Antimicrobial resistance (AMR) poses a critical global health threat. This review systematically examines AMR surveillance technology advances, from conventional culture methods to modern molecular diagnostics (e.g., whole-genome sequencing) and artificial intelligence-assisted approaches. It focuses on the current application of mass spectrometry, machine learning predictive models, and real-time surveillance networks. To address challenges including inadequate technical standardization, clinical translation barriers, and data-sharing limitations, we propose integrated ^"genotype-phenotype^" strategies and global standardization framework, while exploring future applications of CRISPR-based portable detection, single-cell sequencing, and blockchain technologies.

In this paper， by referring to ancient and modern literature， the textual research of Inulae Flos has been conducted to clarify the name， origin， production area， quality evaluation， harvesting， processing and others， so as to provide reference and basis for the development and utilization of famous classical formulas containing this herb. After textual research， it could be verified that the medicinal use of Inulae Flos was first recorded in Shennong Bencaojing of the Han dynasty. In successive dynasties， Xuanfuhua has been taken as the official name， and it also has other alternative names such as Jinfeicao， Daogeng and Jinqianhua. The period before the Song and Yuan dynasties， the main origin of Inulae Flos was the Asteraceae plant Inula japonica， and from the Ming and Qing dynasties to the present， I. japonica and I. britannica are the primary source. In addition to the dominant basal species， there are also regional species such as I. linariifolia， I. helianthus-aquatili， and I. hupehensis. The earliest recorded production areas in ancient times were Henan， Hubei and other places， and the literature records that it has been distributed throughout the country since modern times. The medicinal part is its flower， the harvesting and processing method recorded in the past dynasties is mainly harvested in the fifth and ninth lunar months， and dried in the sun， and the modern harvesting is mostly harvested in summer and autumn when the flowers bloom， in order to remove impurities， dry in the shade or dry in the sun. In addition， the roots， whole herbs and aerial parts are used as medicinal materials. In ancient times， there were no records about the quality of Inulae Flos， and in modern times， it is generally believed that the quality of complete flower structure， small receptacles， large blooms， yellow petals， long filaments， many fluffs， no fragments， and no branches is better. Ancient processing methods primarily involved cleaning， steaming， and sun-drying， supplemented by techniques such as boiling， roasting， burning， simmering， stir-frying， and honey-processing. Modern processing focuses mainly on cleaning the stems and leaves before use. Regarding the medicinal properties， ancient texts describe it as salty and sweet in taste， slightly warm in nature， and mildly toxic. Modern studies characterize it as bitter， pungent， and salty in taste， with a slightly warm nature. Its therapeutic effects remain consistent across eras， including descending Qi， resolving phlegm， promoting diuresis， and stopping vomiting. Based on the research results， it is recommended that when developing famous classical formulas containing Inulae Flos， either I. japonica or I. britannica should be used as the medicinal source. Processing methods should follow formula requirements， where no processing instructions are specified， the raw products may be used after cleaning.

In this paper， by referring to ancient and modern literature， the textual research of Inulae Flos has been conducted to clarify the name， origin， production area， quality evaluation， harvesting， processing and others， so as to provide reference and basis for the development and utilization of famous classical formulas containing this herb. After textual research， it could be verified that the medicinal use of Inulae Flos was first recorded in Shennong Bencaojing of the Han dynasty. In successive dynasties， Xuanfuhua has been taken as the official name， and it also has other alternative names such as Jinfeicao， Daogeng and Jinqianhua. The period before the Song and Yuan dynasties， the main origin of Inulae Flos was the Asteraceae plant Inula japonica， and from the Ming and Qing dynasties to the present， I. japonica and I. britannica are the primary source. In addition to the dominant basal species， there are also regional species such as I. linariifolia， I. helianthus-aquatili， and I. hupehensis. The earliest recorded production areas in ancient times were Henan， Hubei and other places， and the literature records that it has been distributed throughout the country since modern times. The medicinal part is its flower， the harvesting and processing method recorded in the past dynasties is mainly harvested in the fifth and ninth lunar months， and dried in the sun， and the modern harvesting is mostly harvested in summer and autumn when the flowers bloom， in order to remove impurities， dry in the shade or dry in the sun. In addition， the roots， whole herbs and aerial parts are used as medicinal materials. In ancient times， there were no records about the quality of Inulae Flos， and in modern times， it is generally believed that the quality of complete flower structure， small receptacles， large blooms， yellow petals， long filaments， many fluffs， no fragments， and no branches is better. Ancient processing methods primarily involved cleaning， steaming， and sun-drying， supplemented by techniques such as boiling， roasting， burning， simmering， stir-frying， and honey-processing. Modern processing focuses mainly on cleaning the stems and leaves before use. Regarding the medicinal properties， ancient texts describe it as salty and sweet in taste， slightly warm in nature， and mildly toxic. Modern studies characterize it as bitter， pungent， and salty in taste， with a slightly warm nature. Its therapeutic effects remain consistent across eras， including descending Qi， resolving phlegm， promoting diuresis， and stopping vomiting. Based on the research results， it is recommended that when developing famous classical formulas containing Inulae Flos， either I. japonica or I. britannica should be used as the medicinal source. Processing methods should follow formula requirements， where no processing instructions are specified， the raw products may be used after cleaning.

