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 relationship between health literacy and sports injuries among high school students with athletic specialization, so as to provide theoretical basis for the intervention of sports injury occurrence of sports special students. Methods: A total of 443 high school students with athletic specialization aged 16-18 years old from 21 urban and rural areas in Shangrao City, Ganzhou City and Nanchang City of Jiangxi Province were selected by convenient sampling method from March to April 2023, and the health literacy and sports injury incidence were investigated. The assessment of sports injury was based on the Monitoring Method of Child and Adolescent Injury, and the assessment of health literacy level was conducted using the Interactive Health Literacy Questionnaire of Chinese Adolescents. χ 2 test was used to compare the reported rate of sports injuries among the demographic and sports training groups and the correlation between health literacy and sports injuries. Binary Logistic regression analysis was used to infer the correlation between the health literacy level of high school students with athletic specialization and the occurrence of sports injuries. Results: The prevalence of sports injury was 49.4% in high school students with athletic specialization. Binary Logistic regression analysis showed that the risk of sports injury of high school students with athletic specialization with medium and low level of health literacy was significantly increased compared with high level (medium level: OR = 1.98,95% CI =1.12-3.51; low level: OR =2.08, 95% CI =1.18-3.68), high school students with athletic specialization in low level of health awareness of sports injury risk was higher than other dimensions of health literacy level (middle level: OR =2.77, 95% CI =1.48-5.19; low level: OR =2.01, 95% CI =1.04-3.88)( P <0.05). The results of stratified analysis showed that among male students with athletic specialization and students with track and field, students with moderate/low overall health literacy had a higher risk of sports injuries compared to high level students (male students: low level, OR =2.46, 95% CI =1.15-5.28; track and field: intermediate level, OR =2.17, 95% CI =1.06-4.43, low level, OR =2.09, 95% CI =1.02-4.30; P <0.05). Conclusions There is a correlation between the health literacy level of high school students with athletic specialization and the occurrence of sports injuries. Students health awareness should be improved to reduce the risk of sports injuries.

Cinnamomi Ramulus and Alismatis Rhizoma are commonly used in the treatment of external-contraction diseases. Cinnamomi Ramulus is pungent， sweet， and warm， with the effects of ventilating lung and dispersing cold， warming Yang and transforming Qi， and promoting water and liquid flow from Taiyang meridian to remove dampness. Alismatis Rhizoma is sweet and cold， with the effects of draining dampness and promoting urination， regulating the waterway， removing water retention in lung， and promoting urination to remove dampness and heat. Herbal pairs are extracted from the accumulated experience of medical practitioners over the ages in the use of medicines and have been proved by clinical application to be composed of simple and effective combinations for specific diseases. The herb pair Cinnamomi Ramulus-Alismatis Rhizoma is an important part in ancient classic formulas such as Wulingsan. Both herbs play a role in draining dampness and promoting urination， warming Yang and transforming Qi， being a representative herb pair used for treating external-contraction exterior syndrome and water retention inside. The review of ancient medical publications revealed that there were a large number of compound formulas containing Cinnamomi Ramulus-Alismatis Rhizoma for dispersing cold and removing dampness， which were widely used for thousands of years in clinical practice. Modern pharmacological studies have shown that the active pharmacological components of Cinnamomi Ramulus （cinnamaldehyde and cinnamic acid） and Alismatis Rhizoma （alisol A and 23-acetate alisol B） have anti-inflammatory， antiviral， and immunoregulatory effects and high safety. Qingfei Paidu decoction containing this herbal pair played an important role in fighting against COVID-19. Despite the extensive pharmacological studies on Cinnamomi Ramulus and Alismatis Rhizoma， few studies have been carried out regarding this herb pair. This paper summarizes the traditional Chinese medicine knowledge about Cinnamomi Ramulus and Alismatis Rhizoma in the treatment of external-contraction diseases and summarizes the chemical composition， pharmacological effects， toxicology and other aspects of the two herbs before and after compatibility， aiming to provide a reference for further research and clinical application.

