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.

ObjectiveTo investigate the current awareness and optimization recommendations on pneumonia with unknown etiology (PUE) surveillance among Shanghai healthcare professionals, and to provide evidence for the improvement of the surveillance system. MethodsIn December 2024, healthcare professional participants from diverse medical and health institutions were randomly selected within each stratum using a stratified cluster sampling method in all 16 districts of Shanghai for questionnaire surveys on their knowledge and suggestions for optimization of the PUE surveillance system. Descriptive statistics analyses, multivariate logistic regressions, and content analyses were used for data analyses. ResultsA total of 385 survey subjects were involved in the study, including 119 professional staffs from disease prevention and control centers and 266 from medical institutions. Those who understood the case definition for PUE correctly accounted for 67.01% (258/385). Multivariate regression model analyses showed that female gender (aOR=2.22, 95%CI: 1.36‒3.66), professionals from disease control institutions (aOR=2.02, 95%CI: 1.12‒3.72), and those who self-assessed as being familiar with the monitoring system (aOR=2.18, 95%CI:1.23‒3.89) had better understanding of the definition. Overall, 89.61% (345/385) of the survey subjects acknowledged the necessity of the surveillance system. Small number of cases meeting the case definition (54.46%, 214/379), insufficient diagnostic awareness (39.84%, 151/379), and lack of incentives for case reporting (29.29%, 111/379) were major problems for the current surveillance system, with variations observed across different types of institutions. Suggested segments for optimizing the surveillance system included monitoring purpose, case definition, form of monitoring and reporting procedure, organization mechanism, and epidemiological investigation and response. ConclusionThe necessity of PUE surveillance is commonly recognized among healthcare workers. However, the existing system falls short in terms of mastery of case definition and overall surveillance adaptability. There is an urgent need for optimization and improvement to enhance the capacity for preventing and controlling emerging and sudden respiratory infectious diseases.

Background and aims As a ferroptosis inducer, sorafenib, a first-line treatment for hepatocellular carcinoma (HCC), has a significant antitumor effect. Nonetheless, HCC patients frequently develop sorafenib resistance. Here, we investigated the impacts of progestagen-associated endometrial protein (PAEP), which controls sorafenib resistance and tumorigenesis in HCC. Furthermore, we investigated the function of PAEP and the underlying molecular mechanisms that cause HCC ferroptosis when sorafenib is applied. Methods Western blot analysis, cell proliferation, colony formation and animal experiments were performed to investigate the function of PAEP in sorafenib resistance and tumorigenesis in HCC. We detected the levels of reactive oxygen species, iron, and malondialdehyde and preformed transmission electron microscopy to investigate the relationship between PAEP and ferroptosis. Co-immunoprecipitation (co-IP) assay was performed to investigate the underlying molecular mechanisms of PAEP that cause HCC ferroptosis. Results PAEP was significantly overexpressed in HCC tissues, and this was associated with a poor clinical prognosis. PAEP silencing dramatically reduced HCC cells’ malignant phenotype. We found that sorafenib-induced ferroptosis was more sensitive to HCC cells with PAEP knockdown. Furthermore, the orthotopic cell line-derived xenograft mouse model results showed that sorafenib sensitivity can be effectively increased in vivo by PAEP knockdown. We determined that transferrin (TF) was a PAEP target using the String database, and we further supported this finding with Co-IP analysis. Additionally, on sorafenib-induced ferroptosis in HCC cells, TF partially reversed the effects of PAEP knockdown. Conclusions Our research indicates that PAEP may be a potential biomarker for predicting sorafenib resistance in HCC and disruption of PAEP expression may be a potential cancer-directed therapeutic option for HCC.

