Background/Aims: Biliary tract cancer (BTC) frequently harbors KRAS mutations, which are associated with resistance to traditional treatment and a poor prognosis. Synthetic lethality (SL) strategy may provide other targets of KRAS. Therefore, we aim to identify and validate potential therapeutic targets of KRAS for the treatment of BTC via SL. Methods: The dependency (DepMap) projects were used to predict the synthetic lethal gene of KRAS. FDA-approved anticancer drug library was applied to screen potential drugs effective against KRAS-mutant BTC. Furthermore, the synthetic lethal effects or corresponding mechanisms of potential genes and drugs on BTC were investigated using KRAS-mutant and KRAS-wild type BTC cell lines, patient-derived xenografts (PDX), and KRAS oncogene-driven tumor models, as well as other KRAS-mutant cancer cell lines. Results: Initially, we discovered that the loss of GATA2 reduced the viability of KRAS-mutant but not KRAS-wild-type BTC. Subsequently, the drug library screened out disulfiram, which primarily exerts a synthetic lethal effect by inhibiting interleukin-1β (IL-1β) in KRAS-mutant BTC. Mechanistically, GATA2 specifically enhanced the transcription of IL-1β to promote NF-κB signaling in KRAS-mutant BTC. IL-1β inhibition phenocopied GATA2 deficiency, leading to reduced KRAS-mutant BTC viability. These synthetically lethal effects were confirmed using PDX, a KRAS oncogene-driven tumor model, as well as in other KRAS-mutant cancer cell lines. Conclusions In summary, these results indicate that inhibiting GATA2/IL1β could be a therapeutic strategy in KRAS-mutant BTC and potentially other cancers.

Objective To understand the clinical distribution characteristics and drug-resistance genotypes of carbapenem-resistant Pseudomonas aeruginosa(CRPA)in a tertiary hospital in Shanghai,so as to guide the rational use of antibiotics,reduce bacterial resistance and control nosocomial infections.Methods A total of 94 consecutive and non-redundant CRPA strains isolated from clinical specimens were collected in The First Affiliated Hospital of Naval Medical University between Jan.1,and Dec.31,2019.The distribution of CRPA across departments and sample sources were analyzed.Antimicrobial susceptibility testing against 13 antibiotics was determined using the VITEK 2 Compact automated microbiology analyzer and the Kirby-Bauer disk diffusion method.Polymerase chain reaction(PCR)was employed to detect genes encoding extended-spectrum β-lactamase resistance gene,carbapenemase resistance gene,and porin resistance gene.Homology analysis of selected isolates was performed using kSNP3,a k-mer-based software,for single nucleotide polymorphism(SNP)analysis.An evolutionary tree was constructed to assess genetic relationships.Additionally,multilocus sequence typing(MLST)was performed using the Pseudomonas aeruginosa typing scheme from the PubMLST database.Results Among the 94 CRPA isolates,high resistance rates were observed for carbapenems,including imipenem(92.6%,87 strains)and meropenem(87.2%,82 strains).In contrast,low resistance rates were detected for aminoglycosides,such as amikacin(10.6%,10 strains),gentamicin(20.2%,19 strains),and tobramycin(20.2%,19 strains).The top 3 departments in terms of isolate distribution were the Emergency Intensive Care Unit(9.6%,9 strains),Department of Cerebrovascular Surgery(8.5%,8 strains),and Department of Respiratory Medicine(8.5%,8 strains).PCR analysis of 94 CRPA strains detected outer membrane protein D2(OprD2)gene deletion in 47(50.0%)strains,13(13.8%)strains were positive for blaKPC,4(4.3%)strains for blaVIM,2(2.1%)strains for blaIMP,1(1.1%)strain for blaNDM,12(12.8%)strains for blaTEM,4(4.3%)strains for blaPER,and 2(2.1%)strains for blaGES,while blaOXA-48,blaBIC,blaSIM,blaVEB,and blaSHV were not detected.MLST identified 36 different sequence types(STs),with ST463 and ST274 being the most common,and 2 new ST(ST4023 and ST4024)were identified for the first time.Conclusion CRPA strains carry multiple resistance genes and exhibit concurrent resistance to several commonly used clinical antibiotics.The resistance is primarily associated with the presence of blaKPC,blaVIM and blaTEM genes and the deletion of OprD2 gene.Clinical monitoring of CRPA should be strengthened,and rational use of antimicrobial agents is essential to control its spread within the hospital.

