Objective To establish a method for acquisition, perfusion, preservation and transportation of the genetically modified pig kidneys. Methods An eight genetically modified pig was utilized as experimental subject. Prior to kidneys procurement, the health status of the pig was assessed through hematology examination, and the vascular structure of the kidneys was examined using imaging techniques. Following kidneys acquisition, the pig kidneys were perfused and subsequently packaged into the cryogenic storage container labeled "For Organ Transportation Only" for interprovincial transport after communicating the transportation process with transportation department. To evaluate pathological damage to the pig kidneys, a serious of methods were employed such as hematoxylin-eosin (HE) staining, real-time fluorescent quantitative polymerase chain reaction (RT-qPCR), terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) fluorescence staining and enzyme-linked immune absorbent assay (ELISA). Results The preoperative examination of the eight genetically modified pig showed that the serum creatinine was 73.2 μmol/L, blood urea nitrogen was 2.8 mmol/L and hemoglobin was 116 g/L, all within the normal range, indicating normal renal function. CT angiography revealed no lesions in the pig kidneys, and no dilation, stenosis or premature branching of the blood vessels. The total time of obtaining the left and right kidneys from the eight genetically modified pig was (125 ± 10) min, with a blood loss of (20 ± 2) mL. The warm ischemia times were 3 min and 7 min, respectively. The perfusion and trimming times of the left and right kidneys were 36 min and 41 min, respectively. After perfusion, both kidneys were white and moist. The cold preservation and transportation time was 8 h. HE staining showed that some glomeruli were shrunk, and the lumens of the surrounding renal tubules were slightly depressed and swollen with partial inner membrane shedding and microvacuoles formed when the kidneys were preserved for 8 h. The level of cysteinyl aspartate-specific proteinase-3 messenger RNA in the kidneys tissue gradually increased with the extension of cold preservation time after 2 h (P<0.05). TUNEL fluorescence staining showed that only a small number of cells underwent apoptosis after 8 h of cold preservation, which was not significantly different from that at 0 h (P>0.05). ELISA results showed that the contents of lactate dehydrogenase (LDH) and creatinine in the preservation solution remained relatively stable, but the content of kidney injury molecule 1 (KIM-1) gradually increased with the extension of preservation time, suggesting that the pig kidneys had mild injury. Conclusions By establishing methods for acquisition, perfusion, preservation and transportation of the kidneys from genetically modified donor pig, it is possible to effectively and reliably use genetically modified pig kidneys for xenotransplantation.

Objective: To investigate the objective characteristics of tongue manifestation in different stages of damp-heat syndrome in diabetic kidney disease (DKD). Methods: A cross-sectional study enrolled 134 patients with DKD G3-5 stages who met the diagnostic criteria for damp-heat syndrome in DKD. The patients were treated at Dongzhimen Hospital, Beijing University of Chinese Medicine, from May 2023 to January 2024. The patients were divided into three groups: DKD G3, DKD G4, and DKD G5 stage, with 53, 33, and 48 patients in each group, respectively. Clinical general data (gender, age, and body mass index) and damp-heat syndrome scores were collected from the patients. The YZAI-02 traditional Chinese medicine (TCM) AI Tongue Image Acquisition Device was used to capture tongue images from these patients. The accompanying AI Open Platform for TCM Tongue Diagnosis of the device was used to analyze and extract tongue manifestation features, including objective data on tongue color, tongue quality, coating color, and coating texture. Clinical data and objective tongue manifestation characteristics were compared among patients with DKD G3-5 based on their DKD damp-heat syndrome status. Results: No statistically significant difference in gender or body mass index was observed among the three patient groups. The DKD G3 stage group had the highest age (P<0.05). The DKD G3 stage group had a lower score for symptoms of poor appetite and anorexia(P<0.05) than the DKD G5 group. No statistically significant difference was observed in damp-heat syndrome scores among the three groups. Compared with the DKD G5 stage group, the DKD G3 stage group showed a decreased proportion of pale color at the tip and edges of the tongue (P<0.05). The DKD G4 stage group exhibited an increased proportion of crimson at the root of the tongue, a decreased proportion of thick white tongue coating at the root, a decreased proportion of pale color at the tip and edges of the tongue, an increased hue value (indicating color tone) of the tongue color in the middle, an increased brightness value (indicating color lightness) of the tongue coating color in the middle, and an increased thickness of the tongue coating (P<0.05). No statistically significant difference was observed in other tongue color proportions, color chroma values, body characteristics, coating color proportions, coating color chroma values, and coating texture characteristics among the three groups. Conclusion Tongue features differ in different stages of DKD damp-heat syndrome in multiple dimensions, enabling the inference that during the DKD G5 stage, the degree of qi and blood deficiency in the kidneys, heart, lungs, liver, gallbladder, spleen, and stomach is prominent. Dampness is more likely to accumulate in the lower jiao, particularly in the kidneys, whereas heat evil in the spleen and stomach is the most severe. These insights provide novel ideas for the clinical treatment of DKD.

