The "integrated time-space" perspective of life in traditional Chinese medicine holds that structure and function respectively belong to the realms of space and time. The orderly integration of time and space not only supports the existence and development of life， but also serves as the foundation for biological evolution. Based on this standpoint， this paper proposes the spatiotemporal structural theory of visceral manifestation， characterized by the fusion of time and space， the integration of internal and external dimensions， and the absence of primary or secondary distinctions. Using the individual human body as the boundary， it delineates the man and nature spatiotemporal structure， based on the theory of the "correspondence between man and nature"， and the internal spatiotemporal structure of the living body. The man and nature spatiotemporal structure， with yin-yang and the five phases as its core， reveals the correspondence and connectivity between the external environment and the internal zang-fu organs within the spatiotemporal dimension. The internal temporal structure of the body refers to the collective physiological rhythms related to the zang-fu organs， while the spatial structure refers to the tangible forms， spatial positions， and connective structures of the zang-fu organs. The proposal of the spatiotemporal structural theory of visceral manifestation represents a rational reconstruction and practical exploration of the essential connotations of visceral manifestation， aiming to provide new perspectives for subsequent theoretical research and clinical applications.

With the purpose of improving the teaching quality of the course pathogen biology and immunology for students majoring in nursing and with the guidance of the output of talent training goals, this study introduced micro-video assignment in the teaching process as a supplement to the curriculum ideological and political education. This approach included the integration of curriculum ideological and political education in the whole process, the selection of micro-video topics, the distribution of tasks, the tracking and guidance by teacher in the whole process, and evaluation and rewards. This teaching reform used the traditional micro-video to achieve the transformation from "recorded by teachers for students" to "recorded by students for everyone". This improved the awareness and abilities of students in teamwork, lifelong learning, and independent learning. This allowed the development of a scientific questioning attitude and independent thinking ability, guiding students to establish scientific views of life and the world, and expanding the dimension of innovation in teaching the course of microbiology immunology.

To investigate the molecular mechanism of modified Yigong powder(MYG)in the treat-ment of spleen deficiency syndrome based on network pharmacology and analyzed the effect of MYG on gastrointestinal simulated intestinal flora of spleen deficiency dogs and mucosal barrier of Caco-2 cells,as well as the interaction between intestinal flora and mucosal barrier.The molecular mechanism of MYG in the treatment of spleen deficiency syndrome was predicted by network pharmacology.The fecal samples of three canines(12±1)years old with spleen deficiency were collected to establish an in vitro gastrointestinal simulation system,which was divided into the o-riginal fecal sample group,the gastrointestinal simulation group and the gastrointestinal simulation treated by MYG group.The structural changes of the flora in each group were detected by 16S rD-NA sequencing.The metabolites were extracted from the gastrointestinal simulation system trea-ted by MYG group to study its effect on LPS-induced Caco-2 cell mucosal barrier injury model.The cell experiments included the blank control group,LPS model group,modified Yigong metabolite group.The permeability of mucosal was determined by fluorescein sodium,and then relative ex-pression levels of Claudin-1,Occludin and ZO-1 mRNA were determined by qPCR.The correlation between intestinal flora and Caco-2 cell mucosal barrier index after MYG intervention was further analyzed.The results showed that MYG had 76 active ingredients and 45 potential targets for the treatment of spleen deficiency syndrome.Forty key targets were obtained through protein interac-tion analysis,34 items were obtained by GO enrichment analysis,and 16 pathways were obtained by KEGG enrichment analysis.In the gastrointestinal simulation system,compared with the gas-trointestinal simulation group,at the phylum level,the abundance of Firmicutes,Bacteroides and Actinobacteriota increased significantly(P＜0.05),and the abundance of Proteobacteria decreased significantly(P＜0.05).At the genus level,the abundance of Fusobacterium,[Ruminococcus]gna-vus group and Blautia increased significantly(P＜0.05),while the abundance of Escherichia-Shi-gella and Citrobacter decreased significantly(P＜0.05).The diversity index of intestinal flora in the gastrointestinal simulation treated by MYG group was significantly increased(P＜0.05).In cell experiments,compared with the LPS model group,the mucosal permeability of Caco-2 cells in the modified Yigong metabolite group was significantly reduced(P＜0.01),and the expression levels of Claudin-1,Occludin and ZO-1 mRNA were significantly increased(P＜0.01).Correlation analysis showed that there was a certain correlation between bacterial community structure and mucosal barrier indexes.In summary,MYG may act on 40 key targets such as TNF,IL6,IL18,CX-CL8 and AKT1 through 76 active ingredients such as quercetin,arachidonic acid and naringin,and treat spleen deficiency syndrome in dogs through 16 signaling pathways such as AGE-RAGE,FoxO and HIF-l.In addition,the gastrointestinal metabolites of MYG up-regulate tight junction protein mRNA expression,reduce mucosal permeability,and repair mucosal barrier,which may be related to MYG's regulation of flora structure.

