine erythrocyte suspension as a subnormothermic normoxic mechanical perfusate on hypoxic-ischemic brain injury in cynomolgus monkeys caused by traumatic hemorrhage.Methods Cynomolgus monkeys were randomly divided into positive and negative control groups(a total of 3 monkeys,with 3 left cerebral hemispheres as the positive control group and 3 right cerebral hemispheres as the negative control group)and the subnormothermic perfusion group(n=3).The positive control group was directly sampled 1 hour after circulatory arrest,while the negative control group was placed at subnormothermic conditions for 6 hours after circulatory arrest.The subnormothermic perfusion group underwent 6 hours of subnormothermic normoxic mechanical perfusion of the bilateral common carotid arteries of the cynomolgus monkey hypoxic-ischemic brain injury model using genetically modified porcine erythrocyte suspension 1 hour after circulatory arrest.Before perfusion,cross-matching experiments were conducted between the six genetically modified pig and the cynomolgus monkeys.After the start of perfusion,the levels of routine blood indicators in the perfusate were detected at 0,1,2,3,4,5 and 6 hours.Blood oxygen saturation was recorded,and the levels of Na+,K+,Ca2+,glucose and blood pH in the perfusate were measured,as well as the levels of IgG and IgM in the perfusate.After 6 hours of perfusion,the water content of the brain tissue was measured.Nissl staining was performed on the frontal cortex and hippocampal regions,and immunofluorescence staining was used to detect the expression of glial fibrillary acidic protein(GFAP),ionized calcium-binding adapter molecule 1(Iba1)and neuronal nuclear antigen(NEUN).Results The cross-matching results between the six genetically modified pig and the cynomolgus monkeys were negative.The number of red blood cells in the perfusate decreased significantly at 3 hours of perfusion,and the hemoglobin level showed a downward trend at 1,3,5 and 6 hours.The number of white blood cells and platelets decreased at all time points.The blood oxygen saturation in the subnormothermic perfusion group remained stable at 95%-98%,and the levels of blood oxygen saturation,Na+,Ca2+,glucose and pH were stable,while the K+level first increased and then decreased.There was no significant difference in the levels of IgG and IgM before and after perfusion.The water content of brain tissue at the end of perfusion in the subnormothermic perfusion group was significantly higher than that in the positive control group(P<0.001).Nissl staining results showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex of the subnormothermic perfusion group maintained better morphological integrity,with no significant increase in enlarged and deformed cells.In the hippocampal CA1 region,there was a slight increase in enlarged and deformed cells,and a few cells with undamaged structures showed reduced cell size.In the hippocampal dentate gyrus,fewer granule neurons had compromised structural integrity,with increased cell edema.NEUN immunofluorescence staining showed that compared with the positive control group,the pyramidal neurons in the prefrontal cortex and hippocampal CA1 region of the subnormothermic perfusion group had better morphological states,with clear axons.The granule cells in the hippocampal dentate gyrus were well preserved,but the nuclei were less well protected.GFAP immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had sparser protrusions that were more tightly associated with neurons.Iba1 immunofluorescence staining showed that compared with the positive control group,the subnormothermic perfusion group had thicker and fewer protrusions.Conclusions Compared with the positive control group,subnormothermic normoxic mechanical perfusion with genetically modified porcine erythrocyte perfusate increases brain tissue edema in cynomolgus monkeys,but better preserves the morphological integrity of neurons and glial cells.The protective effects may be related to the continuous oxygen and energy supply,maintenance of ion homeostasis and perfusate pH,reduced rejection,and low metabolic state of the whole brain.

