Objective To gain insights into the biological characteristics of different populations of Culex tritaeniorhynchus within and around the Yellow Sea wetland from the perspective of the occurrence of resistance, we investigated the levels of resistance to deltamethrin and kdr gene mutation in the wetland and its peripheral areas. Methods Specimens were collected from Cx. tritaeniorhynchus populations at two monitoring sites in the Rare Bird National Nature Reserve and Tiaozi Ni Wetland Scenic Area, and also from two populations in Yancheng City and the Liuhe District of Nanjing, and the resistance of these mosquitoes to deltamethrin was determined using the CDC biotest bottle method. For each concentration of deltamethrin assessed, a random subset of exposed specimens was selected for amplification of the kdr gene fragment, followed by Sanger sequencing to identify and analyze resistance-associated mutations. Results The LC50 levels of deltamethrin among mosquitoes from the four populations in Luhe, Yancheng, the Rare Bird National Nature Reserve and the Tiaozi Ni Wetland Scenic Area were 2.048 5, 7.798 2, 3.473 3, and 17.695 5 mg/mL, respectively, with corresponding concentrations of deltamethrin ranging from 0.005 to 5.000,0.050 to 50.000,0.050 to 25.000 and 0.050 to 50.000 mg/mL, respectively. Furthermore, the ranges of the KT50 values were 11.76-107.43, 67.05-216.30,29.77-107.43 and 28.40-329.51 min; the 1-h knockdown rates were 34.58%-99.15%, 9.52%-43.80%, 55.09%-73.01%, and 10.09%-68.07%; and the 24-h mortality rates were 12.15%-67.52%,9.52%-79.56%,13.17%-82.21%, and 11.01%-78.99%, respectively. With respect to kdr gene mutation, we assayed a total of 63,70,59, and 57 mosquitoes for the four populations, for which we detected L1014F mutation frequencies of 14.29%, 35.00%, 20.34%, and 31.58%, respectively, with a majority of these mutations being heterozygous for resistance. In addition, five adult mosquitoes were identified has having synonymous mutations at site 1011[i. e. , AAT(asparagine)mutation to AAC(asparagine)]. Conclusions Our findings revealed the clear resistance of Cx. tritaeniorhynchus to deltamethrin in the Yancheng region of the Yellow Sea wetland, and the resistance phenotype and kdr frequency of Cx. tritaeniorhynchus in the wetland environment were comparable to those of Cx. tritaeniorhynchus in the wetland environment, thereby indicating that the resistance of different populations of Cx. tritaeniorhynchus was homogeneous under the pressure of different insecticide selection within and around the wetland. However, the underlying mechanisms need to be further studied.

Objective:To investigate effect and mechanism of hydroxysafflor yellow A(HSYA)activating neuronal autophagy on cerebral ischemia-reperfusion injury through a combination of in vitro and in vivo experiments.Methods:SD rat MCAO/R model was established by improved suture method.Rats were randomly divided into sham surgery(Sham)group,MCAO/R group and MCAO/R+HSYA group,following indicators were detected to determine extent of cerebral ischemia-reperfusion nerve damage:Z-Longa neu-rological function score was detected,TTC staining to measure cerebral infarction area,and TUNEL staining to measure cell apopto-sis;Western blot was used to detect protein expressions of autophagy related markers LC3,Beclin1,P62 and SIRT1 in rat brain tis-sue;immunofluorescence staining was used to observe expression of LC3 co-localization with neurons.OGD/R injury model of SH-SY5Y cells was established and randomly divided into Normal group,OGD/R group,OGD/R+HSYA group,OGD/R+SIRT1 inhibitor(EX-527)group and OGD/R+EX-527+HSYA group.Western blot was used to detect protein expressions of LC3,Beclin1,P62 and SIRT1.Results:Compared with Sham group,model group rats showed impaired neurological function,significantly increased neu-robehavioral scores,widespread cerebral infarction,significantly increased neuronal cell apoptosis,significantly increased autophagy related protein Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,significantly decreased P62 expression,significantly increased LC3/NeuN co-stained cells,and decreased SIRT1 expression;compared with model group,HSYA intervention group showed a significant decrease in neurological functional scores,a significant reduction in cerebral infarction area,a significant decrease in neuronal cell apoptosis,a further increase in Beclin1 expression and LC3-Ⅱ/LC3-Ⅰ,a further decrease in P62 expression,number of LC3/NeuN and P62/NeuN co-stained cells also increased,and SIRT1 expression significantly increased.Expression trends of Beclin1,LC3-Ⅱ/LC3-Ⅰ,P62 and SIRT1 of cells between normal group,model group and HSYA intervention group were same as animal experiment;compared with model group,expressions of SIRT1,Beclin1 and LC3-Ⅱ/LC3-Ⅰ in OGD/R+EX-527 group were significantly reduced,while expression of P62 was significantly increased;compared with OGD/R+EX-527 group,there was no significant change in SIRT1 expression in OGD/R+EX-527+HSYA group,LC3-Ⅱ/LC3-Ⅰ and Beclin1 expression were significantly increased,and P62 expres-sion was significantly decreased.Conclusion:HSYA can significantly improve neurological deficits in rats after cerebral ischemia-reperfusion,reduce cerebral infarction area,and decrease neuronal cell apoptosis rate,whose neuroprotective effect may be related to its activation of SIRT1,which significantly enhances neuronal autophagy.

