Antimicrobial resistance(AMR)has been a difficult problem of globally public health that can threaten human health.If its spread could not be effectively curbed,it will lead to increasing limitation in the options of clinical treatment.In the increasingly complex environment of diagnosis and treatment of infectious diseases,it is particularly urgent to build a rapid,accurate and widely popularized diagnosis system for pathogen,which is the key prerequisite of realizing accurate assessment for susceptibility of antimicrobial drug.This article comprehensively reviewed the main rapid diagnostic techniques currently used in AMR monitoring and detecting for antimicrobial susceptibility on the basis of the above conditions.It not only provided a detailed analysis of molecular typing methods represented by gene sequencing and phenotype identification methods based on conventional culture,but also focused on a new joint diagnostic strategy that combined genomic information with functional verification.Through comparative study for various technical principles,operating procedures and clinical applicability,the purpose of this paper was to provide scientific basis for optimizing the rapid detection scheme of drug susceptibility in clinical microbiology laboratories,and promote a comprehensive improvement of the level of diagnosis and treatment for infectious diseases,and ultimately improve the multi-layered network system of prevention and control for nosocomial infection.

ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

ObjectiveTo investigate the mechanism by which 2，3，5，4'-tetrahydroxyldiphenylethylene-2-O-glucoside （THSG） mitigates cerebral ischemia/reperfusion （CI/R） injury by regulating mitochondrial calcium overload and promoting mitophagy. MethodsSixty male SD rats were randomized into sham， model， SAS （40 mg·kg^-1）， and low-， medium- and high-dose （10， 20， 40 mg·kg^-1， respectively） THSG groups， with 10 rats in each group. The middle cerebral artery occlusion/reperfusion （MCAO/R） model was established by the modified Longa suture method. An oxygen-glucose deprivation/reoxygenation （OGD/R） model was constructed in PC12 cells. Neurological deficits were assessed via Zea Longa scoring， and cerebral infarct volume was measured by 2，3，5-triphenyltetrazolium chloride （TTC） staining. Structural and functional changes of cortical neurons in MCAO/R rats were assessed by hematoxylin-eosin and Nissl staining. PC12 cell viability was detected by cell counting kit-8 （CCK-8） assay， and mitochondrial calcium levels were quantified by Rhod-2 AM. Immunofluorescence was used to detect co-localization of PTEN-induced kinase 1 （PINK1） and leucine zipper/EF-hand-containing transmembrane protein 1 （LETM1） in neurons. Transmission electron microscopy （TEM） was employed to observe mitochondrial morphology in neurons. Western blot was employed to analyze the expression of translocase of outer mitochondrial membrane 20 （TOMM20）， autophagy-associated protein p62， microtubule-associated protein light chain 3 （LC3）， cysteinyl aspartate-specific proteinase-9 （Caspase-9）， B-cell lymphoma 2-associated protein X （Bax）， and cytochrome C （Cyt C）. ResultsCompared with the sham group， the model group exhibited increased infarct volume （P<0.01） and neurological deficit scores （P<0.01）， neuronal structure was disrupted with reduced Nissl bodies. （P<0.01）， mitochondrial swelling/fragmentation， decreased PINK1/LETM1 co-localization （P<0.01）， upregulated protein levels of LC3Ⅱ/LC3Ⅰ， TOMM20， Caspase-9， Bax， and Cyt C （P<0.01）， downregulated protein level of p62 （P<0.05）， weakened PC12 viability （P<0.01）， and elevated mitochondrial calcium level （P<0.01）. Compared with the model group， THSG and SAS groups showed reduced infarct volumes （P<0.05，P<0.01） and neurological deficit scores （P<0.05，P<0.01）， mitigated mitochondrial damage， and increased PINK1/LETM1 co-localization （P<0.01）. Medium/high-dose THSG and SAS alleviated the neurological damage， increased Nissl bodies （P<0.05，P<0.01）， downregulated the protein levels of p62， TOMM20， Caspase-9， Bax， and Cyt C （P<0.05，P<0.01）， and elevated the LC3Ⅱ/LC3Ⅰ level （P<0.05，P<0.01）. High-dose THSG enhanced PC12 cell viability （P<0.01）， increased PINK1/LETM1 co-localization （P<0.01）， and reduced mitochondrial calcium （P<0.01）. ConclusionTHSG may exert the neuroprotective effect on CI/R injury by activating the PINK1-LETM1 signaling pathway， reducing the mitochondrial calcium overload， and promoting mitophagy.