BackgroundPatients with bipolar disorder exhibit persistent cognitive impairment across multiple domains， including executive function， memory， and attention. Furthermore， those with mixed features tend to have a poorer prognosis. Currently， research evidence regarding the characteristics of cognitive function and event-related potential P300， as well as their correlation， in patients with depressive episodes of bipolar disorder with mixed features remains limited. ObjectiveTo explore the correlation between cognitive function and event-related potential P300 in patients with depressive episodes of bipolar disorder with mixed features， analyze the influencing factors of cognitive function， so as to provide references for the improvement of cognitive function in this subtype of patients. MethodsA total of 73 patients who met the diagnostic criteria for depressive episodes of bipolar disorder in the Diagnostic and Statistical Manual of Mental Disorders， fifth edition （DSM-5） were enrolled from the outpatient and inpatient departments of Tianjin Anding Hospital from November 2021 to April 2024. The Hamilton Depression Scale-17 item（HAMD-17） was used to assess the severity of depressive symptoms， and the Clinically Useful Depression Outcome Scale Supplemented with Questions for the DSM-5 Mixed Features Specifier （CUDOS-M） was used to evaluate whether patients presented with mixed features. Based on the CUDOS-M total scores， patients were divided into two groups： the mixed features group （n=34） and the non-mixed features group （n=39）. The MATRICS Consensus Cognitive Battery （MCCB） was used to assess cognitive function， and auditory event-related potential P300 testing was performed. Spearman correlation analysis was used to examine the correlations between P300 indicators and MCCB scores. Multiple linear regression analysis was conducted to investigate the influencing factors of cognitive function. ResultsPatients with depressive episodes of bipolar disorder with mixed features had a longer total illness duration than those without mixed features， and the difference was statistically significant （t=2.030， P<0.05）. Analysis of covariance showed that the MCCB scores for processing speed， attention/vigilance， and reasoning and problem solving were all lower in the mixed features group than those in the non-mixed features group （F=3.650， 3.520， 4.213， P<0.01）. However， no significant difference was found between the two groups in P300 latency or amplitude （P>0.05）. Correlation analysis revealed that the MCCB attention/vigilance score was negatively correlated with P300 latency in patients with depressive episodes of bipolar disorder with mixed features （r_s=-0.576， P<0.01）. Multiple linear regression analysis showed that， in the mixed features group， years of education positively predicted processing speed （β=0.378， P<0.05）， while P300 latency （β=-0.699， P<0.01） and age （β=-0.424， P<0.01） negatively predicted attention/vigilance function. ConclusionPatients with depressive episodes of bipolar disorder with mixed features have a longer illness duration and more severe impairment in three cognitive domains： processing speed， attention/vigilance， and reasoning and problem solving. P300 latency may be an influencing factor of attention/vigilance function in patients with depressive episodes of bipolar disorder with mixed features ［Funded by Tianjin Key Medical Discipline Construction Project （number， TJYXZDXK-3-015B）； Hospital-level Scientific Research Project of Tianjin Anding Hospital （number， ADYKHT2021010）］

Objective:In order to clarify the ABO phenotype and genotype,and explore the molecular biological mechanism,serological detection,genotyping and gene sequencing were performed on an upper gastrointestinal hemorrhage patient with inconsistent forward and reverse ABO blood typing.Methods:ABO forward and reverse blood typing,H antigen identification,capillary centrifugation test and salivary substance detection were performed by classical serological method,moreover,polymerase chain reaction-sequence specific primer(PCR-SSP)was used for ABO genotyping,ABO gene 1-7 exons were sequenced by Sanger analysis in order to identify mutation.Results:Mixed field agglutination with anti-A,anti-AB and no agglutination with anti-A1 were appeared in the forward typing tests,agglutination with B cells but no agglutination with A1 cells and O cells were appeared in the reverse typing tests.3+agglutination strength was showed with anti-H.In capillary centrifugation experiment,erythrocyte after isolation in proximal part and distal end had same strength of agglutination with anti-A.Substances A and H were detected in saliva.The patient was assigned an A3 phenotype according to serological characteristics.Sequencing results of ABO gene 1-7 exons showed c.261delG,c.467C＞T,c.865A＞G,in which,865A＞G was the first discovered mutation,and this new mutation had been submitted to GenBank with accession number PP187306.Conclusion:A novel site mutation c.865A＞G is reported in this study,and this new mutation can result in a replacement of Met with Val at residue 289(p.Met289Val)and lead to an A3 phenotype.

