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.

ObjectiveAccording to the theory of medicinal properties in traditional Chinese medicine （TCM）， Tangye Jingfa Tu， implying that all tonic prescriptions should primarily feature sweetness. However， the "Chart of the Classic Methods of Decoction Therapy" recorded in Mnemonic Aids for Medical Practice （Fuxingjue） proposes that each of the Zangfu organs has a corresponding tonifying flavor， i.e.， pungent for the liver， salty for the heart， sweet for the spleen， sour for the lungs， and bitter for the kidneys. Therefore， deficiency syndromes of different organs should be primarily addressed with specific medicinal flavors. This study applies this theoretical framework to analyze the formulation and compatibility principles and efficacy positioning of commonly used TCM tonic prescriptions， providing a reference for differentiated clinical medication. MethodsA database of tonic prescriptions was established based on the textbook Science of Prescriptions. Excel software was used to separately calculate the number of medicinal types， frequency of use， flavors， and proportional composition of the prescriptions. The prescriptions were categorized to determine their compatibility structures and functional characteristics. ResultsA total of 60 prescriptions were included， classified into six categories， involving 110 medicinal types with 469 instances of use. From the perspective of the "Chart of the Classic Methods of Decoction Therapy"， different tonic prescriptions exhibit distinct dominant medicinal flavors and organ associations. Specifically， 15 Qi-tonifying prescriptions primarily adopted a "sweet flavor" to tonify the spleen. Nine blood-tonifying prescriptions primarily adopted a "pungent-salty flavor" to tonify the liver and heart. Seven Qi-and-blood-tonifying prescriptions primarily featured a "sweet-pungent flavor" to tonify the spleen and liver. Nineteen Yin-tonifying prescriptions primarily adopted a "bitter-sour flavor" to tonify the kidneys and lungs. Seven Yang-tonifying prescriptions primarily featured a "pungent-bitter flavor" to tonify the liver and kidneys. Three Yin-and-Yang-tonifying prescriptions primarily featured a "bitter-pungent-sweet flavor" to tonify the kidneys， liver， and spleen. ConclusionThe "Chart of the Classic Methods of Decoction Therapy" clearly illustrates the formulation and compatibility principles and key differences among various tonic prescriptions， indicating that not all tonic prescriptions are predominantly sweet in flavor. This provides new insights for the clinical modification and application of tonic prescriptions.

ObjectiveAccording to the theory of medicinal properties in traditional Chinese medicine （TCM）， Tangye Jingfa Tu， implying that all tonic prescriptions should primarily feature sweetness. However， the "Chart of the Classic Methods of Decoction Therapy" recorded in Mnemonic Aids for Medical Practice （Fuxingjue） proposes that each of the Zangfu organs has a corresponding tonifying flavor， i.e.， pungent for the liver， salty for the heart， sweet for the spleen， sour for the lungs， and bitter for the kidneys. Therefore， deficiency syndromes of different organs should be primarily addressed with specific medicinal flavors. This study applies this theoretical framework to analyze the formulation and compatibility principles and efficacy positioning of commonly used TCM tonic prescriptions， providing a reference for differentiated clinical medication. MethodsA database of tonic prescriptions was established based on the textbook Science of Prescriptions. Excel software was used to separately calculate the number of medicinal types， frequency of use， flavors， and proportional composition of the prescriptions. The prescriptions were categorized to determine their compatibility structures and functional characteristics. ResultsA total of 60 prescriptions were included， classified into six categories， involving 110 medicinal types with 469 instances of use. From the perspective of the "Chart of the Classic Methods of Decoction Therapy"， different tonic prescriptions exhibit distinct dominant medicinal flavors and organ associations. Specifically， 15 Qi-tonifying prescriptions primarily adopted a "sweet flavor" to tonify the spleen. Nine blood-tonifying prescriptions primarily adopted a "pungent-salty flavor" to tonify the liver and heart. Seven Qi-and-blood-tonifying prescriptions primarily featured a "sweet-pungent flavor" to tonify the spleen and liver. Nineteen Yin-tonifying prescriptions primarily adopted a "bitter-sour flavor" to tonify the kidneys and lungs. Seven Yang-tonifying prescriptions primarily featured a "pungent-bitter flavor" to tonify the liver and kidneys. Three Yin-and-Yang-tonifying prescriptions primarily featured a "bitter-pungent-sweet flavor" to tonify the kidneys， liver， and spleen. ConclusionThe "Chart of the Classic Methods of Decoction Therapy" clearly illustrates the formulation and compatibility principles and key differences among various tonic prescriptions， indicating that not all tonic prescriptions are predominantly sweet in flavor. This provides new insights for the clinical modification and application of tonic prescriptions.