Objective:To compare the efficacy of oliceridine and sufentanil when combined with propofol for painless gastroscopy.Methods:In this randomized controlled trial, 66 patients of either sex, aged 18-64 yr, with a body mass index of 18-26 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰor Ⅱ, scheduled for elective painless gastroscopy from September 2024 to November 2024, were divided into 2 groups ( n=33 each) using a table of computer-generated random numbers: sufentanil combined with propofol group (SP group) and oliceridine combined with propofol group (OP group). Sufentanil 0.1 μg/kg was intravenously injected in group SP, oliceridine 0.02 mg/kg was intravenously injected in group OP, and 1 min later propofol 2 mg/kg was intravenously injected in both groups. When the modified Observer′s Assessment of Alertness and Sedation score ≤ 1, the painless gastroscopy was performed. Heart rate, mean arterial pressure and peripheral oxygen saturation were recorded on admission to the operating room, immediately after insertion of the gastroscope, and at the end of procedure. The success of sedation, time of gastroscopy, emergence time, consumption of propofol and use of vasoactive drugs were recorded. The occurrence of adverse events such as respiratory depression, hypotension, dizziness and nausea was also recorded. Results:Compared with group SP, the incidence of respiratory depression was significantly decreased in group OP ( P<0.05). There was no statistically significant difference in the incidence of hypotension, dizziness and nausea and heart rate, mean arterial pressure and peripheral oxygen saturation at different time points between the two groups ( P>0.05). Conclusions:Oliceridine provides better efficacy than sufentanil when combined with propofol in painless gastroscopy.

Objective To explore the molecular mechanisms by which cucurbitacin B(CuB)regulates the mitochondrial ap-optosis pathway through the nicotinamide phosphoribosyltransferase(NAMPT)/forkhead transcription factor O subfamily member 3a(FoxO3a)axis to affect the biological function of non-small cell lung cancer(NSCLC)cells.Methods Gefitinib(GEF)and CuB were used to intervene in A549 cells.Network pharmacology was employed to analyze the targets and downstream pathways,and different vector-transfected cell groups were set up.Cell viability was detected by CCK-8 assay,cell invasion by Transwell assay,and cell apoptosis by flow cytometry.The mitochondrial membrane potential(MMP)level was measured using a JC-1 kit,and intracellular reactive oxygen species(ROS)levels were detected using a kit.Western blot was used to detect the expression of mitochondrial dynamics-related proteins(DRP1,Mfn1),as well as the protein levels of NAMPT and FoxO3a in A549 cells.The mRNA expression of NAMPT was detected by qRT-PCR.Additionally,a xenograft model in nude mice was es-tablished to verify the therapeutic effects of CuB in vivo.Results Compared with the control group,GEF and CuB intervention significantly inhibited the viability and invasion of A549 cells and induced cell apoptosis.Moreover,CuB intervention significant-ly decreased the MMP level,increased ROS concentration,and upregulated the expression of DRP1 while downregulating Mfn1 in A549 cells(P＜0.05).NAMPT,a therapeutic target,was highly expressed in A549 cells and was inhibited by CuB.Compared with the vector group,overexpression of NAMPT significantly promoted the growth of A549 cells and inhibited mitochondrial apoptosis.Compared with the vector+CuB(100 nmol/L)group,overexpression of NAMPT reversed the effects of CuB on A549 cells(P＜0.05).Compared with the si-NAMPT-NC group,silencing NAMPT significantly inhibited the growth of A549 cells and promoted mitochondrial apoptosis,while knockdown of FoxO3a enhanced the viability and invasion of A549 cells and re-versed the damaging effects of si-NAMPT on cancer cells(P＜0.05).In vivo experimental results showed that CuB inhibited tumor weight and volume in a dose-dependent manner.Conclusion CuB regulates the NAMPT-FoxO3a axis through the mito-chondrial apoptosis pathway to inhibit the progression of NSCLC both in vitro and in vivo.