Objective To prepare decellularized scaffolds from rabbit cartilage at various concentrations and assess their physicochemical properties and compatibility with stem cells to provide an experimental basis for cartilage repair.Methods Bone marrow mesenchymal stem cells(BMSCs)were cultured using the Percoll density gradient separation method,and this was followed by flow cytometric analysis and testing of their osteogenic and chondrogenic differentiation capabilities.Cartilage pieces were excised from rabbit knees and hip joints and subjected to physical crushing,repeated freeze-thaw cycles,and mixed enzymatic digestion for decellularization.To compare and observe the physicochemical properties of the decellularized scaffolds at different concentrations,three groups of scaffolds(labelwd A,B,and C)were designed with concentrations of 100％,50％and 30％,with three replicates each.Third-generation PKH26-labeled BMSCs were seeded onto optimally concentrated scaffolds and cultured for 1 week to observe cell growth.Results Flow cytometry detected BMSC surface antigens with positive expression of CD44 and CD90 and negative expression of CD45.Osteogenic induction stained with alizarin red showed red calcific nodules,and chondrogenic induction stained with alcian blue showed blue cartilaginous nodules.No apparent cell morphology was observed in the three groups of scaffolds stained with hematoxylin-eosin,and toluidine blue.There was a significant difference in DNA concentration between decellularized samples and non-decellularized scaffolds(P＜0.05).The content of glycosaminoglycans was slightly lower than the normal values.Significant differences were observed between the three groups of scaffolds in terms of pore size,water absorption,porosity,tensile strength,and Young's modulus(P＜0.05).After co-cultivation of stem cells with the scaffolds,cell adhesion was found to be good.Conclusions Percoll density gradient separation can obtain high-purity rabbit BMSCs,and the mixed decellularization method is superior.Group B scaffolds were the most suitable for tissue-engineered cartilage repair.BMSCs cultured in vitro grew well on Group B scaffolds.

Background In recent years, ozone (O₃) has gradually become a key air pollutant affecting public health. Studies have been conducted to evaluate O₃ pollution-associated disease burden for general population, but there is a lack of research on O₃ pollution-associated disease burden of gestational complications. Objective To assess the disease burden of O₃ pollution on pregnancy complications in the Yangtze River Delta region and the changes in the disease burden resulting from the improvement of O₃ pollution levels. Methods Through Chinese databases such as China National Knowledge Infrastructure (CNKI) and Wanfang, as well as English databases including PubMed and Web of Science, a systematic search was conducted to retrieve epidemiological studies investigating the association between O₃ exposure and pregnancy complications from January 1, 2010 to February 28, 2023. Using predetermined inclusion and exclusion criteria, retrieved studies were screened and assessed for heterogeneity using the Higgins I² statistic. The exposure increment was standardized to 10 µg·m⁻³, and a meta-analysis was conducted to evaluate the exposure-response relationship. Spatial analysis and environmental health risk assessment were then employed, using O₃ monitoring data at national air monitoring stations, population data, and related disease burden parameters in the Yangtze River Delta region, to evaluate changes in the disease burden of pregnancy complications associated with improvements in O₃ pollution levels. Results The meta-analysis revealed a significant association between maternal O₃ exposure and increased risks of gestational diabetes mellitus (GDM) and hypertensive disorders of pregnancy (HDP), particularly in early pregnancy, and the related odds ratios (OR) and 95% confidence intervals (CI) were 1.028 (1.002, 1.055) and 1.031 (1.023, 1.040), respectively. The cities with higher attribution fractions (AF) of GDM and HDP related to O₃ exposure in 2017 and 2020 in the Yangtze River Delta region were Xuzhou, Huaibei, Suzhou (Anhui Province), Changzhou, and Wuxi, with most cities showing a significant decrease in AF in 2020. Except for Hefei, the cases of GDM and HDP attributable to O₃ exposure in the remaining cities of the Yangtze River Delta region decreased by 16.31% to 57.41% in 2020 compared to 2017. In addition, except for Hefei, Suqian, Anqing, and Wuxi, the direct medical costs attributed to O₃ exposure in the remaining cities decreased by 1.12% to 45.36% in 2020 compared to 2017. Conclusion Exposure to O₃ during pregnancy is associated with increased risks of GDM and HDP. There are differences in the disease burden of GDM and HDP related to O₃ pollution among cities in the Yangtze River Delta region in 2017 and 2020. In 2020, compared to 2017, the disease burden in most cities is decreased.