Background and purpose:In recent years,chimeric antigen receptor T(CAR-T)cell therapy has achieved breakthrough progress in cancer treatment.γδ T cells,with their non-major histocompatibility complex(MHC)-restricted antigen recognition,broad antitumor activity,and low risk of graft-versus-host disease(GVHD),have garnered significant interest in CAR-γδ T cell therapy.However,critical challenges including suboptimal in vitro expansion and low viral transduction efficiency severely hinder the research and clinical application of CAR-γδ T cells.This study aimed to establish an efficient platform for preparing CAR-γδ T cells by optimizing the in vitro expansion conditions of γδ T cells and refining lentiviral transduction strategies.Methods:We first optimized the expansion protocol for γδ T cells by screening various cytokine combinations and the concentrations of individual cytokines within combination,and evaluating cell purity,viability,fold expansion,and expressions of cytotoxicity and exhaustion markers to identify the optimal culture conditions.Subsequently,the transduction conditions for CAR-γδ T cells were improved by determining the optimal activation duration of γδ T cells prior to gene transfer,as well as the optimal multiplicity of infection(MOI)for lentiviral transduction.Finally,CAR-γδ T cells were successfully generated using the optimized protocol,and their cytotoxic activity against target cells was validated via calcein-release assay and flow cytometry,with a preliminary assessment of the potential risk of GVHD induction.Results:Experimental data demonstrated that,compared with the interleukin(IL)-2-only culture,the IL-2+IL-7+IL-15 combination significantly enhanced the expansion capacity of γδ T cells(876.50±238.35-fold vs 1 627.50±472.15-fold),cell purity(73.67%±1.53%vs 90.69%±2.00%),and cell viability(63.01%±7.05%vs 89.00%±3.61%).It also increased the expression of the cytotoxicity marker CD16(4.20%±1.73%vs 14.66%±0.58%)and reduced the expression of the exhaustion marker programmed death-1(PD-1)(35.67%±6.26%vs 21.10%±6.49%).A cytokine concentration gradient orthogonal assay further identified 10 ng/mL IL-7 and 10 ng/mL IL-15 as the optimal concentrations within the IL-2+IL-7+IL-15 combination.Gene transduction performed 96-120 h after activation using a multiplicity of infection(MOI)of 5-10 resulted in the highest transduction efficiency for CAR-γδ T cells(96 h:12.87%±4.35%;120 h:11.37%±2.35%).CAR-γδ T cells generated using the optimized system exhibited specific cytotoxic effects against tumor cells expressing the target antigen,and no evidence of GVHD induction was observed.Conclusion:CAR-γδ T cells produced using the IL-2+IL-7(10 ng/mL)+IL-15(10 ng/mL)regimen combined with a 96-120 h activation period prior to transduction using a multiplicity of infection(MOI)of 5-10 significantly outperformed conventional methods in terms of expansion efficiency,cell purity,and transduction efficiency.The synergistic antitumor effects mediated by both natural immune receptors and CAR-specific recognition,along with the initial absence of GVHD risk,provide critical technical support for the clinical translation of CAR-γδ T cell therapy,establishing a solid theoretical and practical foundation.

Background and purpose:Salivary duct carcinoma(SDC)is a group of rare and highly heterogeneous diseases.It predominantly arises in the parotid glands of middle-aged and elderly males,with high rates of recurrence and metastasis,as well as a poor prognosis.Currently,there is a lack of clinical data on SDC.This study aimed to evaluate the clinical characteristics of SDC patients and explore high-risk factors affecting prognosis,so as to provide clinical references for physicians.Methods:Clinical data of patients with primary SDC who were admitted to Tianjin Medical University Cancer Institute and Hospital and Tianjin Stomatological Hospital Affiliated to Nankai University School of Medicine from 2012 to 2024,were collected retrospectively.Inclusion criteria:① patients diagnosed with primary SDC;② availability of American Joint Committee on Cancer(AJCC)staging data.Exclusion criteria:① concurrent other malignant tumors;② incomplete or missing medical records;③ death due to non-SDC causes;④ duplicate cases from the two participating hospitals.Data retrieved encompassed epidemiological information(gender,age)and clinical details(time of diagnosis,tumor characteristics,treatment regimen,recurrence and metastasis status,and pathological data).Survival analysis was performed using the Kaplan-Meier method,and factors related to prognosis were explored through univariate COX proportional hazards regression model analysis.This study was approved by the Ethics Committee of Tianjin Stomatological Hospital Affiliated to Nankai University School of Medicine(ethics number:PH2023-B-016),and patient informed consent was waived.Results:A total of 87 patients with primary SDC were included in this study,among whom 77%were male,69%had primary lesions in the parotid gland,29.9%in the submandibular gland,and one patient had a primary lesion in the minor salivary gland of the nasal cavity.49.3%of the patients had concurrent cervical lymph node metastasis.The median overall survival(OS)of the entire group was 31.2 months,the median progression-free survival(PFS)was 20.3 months,and the 5-year OS rate was 52.6%.The 5-year OS rate for tumors originating from the parotid gland was 60%,which was better than the 32.9%for those originating from the submandibular gland.Among the 85 patients who received surgical treatment,65.9%underwent both resection of the primary tumor and neck dissection.Postoperative radiotherapy was administered to 49 patients.During the follow-up period,46%of the patients developed recurrence or metastasis,with lung and bone metastases being the most common.The median OS and local progression-free time in the postoperative radiotherapy group were significantly longer compared with those in the group without radiotherapy,however,the difference was not statistically significant.Conclusion:SDC is a malignant and aggressive disease that predominantly occurs in the parotid glands of middle-aged and elderly males,with a high rate of lymph node metastasis and poor prognosis.Clinically,it is recommended that patients with SDC undergo radical resection of the primary lesion and cervical lymph node dissection,combined with postoperative adjuvant radiotherapy.Targeted therapy and immunotherapy are worthy of further exploration.