Objective To explore the safety of establishing the veno-arterial extracorporeal membrane oxygenation(VA-ECMO)pipeline through percutaneous puncture and the effectiveness of high-risk percutaneous coronary intervention(HR-PCI)under VA-ECMO assistance.Methods In this single-center retrospective study,patients who underwent HR-PCI assisted by VA-ECMO at Wuhan Asian Heart Hospital from December 2022 to December 2023 were included.The primary endpoint was the safety of establishing the VA-ECMO pipeline through percutaneous puncture,mainly characterized by extracorporeal membrane oxygenation(ECMO)-related complications,including severe bleeding/hematoma at the wound,arteriovenous fistula,pseudoaneurysm,peripheral arterial dissection,etc.The secondary endpoint was the effectiveness of HR-PCI under VA-ECMO assistance,defined as major adverse cardiovascular and cerebrovascular events(MACCE)at 6-month follow-up,including the composite events of all-cause death,non-fatal myocardial infarction,repeat revascularization,and ischemic stroke.Results Among the 189 patients included in the study,15(7.9％)had severe bleeding/hematoma at the puncture wound caused by VA-ECMO,8(4.2％)had wound infection,1(0.5％)had arteriovenous fistula,4(2.1％)had pseudoaneurysm,3(1.6％)had peripheral arterial dissection,8(4.2％)had systemic embolism,8(4.2％)had acute kidney injury,3(1.6％)had acute neurological injury,and 1(0.5％)died.percutaneous coronary intervention(PCI)was successful in 182 cases(96.3％),and a total of 36 cases(19.0％)had PCI-related complications.164 patients completed 6-month follow-up,among which 4(2.4％)had all-cause death,11(6.7％)had non-fatal myocardial infarction,11(6.7％)had unplanned repeat revascularization,and 2(1.2％)had the composite event of ischemic stroke.Conclusions Ultrasound-guided percutaneous puncture for establishing the VA-ECMO pipeline has high safety,and HR-PCI patients assisted by VA-ECMO have good short-term effects.

Systemic lupus erythematosus(SLE)complicated refractory immune thrombocytopenia(ITP)is mainly characterized by persistent severe thrombocytopenia and bleeding.Currently,there are no guidelines for treating such ITP.Hereby we reported a patient diagnosed with SLE with more than 10 years history of anemia and thrombocytopenia.This patient had been treated by various immunity inhibitors,such as corticosteroids,cyclosporine,mycophenolate mofetil,azathioprine,tofacitinib and telitacicept,but there was no obvious effect.The platelet count continued to be below 50×109/L.However,when serum cyclosporine trough level between 100 and 150 ng/mL,the clinical efficacy was improved and the platelet count was above 100×109/L.The patient was followed up for more than 6 months,and no recurrence of disease was found.Cyclosporine combined with low-dose glucocorticoid is safe and effective for SLE associated ITP.Therapeutic drug monitoring is a powerful tool for improving clinical efficacy and ensuring drug safety.

