The theory of constitution in traditional Chinese medicine （TCM） has emerged as a new discipline in recent years. Constitution plays a vital role in the onset，progression，transformation，and prognosis of diseases. At present，some clinical scholars have adopted a novel diagnostic and treatment model of "constitution differentiation-disease identification-syndrome differentiation"，in which constitution is regarded as a core element throughout the diagnostic and therapeutic process. Constitution is closely associated with etiology，onset，pathogenesis，syndrome differentiation，and treatment. Against this background，the construction of animal models based on constitution holds far-reaching significance for advancing clinical research. This paper focuses on the construction and evaluation of rodent models with Qi-deficiency constitution，aiming to explore how to further induce Qi-deficiency syndromes and related disease states on the basis of Qi-deficiency constitution models，thereby developing an integrated animal model that embodies the trinity of "constitution-disease-syndrome". The establishment of this model not only provides a solid experimental foundation for the development of new therapies and drugs in TCM targeting specific constitutions，diseases，and syndromes，but also greatly promotes the modernization and scientific advancement of TCM theory. By comprehensively applying multidisciplinary technologies and methods，the study evaluates the model's validity，reliability，and practicality，with the aim of opening new avenues for future research in TCM and promoting the development of the field.

The theory of constitution in traditional Chinese medicine （TCM） has emerged as a new discipline in recent years. Constitution plays a vital role in the onset，progression，transformation，and prognosis of diseases. At present，some clinical scholars have adopted a novel diagnostic and treatment model of "constitution differentiation-disease identification-syndrome differentiation"，in which constitution is regarded as a core element throughout the diagnostic and therapeutic process. Constitution is closely associated with etiology，onset，pathogenesis，syndrome differentiation，and treatment. Against this background，the construction of animal models based on constitution holds far-reaching significance for advancing clinical research. This paper focuses on the construction and evaluation of rodent models with Qi-deficiency constitution，aiming to explore how to further induce Qi-deficiency syndromes and related disease states on the basis of Qi-deficiency constitution models，thereby developing an integrated animal model that embodies the trinity of "constitution-disease-syndrome". The establishment of this model not only provides a solid experimental foundation for the development of new therapies and drugs in TCM targeting specific constitutions，diseases，and syndromes，but also greatly promotes the modernization and scientific advancement of TCM theory. By comprehensively applying multidisciplinary technologies and methods，the study evaluates the model's validity，reliability，and practicality，with the aim of opening new avenues for future research in TCM and promoting the development of the field.

Objective:To screen out the disinfection procedure and disinfectant suitable for the actual production of specific pathogen free(SPF)chicken embryo eggs,so as to ensure the disinfection effect of specific pathogen free(SPF)chicken embryo eggs in vaccine production.Methods:This study compares the microbial counting methods of soaking,swabbing with cotton swabs and pouring after thin-film filtration for SPF chicken embryo eggs in a GMP production workshop,and selects the most suitable method for SPF chicken embryo egg microbial counting.Experi-mental groups A,B,and C use self-prepared concentrations of 1∶50 sporicidal agent dilution solution,1∶128 alka-line phenol salt dilution solution,and ready-to-use compound quaternary ammonium disinfectant(sterile),respec-tively,and follow the actual disinfection procedures in the workshop to disinfect and sample the surfaces of SPF chicken embryo eggs entering different cleanliness grades,while using sterile water instead of disinfectant as the control group.The average bactericidal rate is calculated by recording the number of colonies and monitoring the viability of chicken embryo cells using microbial culture,and the disinfection effect of the three disinfectants on SPF chicken embryo eggs is evaluated.Results:The comparison of the results from the three methods shows that the method of sampling SPF chicken embryo eggs by immersion and then counting the microbial colonies through membrane filtration is superior to the other two methods.The final cleaning rate of the control group,which used sterilized injection water to clean the SPF chicken embryo eggs,was 91.67％to 96.97％,while the final steriliza-tion rate of the experimental group,which used the above three disinfectants to disinfect the SPF chicken embryo eggs,was 100.00％.By comparing the cell counts of the experimental group and the control group,it was found that the live cell density of the control group was(6.03-6.25)× 105 cells·mL-1,and that of the experimental groups A-C was(6.08-6.17)× 105 cells·mL-1,(5.99-6.25)× 105 cells·mL-1,and(5.87-6.21)× 105 cells·mL-1 respectively;the cell viability of the control group was 90.33％to 91.35％,and that of the experi-mental groups A-C was 88.25％to 92.12％,89.45％to 93.59％,and 88.02％to 92.89％respectively.Through statistical analysis,it was found that the P values of all experimental groups compared with the control group were greater than 0.05,indicating no statistically significant difference.Conclusion:By comparing the dis-infection effects,cell density and cell viability of the three disinfectants and comprehensively considering factors such as cost and risk of the three disinfectants,1∶50 sporicide dilution,1∶128 alkaline phenolate dilution and ready-to-use compound quaternary ammonium salt disinfectant(sterile)can all be used for the daily surface disin-fection of SPF chicken embryo eggs in the production workshop.The selection of an appropriate disinfectant should be based on specific application scenarios and requirements.