Objective To analyze the characteristics of viral hepatitis mortality and life loss among residents in Pudong New Area from 2003 to 2023, and to provide a basis for related prevention and control work. Methods Viral hepatitis mortality data were obtained from the Pudong New Area mortality monitoring system. The crude mortality rate (CMR), standardized mortality rate (SMR), potential years of life lost (PYLL), average years of life lost (AYLL), and standardized potential years of life lost (SPYLL) were calculated to analyze viral hepatitis deaths. The average annual change (AAPC) and annual percentage change (APC) of the mortality rate were calculated by Joinpoint regression analysis to analyze the trend of mortality. Results The CMR and SMR of viral hepatitis among residents in Pudong New Area from 2003 to 2023 were 3.89/100000 and 1.98/100000, respectively. Both CMR and SMR of viral hepatitis showed a decreasing trend over time (CMR:APC=-5.476, t=-13.581, P<0.001; SMR:APC=- 7.624, t= -21.253, P<0.001). The CMR for males was 4.75/100000 and the SMR for males was 2.65/100000; the CMR for females was 3.04/100000 and the SMR for females was 1.32/100000, with a higher mortality rate for males than for females（Z_CME=12.094，P<0.001; Z_SMR=-14.718，P<0.001). Deaths were concentrated in the age groups of 45-64 years old and 65 years old and above, accounting for 91.62% of the total deaths. The PYLL of deaths due to viral hepatitis among residents in Pudong New Area from 2003 to 2023 was 26912 person-years, with a PYLLR of 0.45% and an AYLL of 8.88 years per person. Conclusion The mortality rate of viral hepatitis among the residents of Pudong New Area in 2003-2023 shows a decreasing trend over time. The mortality rate of males is higher than that of females, and the deaths of middle-aged and elderly people account for a large proportion of the total deaths. Chronic hepatitis B is the main cause of death.

Background The crude mortality rate of sudden death has shown a steady upward trend. Pudong New Area, Shanghai, is a highly urbanized and densely populated district that is substantially affected by the urban heat island effect and has experienced frequent extreme temperature events in recent years. However, evidence on the association between ambient temperatures and sudden death among local residents remains limited. Objective To examine the association between daily mean temperature and sudden death in Pudong New Area, and to provide evidence for reducing temperature-related risks of sudden death. Methods Data on sudden death, meteorological factors (daily average temperature, daily average relative humidity, and daily average air pressure), and air pollutants [inhalable particulate matter (PM₁₀), sulfur dioxide (SO₂), and nitrogen dioxide (NO₂)] in Pudong New Area from 2016 to 2021 were collected. A distributed lag non-linear model was used to examine the associations of daily mean temperature with total sudden death and sudden death due to cardiovascular and cerebrovascular diseases, including lagged and cumulative effects (lag0, 0-3, 0-7, 0-14, and 0-21 d). Stratified analyses were performed by sex, age, and education level to identify vulnerable populations. The temperature attributable burden of sudden death was assessed by calculating attributable number (AN) and attribute fraction (AF). Results A total of 19458 sudden deaths were reported in Pudong New Area from 2016 to 2021, of which 11082 (56.95%) were due to cardiovascular and cerebrovascular diseases. Both low (P₅, 4.6 ℃) and high (P₉₅, 30.2 ℃) temperatures were associated with increased risks of sudden death. The cumulative risks associated with low temperature peaked at lag 0-21 d for both total sudden death and sudden death due to cardiovascular and cerebrovascular diseases. In contrast, the cumulative risks associated with high temperature peaked at lag 0-7 d for total sudden death and at lag 0-3 d for sudden death due to cardiovascular and cerebrovascular diseases. Stratified analyses showed that females, individuals aged ≥65 years, and those with primary school education or below were more sensitive to low temperature; whereas females, individuals aged <65 years, and those with secondary school education or above were more sensitive to high temperature. Using 21.5 ℃ as the minimum mortality temperature, the AN of total sudden deaths attributable to non-optimal temperature was 3791 (95%CI: 1853, 5587), with an AF of 20.41% (95%CI: 9.47%, 28.94%); the AN attributable to low temperature was 3358 (95%CI: 1087, 5062), with an AF of 17.26% (95%CI: 5.89%, 26.01%), whereas the AN attributable to high temperature was 225 (95%CI: −274, 655), with an AF of 2.03% (95%CI: −2.53%, 5.78%). For sudden deaths due to cardiovascular and cerebrovascular diseases, the AN attributable to non-optimal temperature was 2721 (95%CI: 1747, 5575), with an AF of 24.55% (95%CI: 9.59%, 28.65%); the AN attributable to low temperature was 2496 (95%CI: 988, 3562), with an AF of 22.53% (95%CI: 8.97%, 32.41%), whereas the AN attributable to high temperature was 614 (95%CI: −15, 1176), with an AF of 3.15% (95%CI: −0.08%, 6.02%). Conclusions Both low (P₅, 4.6 ℃) and high (P₉₅, 30.2 ℃) temperatures are associated with increased risks of sudden death in Pudong New Area, Shanghai. High temperature shows a relative short-term effect, whereas low temperature shows a stronger cumulative effect. Females, people ≥65 years, and individuals with primary school educational attainment or below are more vulnerable to low temperature, while females, people <65 years, and those with secondary school educational attainment or above are more vulnerable to high temperature. The attributable burden of low temperature is greater than that of high temperature. Targeted health protection measures should be strengthened for vulnerable populations during extreme temperature events.