Systemic lupus erythematosus(SLE)complicated refractory immune thrombocytopenia(ITP)is mainly characterized by persistent severe thrombocytopenia and bleeding.Currently,there are no guidelines for treating such ITP.Hereby we reported a patient diagnosed with SLE with more than 10 years history of anemia and thrombocytopenia.This patient had been treated by various immunity inhibitors,such as corticosteroids,cyclosporine,mycophenolate mofetil,azathioprine,tofacitinib and telitacicept,but there was no obvious effect.The platelet count continued to be below 50×109/L.However,when serum cyclosporine trough level between 100 and 150 ng/mL,the clinical efficacy was improved and the platelet count was above 100×109/L.The patient was followed up for more than 6 months,and no recurrence of disease was found.Cyclosporine combined with low-dose glucocorticoid is safe and effective for SLE associated ITP.Therapeutic drug monitoring is a powerful tool for improving clinical efficacy and ensuring drug safety.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Objective By observing the regulatory effect of Strengthening Spleen and Eliminating Cancer Formula on N6-methyladenosine(m6A)methyltransferase,To explore the effect of Strengthening Spleen and Eliminating Cancer Formula on inhibiting the IRX5 m6A level in colorectal cancer(CRC),the regulatory effect on N6-methyladenosine(m6A)methyltransferase was observed.Methods Clinically,m6A hypermethylated genes in colorectal cancer was analyzed by m6A sequencing of pathological tissues from five CRC patients after radical surgery,looking for protein detection indexes for validation.23 BALB/c nude mice were selected and injected with HCT116 cells to establish a nude-mouse transplantation model of human colorectal cancer.They were divided into Model group,Western medicine group(5-fluorouracil group),Chinese medicine group(Jianpi Xiaoai Formula low-dose group,Jianpi Xiaoai Formula high-dose group),with 6 rats in each group,5 rats in control group.The tumor volume of all groups was compared.The overall methylation level of m6A was detected by colorimetric method.The protein expression levels of METTL3,METTL14,and WTAP,in tumor were detected by Western blot.The SRAMP website was used to predict the m6A sites of IRX5.HCT116 cells were treated with oe-NC,oe-METTL3,sh-NC,and sh-METTL3.The expression of IRX5 protein was detected by Western blot.HCT116 cell line was treated with Jianpi Xiaoai Formula drug-containing serum,and transfected with oe-METTL3 and oe-IRX5.The group was set as followed:control group,Jianpi Xiaoai Formula drug-containing serum group,Jianpi Xiaoai Formula drug-containing serum group+oe-NC,Jianpi Xiaoai Formula drug-containing serum group+oe-METTL3,Jianpi Xiaoai Formula drug-containing serum group+oe-IRX5,cell cloning experiment and Transwell experiment were performed to detect cell proliferation,migration and invasion ability of each group.The protein expression levels of METTL3 and IRX5 were detected by Western blot.Results The results of m6A sequencing of genes showed that the m6A methylation level increased in patients with CRC,and the m6A methylation levels of SOX1 and IRX5 were significantly elevated.Compared with the model group,the tumor volume of Jianpi Xiaoyou Formula high-dose group,low-dose group and 5-Fu group decreased significantly(P<0.01),and the tumor inhibition effect was more obvious with the increase of Jianpi Xiaoai Formula concentration(P<0.01).The methylation level of m6A in Jianpi Xiaoai Formula high dose group,low dose group and 5-Fu group decreased significantly(P<0.01).The SRAMP website predicted that IRX5 contained multiple m6A sites.Overexpression of METTL3 promoted the expression of IRX5 protein(P<0.001),while knockdown of METTL3 inhibited the expression of IRX5 protein(P<0.001).The drug-containing serum of Jianpi Xiaoai Formula could inhibit the protein expression of METTL3 and IRX5(P<0.05)and inhibit the proliferation,migration and invasion of HCT116(P<0.01).Overexpression of METTL3 and IRX5 reversed the inhibitory effect of Jianpi Xiaoai Formula on HCT116 evil phenotype(P<0.01).Conclusion Jianpi Xiaoai Formula may inhibit METTL3 expression mediated IRX5 low expression to inhibit the progression of colorectal cancer.

Over 40%of critically ill patients will develop coagulopathy.Once critically ill patients are complicated with coagulopathy,the incidence of bleeding and mortality can increase by more than 4 times.Early identification of coagulopathy and accurate evaluation of coagulation function are essential for correcting coagulopathy as soon as possible.Therefore,Chinese Society of Thrombosis,Hemostasis and Critical Care,Chinese Medicine Education Association,together with Chinese People's Liberation Army Professional Committee of Critical Care Medicine updated the"Chinese expert consensus on standardized assessment of severe coagulopathy(2025 Edition)"on the basis of the"Consensus of Chinese experts on standardized evaluation of coagulation dysfunction in severe patients"formulated in 2022.This consensus includes four parts:classification and typing,etiology and mechanism,assessment methods,and diagnostic criteria of severe coagulopathy,with a total of 14 recommendations,aiming to provide corresponding guidance for clinical practice.