Objective Through the safety evaluation of Chinese patent medicines containing cinnabar,the correlation between the evaluation score and the content of drug instructions was analyzed.And the methods and suggestions for improving the label quality of Chinese patent medicines were proposed.Methods Establish a database of Chinese patent medicines and sim-ple preparations containing cinnabar that meet the screening criteria in the 2020 edition of the Chinese Pharmacopoeia,and quan-titatively score such Chinese patent medicines and simple preparations from the two first level dimensions of safety and toxicity characteristics of Chinese patent medicines and simple preparations,and at the same time,grade the level of safety evidence.Results From the perspective of safety evaluation dimension,Chinese patent medicines and simple preparations with relatively high safety score(safety score ≥ 5 points)has a certain correlation with the content of adverse drug reactions clearly reported in the instructions of this kind of medicine,and 54.55％of the evidence risk of this kind of Chinese patent medicines and simple preparations is controllable,with good safety.From the perspective of the evaluation dimension of toxicity characteristics of Chi-nese patent medicines and simple preparations,the types of toxic ingredients in the compatibility of prescriptions and the scores of daily maximum dosage have certain guiding significance for clinical medication precautions.Conclusion For improving the ad-verse reaction information in the instructions of Chinese patent medicines and simple preparations containing cinnabar,and remin-ding the special population of drug use restrictions,it not only has an important impact on drug safety evaluation and grading,but also provides warning and reference for clinical and patient safety in drug use.

AIM:To investigate the effect of cryptotanshinone(CPT)on myocardial hypertrophy induced by isoprenaline(ISO)in rats and explore its potential mechanism.METHODS:The experimental design consisted of two parts.The first aimed to investigate the effects of CPT on cardiac function,pathological manifestations,and the Janus ki-nase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in rats with myocardial hyper-trophy.The rats were divided into six groups,namely the control,CPT control,and model groups and low-(15 mg·kg-1·d-1),medium-(30 mg·kg-1·d-1),and high-dose(60 mg·kg-1·d-1)CPT treatment groups,with six rats per group.The sec-ond part aimed to validate the role of the JAK2/STAT3 signaling pathway in the CPT-mediated myocardial hypertrophy treatment.Rats were divided into four groups,namely the control,model,high-dose CPT treatment,and coumermycin A1(CA1,a JAK2/STAT3 agonist)intervention(rats received ISO injection followed by 60 mg·kg-1·d-1 of high-dose CPT and 1 mg·kg-1·d-1 of CA1 for 15 d)groups,with five rats per group.Myocardial hypertrophy was induced in rats via intra-peritoneal injection of ISO(5 mg/kg),and CPT intervention lasted for 15 days.Cardiac function-related parameters were assessed using echocardiography,and pathological changes were evaluated through hematoxylin-eosin,Masson,and wheat germ agglutinin staining.Protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and JAK2/STAT3 signaling pathway-related proteins were detected by Western blot analysis.RESULTS:Compared with the model group,CPT administration improved cardiac dysfunction-related ul-trasound markers and significantly reduced ISO-induced cardiomyocyte hypertrophy and myocardial fibrosis in rats with hy-pertrophy in a dose-dependent manner(P<0.05).Additionally,CPT decreased the levels of ANP,BNP,and β-MHC in-duced by ISO modeling(P<0.05),and inhibiting the phosphorylation of JAK2 and STAT3(P<0.05).Furthermore,a partial reversal of the therapeutic effect on myocardial hypertrophy induced by ISO modeling was observed when CA1 was administered(P<0.05).CONCLUSION:The CPT exhibits potential as a therapeutic agent for cardiac hypertrophy by effectively attenuating ISO-induced cardiac hypertrophy in rats through JAK2/STAT3 signaling inhibition.