ObjectiveTo screen and identify uridine diphosphate （UDP）-rhamnose synthase （RHM） genes from Ornithogalum saundersiae and verify their UDP-rhamnose （UDP-Rha） biosynthetic functions， thereby providing reliable candidate genes for the efficient biosynthesis of neohesperidin and other rhamnoside natural products. MethodsBased on previously reported plant RHM sequences， two candidate RHM genes， OsRHM1 and OsRHM2， were identified from O. saundersiae through transcriptome screening and sequence alignment. The two genes were cloned and subjected to bioinformatics analysis and multiple sequence alignment. An in vitro RHM-Cm1，2RhaT cascade reaction was established， and the ability of OsRHM1 and OsRHM2 to catalyze the conversion of UDP-glucose （UDP-Glc） to UDP-Rha was evaluated based on neohesperidin production. Real-time fluorescence quantitative polymerase chain reaction PCR （Real-time PCR） was performed to analyze the relative expression levels of the two genes in roots， bulbs， and leaves. ResultsThe open reading frame （ORF） of OsRHM1 was 2 076 bp and encoded 692 amino acids， with a predicted protein molecular mass of 77.97 kDa and a theoretical isoelectric point of 7.25. The ORF of OsRHM2 was 2 019 bp and encoded 673 amino acids， with a predicted protein molecular mass of 75.82 kDa and a theoretical isoelectric point of 7.24. Multiple sequence alignment showed that OsRHM1 and OsRHM2 contained the typical conserved motifs GxxGxxG/A and YxxxK of the plant RHM family and shared high amino acid sequence identity with previously reported RHMs. The in vitro cascade reaction demonstrated that both OsRHM1 and OsRHM2 were capable of catalyzing the conversion of UDP-Glc to UDP-Rha. The neohesperidin production in the OsRHM2 cascade reaction system was approximately 2.58-fold that in the OsRHM1 system. Real-time PCR analysis showed that the relative expression levels of OsRHM1 and OsRHM2 were higher in roots than in bulbs and leaves. ConclusionOsRHM2 exhibited a relatively high capacity for UDP-Rha supply and may serve as a candidate enzyme resource for the subsequent biosynthesis of rhamnoside natural products and the construction of UDP-Rha supply systems.