According to the codon characteristics of the prokaryotic system,we synthesized the optimized Rv2628c-Rv1737c nucleic acid sequence and constructed the expression plasmid pET24a-Rv2628c-Rv1737c.After sequencing,expression in E.coli was induced.The fusion protein was found in inclusion bodies.After renaturation and affinity chromatography purifi-cation,the Rv2628c-Rv1737c recombinant protein,with a molecular weight of 57 kDa and a purity exceeding 90％,was ob-tained.The recombinant protein was used to stimulate PBMC cells from different patients,and the differences in IFN-γ mRNA expression were analyzed with q-PCR.The Rv2628c-Rv1737c fusion protein stimulated patients with TB,particularly latent TB infection(LTBI).The level of IFN-γ mRNA in PBMC cells was higher than that in healthy controls(P＜0.05).Mice immu-nized with BCG+Rv2628c-Rv1737c/DMT showed significant induction of high levels of IgG antibodies.Rv2628c-Rv1737c re-combinant protein,as a latent infection antigen,is recognized by PBMCs cells infected with TB,with strong immunogenicity,and thus may serve as a potential TB subunit vaccine target antigen.This protein may be used to prevent TB infection,particu-larly latent infection,and to perform laboratory diagnosis.

A rapid quantitative immunochromatographic assay for procalcitonin(PCT)using metal-organic frameworks modified with gold and platinum nanoparticles(MAPs)as labels was established in this work.The detection probe was prepared by conjugating MAPs with anti-PCT monoclonal antibody via an electrostatic adsorption method.Anti-PCT polyclonal antibody and sheep anti-mouse IgG were sprayed onto the nitrocellulose(NC)membrane as the test line and quality control line,respectively,to construct immunochromatographic strip for PCT quantitative detection via signal-amplification-based sandwich immunoassay.The results showed that the MAP-based immunochromatographic test had high sensitivity,high specificity,and good stability.The dynamic range for detection of PCT was 0.61 pg/mL-320 ng/mL,the detection limit was 0.25 pg/mL,and the intra-day and inter-day precision(Relative standard deviation)were less than 15%.The results of real sample analysis showed that a quite low volume of sample was required for detection of PCT in whole blood,which was of great significance for the early diagnosis,monitoring and treatment,and prognosis of inflammation.

Objective:To explore the compatibility and medication law of Cinnamomi Ramulus-Alismatis Rhizoma medicinal pair in ancient and modern prescriptions.Methods:Ancient prescriptions and Chinese patent medicines containing Cinnamomi Ramulus-Alismatis Rhizoma medicinal pair were retrieved from the database of ancient classic famous prescriptions 1.0 and the database of listed Chinese patent medicines 1.0 developed by the Institute of Information on Traditional Chinese Medicine, China Academy of Chinese Medical Sciences. Excel 2019 was used to establish a database. The ancient and modern medical record cloud platform V2.3.5 and SPSS Modeler 18.0 software were used to perform frequency statistics, association rule analysis, clustering analysis, etc. on the data.Results:Totally 79 ancient articles with Cinnamomi Ramulus-Alismatis Rhizoma medicinal pair were obtained, including 76 ancient prescriptions, involving 250 kinds of Chinese materia medica; 25 kinds of Chinese patent medicine were obtained, involving 186 kinds of Chinese materia medica. The drug properties of ancient prescriptions and modern TCM patent medicines were both mainly warm, cold and neutral. The main tastes of ancient prescriptions and modern Chinese patent medicines were pungent, sweet and bitter. And the drugs mainly belong to spleen, lung, liver and kidney meridians. Correlation analysis suggested the same high-frequency association compatibility of ancient and modern prescriptions, Poria-Cinnamomi Ramulus-Alismatis Rhizoma, Atractylodis Rhizoma-Cinnamomi Ramulus-Alismatis Rhizoma, Atractylodis Macrocephalae Rhizoma-Cinnamomi Ramulus-Alismatis Rhizoma. Both clinical symptoms and diseases associated with medicinal compatibility of ancient prescriptions were intestinal flora, edema and vomiting. The syndrome types included bladder impoundment, dampness trapped in the guardian surface, internal retention of phlegm and morbid fluid. The clinical symptoms associated with medicinal compatibility of modern TCM patent medicine were limb joints pain and edema. The diseases included rheumatic arthritis (RA) and kidney disease. The syndrome types included wind-cold-dampness RA, stagnation of collaterals and kidney yang deficiency. High frequency drug clustering yielded 4 clustered squares.Conclusion:The core indications treated by Cinnamomi Ramulus-Alismatis Rhizoma are exogenous diseases with dampness caused by syndrome types including internal storage of water-dampness, cold-dampness obstruction and so on, which can provide reference for further in-depth research and guidance on clinical medication.