Endothelial dysfunction(ED)is a crucial pathological feature and a key initiating factor in diabetic angiopathy.A major pathway contributing to ED is the reduction of nitric oxide(NO)production.NO is synthesized by endothelial nitric oxide synthase(eNOS)using L-arginine as a substrate.However,arginase(Arg)can compete with eNOS for this substrate.Arg has subtype 1 and 2.In diabetes mellitus(DM),the abnormal increase in Arg1 can lead to excessive consumption of the substrate in the vascular endothelium,which causes a decrease in NO levels,leading to diabetes-induced vascular ED.This article reviews the research progress of arginase 1 in diabetes-induced vascular ED.

Objective:To screen out the disinfection procedure and disinfectant suitable for the actual production of specific pathogen free(SPF)chicken embryo eggs,so as to ensure the disinfection effect of specific pathogen free(SPF)chicken embryo eggs in vaccine production.Methods:This study compares the microbial counting methods of soaking,swabbing with cotton swabs and pouring after thin-film filtration for SPF chicken embryo eggs in a GMP production workshop,and selects the most suitable method for SPF chicken embryo egg microbial counting.Experi-mental groups A,B,and C use self-prepared concentrations of 1∶50 sporicidal agent dilution solution,1∶128 alka-line phenol salt dilution solution,and ready-to-use compound quaternary ammonium disinfectant(sterile),respec-tively,and follow the actual disinfection procedures in the workshop to disinfect and sample the surfaces of SPF chicken embryo eggs entering different cleanliness grades,while using sterile water instead of disinfectant as the control group.The average bactericidal rate is calculated by recording the number of colonies and monitoring the viability of chicken embryo cells using microbial culture,and the disinfection effect of the three disinfectants on SPF chicken embryo eggs is evaluated.Results:The comparison of the results from the three methods shows that the method of sampling SPF chicken embryo eggs by immersion and then counting the microbial colonies through membrane filtration is superior to the other two methods.The final cleaning rate of the control group,which used sterilized injection water to clean the SPF chicken embryo eggs,was 91.67％to 96.97％,while the final steriliza-tion rate of the experimental group,which used the above three disinfectants to disinfect the SPF chicken embryo eggs,was 100.00％.By comparing the cell counts of the experimental group and the control group,it was found that the live cell density of the control group was(6.03-6.25)× 105 cells·mL-1,and that of the experimental groups A-C was(6.08-6.17)× 105 cells·mL-1,(5.99-6.25)× 105 cells·mL-1,and(5.87-6.21)× 105 cells·mL-1 respectively;the cell viability of the control group was 90.33％to 91.35％,and that of the experi-mental groups A-C was 88.25％to 92.12％,89.45％to 93.59％,and 88.02％to 92.89％respectively.Through statistical analysis,it was found that the P values of all experimental groups compared with the control group were greater than 0.05,indicating no statistically significant difference.Conclusion:By comparing the dis-infection effects,cell density and cell viability of the three disinfectants and comprehensively considering factors such as cost and risk of the three disinfectants,1∶50 sporicide dilution,1∶128 alkaline phenolate dilution and ready-to-use compound quaternary ammonium salt disinfectant(sterile)can all be used for the daily surface disin-fection of SPF chicken embryo eggs in the production workshop.The selection of an appropriate disinfectant should be based on specific application scenarios and requirements.