Objective To explore the molecular mechanisms by which cucurbitacin B(CuB)regulates the mitochondrial ap-optosis pathway through the nicotinamide phosphoribosyltransferase(NAMPT)/forkhead transcription factor O subfamily member 3a(FoxO3a)axis to affect the biological function of non-small cell lung cancer(NSCLC)cells.Methods Gefitinib(GEF)and CuB were used to intervene in A549 cells.Network pharmacology was employed to analyze the targets and downstream pathways,and different vector-transfected cell groups were set up.Cell viability was detected by CCK-8 assay,cell invasion by Transwell assay,and cell apoptosis by flow cytometry.The mitochondrial membrane potential(MMP)level was measured using a JC-1 kit,and intracellular reactive oxygen species(ROS)levels were detected using a kit.Western blot was used to detect the expression of mitochondrial dynamics-related proteins(DRP1,Mfn1),as well as the protein levels of NAMPT and FoxO3a in A549 cells.The mRNA expression of NAMPT was detected by qRT-PCR.Additionally,a xenograft model in nude mice was es-tablished to verify the therapeutic effects of CuB in vivo.Results Compared with the control group,GEF and CuB intervention significantly inhibited the viability and invasion of A549 cells and induced cell apoptosis.Moreover,CuB intervention significant-ly decreased the MMP level,increased ROS concentration,and upregulated the expression of DRP1 while downregulating Mfn1 in A549 cells(P＜0.05).NAMPT,a therapeutic target,was highly expressed in A549 cells and was inhibited by CuB.Compared with the vector group,overexpression of NAMPT significantly promoted the growth of A549 cells and inhibited mitochondrial apoptosis.Compared with the vector+CuB(100 nmol/L)group,overexpression of NAMPT reversed the effects of CuB on A549 cells(P＜0.05).Compared with the si-NAMPT-NC group,silencing NAMPT significantly inhibited the growth of A549 cells and promoted mitochondrial apoptosis,while knockdown of FoxO3a enhanced the viability and invasion of A549 cells and re-versed the damaging effects of si-NAMPT on cancer cells(P＜0.05).In vivo experimental results showed that CuB inhibited tumor weight and volume in a dose-dependent manner.Conclusion CuB regulates the NAMPT-FoxO3a axis through the mito-chondrial apoptosis pathway to inhibit the progression of NSCLC both in vitro and in vivo.

Objective:To compare the efficacy of oliceridine and sufentanil when combined with propofol for painless gastroscopy.Methods:In this randomized controlled trial, 66 patients of either sex, aged 18-64 yr, with a body mass index of 18-26 kg/m 2, of American Society of Anesthesiologists Physical Status classification Ⅰor Ⅱ, scheduled for elective painless gastroscopy from September 2024 to November 2024, were divided into 2 groups ( n=33 each) using a table of computer-generated random numbers: sufentanil combined with propofol group (SP group) and oliceridine combined with propofol group (OP group). Sufentanil 0.1 μg/kg was intravenously injected in group SP, oliceridine 0.02 mg/kg was intravenously injected in group OP, and 1 min later propofol 2 mg/kg was intravenously injected in both groups. When the modified Observer′s Assessment of Alertness and Sedation score ≤ 1, the painless gastroscopy was performed. Heart rate, mean arterial pressure and peripheral oxygen saturation were recorded on admission to the operating room, immediately after insertion of the gastroscope, and at the end of procedure. The success of sedation, time of gastroscopy, emergence time, consumption of propofol and use of vasoactive drugs were recorded. The occurrence of adverse events such as respiratory depression, hypotension, dizziness and nausea was also recorded. Results:Compared with group SP, the incidence of respiratory depression was significantly decreased in group OP ( P<0.05). There was no statistically significant difference in the incidence of hypotension, dizziness and nausea and heart rate, mean arterial pressure and peripheral oxygen saturation at different time points between the two groups ( P>0.05). Conclusions:Oliceridine provides better efficacy than sufentanil when combined with propofol in painless gastroscopy.

Antimicrobial resistance(AMR)has been a difficult problem of globally public health that can threaten human health.If its spread could not be effectively curbed,it will lead to increasing limitation in the options of clinical treatment.In the increasingly complex environment of diagnosis and treatment of infectious diseases,it is particularly urgent to build a rapid,accurate and widely popularized diagnosis system for pathogen,which is the key prerequisite of realizing accurate assessment for susceptibility of antimicrobial drug.This article comprehensively reviewed the main rapid diagnostic techniques currently used in AMR monitoring and detecting for antimicrobial susceptibility on the basis of the above conditions.It not only provided a detailed analysis of molecular typing methods represented by gene sequencing and phenotype identification methods based on conventional culture,but also focused on a new joint diagnostic strategy that combined genomic information with functional verification.Through comparative study for various technical principles,operating procedures and clinical applicability,the purpose of this paper was to provide scientific basis for optimizing the rapid detection scheme of drug susceptibility in clinical microbiology laboratories,and promote a comprehensive improvement of the level of diagnosis and treatment for infectious diseases,and ultimately improve the multi-layered network system of prevention and control for nosocomial infection.