Objective:To identify risk factors influencing the incidence of hypertension among steelworkers (Homo sapiens) and establish an effective and easily implementable hypertension prediction model.Methods:In September 2023, 2214 steelworkers (Homo sapiens) were selected as study subjects. Basic demographic information, lifestyle, and occupational exposure data were collected, along with physiological measurements such as height, weight, and blood pressure. Multivariate unconditional logistic regression analysis was employed based on relevant literature to determine influencing factors for hypertension among steelworkers (Homo sapiens). Python 3.9 software was used to construct and compare logistic regression, support vector machine (SVM), random forest, extreme gradient boosting tree (XGBoost), and LGBM models. Model performance was evaluated using metrics such as receiver operating characteristic (ROC) curves, accuracy, calibration curves, and F1 scores. The Shapley Additive Explanations (SHAP) model was introduced for feature importance analysis to enhance the interpretability of the prediction model.Results:A total of 432 cases of hypertension were detected among 2214 study subjects, with a detection rate of 19.51%. Age, smoking status, salt intake, use of cooling equipment, carbon monoxide exposure, family history of hypertension, fasting blood glucose, triglycerides, and hemoglobin were identified as independent risk factors for hypertension ( P<0.05). A comparison of the five models revealed the following performance metrics: logistic regression achieved an accuracy of 0.853, F1 score of 0.680, Brier score of 0.108, and AUC of 0.907; SVM demonstrated an accuracy of 0.863, F1 score of 0.687, Brier score of 0.081, and AUC of 0.910; random forest showed an accuracy of 0.857, F1 score of 0.603, Brier score of 0.105, and AUC of 0.861; XGBoost yielded an accuracy of 0.850, F1 score of 0.684, Brier score of 0.117, and AUC of 0.899; and the LGBM model exhibited an accuracy of 0.838, F1 score of 0.625, Brier score of 0.112, and AUC of 0.870. Conclusion:The SVM model demonstrated strong predictive performance, effectively assessing the risk of hypertension among steelworkers (Homo sapiens) and facilitating targeted health management interventions.