Endothelial dysfunction(ED)is a crucial pathological feature and a key initiating factor in diabetic angiopathy.A major pathway contributing to ED is the reduction of nitric oxide(NO)production.NO is synthesized by endothelial nitric oxide synthase(eNOS)using L-arginine as a substrate.However,arginase(Arg)can compete with eNOS for this substrate.Arg has subtype 1 and 2.In diabetes mellitus(DM),the abnormal increase in Arg1 can lead to excessive consumption of the substrate in the vascular endothelium,which causes a decrease in NO levels,leading to diabetes-induced vascular ED.This article reviews the research progress of arginase 1 in diabetes-induced vascular ED.

The development of acute liver injury can result in liver cirrhosis, liver failure, and even liver cancer, yet there is currently no effective therapy for it. The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lyciumbarbarum polysaccharides (LBPs) on acute liver injury induced by carbon tetrachloride (CCl₄). To create a model of acute liver injury, experimental canines received an intraperitoneal injection of 1 mL/kg of CCl₄ solution. The experimental canines in the therapy group were then fed LBPs (20 mg/kg). CCl₄-induced liver structural damage, excessive fibrosis, and reduced mitochondrial density were all improved by LBPs, according to microstructure data. By suppressing Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1), promoting the production of sequestosome 1 (SQSTM1)/p62, nuclear factor erythroid 2-related factor 2 (Nrf2), and phase II detoxification genes and proteins downstream of Nrf2, and restoring the activity of anti-oxidant enzymes like catalase (CAT), LBPs can restore and increase the antioxidant capacity of liver. To lessen mitochondrial damage, LBPs can also enhance mitochondrial respiration, raise tissue adenosine triphosphate (ATP) levels, and reactivate the respiratory chain complexes I‒V. According to serum metabolomics, the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism. 9-Hydroxyoctadecadienoic acid (9-HODE), lysophosphatidylcholine (LysoPC/LPC), and phosphatidylethanolamine (PE) may be potential indicators of acute liver injury. This study confirmed that LBPs, an effective hepatoprotective drug, may cure acute liver injury by lowering oxidative stress, repairing mitochondrial damage, and regulating metabolic pathways.
Animals ; Dogs ; Antioxidants/metabolism* ; Carbon Tetrachloride ; Chemical and Drug Induced Liver Injury/drug therapy* ; Kelch-Like ECH-Associated Protein 1/metabolism* ; Liver ; Metabolic Networks and Pathways ; Mitochondria/metabolism* ; NF-E2-Related Factor 2/metabolism* ; Oxidative Stress ; Polysaccharides/pharmacology* ; Lycium/chemistry*

The development of acute liver injury can result in liver cirrhosis, liver failure, and even liver cancer, yet there is currently no effective therapy for it. The purpose of this study was to investigate the protective effect and therapeutic mechanism of Lycium barbarum polysaccharides (LBPs) on acute liver injury induced by carbon tetrachloride (CCl4). To create a model of acute liver injury, experimental canines received an intraperitoneal injection of 1 mL/kg of CCl4 solution. The experimental canines in the therapy group were then fed LBPs (20 mg/kg). CCl4-induced liver structural damage, excessive fibrosis, and reduced mitochondrial density were all improved by LBPs, according to microstructure data. By suppressing Kelch-like epichlorohydrin (ECH)-associated protein 1 (Keap1), promoting the production of sequestosome 1 (SQSTM1)/p62, nuclear factor erythroid 2-related factor 2 (Nrf2), and phase Ⅱ detoxification genes and proteins downstream of Nrf2, and restoring the activity of anti-oxidant enzymes like catalase (CAT), LBPs can restore and increase the antioxidant capacity of liver. To lessen mitochondrial damage, LBPs can also enhance mitochondrial respiration, raise tissue adenosine triphosphate (ATP) levels, and reactivate the respiratory chain complexes I?V. According to serum metabolomics, the therapeutic impact of LBPs on acute liver damage is accomplished mostly by controlling the pathways to lipid metabolism. 9-Hydroxyoctadecadienoic acid (9-HODE), lysophosphatidylcholine (LysoPC/LPC), and phosphatidylethanolamine (PE) may be potential indicators of acute liver injury. This study confirmed that LBPs, an effective hepatoprotective drug, may cure acute liver injury by lowering oxidative stress, repairing mitochondrial damage, and regulating metabolic pathways.