Objective:To explore the impact of a comprehensive intervention program that integrates transitional care with personalized discharge preparation services on discharge preparedness on the growth, development, and motor development in premature infants, providing guidance and reference for clinical practice.Methods:The 90 pairs of premature infants and their main caregivers who were treated in the neonatal department, Children ′s Hospital, Quanzhou Maternal and Child Health Hospital were studied from February 2023 to February 2024 by randomized control method. Used the table of random numbers, they were divided into the control group and the observation group, with 45 pairs in each group. The control group routinely administered care, while the observation group was implemented a transitional care combined with personalized discharge preparation services. The discharge preparedness, growth and motor development, and the disease uncertainty of caregivers were observed between the 2 groups. Results:There were 27 males and 18 females of the 45 preterm infants,with gestational age of 30.86 (29.36, 31.50) weeks in the control group, 24 males and 21 females with gestational age of 30.29(29.00, 31.07) weeks in the observation group. The main caregiver identities 43 were mothers and 2 were other identities in the control group, 42 were mothers and 3 were other identities in the observation group, with them being 31.00(28.00, 35.00) years old. There were 97.78% (44 /45) caregivers who thought the child was ready to go home in the observation group, while the control group were 84.44% (38 /45), these differences were statistically significant ( χ2=4.88, P<0.05). The total score of discharge readiness in the observation group were 240.00(237.00, 242.50) points, higher than in the control group 226.00(219.00, 229.50) points, these differences were statistically significant ( Z=-6.23, P<0.05). The head circumference and body weight of the observation group were (34.82 ± 1.14) cm and (3.60 ± 0.55) kg, while the control group were (34.25 ± 1.22) cm and (3.35 ± 0.53) kg, there were statistically significant between the two groups ( t=-2.29, -2.22, all P<0.05). The Test of Infant Motor Performance score in the observation group was 50.00(46.00, 52.00) points, while the control group was 45.00(42.00, 48.00) points, there were statistically significant between the two groups ( Z=-3.65, P<0.05). The total score of disease uncertainty in the observation was 52.00(45.50, 60.00) points, while the control group was 61.00(58.50, 65.00) points, there was statistically significant between the two groups ( Z=-4.62, P<0.05). Conclusions:The discharge preparedness of the caregivers of preterm infants was improved because of the use of transitional care combined with personalized discharge preparation services, and the growth and motor development of preterm infants were promoted, and the uncertainty of the family caregivers of preterm infants about the disease was reduced.

Objective To investigate the susceptibility and infection characteristics of dengue virus(DENV)in cells derived from diverse tissues of tree shrews and to provide a basis for expanding the repertoire of DENV-permissive cell models in this species.Methods DENV was inoculated at a multiplicity of infection(MOI)of 0.02 into tree shrew skin fibroblasts(TSFs),primary tree shrew renal epithelial cells(pTRECs),tree shrew aortic endothelial cells(TAECs),tree shrew aortic smooth muscle cells(TASMCs),tree shrew hepatocytes(THs),tree shrew corneal stromal cells(TCSCs),tree shrew brain microvascular endothelial cells(TBMECs),and tree shrew retinal microvascular endothelial cells(TRMECs).C6/36,Vero,A549,and BHK-21 cells(commonly used for DENV propagation)were used as positive controls.Over 6 days post-infection,cellular cytopathic effects were monitored at 12-hour intervals using an inverted microscope,viral RNA loads in cell lysates were quantified by real-time fluorescent quantitative PCR to generate proliferation curves,and viral titers were determined by plaque assay.Results Seven types of tree shrew cells,except TRMECs,were susceptible to DENV.Prolonged infection induced pronounced cytopathic effects,including cell rounding,detachment,necrosis,and lysis,across all susceptible cells.The viral RNA loads detected in lysates of pTRECs,TBMECs,TASMCs,TAECs and THs,approached those of positive controls(≥4×107 copies/μL).Infectious progeny viruses were produced by these five cell types,with three(TAECs,3.13×105 PFU/mL;THs,2.03×105 PFU/mL;pTRECs,1.58×105 PFU/mL)exhibiting titers comparable to C6/36(3.85×10 5 PFU/mL)and earlier viral harvests.Conclusion DENV exhibits broad susceptibility to tree shrew cells of multiple tissue origins,with proliferation rates surpassing those of conventional cell lines sourced from other species.TAECs,THs,and pTRECs are particularly suitable for large-scale DENV proliferation,suggesting their potential involvement in in vivo infection.

