Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Objective Based on the general belief that the tongue characteristics of blood stasis constitution is"purple tongue,tortuous sublingual vessels",this study systematically summarized blood stasis tongue characteristics using artificial intelligence image processing combined with expert evaluation.Methods A physical questionnaire survey was conducted on 1275 college students,and 100 cases were randomly selected from the blood stasis constitution group and the mild constitution group.After collecting the tongue images,Unet,OpenCV,YOLOv5 and YOLOv8 algorithms combined with expert evaluation method were used to extract and analyze the tongue features of the two groups.By comparing the differences of tongue image characteristics between the blood stasis constitution group and the mild constitution group,the tongue characteristics of blood stasis constitution were systematically summarized.Results Compared with the mild constitution group,the proportion of light dark tongue and light purple tongue in the blood stasis constitution group was significantly higher(P＜0.05).The I value,L value and a value of the tongue area 1 in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).The S value and a value of area 2 and area 3 were significantly larger(P＜0.05),and the I value,L value,a value and b value of area 5 were significantly smaller(P＜0.01,P＜0.05).Compared with the mild constitution group,the proportion of tooth marks tongue in the blood stasis constitution group was significantly higher(P＜0.01),the proportion of cracked tongue in the blood stasis constitution group was significantly larger(P＜0.05),and the number of tooth marks,crack area,relative height of crack and relative width of crack in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).Compared with the mild constitution group,the I value,L value,a value and b value of tongue coating area 1 in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05),the I value,L value,S value,a value and b value of tongue coating area 2 were significantly larger(P＜0.01,P＜0.05),the I value,S value,a value and b value of tongue coating area 3 were significantly larger(P＜0.01,P＜0.05),the b value of tongue coating area 4 was significantly larger(P＜0.05),the S value,a value and b value of tongue coating area 5 were significantly larger(P＜0.01,P＜0.05).The proportion of thick tongue coating and greasy tongue coating in the blood stasis group was significantly higher than that in the gentleness group(P＜0.05).Compared with the mild constitution group,the proportion of tortuous,thickened and nodular sublingual collaterals in the blood stasis constitution group was significantly larger(P＜0.01,P＜0.05).The a value,I value,S value,b value and relative width of sublingual collaterals in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).Conclusion On the basis of the general belief that the characteristics of blood stasis constitution tongue are dark purple tongue and dark purple sublingual collaterals/varicose veins/thickening,blood stasis constitution people also have new tongue characteristics,such as redder tips and edges,more teeth marks and fissures,thicker and greasier coatings,and yellow fur.These findings offer new insights for"to identify the constitution of traditional Chinese medicine and prevent diseases by observing the tongue manifestation".

Objective Based on the general belief that the tongue characteristics of blood stasis constitution is"purple tongue,tortuous sublingual vessels",this study systematically summarized blood stasis tongue characteristics using artificial intelligence image processing combined with expert evaluation.Methods A physical questionnaire survey was conducted on 1275 college students,and 100 cases were randomly selected from the blood stasis constitution group and the mild constitution group.After collecting the tongue images,Unet,OpenCV,YOLOv5 and YOLOv8 algorithms combined with expert evaluation method were used to extract and analyze the tongue features of the two groups.By comparing the differences of tongue image characteristics between the blood stasis constitution group and the mild constitution group,the tongue characteristics of blood stasis constitution were systematically summarized.Results Compared with the mild constitution group,the proportion of light dark tongue and light purple tongue in the blood stasis constitution group was significantly higher(P＜0.05).The I value,L value and a value of the tongue area 1 in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).The S value and a value of area 2 and area 3 were significantly larger(P＜0.05),and the I value,L value,a value and b value of area 5 were significantly smaller(P＜0.01,P＜0.05).Compared with the mild constitution group,the proportion of tooth marks tongue in the blood stasis constitution group was significantly higher(P＜0.01),the proportion of cracked tongue in the blood stasis constitution group was significantly larger(P＜0.05),and the number of tooth marks,crack area,relative height of crack and relative width of crack in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).Compared with the mild constitution group,the I value,L value,a value and b value of tongue coating area 1 in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05),the I value,L value,S value,a value and b value of tongue coating area 2 were significantly larger(P＜0.01,P＜0.05),the I value,S value,a value and b value of tongue coating area 3 were significantly larger(P＜0.01,P＜0.05),the b value of tongue coating area 4 was significantly larger(P＜0.05),the S value,a value and b value of tongue coating area 5 were significantly larger(P＜0.01,P＜0.05).The proportion of thick tongue coating and greasy tongue coating in the blood stasis group was significantly higher than that in the gentleness group(P＜0.05).Compared with the mild constitution group,the proportion of tortuous,thickened and nodular sublingual collaterals in the blood stasis constitution group was significantly larger(P＜0.01,P＜0.05).The a value,I value,S value,b value and relative width of sublingual collaterals in the blood stasis constitution group were significantly larger(P＜0.01,P＜0.05).Conclusion On the basis of the general belief that the characteristics of blood stasis constitution tongue are dark purple tongue and dark purple sublingual collaterals/varicose veins/thickening,blood stasis constitution people also have new tongue characteristics,such as redder tips and edges,more teeth marks and fissures,thicker and greasier coatings,and yellow fur.These findings offer new insights for"to identify the constitution of traditional Chinese medicine and prevent diseases by observing the tongue manifestation".