Objective:To explore the risk factors and etiological characteristics of urinary tract infection caused by urinary calculi in eastern Fujian region,in order to attract clinical attention and improve the prevention and treatment of urinary tract infection caused by urinary calculi.Methods:A total of 154 patients with urinary calculi admitted to Ningde People's Hospital(n=80)and Ningde Mindong Hospital(n=74)from November 2022 to October 2023 were selected as the main research objects.According to whether the patients had urinary tract infection,they were divided into infection group and non-infection group.The baseline data of the two groups were analyzed in detail,and the risk factors and pathogen distribution of urinary tract infection in patients with urinary calculi were analyzed.Results:There were 33 cases of urinary tract infection in 154 patients with urinary calculi,accounting for 21.43％.Univariate analysis showed that the urinary white blood cell count in the infection group was higher than that in the non-infection group,and the proportion of patients with effusion,urinary tract obstruction,calculi in the upper urinary tract,staghorn calculi,smoking history,diabetes,and urinary nitrite positive was higher than that in the uninfected group(P＜0.05).The results of binary logistic regression analysis showed that effusion,urinary tract obstruction,staghorn calculi,smoking history,diabetes,high urine white blood cell count and positive urine nitrite were independent risk factors for urinary tract infection in patients with urinary calculi(OR＞1,P＜0.05).A total of 33 strains of pathogenic bacteria were isolated from 33 patients in the infection group.Among them,23 strains(69.70％)were gram-negative bacteria,8 strains(24.24％)were gram-positive bacteria,and 2 strains(6.06％)were fungi.Among gram-negative bacteria,escherichia coli accounted for the highest proportion(48.48％),followed by klebsiella pneumoniae(9.09％).Among gram-positive bacteria,enterococcus faecalis accounted for the highest proportion(12.12％),followed by enterococcus faecium(6.06％).Candida and candida tropicalis in fungi was the same,accounted for 3.03％.Conclusion:The risk of urinary tract infection in patients with urinary calculi in eastern Fujian region is high.Effusion,urinary tract obstruction,staghorn calculi,smoking history,diabetes,high urine white blood cell count and positive urine nitrite are independent risk factors for urinary tract infection in patients with urinary calculi.The main urinary tract pathogens are gram-negative bacteria.

Objective:To compare the application of cloning sequencing and third-generation nanopore sequencing technology in the identification of raw materials mixed in Tibetan patent medicine Shiliujianwei powder,and to pro-vide a reference for the establishment of the specific identification method for the formulation of medicinal prepara-tions using the raw powder of medicinal herbs or herbal pieces.Methods:By investigating the different concentra-tions of ExTaq enzyme,genomic DNA,upstream and downstream primers in the PCR amplification system,the suit-able PCR amplification system of genomic DNA universal primers for self-made Tibetan medicine Shiliujianwei powder was determined.The amplified products of Shiliujianwei powder were sequenced by two methods,the first was cloned and sequenced by Sanger sequencing technology,and the second was sequenced by third generation nanopore sequen-cing technology.Results:The addition of ExTaq enzyme,upstream and downstream primers and genomic DNA in 20 μL PCR amplification system of Shiliujianwei powder and single medicinal samples were determined to be 0.4,1.5 and 1 μL,respectively.The amplification products of the DNA from self-made Shiliujianwei powder were cloned and sequenced by Sanger sequencing technology,then only the ITS2 sequence of Carthami flos was obtained.The amplified products of self-made pomegranate Jianwei powder DNA were sequenced by three-generation nanopore se-quencing technology,and the ITS2 sequences of Granati semen,Carthami flos,Piperis longi fructus and Amomi fruc-tus rotundus were finally obtained,but the ITS2 sequence of cinnamomi cortex was not obtained.Conclusion:Both cloning sequencing and the third generation nanopore sequencing could solve the problem of overlapping peaks in the direct sanger sequencing for the universal primer amplification products of patent drugs.The third generation nano-pore sequencing was better than cloning sequencing in the sequencing of the universal primer amplification products of patent drugs,but there were still one raw material medicine that have not been detected.It is of certain reference val-ue for the molecular biological identification and DNA barcoding study of different raw materials in the patent medi-cine to establish the specific identification method of Tibetan patent medicine Shiliujianwei powder.