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 changes of OAT expression in bone cells in chronic renal failure(CRF)and involved mechanism,and to explore the effect of OAT expression on bone metabolism.Methods Ra-ndomly divide the rats into a control group(n=6)and a model group(n=6).The model group established a rat chronic renal failure model using"single nephrectomy+adenine gavage"method,and the red blood cell(RBC)and hemoglobin(HGB)of the rat body were measured using a blood routine analyzer;Measure indicators such as creatinine(Cr),urea nitrogen(BUN),uric acid(UA),blood calcium(Ca2+),and blood phosphorus(P3+)using a fully automated biochemical analyzer;Pathological examination of rat kidneys;X-ray examination of rat tibia;Immunohistochemical examination of bone tissue OAT1 level.Results The bone density of the model group rats is lower than that of the control group;The calcium and phosphorus metabolism of rats in the model group was in metabolic disorder,and the OAT1 value of bone tissue binding was much lower than that of the control group(P= 0.0018),which was statistically significant.(P=0.0018)Conclusion Chronic renal failure affected the binding ability of OAT1 in bone tissue,leading to the metabolic disorder for calcium absorption and phosphorus metabolism,thus aggravating renal osteodystrophy(P<0.05).

Objective:To investigate the application value of modified multivisceral trans-plantation (MMT) in chronic intestinal pseudo-obstruction (CIPO) secondary to autoimmune enteril leiomyositis (AEL).Methods:The retrospective and descriptive study was conducted. The clinico-pathological data of a recipient who was admitted to Beijing Tsinghua Changgung Hospital Affiliated to Tsinghua University on February 2022 and underwent MTT for CIPO secondary to AEL were collected. The recipient was a male, aged 29 years old. Results of preoperative histopathological examination showed that there were muscle plexus and ganglion cells in the rectum, sigmoid colon, ascending colon, intrinsic muscle layer of ileum, and a small amount of submucosal layer. There was also a small amount of chronic inflammatory cell infiltration in the muscle, indicating a high possi-bility of diagnosis of neurogenic CIPO.Results:(1) Surgical situations. The operation time was 14 hours and 30 minutes, and the cold ischemia time was 9 hours and 30 minutes. The intra-operative blood product dosage included 14 U of red blood cells, 1 400 mL of fresh frozen plasma, and two therapeutic doses of platelets. (2) Postoperative histopathological examination. Results of postoperative histopathological examination showed chronic inflammation and local erosion of the small intestine and duodenal mucosa, with scattered disappearance of the focal mucosal muscle layer; There is a large infiltration of CD3 + and CD8 + lymphocytes in the lamina propria, especially in the muscularis propria. In severe lesions, there is infiltration of ribbon lymphocytes in the subserosal and muscular layers; Muscle fiber degeneration, reduction, and fibrosis. Deposition of pigment granules in the cytoplasm of smooth muscle cells; No abnormalities were found in the intermuscular, submu-cosal ganglia, and Cajal cells; Fibrosis of the serosal layer with local cellulose exudation; Chronic inflammation of the colonic mucosa, scattered and focal lymphocyte infiltration in the local muscle layer, and myositis related changes. Pathological diagnosis was secondary CIPO induced by AEL. (3) Postoperative immune rejection, recurrence and treatment. Results of colonoscopy and histopatholo-gical examination at postoperative 8 days showed acute cellular rejection. The cell count of reci-pient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 27.00×10 3, 373.00×10 3, 179.00×10 3 and 142.00×10 3 cell/mL, respectively. Anti-immune rejection treatment was performed using tacrolimus, rabbit anti-human thymocyte immunoglobulin, methylprednisolone mycophenolate mofetil, and monoclonal antibodies against basil. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes at postoperative 57 days were 0.72×10 3, 239.59×10 3, 89.28×10 3 and 91.53×10 3 cell/mL, respectively. Results of colonoscopy and histopathological examination at postoperative 79 days showed the recurrence of AEL. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lymphocytes, and CD8 + lymphocytes were 0.32×10 3, 264.92×10 3, 46.95×10 3 and 169.54×10 3 cell/mL, respectively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examina-tion at postoperative 89 days showed AEL recurrence without remission. The cell count of recipient′s B lymphocytes, CD3 + lymphocytes, CD4 + lympho-cytes, and CD8 + lymphocytes were 0.28×10 3, 187.00×10 3, 55.52×10 3 and 92.45×10 3 cell/mL, respec-tively. The tacrolimus and methylprednisolone were used for treatment. Results of colonoscopy and histopathological examination at postoperative 92 days showed the intestinal mucosa had returned to a normal state. (4) Postoperative oral feeding time and time to get rid of parenteral nutrition. The recipient began oral feeding at postoperative 28 days and eliminated parenteral nutrition at postoperative 35 days. (5) Follow-up. The recipient was discharged 114 days after surgery and as of the follow-up deadline, the graft function was good. The recipient maintained a low-fat, high sugar, and high protein diet, completely consumed orally, with a body mass index of 22 kg/m 2, and has returned to normal work. Conclusion:MMT can be used for the treatment of CIPO secondary to AEL.

Objective To investigate the awareness of the Radiation Shielding Requirements for Radiotherapy Room–Part 2: Radiotherapy Room of Electron Linear Accelerators (GBZ/T 201.2—2011) among relevant practitioners in medical institutions as well as its implementation and application situation and collect relevant problems and suggestions for an evaluation of the scientificalness, standardization, and timeliness of the standard, and to provide a scientific basis for the further revision and implementation of the standard. Methods An online questionnaire survey was conducted among relevant employees in medical institutions providing medical linear accelerator radiotherapy across 22 provinces of China, which investigated the awareness, training, application, and revision suggestions related to GBZ/T 201.2—2011. The questionnaires were collected and analyzed. Results A total of 340 relevant practitioners filled out the questionnaire. Of the participants, 66.80% were physicists; 79.11% had an awareness of the standard; 56.18% ever participated in the standard-related training; but the survey results showed that the practitioners did not have a good knowledge of the standard’s content, and the training and promotion were not enough; 83.24% thought that the standard had been widely used; 17.60% thought that the standard needed to be revised; 76.76% thought that there was a need to add calculation examples; 88.82% thought that neutron shielding needed to be considered for the 10 MV X-ray accelerator room. Conclusion The standard has been widely known in the field of radiotherapy protection. With the development of radiotherapy technology, the standard should be revised to add calculation examples and consider neutron shielding in the 10 MV X-ray accelerator room. The standard is highly technical and difficult to grasp, so the promotion and implementation goals should be appropriate for different personnel groups, the training for employees at key posts should be strengthened, and the methods recommended in the standard should be uniformly used throughout the country.