African swine fever(ASF),caused by the African swine fever virus(ASFV),is a highly contagious infectious disease of pigs.This disease has been spread rapidly in China since 2018,po-sing a huge threat to China's pig farming industry.To rapid detect the ASFV,a loop-mediated iso-thermal amplification(LAMP)combined with the disposable nucleic acid visualization test strip was established for visual detection of the nucleic acid of ASFV B646L gene.The method was easy to operate without special instruments and equipment,while it effectively avoided the disadvantage of false positives caused by aerosol contamination.The method was able to detect 1.16 copies/μL of the recombinant plasmid in 50 min at 65 ℃.In addition,the method was specific with no cross-re-action with classical swine fever virus,porcine reproductive and respiratory syndrome virus,por-cine parvovirus,transmissible gastroenteritis virus.The results in this study provides a rapid,con-venient,sensitive and reliable method for early diagnosis and screening for ASFV suspected infec-tion cases.

To establish a simple,convenient,sensitive,and specific method for rapid detection of Zi-ka virus(ZIKV),the whole genome sequences of ZIKV isolated from different times and regions were analyzed.The specific primers and probes were designed based on the screened target se-quences located in the conserved region of the ZIKV NS5 gene.By combining RT-LAMP isother-mal amplification technology and immunochromatography technology,a reverse transcription loop mediated isothermal amplification nucleic acid and flow visualization strip(RT-LAMP-VF)detec-tion method for ZIKV was established.The results showed that the method had good specificity and sensitivity.When the ratio of inner,outer,and ring primers(FIP∶LF∶F3)was 4∶2∶1,the detection method can specifically detect 102 copies/pL RNA transcripts or 2.15 pfu ZIKV at 61 ℃for 45 minutes,with no cross reaction with other flaviviruses such as Japanese encephalitis virus and classical swine fever virus.Other RNAs in blood tissue samples did not affect the sensitivity and specificity of RT-LAMP-VF,indicating that the method can be applied to clinical practice.The ZIKV RT-LAMP-VF detection method established in this study is easy to perform and does not require special instruments and equipment.It is particularly suitable for the rapid detection of ZIKV in grassroots units,providing technical support and material support for the establishment of on-site rapid detection and early warning and prediction systems for ZIKV disease.