Objective To explore the safety of establishing the veno-arterial extracorporeal membrane oxygenation(VA-ECMO)pipeline through percutaneous puncture and the effectiveness of high-risk percutaneous coronary intervention(HR-PCI)under VA-ECMO assistance.Methods In this single-center retrospective study,patients who underwent HR-PCI assisted by VA-ECMO at Wuhan Asian Heart Hospital from December 2022 to December 2023 were included.The primary endpoint was the safety of establishing the VA-ECMO pipeline through percutaneous puncture,mainly characterized by extracorporeal membrane oxygenation(ECMO)-related complications,including severe bleeding/hematoma at the wound,arteriovenous fistula,pseudoaneurysm,peripheral arterial dissection,etc.The secondary endpoint was the effectiveness of HR-PCI under VA-ECMO assistance,defined as major adverse cardiovascular and cerebrovascular events(MACCE)at 6-month follow-up,including the composite events of all-cause death,non-fatal myocardial infarction,repeat revascularization,and ischemic stroke.Results Among the 189 patients included in the study,15(7.9％)had severe bleeding/hematoma at the puncture wound caused by VA-ECMO,8(4.2％)had wound infection,1(0.5％)had arteriovenous fistula,4(2.1％)had pseudoaneurysm,3(1.6％)had peripheral arterial dissection,8(4.2％)had systemic embolism,8(4.2％)had acute kidney injury,3(1.6％)had acute neurological injury,and 1(0.5％)died.percutaneous coronary intervention(PCI)was successful in 182 cases(96.3％),and a total of 36 cases(19.0％)had PCI-related complications.164 patients completed 6-month follow-up,among which 4(2.4％)had all-cause death,11(6.7％)had non-fatal myocardial infarction,11(6.7％)had unplanned repeat revascularization,and 2(1.2％)had the composite event of ischemic stroke.Conclusions Ultrasound-guided percutaneous puncture for establishing the VA-ECMO pipeline has high safety,and HR-PCI patients assisted by VA-ECMO have good short-term effects.

Objective To investigate the distribution and antimicrobial resistance profiles of clinically isolated Haemophilus influenzae and Moraxella catarrhalis in hospitals across China from 2015 to 2021,and provide evidence for rational use of antimicrobial agents.Methods Data of H.influenzae and M.catarrhalis strains isolated from 2015 to 2021 in CHINET program were collected for analysis,and antimicrobial susceptibility testing was performed by disc diffusion method or automated systems according to the uniform protocol of CHINET.The results were interpreted according to the CLSI breakpoints in 2022.Beta-lactamases was detected by using nitrocefin disk.Results From 2015 to 2021,a total of 43 642 strains of Haemophilus species were isolated,accounting for 2.91％of the total clinical isolates and 4.07％of Gram-negative bacteria in CHINET program.Among the 40 437 strains of H.influenzae,66.89％were isolated from children and 33.11％were isolated from adults.More than 90％of the H.influenzae strains were isolated from respiratory tract specimens.The prevalence of β-lactamase was 53.79％in H.influenzae strains.The H.influenzae strains isolated from children showed higher resistance rate than the strains isolated from adults.Overall,779 strains of H.influenzae did not produce β-lactamase but were resistant to ampicillin(BLNAR).Beta-lactamase-producing strains showed significantly higher resistance rates to these antimicrobial agents than the β-lactamase-nonproducing strains.Of the 16 191 M.catarrhalis strains,80.06％were isolated from children and 19.94％isolated from adults.M.catarrhalis strains were mostly susceptible to both amoxicillin-clavulanic acid and cefuroxime,evidenced by resistance rate lower than 2.0％.Conclusions The emergence of antibiotic-resistant H.influenzae due to β-lactamase production poses a challenge for clinical anti-infective treatment.Therefore,it is very important to implement antibiotic resistance surveillance for H.influenzae and guide rational antibiotic use.All local clinical microbiology laboratories should actively improve antibiotic susceptibility testing and strengthen antibiotic resistance surveillance for H.influenzae.