Objective To establish an inhalation infection pneumonia model of C57BL/6J mice with highly virulent and multi-drug resistant Pseudomonas aeruginosa(PA)strain F291007,and to study the microbiological,pathological and immunological characteristics of this model.Methods The strain F291007 was isolated and identified before the bacterial suspension was administered to the mice via aerosolized intratracheal inoculation to establish the pneumonia infection model.In the course of infection,the conditions and survival of the mice were observed,and the bacterial loads,the histopathological states and the cytokine expression levels in the major organs were detected.Finally,three key cytokines were blocked to observe the survival of mice.Results The strain F291007 was isolated and identified.After lethal dose infection,all the mice died within 24 h.After sub-lethal dose infection,a large number of immune cells in the body were capable of phagocytosis and killing of invading pathogens,which was manifested as rapid clearance of bacteria in lungs and the exponential decrease of bacterial load with the passage of time.The pathological changes in lungs were most severe at 1 to 3 days but gradually recovered.After infection,interleukin-6(IL-6),IL-17A and tumor necrosis factor-α(TNF-α)in alveolar lavage fluid and serum were significantly increased at 1 to 3 days.After blocking of these three cytokines with specific antibodies,the survival rates of infected mice decreased significantly.Conclusion A mouse model of gradually-recovered pneumonia infection caused by PA inhalation has been established,suggesting that the first one to three days are critical to immune response after infection through multiple indicators.This mouse model can be used for research on the pathogenesis,immunoregulation and treatment evaluation of highly virulent and multi-drug resistant PA inhalation pneumonia infection.

Objective To explore the role of protective function of Sestrin2(Sesn2)to mitochondria in alleviating cognitive dysfunction in mice with sepsis-associated encephalopathy(SAE).Methods 6-week-old male C57BL/6J mice were randomly divided into three groups:sham group,CLP group and CLP plus eupatilin group,40 mice in each group.A sepsis model was induced by cecal ligation and perforation(CLP).The CLP plus eupatilin group was treated with eupatilin.Neurobehavioral test and Morris water maze(MWM)were used to deter-mine neurobehavior and spatial learning and memory function in mice.The number of neurons in hippocampal CA1 area was counted by Nissl staining.HT22 cells were randomly divided into a control group(Con),lipopolysaccha-ride group(LPS),LPS plus eupatilin treatment group(LPS plus eupatilin)and LPS plus eupatilin and Nrf2 siRNA treatment group(LPS plus eupatilin and si-Nrf2).Apoptosis was analyzed by terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)staining,Mitochondrial membrane potential(MMP)was used to analyze mitochondrial damage.Results Seven days after CLP,as compared with sham mice,Sesn2 in hippocampus and cortex decreased significantly in CLP mice(P<0.01).As compared with CLP group,the survival rate in CLP plus eupatilin group increased significantly(P<0.05).As compared with sham group,the mice in CLP group showed a relatively high nerve injury score(P<0.05),and had fewer platform crossings and shorter target stay time,while the mice in CLP plus eupatilin group exhibited a lower injury score(P<0.05),and stayed in the target area for a longer time(P<0.05).As compared with sham group,the co-localization rate of neurons,Sesn2 and Nrf2 in CLP group decreased significantly(P<0.05),and the number of CD68/Iba-1 positive microglia increased significantly(P<0.05),while CLP plus eupatilin group reversed these changes.As compared with Con group,apoptosis and MMP level in LPS group increased significantly(P<0.01),while apoptosis and MMP level in LPS plus eupatilin group were lower than those in LPS group(P<0.05).However,Nrf2 knockdown(LPS plus eupatilin and si-Nrf2 group)reversed the anti-apoptosis and mitochondrial protection of eupatilin.Conclusions Eupatilin can alleviate cognitive dysfunction and neurological deficit in SAE mice by activating Sesn2-Nrf2 pathway,and improve inflammatory microenvironment by alleviating mitochondrial dysfunction.

ObjectiveTo investigate the contamination status of Staphylococcus aureus in food and the presence of enterotoxin genes in Jiading District， Shanghai， and to provide a basis for the prevention and treatment of foodborne Staphylococcus aureus disease. MethodsFrom 2021 to 2023， 15 types of food were sampled for S. aureus testing， and the presence of five enterotoxin genes， including sea⁃see， was tested in the strains. ResultsOut of 705 food samples， 88 （12.48%） were positive for S. aureus. S. aureus was detected in 12 of the 15 food types， with the three food types with the highest positive rates being cold noodles （45.00%）， raw poultry （26.25%）， and vegetable salads （20.00%）. The enterotoxin gene carriage rate was 32.95% in food strains. The carriage rates for sea， seb， and sec were 7.95%， 12.50%， and 14.77%， respectively. Neither sed nor see was detected. The detection rate of strains carrying two types of enterotoxin genes was 2.27%. The enterotoxin carriage rates in strains from vegetables， beverages， and raw meat were 57.14%， 40.00%， and 30.00%， respectively. ConclusionThe S. aureus detection rate in food in Jiading District is much higher than the national average. The enterotoxin gene carriage rates are high， with food strains carrying sea， seb， and sec， with sec being the most prevalent. There is a need to enhance monitoring of S. aureus and enterotoxins， especially in high-risk foods such as noodles， vegetables， and non-packaged beverages.