Liumotang comes from the Yuan dynasty's Effective Prescription Handed Down for Generations of Physicians. It is composed of six medicinal materials： Arecae Semen， Aquilariae Lignum Resinatum， Aucklandiae Radix， Linderae Radix， Rhei Radix et Rhizoma， and Aurantii Fructus. It is a classical formula for treating abdominal pain due to Qi stagnation and constipation accompanied by heat. This study systematically collated the records of Liumotang in ancient medical books and modern clinical literature and conducted in-depth analysis and textual research on its formula source， main diseases， composition， dosage， medical books， container capacity， processing， preparation method， usage， drug basis， formula meaning， and other key information， so as to provide a powerful reference for the development and clinical application of compound preparations of the classical formula Liumotang. The results show that Liumotang was first seen in Effective Prescription Handed Down for Generations of Physicians， and many medical books of the past dynasties have imitated this. In terms of drug basis， the dried and mature seeds of the palm plant Areca catechu， resin-containing wood of the Daphneaceae plant Aquilaria sinensis， the dried roots of the Asteraceae plant woody Aucklandia lappa， the dried tuber root of the Lauraceae plant Lindera aggregata， the dried roots and rhizomes of the knotweed plant， R. palmatum， R.tangutikum， and R. officinale， and the dried and unripe fruits of the citrus genus C. aurantium and its cultivated varieties from the family Rutaceae were selected. In terms of dosage， through the textual research on bowls in the Ming and Qing dynasties， combined with the conversion of medicines and bowl capacity in the Qing dynasty， it was estimated that the dosage of each drug in the Yuan dynasty was 10.86 g. In the Ming and Qing dynasties， the dosage of drugs was mostly equal， but the dosage of drugs was somewhat different. In terms of processing， preparation method， and usage， in the medical books of the past dynasties， the processing of drugs has slightly changed， but raw drugs are used in all preparations. The preparation method and usage did not change much during the Yuan， Ming， and Qing dynasties， except for certain differences in dosage. In terms of syndrome， Liumotang was first used to treat abdominal pain due to Qi stagnation and constipation accompanied by heat. Medical books of the past dynasties often omit the symptoms of heat. In modern clinical practice， Liumotang is mainly used in the digestive system and urinary system diseases and is mostly used to treat constipation-predominant irritable bowel syndrome， biliary reflux gastritis， functional constipation， slow transit constipation， and other diseases， with no adverse reactions found yet. The above results provide a reliable scientific basis for the development and clinical treatment of Liumotang compound preparations.

Objective:To summarize the surgical strategies and to evaluate the clinical outcomes of up-per urinary tract reconstruction in patients with stone-related ureteral strictures.Methods:This retro-spective study included 71 patients diagnosed with ureteral strictures secondary to urinary stones who un-derwent upper urinary tract reconstructive surgery at Peking University First Hospital between March 2014 and November 2023.Patient data were collected,including demographic characteristics,clinical presen-tation,laboratory results,imaging findings,surgical procedures,and follow-up outcomes.Ureteral stric-tures were classified according to anatomical location into upper,middle,lower,or multiple segments.Surgical procedures were carried out depending on the stricture characteristics.Surgical success was de-fined as resolution or improvement of clinical symptoms,radiographic improvement or stabilization of hy-dronephrosis,and maintenance of normal and stable renal function.Results:Among the 71 patients,36(50.7％)had strictures in the upper ureter,9(12.7％)in the middle ureter,15(21.1％)in the lower ureter,and 11(15.5％)had multifocal ureteral strictures.The median stricture length was 5.0 cm(interquartile range:3.0-15.0 cm).Surgical approach selection was individualized based on the location and extent of the stricture.For upper ureteral strictures,the most frequently employed techniques were oral mucosal graft ureteroplasty(13/36,36.1％)and appendiceal flap ureteroplasty(8/36,22.2％).Other options included ureteroureterostomy and ileal ureter replacement for longer or more complex strictures.In middle ureteral strictures,treatment was stratified by length:balloon dilation(1/9,11.1％)and ureteroureterostomy(1/9,11.1％)were applied in shorter strictures,while oral mucosal graft ureteroplasty(3/9,33.3％)and ileal ureter replacement(4/9,44.4％)were reserved for longer segments.For lower ureteral strictures,ureteral reimplantation into the bladder was the most common approach(10/15,66.7％),often combined with a psoas hitch or Boari flap when necessary.All the patients with multiple segmental strictures underwent ileal ureter replacement due to the extensive nature of the disease.The median follow-up period was 14.2 months(range:6.1-107.1 months).During follow-up,69 of 71 patients(97.2％)achieved surgical success.Conclusion:Stone-related ure-teral strictures present with considerable heterogeneity in terms of anatomical location,length,and com-plexity.Careful preoperative evaluation and individualized surgical planning are critical to successful re-construction.With appropriate selection of surgical methods,favorable long-term clinical outcomes can be achieved in the majority of patients.