OBJECTIVE To explore the differences in the clinical laboratory test indexes between the patients with two different TCM syndrome types of eczema(dampness-heat infiltration type and non-dampness-heat infiltration type)and observe the distribution and drug resistance rate of pathogens isolated from the skin lesions so as to pro-vide bases for syndrome differentiation and reasonable use of antibiotics.METHODS A total of 180 patients with eczema who were positive for bacterial culture of skin secretions and were treated in Beijing Hospital of Traditional Chinese Medicine from Jan.2021 to Dec.2023 were enrolled in the study and were divided into the dampness-heat infiltration group with 134 cases and the non-dampness-heat infiltration group with 46 cases.The data of clini-cal laboratory test indexes were collected from the two groups of patients,the secretion specimens were sampled from the skin lesion sites,the isolated pathogens were identified by VITEK 2 Compact automatic microorganism analysis system,and the drug susceptibility testing was performed.RESULTS The direct bilirubin level of the dampness-heat infiltration group was(3.99±1.62)umol/L,higher than(3.46±1.12)umol/L of the non-damp-ness-heat infiltration group(P＜0.05),but both were in the normal range;there were no significant differences in other test indexes between the two groups.Totally 180 strains of pathogens were isolated from the 180 patients with eczema,156(86.67％)of which were gram-positive bacteria;Staphylococcus aureus(98 strains),Staphy-lococcus epidermidis(29 strains)and Staphylococcus haemolyticus(15 strains)were the predominant species of the gram-positive bacteria.The gram-negative bacteria accounted for 12.22％(22 strains).The drug resistance rate of the S.aureus strains to penicillin was up to 82.65％,and the isolation rate of methicillin-resistant Staphy-lococcus aureus(MRSA)was 12.24％(12/98);the drug resistance rates of the S.epidermidis strains to penicil-lin and erythromycin were 75.86％,and the drug resistance rate of the S.haemolyticus strains to erythromycin was 100.00％.CONCLUSIONS The gram-positive bacteria(dominated by the S.aureus)are dominant among the pathogens isolated from the skin lesion specimens of the eczema patients and are highly resistant to penicillin and erythromycin.The isolation rate of MRSA is relatuvely low.There is limited clinical significant difference in the direct bilirubin between the two groups.It is necessary to further explore more reliable indexes for syndrome dif-ferentiation.

Objective Under the theory guidance of the Fuxingjue and the Tang-Ye-Jing-Fa map,this study aims to explore the common patterns and unique characteristics of five-flavor combinations in formulas for treating different syndromes of cough.Methods Chinese herbal formulas for cough treatment were collected from traditional Chinese medicine(TCM)clinical practice guidelines,consensuses,and textbooks,and their flavor combinations and distribution were clarified.Based on the theory of the Tang-Ye-Jing-Fa map regarding the five flavors for lung diseases(sour for tonifying,salty for purging,and pungent for transforming),the dominant flavors in each formula were statistically analyzed.Further classification was conducted according to the cold-heat properties and the amount of phlegm,and the characteristics and five-flavor combination patterns of formulas for different syndromes were analyzed.Results A total of 21 formulas for treating different syndromes of cough were statistically analyzed,among which 85.7%of the formulas had dominant flavors of"sour,salty,and pungent".For hot cough(9 formulas),sour was mainly used to tonify the lung and bitter to purge the heart(e.g.,Sangxing Tang and Qingjin Huatan Tang);for cold cough(4 formulas),pungent was mainly used to tonify the liver and warm yang(e.g.,San'ao Tang combined with Zhike San and Xiaoqinglong Tang);for cough with abundant phlegm(7 formulas),the focus was on using pungent to purge the spleen and eliminate dampness(e.g.,Erchen Tang combined with Sanzi Yangqin Tang);for cough with little or no phlegm(6 formulas),sour was used as the core to tonify the lung and nourish yin(e.g.,Sha Shen Mai Dong Tang).Conclusion From the perspective of the Tang-Ye-Jing-Fa map,the flavors of"sour,salty,and pungent"play an important role in the composition of formulas for treating cough.Although the five-flavor combinations of formulas for different syndromes of cough vary,they share common characteristics when classified according to cold-heat properties and phlegm amount.These findings provide a theoretical basis for optimizing the combination of Chinese herbal formulas for cough treatment by adjusting the proportion of the five flavors in clinical practice.

Objective Under the theory guidance of the Fuxingjue and the Tang-Ye-Jing-Fa map,this study aims to explore the common patterns and unique characteristics of five-flavor combinations in formulas for treating different syndromes of cough.Methods Chinese herbal formulas for cough treatment were collected from traditional Chinese medicine(TCM)clinical practice guidelines,consensuses,and textbooks,and their flavor combinations and distribution were clarified.Based on the theory of the Tang-Ye-Jing-Fa map regarding the five flavors for lung diseases(sour for tonifying,salty for purging,and pungent for transforming),the dominant flavors in each formula were statistically analyzed.Further classification was conducted according to the cold-heat properties and the amount of phlegm,and the characteristics and five-flavor combination patterns of formulas for different syndromes were analyzed.Results A total of 21 formulas for treating different syndromes of cough were statistically analyzed,among which 85.7%of the formulas had dominant flavors of"sour,salty,and pungent".For hot cough(9 formulas),sour was mainly used to tonify the lung and bitter to purge the heart(e.g.,Sangxing Tang and Qingjin Huatan Tang);for cold cough(4 formulas),pungent was mainly used to tonify the liver and warm yang(e.g.,San'ao Tang combined with Zhike San and Xiaoqinglong Tang);for cough with abundant phlegm(7 formulas),the focus was on using pungent to purge the spleen and eliminate dampness(e.g.,Erchen Tang combined with Sanzi Yangqin Tang);for cough with little or no phlegm(6 formulas),sour was used as the core to tonify the lung and nourish yin(e.g.,Sha Shen Mai Dong Tang).Conclusion From the perspective of the Tang-Ye-Jing-Fa map,the flavors of"sour,salty,and pungent"play an important role in the composition of formulas for treating cough.Although the five-flavor combinations of formulas for different syndromes of cough vary,they share common characteristics when classified according to cold-heat properties and phlegm amount.These findings provide a theoretical basis for optimizing the combination of Chinese herbal formulas for cough treatment by adjusting the proportion of the five flavors in clinical practice.