AIM: To investigate whether rs7903146 polymorphisms in transcription factor 7-like 2 (TCF7L2) gene are associated with susceptibility to type 2 diabetes mellitus (T2DM) in Chinese Uygur population.METHODS: In this case-control study, 935 cases of T2DM patients were recruited in T2DM group, and 971 healthy examination individuals were selected as normal control.The TCF7L2 gene polymorphism was detected by matrix-assisted laser desorption/ionization time-of-flight mass spectrum.RESULTS: Significant differences in the frequencies of CC, CT and TT genotypes and the frequencies of C and T alleles on TCF7L2 rs7903146 were found between T2DM group and control group(P<0.05).As compared with C allele, the patients with T allele had a significantly higher risk of T2DM with OR of 1.190 (95% CI: 1.034~1.371).As compared with CC genotype, the patients with CT genotype had a significantly higher risk of T2DM with OR of 1.374 (95% CI: 1.122~1.683), and the patients with CT+TT genotype had a significantly higher risk of T2DM with OR of 1.307 (95% CI: 1.090~1.567).The levels of fasting plasma glucose, serum creatinine and blood urea nitrogen were higher in all participants with CT+TT genotype of rs7903146 than those with CC genotype, which showed a significant difference (P<0.05).CONCLUSION: The polymorphisms of rs7903146 in TCF7L2 gene may be associated with T2DM in Uygur population from Xinjiang region.The T allele and CT genotype of rs7903146 are the risk factors for T2DM.

Objective To investigate the correlation between TCF7L2 gene rs3814570 polymorphisms with type 2 diabetes mellitus(T2DM) in Uygur population of Xinjiang area.Methods By adopting the case-control study design,949 cases of T2DM were recruited as the observation group and 963 individuals Undergoing healthy physical examination were selected as the control group.The TCF7L2 gene polymorphism was detected by matrix-assisted laser desorption/ionization-time of flight(MALDI-TOF).Results The statistical differences in frequencies of CC,CT and TT genotypes and the C and T allele frequencies on TCF7L2 rs3814570 were found between the T2DM group and control group(P＜0.05).The risk of suffering from T2DM in the carriers of CT genotype was 0.331 times of that in the carriers of CC genotype(OR =0.331,95 ％ CI:0.166-0.661,P =0.002),the risk of suffering from T2DM in the carriers of TT genotype was 0.539 times of that in the carriers of CC genotype(OR=0.539,95％CI:0.348-0.834,P=0.005),and the risk of suffering from T2DM in the carriers of T allele was 0.501 times of that in the carriers of C allele(OR=0.501,95 ％ CI:0.377-0.664,P＜ 0.01).Among all subjects,the FPG level of the CT + TT genotype group on TCF7L2 gene rs3814570 locus was significantly lower than that of the CC genotype group(P＜0.05).Conclusion The rs3814570 locus in TCF7L2 gene may be associated with T2DM occurrence in Uygur population of Xinjiang area,the T allele and TT genotype might be protective factors of T2DM.