ObjectiveTo develop a nomogram-based risk prediction model for chronic obstructive pulmonary disease (COPD) incidence among the community-dwelling population aged 40 years old and above, so as to provide targeted references for the screening and prevention of COPD. MethodsBased on a natural population cohort in suburban Shanghai, a total of 3 381 randomly selected participants aged ≥40 years underwent pulmonary function tests between July and October 2021. Cox stepwise regression analysis was used to develop overall and gender-specific risk prediction models, along with the construction of corresponding risk nomograms. Model predictive performance was evaluated using the C-indice, area under the curve (AUC) values, and Brier score. Stability was assessed through 10-fold cross-validation and sensitivity analysis. ResultsA total of 3 019 participants were included, with a median follow-up duration of 4.6 years. The COPD incidence density was 17.22 per 1 000 person-years, significantly higher in males (32.04/1 000 person-years) than that in females (7.38/1 000 person-years) (P<0.001). The overall risk prediction model included the variables such as gender, age, education level, BMI, smoking, passive smoking, and respiratory comorbidities. The male-specific model incorporated the variables such as age, BMI, respiratory comorbidities, and smoking, while the female-specific model included age, marital status, respiratory comorbidities, and pulmonary tuberculosis history. The C-indices for the overall, male-specific, and female-specific models were 0.829, 0.749, and 0.807, respectively. The 5-year AUC values were 0.785, 0.658, and 0.811, with Brier scores of 0.103, 0.176, and 0.059, respectively. Both 10-fold cross-validated C-indices and sensitivity analysis (excluding participants with a follow-up duration of <6 months) yielded C-indices were above 0.740. ConclusionThis study developed concise and practical overall and gender-specific COPD risk prediction models and corresponding nomograms. The models demonstrated robust performance in predicting COPD incidence, providing a valuable reference for identifying high-risk populations and formulating targeted screening and personalized management strategies.

Background and purpose:In recent years,chimeric antigen receptor T(CAR-T)cell therapy has achieved breakthrough progress in cancer treatment.γδ T cells,with their non-major histocompatibility complex(MHC)-restricted antigen recognition,broad antitumor activity,and low risk of graft-versus-host disease(GVHD),have garnered significant interest in CAR-γδ T cell therapy.However,critical challenges including suboptimal in vitro expansion and low viral transduction efficiency severely hinder the research and clinical application of CAR-γδ T cells.This study aimed to establish an efficient platform for preparing CAR-γδ T cells by optimizing the in vitro expansion conditions of γδ T cells and refining lentiviral transduction strategies.Methods:We first optimized the expansion protocol for γδ T cells by screening various cytokine combinations and the concentrations of individual cytokines within combination,and evaluating cell purity,viability,fold expansion,and expressions of cytotoxicity and exhaustion markers to identify the optimal culture conditions.Subsequently,the transduction conditions for CAR-γδ T cells were improved by determining the optimal activation duration of γδ T cells prior to gene transfer,as well as the optimal multiplicity of infection(MOI)for lentiviral transduction.Finally,CAR-γδ T cells were successfully generated using the optimized protocol,and their cytotoxic activity against target cells was validated via calcein-release assay and flow cytometry,with a preliminary assessment of the potential risk of GVHD induction.Results:Experimental data demonstrated that,compared with the interleukin(IL)-2-only culture,the IL-2+IL-7+IL-15 combination significantly enhanced the expansion capacity of γδ T cells(876.50±238.35-fold vs 1 627.50±472.15-fold),cell purity(73.67%±1.53%vs 90.69%±2.00%),and cell viability(63.01%±7.05%vs 89.00%±3.61%).It also increased the expression of the cytotoxicity marker CD16(4.20%±1.73%vs 14.66%±0.58%)and reduced the expression of the exhaustion marker programmed death-1(PD-1)(35.67%±6.26%vs 21.10%±6.49%).A cytokine concentration gradient orthogonal assay further identified 10 ng/mL IL-7 and 10 ng/mL IL-15 as the optimal concentrations within the IL-2+IL-7+IL-15 combination.Gene transduction performed 96-120 h after activation using a multiplicity of infection(MOI)of 5-10 resulted in the highest transduction efficiency for CAR-γδ T cells(96 h:12.87%±4.35%;120 h:11.37%±2.35%).CAR-γδ T cells generated using the optimized system exhibited specific cytotoxic effects against tumor cells expressing the target antigen,and no evidence of GVHD induction was observed.Conclusion:CAR-γδ T cells produced using the IL-2+IL-7(10 ng/mL)+IL-15(10 ng/mL)regimen combined with a 96-120 h activation period prior to transduction using a multiplicity of infection(MOI)of 5-10 significantly outperformed conventional methods in terms of expansion efficiency,cell purity,and transduction efficiency.The synergistic antitumor effects mediated by both natural immune receptors and CAR-specific recognition,along with the initial absence of GVHD risk,provide critical technical support for the clinical translation of CAR-γδ T cell therapy,establishing a solid theoretical and practical foundation.