Objective:To compare the efficiency and characteristics of different carrier proteins and signal sequences in delivering antigens into Escherichia coli outer membrane vesicles (OMVs). Methods:The fusion protein F1V, which consisted of the main protective antigen of Yersinia pestis F1 and LcrV, was expressed using the carrier proteins such as cytolysin A (ClyA), outer membrane protein A (OmpA), or β-lactamases (Bla) signal sequence as a carrier protein. The expression, localization, and content of F1V protein in OMVs were compared and analyzed. Results:All three delivery methods successfully incorporated F1V protein into OMVs and localized it on the surface of OMVs. Notably, when OmpA was used as the carrier protein, the F1V fusion protein constituted up to 30% of the total protein in OMVs. The highest yield of OMVs, reaching 4.2 mg/L, was achieved when Bla signal sequence was used as the carrier.Conclusions:There is a significant difference in the efficiency of different carrier proteins in delivering the F1V antigen into OMVs of Escherichia coli. Considering both the yield of OMVs and the proportion of antigen in the total protein of OMVs, the carrier Bla signal sequence demonstrated the highest efficiency in delivering F1V into OMVs, showing a potential for the future development of OMVs-based plague vaccines.

Tongue diagnosis is an important method of TCM diagnosis and treatment.Tongue is the key link of auxiliary diagnosis of tongue feature extraction and processing,and also is the bottleneck of intelligent tongue diagnosis in traditional Chinese medicine.Using image processing,artificial intelligence technology to the tongue as a quantitative and identify characteristics of traditional Chinese medicine,looking for both conforms to the original thinking of TCM,and TCM tongue diagnosis method of accurately,has become a common concern of traditional Chinese medicine and computer field.From the mobile terminal tongue as auxiliary diagnostic system of traditional Chinese medicine tongue acquisition basic attribute,tongue diagnosis and image information building,tongue like features are required for accurate extraction and so on related key technology is analyzed,and build overall architecture,so as to provide technical reference for the tongue like intelligent diagnosis,promote the development of technology of tongue diagnosis in traditional Chinese medicine modernization.

Photodynamic therapy, a new type of non-invasive treatment, is based on the principle that the photosensitizer excited by laser can transfer energy to oxygen, which generates cytotoxic singlet oxygen and thus induce tumor cell apoptosis or necrosis. As an oxygen-dependent therapy, the antitumor effect of photodynamic therapy is obviously limited by hypoxia environment of solid tumor tissue. Therefore, reversing and improving the hypoxia of tumor tissue can significantly enhance the efficacy of photodynamic therapy. This review focuses on the progress of tumor oxygenation strategy mediated by nano-delivery system, including direct oxygen delivery strategies, catalytic oxygen production strategies, responsive material in situ oxygen supply strategies and microorganism oxygen supply strategies, aiming to improve the antitumor effect of photodynamic therapy. It provides new ideas and new approaches for further study of oxygen-enchancing nano-delivery system for photodynamic therapy.

BACKGROUND:Restoration of neurological functions after the damaged peripheral nerve is reconstructed is a hot topic in existing research. Within a short term fol owing peripheral nerve injury, nerve and muscle begin to develop irreversible degeneration. Restoration of the damaged nerve requires delayed degeneration and basic microenvironment. OBJECTIVE:To investigate the protective effect of cardiotrophin-1 on PC12 cells and Schwann cells. METHODS:Schwann cells and PC12 cells were obtained and cultured in complete medium, serum-free medium and 50 ℃ medium, respectively. cells in cardiotrophin-1 group were treated with exogenous cardiotrophin-1 solvent, while those in the control group were treated with equivalent Dulbecco’s modified Eagle’s medium, for 24 hours. The survival rate for PC12 cells and Schwann cells was determined using cellCounting Kit-8 colorimetric method. The lactate dehydrogenase activity in supernatant was detected by lactate dehydrogenase kit, and the malondialdehyde content and superoxide dismutase activity were measured by thiobarbituricaicd and xanthine oxidese method respectively. RESULTS AND CONCLUSION:The survival rate of PC12 cells and Schwann cells in cardiotrophin-1 group was obviously increased, lactate dehydrogenase releasing and malondialdehyde content were obviously decreased, superoxide dismutase activity was dramatical y improved compared with control group. Exogenous cardiotrophin-1 reduces the injury caused by ischemia and heat stress stimulation for PC12 cells and Schwann cells. The mechanism of protection may be related to the expression of anti-apoptosis protein activated by the combination of cardiotrophin-1 and its receptors.