Syncope is a common emergency in children and adolescents, which has serious influence on the quality of life.After nearly 30 years′ exploration, great progress has been made in the diagnosis and treatment of children′s syncope in China.We have established the diagnosis procedure of syncope in line with health economics.We have also proposed the concept of individualized precise treatment of syncope in children for the first time.The disease spectrum of syncope in children has been continuously enriched.Relevant progresses have been cited by the guidelines of the United States, Europe and Canada, which has also been unanimously recognized by international experts.

To develop a quality control checklist for the prevention and control of coronavirus disease 2019 （COVID-19） in fever clinic and isolation ward of the general hospital and to assess its application. Based on the relevant prevention and control plans and technical guidelines for COVID-19，Delphi method was used to identity items for evaluation，and a quality control checklist for the prevention and control of COVID-19 in the fever clinic and isolation ward was developed in Sir Run Run Shaw Hospital. The checklists included 8 dimensions and 32 items for fever clinic，7 dimensions and 27 items for the isolation ward. The appointed inspectors conducted daily quality control for each shift with this checklist. The expert authority coefficient was 0.88，the mean of the importance of each index in the quality control table was not less than 4.8，and the coefficient of variation was not more than 0.07. During the entire February 2020，8 problems were found and rectified on-the-spot with the application of the checklist. Quality inspection rate was 100% in both isolation wards and fever clinic. The compliance rate and accuracy rate of hand hygiene were 100%; the correct rate of wearing and removing protective equipment increased from 96% to 100%. During the same period，a total of 1915 patients were admitted to the fever clinic，including 191 suspected patients （all were isolated in the hospital，3 were confirmed）. There were no medical staff infected with COVID-19，no cross infection of patients and their families in the hospital. A quality control checklist for the prevention and control of COVID-19 has been developed and applied in the isolation wards and fever clinic，which plays an important role in preventing nosocomial infection.
COVID-19 ; Checklist ; Fever ; Hospitals, General ; Humans ; SARS-CoV-2

Objective:To explore the application value of dynamic monitoring of serum soluble interleukin-2 receptor (sIL-2R) and stromal cell-derived factor 1 α (SDF-1α) in patients with acute leukemia (AL).Methods:A total of 187 patients with AL admitted to the Second Hospital of Shanxi Medical University from March 2017 to August 2018 were selected and 90 healthy subjects at the same period were selected as the healthy controls. The levels of serum sIL-2R and SDF-1α were detected at the initial diagnosis, remission phase and relapse phase, respectively. The clinical value of dynamic monitoring of sIL-2R and SDF-1α was analyzed.Results:Among 187 patients with AL, 135 patients (72.19%) had complete remission (CR) after chemotherapy, 52 patients (27.81%) had partial remission, and 43 patients (31.85%) relapsed. The level of serum sIL-2R at the AL initial diagnosis period was (533±32) U/ml, which was higher than that in the healthy controls [(247±30) U/ml], and the difference was statistically significant ( t = 71.976, P < 0.01); the level of serum SDF-1α at the AL initial diagnosis was (2 968±305) pg/ml, which was higher than that in the healthy controls [(1 358±160) pg/ml], and the difference was statistically significant ( t = 47.043, P < 0.01). The levels of serum sIL-2R [(308±30) U/ml] and SDF-1α [(1 576±184) pg/ml] in the CR phase were lower than those in the initial diagnosis of patients with AL; and the levels of sIL-2R [(599±36) U/ml] and SDF-1α [(2 894±301) pg/ml] in the relapse phase were higher than those in the CR phase of patients with AL (all P < 0.01). Conclusions:The levels of serum sIL-2R and SDF-1α in patients with AL are increased, and there are big differences in the levels of sIL-2R and SDF-1α at the initial diagnosis, remission phase, and relapse phase. Dynamic monitoring of both can provide the data support for early clinical intervention.