Objective Through the safety evaluation of Chinese patent medicines containing cinnabar,the correlation between the evaluation score and the content of drug instructions was analyzed.And the methods and suggestions for improving the label quality of Chinese patent medicines were proposed.Methods Establish a database of Chinese patent medicines and sim-ple preparations containing cinnabar that meet the screening criteria in the 2020 edition of the Chinese Pharmacopoeia,and quan-titatively score such Chinese patent medicines and simple preparations from the two first level dimensions of safety and toxicity characteristics of Chinese patent medicines and simple preparations,and at the same time,grade the level of safety evidence.Results From the perspective of safety evaluation dimension,Chinese patent medicines and simple preparations with relatively high safety score(safety score ≥ 5 points)has a certain correlation with the content of adverse drug reactions clearly reported in the instructions of this kind of medicine,and 54.55％of the evidence risk of this kind of Chinese patent medicines and simple preparations is controllable,with good safety.From the perspective of the evaluation dimension of toxicity characteristics of Chi-nese patent medicines and simple preparations,the types of toxic ingredients in the compatibility of prescriptions and the scores of daily maximum dosage have certain guiding significance for clinical medication precautions.Conclusion For improving the ad-verse reaction information in the instructions of Chinese patent medicines and simple preparations containing cinnabar,and remin-ding the special population of drug use restrictions,it not only has an important impact on drug safety evaluation and grading,but also provides warning and reference for clinical and patient safety in drug use.

AIM:To investigate the effect of cryptotanshinone(CPT)on myocardial hypertrophy induced by isoprenaline(ISO)in rats and explore its potential mechanism.METHODS:The experimental design consisted of two parts.The first aimed to investigate the effects of CPT on cardiac function,pathological manifestations,and the Janus ki-nase 2(JAK2)/signal transducer and activator of transcription 3(STAT3)signaling pathway in rats with myocardial hyper-trophy.The rats were divided into six groups,namely the control,CPT control,and model groups and low-(15 mg·kg-1·d-1),medium-(30 mg·kg-1·d-1),and high-dose(60 mg·kg-1·d-1)CPT treatment groups,with six rats per group.The sec-ond part aimed to validate the role of the JAK2/STAT3 signaling pathway in the CPT-mediated myocardial hypertrophy treatment.Rats were divided into four groups,namely the control,model,high-dose CPT treatment,and coumermycin A1(CA1,a JAK2/STAT3 agonist)intervention(rats received ISO injection followed by 60 mg·kg-1·d-1 of high-dose CPT and 1 mg·kg-1·d-1 of CA1 for 15 d)groups,with five rats per group.Myocardial hypertrophy was induced in rats via intra-peritoneal injection of ISO(5 mg/kg),and CPT intervention lasted for 15 days.Cardiac function-related parameters were assessed using echocardiography,and pathological changes were evaluated through hematoxylin-eosin,Masson,and wheat germ agglutinin staining.Protein expression levels of atrial natriuretic peptide(ANP),brain natriuretic peptide(BNP),β-myosin heavy chain(β-MHC),and JAK2/STAT3 signaling pathway-related proteins were detected by Western blot analysis.RESULTS:Compared with the model group,CPT administration improved cardiac dysfunction-related ul-trasound markers and significantly reduced ISO-induced cardiomyocyte hypertrophy and myocardial fibrosis in rats with hy-pertrophy in a dose-dependent manner(P<0.05).Additionally,CPT decreased the levels of ANP,BNP,and β-MHC in-duced by ISO modeling(P<0.05),and inhibiting the phosphorylation of JAK2 and STAT3(P<0.05).Furthermore,a partial reversal of the therapeutic effect on myocardial hypertrophy induced by ISO modeling was observed when CA1 was administered(P<0.05).CONCLUSION:The CPT exhibits potential as a therapeutic agent for cardiac hypertrophy by effectively attenuating ISO-induced cardiac hypertrophy in rats through JAK2/STAT3 signaling inhibition.