Objective:To investigate the effect of low expression of silencing information regulator-6 (SIRT-6) on inflammatory reaction and autophagy in monocytes.Methods:Human acute monocytic leukemia cell line THP-1 was transfected with si-SIRT6 to establish THP-1 cell line with low expressed SIRT-6. The cells were divided into control group, MUS group and MUS+ RAPA group. Cells in control group were cultured with medium added with PBS, cells in MUS Group were cultured with medium added with MUS, and cells in MUS+ RAPA Group were added with MUS and Rapamycin. Cells in each group were cultured for 48 hours. The levels of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in the supernatant of each group were detected by enzyme-linked immunosorbent assay (ELISA). The gene expression levels of autophagy-associated protein-5 (ATG-5), Beclin-1, lysosomal-associated membrane protein-1 (LAMP-1), microtubule-associated protein-1 light chain 3B (LC3B) and p62 in cells of each group were detected by Q-PCR. The protein expression levels of p62, ATG-5 and LC3B Ⅱ/LC3BⅠ in cells of each group. The one-way analysis of variance (ANOVA) was used for the measurement data in multi-groups, and the LSD- t test was used for the measurement data in both groups. Results:The gene and protein expression of SIRT-6 in THP-1 cells decreased significantly after si-SIRT6 transfection (Gene: 1.09±0.08 vs. 0.57±0.03, t=14.91, P<0.001; Protein: 0.21±0.04 vs. 0.12±0.03, t=4.41, P=0.070). The levels of IL-1β, IL-6, and TNF-α in the supernatant of si-SIRT6/si-SIRT6 NC-transfected THP-1 cells increased significantly by MUS ( P<0.05), and the levels of IL-1β, IL-6, and TNF-α in the supernatant of cells further increased by MUS ( P<0.05). The levels of IL-1β, IL-6 and TNF-α in the supernatant of si-SIRT6-transfected THP-1 cells increased significantly compared with those of si-SIRT6 NC-transfected THP-1 cells ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The gene expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA ( P<0.05), the gene expression of ATG-5, Beclin-1, LAMP-1 and LC3B in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The gene expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the gene expression level of ATG-5, LC3B, Beclin-1 and LAMP-1 significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly decreased by MUS ( P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ protein in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells significantly increased by MUS ( P<0.05). The protein expression of p62 in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further decreased by RAPA P<0.05), the protein expression of ATG-5 and LC3B Ⅱ/LC3BⅠ in si-SIRT6/si-SIRT6 NC-transfected THP-1 cells further increased by RAPA ( P<0.05). The protein expression level of p62 in si-SIRT6 transfected THP-1 cells significantly decreased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05), and the protein expression level of ATG-5 and LC3B Ⅱ/LC3BⅠ significantly increased than that in si-SIRT6 NC transfected THP-1 cells ( P<0.05). Conclusion:Low expression of SIRT-6 gene can promote inflammatory reaction and autophagy in monocytes, and Monosodium urate and autophagy agonist rapamycin can aggravate inflammatory reaction and autophagy.

AIM:To compare the recovery of psy-chomotor function after intravenous anesthesia with remazolam or propofol compound alfentanil in patients undergoing painless gastrointestinal en-doscopy.METHODS:78 patients undergoing pain-less gastrointestinal endoscopy were randomly di-vided into group RA and group PA.Remiazolam or propofol combined with alfentanil were given intra-venously in group RA or group PA.The blood pres-sure,heart rate,respiratory rate and saturation of puls oxygen were recorded before procdure(T1),during checking(T2),awaking from anaesthesia(T3)and at discharging from PACU(T4).Psychomo-tor function,as measured by the Trieger's dot test(TDT)and digit symbol substitution test(DSST),were evaluated before anesthesia(T1),at discharg-ing from PACU(T4),1 h(T5)and 2 h(T6)after checking.RESULTS:From assessment of the TDT,number of dots missed(NDM),maximum distance of dots missed(MDDM)and average distance of dots missed(ADDM)at T4,T5 were significantly lower than those at T1 in two groups.The comple-tion rates and accuracy rates of DSST at T4,T5 were significantly lower than those at T1.Results of TDT and DSST at T6 were not significantly differ-ent to those at T1.The results of NDT,MDDM and ADDM at T4,T5 in group RA were significantly low-er than those in group PA.The completion rates and accuracy rates of DSST at T4,T5 in group RA increased significantly compared with group PA.Compared to group PA,the incidence of hypoten-sion was significantly lower in group RA.There was no significant difference in the incidence of respira-tory depression between the two groups.CONCLU-SION:Psychomotor function was fully recovered 2 h after surgery when remazolam compound alfent-anil was used for painless gastrointestinal endosco-py.Compared with propofol,psychomotor function recovery in the remazolam group was faster and there were fewer adverse effects after surgery in group RA.