AIM:Regulatory T cells(Tregs)are a specialized subset of CD4+T cells primarily involved in im-munosuppressive functions.AMP-activated protein kinase(AMPK)serves as a metabolic sensor that governs the differen-tiation,maturation,and immune functions of Tregs through metabolic reprogramming.However,the impact of AMPKα1(the catalytic subunit of AMPK)knockout specifically in Tregs on the host's immune microenvironment remains largely un-explored.METHODS:Histological changes in immune organs were assessed using HE staining.The types of immune cells and their relative population percentages in immune organs and blood were quantified through flow cytometry in both AMPKα1flox/flox(AMPKα1fl/fl)mice and Treg-specific AMPKα1 knockout mice(AMPKα1fl/flFoxp3cre mice).RESULTS:Compared to AMPKα1fl/fl mice,the percentage of eosinophils in the bone marrow of AMPKα1fl/flFoxp3cre mice was significant-ly reduced.Additionally,while the thymus of AMPKα1fl/flFoxp3cre mice exhibited normal structure,both its size and the ra-tio of thymus weight to body weight were significantly decreased.The knockout of AMPKα1 in Tregs led to a notable reduc-tion in the total percentage of immature double-negative(DN)cells.Consequently,the percentage of CD4+T cells derived from these DN cells also decreased,even though the percentages of DN1 and DN4 cells were higher in the thymus of AMPKα1fl/flFoxp3cre mice compared to AMPKα1fl/fl mice.Importantly,the proportion of Siglec-F+CD11b+eosinophils in the thymus was significantly lower in AMPKα1fl/flFoxp3cre mice.Knockout of AMPKα1 in Tregs resulted in a marked increase in the percentage of CD4+T cells in peripheral blood,alongside a decrease in the proportion of mature CD8+T cells.Similar-ly,the proportion of CD4+T cells in the spleen of AMPKα1fl/flFoxp3cre mice was elevated compared to AMPKα1fl/fl mice.In contrast,the proportion of neutrophils significantly decreased,while mononuclear cell proportions increased in the spleen of AMPKα1fl/flFoxp3cre mice.In lymph nodes,the medullary boundaries in AMPKα1fl/flFoxp3cre mice were blurred,and the lymphoid follicles were missing,a feature not observed in AMPKα1fl/fl mice.Furthermore,the knockout of AMPKα1 in Tregs reduced the CD3+T cell population,particularly the CD8+T cell population,in lymph nodes.Although the mature Treg cell population was significantly lower in AMPKα1fl/flFoxp3cre mice,the percentage of CD4+T cells was markedly in-creased.In contrast,there was no statistically significant difference in granulocyte populations between AMPKα1fl/flFoxp3cre and AMPKα1fl/fl mice.CONCLUSION:The populations of mature Tregs,CD8+T cells and eosinophils in various im-mune organs were significantly altered in mice with Treg-specific AMPKα1 knockout,suggesting a potential remodeling of the host immune microenvironment in response to inflammatory stimuli.

Objective To investigate the susceptibility and infection characteristics of dengue virus(DENV)in cells derived from diverse tissues of tree shrews and to provide a basis for expanding the repertoire of DENV-permissive cell models in this species.Methods DENV was inoculated at a multiplicity of infection(MOI)of 0.02 into tree shrew skin fibroblasts(TSFs),primary tree shrew renal epithelial cells(pTRECs),tree shrew aortic endothelial cells(TAECs),tree shrew aortic smooth muscle cells(TASMCs),tree shrew hepatocytes(THs),tree shrew corneal stromal cells(TCSCs),tree shrew brain microvascular endothelial cells(TBMECs),and tree shrew retinal microvascular endothelial cells(TRMECs).C6/36,Vero,A549,and BHK-21 cells(commonly used for DENV propagation)were used as positive controls.Over 6 days post-infection,cellular cytopathic effects were monitored at 12-hour intervals using an inverted microscope,viral RNA loads in cell lysates were quantified by real-time fluorescent quantitative PCR to generate proliferation curves,and viral titers were determined by plaque assay.Results Seven types of tree shrew cells,except TRMECs,were susceptible to DENV.Prolonged infection induced pronounced cytopathic effects,including cell rounding,detachment,necrosis,and lysis,across all susceptible cells.The viral RNA loads detected in lysates of pTRECs,TBMECs,TASMCs,TAECs and THs,approached those of positive controls(≥4×107 copies/μL).Infectious progeny viruses were produced by these five cell types,with three(TAECs,3.13×105 PFU/mL;THs,2.03×105 PFU/mL;pTRECs,1.58×105 PFU/mL)exhibiting titers comparable to C6/36(3.85×10 5 PFU/mL)and earlier viral harvests.Conclusion DENV exhibits broad susceptibility to tree shrew cells of multiple tissue origins,with proliferation rates surpassing those of conventional cell lines sourced from other species.TAECs,THs,and pTRECs are particularly suitable for large-scale DENV proliferation,suggesting their potential involvement in in vivo infection.