Tinnitus is a clinically refractory disease with a high incidence.LAI's Tongyuan acupuncture method believes that tinnitus is nothing more than the two ends of deficiency and excess.The deficiency is closely related to original spirit,and the excess is related to the pathological factors such as externally-contracted six pathogenic factors,phlegm-damp and blood stasis,and qi stagnation and yang constraint.The Tongyuan acupuncture method is based on original spirit,takes the two vessels of conception vessel(CV)and governor vessel(GV)as the general outline,takes the regulation of yin and yang as the main method,pays attention to the combination of local and overall,and emphasizes the application of tonification and purgation,and has the clinical effect of reinforcing healthy qi and dispelling pathogen,cultivating the vital essence.In clinical application,the method of'unblocking governor vessel and nourishing spirit'is used to play the role of heart and brain nourishing spirit and warming and supporting yang qi.The method of'conducting qi back to its source'has the effect of cultivating the vital essence and regulating qi movement.The treatments should be cooperated according to the syndromes,so that the pathogens can be expelled and healthy qi can be settled.Qi and blood have origins of generation and transformation and normally nourishing in the ear,so as to effectively alleviate the patient's tinnitus symptoms.This paper summarizes and analyzes Professor LAI Xin-Sheng's Experience in the diagnosis and treatment of tinnitus by Tongyuan acupuncture method,and provides a new acupuncture treatment plan for clinical application.

Objective To explore the diagnostic efficacy of serum 14-3-3β protein combined with fractional exhaled nitric oxide(FeNO)and conventional ventilatory lung function parameters in diagnosing bronchial asthma(referred to as"asthma")in children.Methods A prospective study included 136 children initially diagnosed with asthma during an acute episode as the asthma group,and 85 healthy children undergoing routine health checks as the control group.The study compared the differences in serum 14-3-3β protein concentrations between the two groups,analyzed the correlation of serum 14-3-3β protein with clinical indices,and evaluated the diagnostic efficacy of combining 14-3-3β protein,FeNO,and conventional ventilatory lung function parameters for asthma in children.Results The concentration of serum 14-3-3β protein was higher in the asthma group than in the control group(P<0.001).Serum 14-3-3β protein showed a positive correlation with the percentage of neutrophils and total serum immunoglobulin E,and a negative correlation with conventional ventilatory lung function parameters(P<0.05).Cross-validation of combined indices showed that the combination of 14-3-3β protein,FeNO,and the percentage of predicted value of forced expiratory flow at 75%of lung volume had an area under the curve of 0.948 for predicting asthma,with a sensitivity and specificity of 88.9%and 93.7%,respectively,demonstrating good diagnostic efficacy(P<0.001).The model had the best extrapolation.Conclusions The combination of serum 14-3-3β protein,FeNO,and the percentage of predicted value of forced expiratory flow at 75%of lung volume can significantly improve the diagnostic efficacy for asthma in children.

Objective To observe and compare the changes of vertebral bone mineral density(BMD)in the early stages of spinal Mycobacterium tuberculosis infection.Methods Patients who underwent spinal surgery at Xiangya Hospital,Central South University from January 1 to December 31,2023 were continuously enrolled(spinal tuber-culosis group),based on gender matching,non-spinal tuberculosis surgical patients treated for spinal stenosis were selected as the control group.Dual-energy X-ray scans were performed on the enrolled patients,difference in verte-bral BMD between two groups of patients was compared.An animal model of spinal Mycobacterium tuberculosis in-fection(referred to as the animal model)was constructed,differences in microstructure of trabecular bone between spinal tuberculosis group and control group was compared,and the bone volume/tissue volume(BV/TV),the thickness of trabecular bone(Tb.Th),the number of trabecular bone(Tb.N),and sparse density of trabecular(Tb.Sp)were used as evaluation indexes to further analyze the bone quality differences between the diseased verte-brae and the neighboring vertebrae.Results 69 patients were included in the spinal tuberculosis group and the con-trol group,respectively.The BMD of patients in the spinal tuberculosis group(0.793[0.712,0.869]g/cm2)was lower than that of the control group(0.907[0.800,1.020]g/cm2),difference was statistically significant(P＜0.05).Microstructure of trabecular bone BV/TV([18.4±5.4]％),Tb.Th([0.124±0.010]mm)in the spinal tuberculosis group of animal model were significantly altered compared with BV/TV([22.6±3.2]％),Tb.Th([0.160±0.017]mm)in the control group(both P＜0.05).In the spinal tuberculosis group,microstructure of diseased vetebral trabecular bone BV/TV([25.5±6.7]％)and Tb.N([1.871±0.443]/mm)were significantly lower than BV/TV([26.6±6.8]％)and Tb.N([1.969±0.454]/mm)in the neighboring vertebrae,both with statistically difference(both P＜0.05).Conclusion In the early stages of spinal Mycobacterium tuberculosis infec-tion,microstructure of vertebral trabecular bone can be altered,leading to a decrease in BMD.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.