The PCR-amplified severe fever with thrombocytopenia syndrome virus(SFTSV)Gn-DⅢ-Ⅲ gene was inserted into the pET-30a(+)prokaryotic expression vector to generate the re-combinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ.The plasmid was transformed into E.coli BL21(DE3)for Gn-DⅢ-m protein expression and the expression conditions were optimized.The Gn-DⅢ-Ⅲ protein purified with Ni-NTA column affinity chromatography was applied as the captured antigen to establish an indirect ELISA method for the detection of SFTSV antibody.The results demonstrated that the recombinant plasmid pET-SFTSV-Gn-D Ⅲ-Ⅲ was successfully constructed as identified by PCR and sequencing.The recombinant protein SFTSV Gn-D m-Ⅲ was soluble ex-pression in E.coli under the optimal induction conditions of 0.4 mmol/L IPTG at 25 ℃ for 4 h,and the protein purity was 91.77％after purification by Ni-NTA column.The optimal reaction con-ditions for the indirect ELISA of SFTSV antibody were as follows:coating antigen concentration(5 μg/mL),primary antibody(incubation at 37 ℃ for 1.5 h),and secondary antibody(diluted 1:10 000 and incubated at 37 ℃ for 1 h).The established method had no cross-reactivity with Rift Valley fever virus(RVFV),Ebola virus(EBOV),and tick-borne encephalitis virus(TBEV)posi-tive sera.The method had a high sensitivity,with P/N＞2.1 for SFTSV-positive sera diluted to 81920.Coefficients of variation for intra-and inter-batch reactions were less than 10％.Detection of four SFTSV-infected human clinical serum samples showed the serum samples from patients in re-mission were tested as positive(P/N＞2.1),while serum samples from patients with multiple or-gan failure were detected as negative(P/N＜2.1).The results indicated that the SFTSV Gn-D Ⅲ-Ⅲ protein was successfully expressed and purified,and it was used as the coating protein to estab-lish an indirect ELISA assay for SFTSV antibody,which possesses good specificity,sensitivity and reproducibility.This method might be applied to detect human SFTSV clinical serum samples.

To reduce the subjectivity and uncertainty present in the current international methods of drug value pricing when converting value into monetary prices,based on the hedonic pricing theory,it considers the post-negotiation price between manufacturers and payers as a reasonable price reference in the value pricing of Chinese patent medicine.By constructing an indicator system for the characteristics of Chinese patent medicine,it selects and measures the value characteristic variables that affect the price of Chinese patent medicine.It serves as the theoretical foundation and research basis for establishing a Hedonic price model between characteristic price variables and negotiation prices,thereby promoting the enhancement of rationality and objectivity in value-guided pricing of Chinese patent medicine.

Objective The purpose is to sort out the key steps and core indicators of priority setting for health technology assessment,and provide references for the research of priority setting for health technology assessment in China.Methods To search information from the official website of the World Health Organization,the websites of international health technology assessment agencies/organizations,and CNKI,Wanfang,Pubmed,Embase and other databases related for the setting of health technology assessment priority topics,and the key steps and core indicators of the setting of priority topics were analyzed by descriptive statistical analysis.Results 21 priority setting schemes for health technology assessment were finally incorporated,and the key steps were extracted to set indicators for collecting evaluations.Ratings and rankings and review decisions.The core indicators are disease burden,economic impact and clinical/health impact.Conclusion The key steps and core indicators of international priority setting provide rich practical experience for China's health technology assessment priority setting,which should be actively used for reference to promote evidence-based and scientific decision-making of health technology assessment in China.

To reduce the subjectivity and uncertainty present in the current international methods of drug value pricing when converting value into monetary prices,based on the hedonic pricing theory,it considers the post-negotiation price between manufacturers and payers as a reasonable price reference in the value pricing of Chinese patent medicine.By constructing an indicator system for the characteristics of Chinese patent medicine,it selects and measures the value characteristic variables that affect the price of Chinese patent medicine.It serves as the theoretical foundation and research basis for establishing a Hedonic price model between characteristic price variables and negotiation prices,thereby promoting the enhancement of rationality and objectivity in value-guided pricing of Chinese patent medicine.

Objective The purpose is to sort out the key steps and core indicators of priority setting for health technology assessment,and provide references for the research of priority setting for health technology assessment in China.Methods To search information from the official website of the World Health Organization,the websites of international health technology assessment agencies/organizations,and CNKI,Wanfang,Pubmed,Embase and other databases related for the setting of health technology assessment priority topics,and the key steps and core indicators of the setting of priority topics were analyzed by descriptive statistical analysis.Results 21 priority setting schemes for health technology assessment were finally incorporated,and the key steps were extracted to set indicators for collecting evaluations.Ratings and rankings and review decisions.The core indicators are disease burden,economic impact and clinical/health impact.Conclusion The key steps and core indicators of international priority setting provide rich practical experience for China's health technology assessment priority setting,which should be actively used for reference to promote evidence-based and scientific decision-making of health technology assessment in China.