Objective To evaluate the efficacy and safety of enteric-coated metformin hydrochloride capsules(Junlida?)in patients with T2DM and poor glycemic control under lifestyle interventions.Methods In this study,419 patients with T2DM were recruited from 15 research centers from July 2020 to March 2022,and randomly divided into observation(Obs)group(n=209)and control group(Con,n=210)using a multicenter,randomized,double-blind,non-inferiority trial design.Patients in the Obs group were treated with enteric-coated Metformin hydrochloride capsules(Junlida?),and patients in the Con group were treated with Metformin hydrochloride tablets(Glucophage?).The optimal effective dose of 2 g/d was achieved within 4 weeks,and the reasonable dose was maintained until the end of treatment.The treatment period was 24 weeks.HbA1c and its compliance rate,FPG,and body weight were compared between the two groups in full analysis set(FAS)and protocol set(PPS).Safety and adverse events(AE)were evaluated in safety set(SS).Results A total of 414 participants were randomized(207 cases in Obs group and 207 cases in Con group).414 cases in FAS population(207 cases in Obs group and 207 cases in Con group),and 328 cases in PPS population(164 cases in Obs group and 164 cases in Con group),and 414 cases in SS population(207 cases in Obs group and 207 cases in Con group).After treatment,HbA1c,FPG and body weight were lower in both groups(P<0.05)in FAS and PPS.HbA1c compliance rate was not significantly different between the two groups in FAS and PPS(P>0.05).The results of non-inferiority test showed that the lower limit was>-0.4%in both FAS(-0.154,95%CI-0.384～0.069)and PPS(-0.139,95%CI-0.390～0.112),and the Obs group reached non-inferiority end point.The achievement rate,compliance rate,safety index and incidence of AE were not significantly different between the two groups(P>0.05).Conclusions Junlida? demonstrated non-inferiority to Glucophage? in glycemic control and can be safely and effectively used in patients with diabetes.

Aim To elucidate the underlying mecha-nism by which total triterpenoids extracted from Hove-nia dulcis(H-TP)enhance the sensitivity of A549/DDP cells to cisplatin.Methods The ARE-Nrf2 lu-ciferase reporter assay was applied to investigate the impact of H-TP on Nrf2 expression.Western blot was used to detect the protein levels of Keap-1/Nrf2/HO-1,Nrf2-GPX4 signaling pathway,apoptosis-related proteins of Bcl-2 and Bax.Further validation of its effects on Nrf2 was conducted by using Nrf2 activator/inhibitor.Results H-TP could enhance the sensitivi-ty of A549/DDP cells to cisplatin by modulating the expression of apoptosis-related proteins Bax and Bcl-2,inhibiting the Keap-1/Nrf2/HO-1/GPX4 signating pathway in A549/DDP cells,and inducing ferroptosis.Conclusion H-TP enhances the sensitivity of A549/DDP cells to cisplatin by inducing the Nrf2-mediated ferroptosis pathway.

Aim To elucidate the underlying mecha-nism by which total triterpenoids extracted from Hove-nia dulcis(H-TP)enhance the sensitivity of A549/DDP cells to cisplatin.Methods The ARE-Nrf2 lu-ciferase reporter assay was applied to investigate the impact of H-TP on Nrf2 expression.Western blot was used to detect the protein levels of Keap-1/Nrf2/HO-1,Nrf2-GPX4 signaling pathway,apoptosis-related proteins of Bcl-2 and Bax.Further validation of its effects on Nrf2 was conducted by using Nrf2 activator/inhibitor.Results H-TP could enhance the sensitivi-ty of A549/DDP cells to cisplatin by modulating the expression of apoptosis-related proteins Bax and Bcl-2,inhibiting the Keap-1/Nrf2/HO-1/GPX4 signating pathway in A549/DDP cells,and inducing ferroptosis.Conclusion H-TP enhances the sensitivity of A549/DDP cells to cisplatin by inducing the Nrf2-mediated ferroptosis pathway.