Background and purpose:Salivary duct carcinoma(SDC)is a group of rare and highly heterogeneous diseases.It predominantly arises in the parotid glands of middle-aged and elderly males,with high rates of recurrence and metastasis,as well as a poor prognosis.Currently,there is a lack of clinical data on SDC.This study aimed to evaluate the clinical characteristics of SDC patients and explore high-risk factors affecting prognosis,so as to provide clinical references for physicians.Methods:Clinical data of patients with primary SDC who were admitted to Tianjin Medical University Cancer Institute and Hospital and Tianjin Stomatological Hospital Affiliated to Nankai University School of Medicine from 2012 to 2024,were collected retrospectively.Inclusion criteria:① patients diagnosed with primary SDC;② availability of American Joint Committee on Cancer(AJCC)staging data.Exclusion criteria:① concurrent other malignant tumors;② incomplete or missing medical records;③ death due to non-SDC causes;④ duplicate cases from the two participating hospitals.Data retrieved encompassed epidemiological information(gender,age)and clinical details(time of diagnosis,tumor characteristics,treatment regimen,recurrence and metastasis status,and pathological data).Survival analysis was performed using the Kaplan-Meier method,and factors related to prognosis were explored through univariate COX proportional hazards regression model analysis.This study was approved by the Ethics Committee of Tianjin Stomatological Hospital Affiliated to Nankai University School of Medicine(ethics number:PH2023-B-016),and patient informed consent was waived.Results:A total of 87 patients with primary SDC were included in this study,among whom 77%were male,69%had primary lesions in the parotid gland,29.9%in the submandibular gland,and one patient had a primary lesion in the minor salivary gland of the nasal cavity.49.3%of the patients had concurrent cervical lymph node metastasis.The median overall survival(OS)of the entire group was 31.2 months,the median progression-free survival(PFS)was 20.3 months,and the 5-year OS rate was 52.6%.The 5-year OS rate for tumors originating from the parotid gland was 60%,which was better than the 32.9%for those originating from the submandibular gland.Among the 85 patients who received surgical treatment,65.9%underwent both resection of the primary tumor and neck dissection.Postoperative radiotherapy was administered to 49 patients.During the follow-up period,46%of the patients developed recurrence or metastasis,with lung and bone metastases being the most common.The median OS and local progression-free time in the postoperative radiotherapy group were significantly longer compared with those in the group without radiotherapy,however,the difference was not statistically significant.Conclusion:SDC is a malignant and aggressive disease that predominantly occurs in the parotid glands of middle-aged and elderly males,with a high rate of lymph node metastasis and poor prognosis.Clinically,it is recommended that patients with SDC undergo radical resection of the primary lesion and cervical lymph node dissection,combined with postoperative adjuvant radiotherapy.Targeted therapy and immunotherapy are worthy of further exploration.

'Injury but not disability, disability but not useless’ is the guiding principle behind the establishment of plastic and reconstructive surgery, making it a unique specialty in modern medicine. Its development relies not only on the optimized use of donor resources but also on technological advancements and innovations in concepts. However, in the face of severe tissue damage caused by burns, trauma, and congenital deformities, traditional repair method often fall short, necessitating more effective solutions. Against this backdrop, progress in donor sourcing and interdisciplinary innovation will inject new vitality into reconstructive surgery. In this context, we take facial transplantation reconstruction as a starting point, reviewing and organizing the technological development, issues, and challenges of autologous and allogeneic facial transplantation reconstruction from the perspective of donors. This exploration aims to shed light on the future direction of this field and gain a deeper understanding of the present and future of donor-driven reconstructive surgery.