BACKGROUND:It is reported that tailless-like protein (TLX) plays critical roles in the regulation of early developmental processes in vertebrates, and it plays a key role in stem cells proliferation and differentiation into neurons. OBJECTIVE: To construct recombinant plasmid pEGFPN1-TLX and study the transfection into dermal multipotential stem cells. DESIGN, TIME AND SETTING: Cytogene experiment was performed at the Department of Pathogen Biology, School of Basic Medical Science, the Third Military Medical University of Chinese PLA from March to December 2007. MATERIALS: An adult SD was obtained from the Experimental Animal Center of the Third Military Medical University of Chinese PLA; dermal moltipotential stem cells (DMSCs) were cultured by the Institute of Combined Injury of the Third Military Medical University of Chinese PLA; pEGFPN1 and DH5α was gifted by professor Xu.METHODS: Total RNA was extracted from rat brain tissue to amplify TLX-coded cDNA sequence using RT-PCR. T/A was cloned on pMD18-T vector and determined using BamHI and Hindlll. The products were positive recombinant plasmid pMD18-T-TLX segments, which were sub-cloned in pEGFPN1 to construct recombinant plasmid pEGFPN1-TLX. Finally, pEGFPN1-TLX was transfected into DMSCs.MAIN OUTCOME MEASURES: The fluorescence protein expression was observed under fluorescence microscope at 24 hours after transfection; TLX mRNA expression was detected using RT-PCR; neuronal differentiation was observed using immunohistochemical staining.RESULTS: TLX full length cDNA was successfully cloned into pEGFPN1, and pEGFPN1-TLX was successfully constructed by means of sequence analysis and enzyme cutting identification. As compared with non-transfected DMSCs, pEGFPN1-TLX transfected DMSCs were observed after 10 days, formed resistant clones after 15 days, and shown a green fluorescent protein expression. However, non-transfected DMSCs died at day 10. RT-PCR indicated that pEGFPN1-TLX transfected DMSCs could express TLX mRNA. At day 3 after induction, NF200 positive cells were increased, but glial fibrillary acidic protein positive cells were decreased after induction of pEGFPN1-TLX transfected DMSCs.CONCLUSION: TLX was successfully constructed and transfected into DMSCs. After transfection, neuronal differentiation of DMSCs was enhanced, and the differentiation to gliocytes was inhibited.

Objective To investigate the relationship between the immune defence reaction against Plasmodium infection and the prophenoloxidase (PPO) of the midgut by comparative analysis of the distributions and the changes of PPO in the midgut of Anopheles stephensi and Anopheles dirus before and after infection with Plasmodium yoelii . Methods Midguts were dissected out from both Anopheles stephensi and Anopheles dirus at 3, 5, 7, 11 and 15 d before and after infection with Plasmodium yoelii . Immunohistochemistry and Western blotting were performed respectively on the collected midguts using Manduca Sexta PPO IgG polyantibody. Results PPO in the midguts from both Anopheles stephensi and Anopheles dirus was mainly located in the circulation conduit of midgut before infection with Plasmodium yoelii , but aggregated and distributed at the interspace of midguts as pieced or/and stripped forms after infection. Furthermore, PPO in the midgut of Anopheles dirus was more concentrated than that of Anopheles stephensi . Western blotting revealed that the PPO band with about molecular weight of 67?10 3 was detected in the midguts of both Anopheles stephensi and Anopheles dirus before and after Plasmodium yoelii infection. There was significant difference before and after infection, and the PPO band was obviously enhanced after infection. PPO bands in the midgut of Anopheles dirus were more prominent than those of Anopheles stephensi . Conclusion PPO in the midgut of Anopheles mosquitoes may come from the hemolymph by the circulation conduit before Plasmodium yoelii infection. However, the different distributions and changes of PPO in the midguts resulted from the Anopheles mosquitoes infected with Plasmodia may be closely correlated with Plasmodia infection, which may be of important physiological significance and may be involved in the immune defensive reaction against Plasmodium .