ObjectiveTo investigate the prevalence of Staphylococcus aureus in patients with diarrhea, and to analyze the genes carriage of enterotoxin in the strains of these patients in Jiading District, Shanghai. MethodsFrom 2021 to 2023, anal swabs of diarrhea outpatients from one sentinel hospital and nine community health service centers in different townships in Jiading District, Shanghai, were tested for Staphylococcus aureus, from which five enterotoxin virulence genes such as SEA, SEB, SEC, SED, and SEE were tested simultaneously. ResultsA total of 1 080 anal swabs were collected, 81 of which were tested positive for S. aureus, with a detection rate of 7.50%, and the detection rate of S. aureus was similar in patients with diarrhea from 2021‒2023. There was no statistically significant difference in detection rates between males and females (χ²=0.821, P=0.365). S. aureus detection rate was highest in infants and young children with diarrhea (29.51%), followed by 14.06% in the people aged between 4‒<31 years, and 2.99% in those aged ≥31 years. Significant differences were observed in the detection rate of S. aureus in the diarrhoeal patients from different townships of Jiading District(χ²=66.134,P<0.05). The carriage rates of the 5 enterotoxin genes, namely SEA, SEB, SEC, SED, SEE, were 13.58%, 14.81%, 11.11%, 7.41%, and 0, respectively. ConclusionThe prevalence of S. aureus among the patients with diarrhea in Jiading District is relatively stable but with distinct geographical patterns. Children and adolescents are high-risk groups. SEB were the dominant gene, followed by SEA.

Objective:To explore the economic applicability and safetyof the novel uterovaginal pubic comb suspension(UPCS)surgery with Mersilene tape in the treatment of pelvic organ prolapse(POP).Methods:A ret-rospective analysis was conducted on the clinical data of patients who underwent UPCS surgery due to POP from January 1st,2021 to February 28,2023.They were divided to the UPCS surgery with Mersilene tape group(group A)and suspension surgery with Y-shaped mesh group(group B)respectively.The POP-Q indication points,sus-pension surgery duration,intraoperative bleeding volume,material expense,postoperative catheter retention time,anal exhaust time and hospitalization duration were recorded for both groupbefore and after surgery.Evaluate the severity of POP related symptoms in patients before and after surgery using the pelvic Floor Distress Invento-ry-short Form 20(PFDI-20)and Pelvic Organ Prolapsed/Urinary Incontinence Sexual Questionnaire-12(PISQ-12),and follow up and observe the patients and analyze the complications.Results:A total of 17 POP patients were included in the study.There were 12 patients in group A while 5 patients in group B.The suspension material expense of group A was considerably lower than that of group B(P＜0.05).There was no significant difference between the two groups in preoperative PFDI-20 score,preoperative PISQ-12 score,UPCS surgery duration,intr-aoperative bleeding volume,postoperative urinary catheter retention time,postoperative anal exhaust time and hospitalization duration.All patients showed stable vital signs during the surgery and no severe complications were reported.Compared with the preoperative status,the positions of the Aa,Ba,and C indicatorpoint in group A and group B were all increased significantly(P＜0.05).The PFDl-20 and PISQ-12 scores of the two groups at the last follow-up after surgery showed significant improvement compared to those before surgery(P＜0.05).No signifi-cant difference was found in the PFDI-20 and PISQ-12 scores between the two groups after surgery(P＞0.05).There were no significant differences in the postoperative complications between the two groups(P＞0.05).Con-clusions:Compared with suspension surgery with Y-shaped mesh,UPCS surgery with Mersilene tape is safe and effective in the treatment of POP.The UPCS surgery with Mersilene tape showed better cost-effectiveness in the treatment of POP,and the surgical steps are relatively simple.Therefore,UPCS surgery with Mersilene tape was worthy of promotion in clinical practice.

【Objective】 To explore the timing of surgical drainage for ureteral calculi with upper urinary tract infection. 【Methods】 Clinical data of 117 cases of ureteral calculi with upper urinary tract infection treated in our hospital during Jan.2018 and Jan.2020 were retrospectively analyzed. According to different treatment methods, the patients were divided into surgical drainage group and non-surgical drainage group. The patients’ age, gender, side of calculi, peak body temperature, time of onset, white blood cell (WBC) count, C-reactive protein (CRP) and other clinical indicators were compared between the two groups. The cutoff value of surgical drainage was determined with receiver operator characteristic (ROC) curve. 【Results】 The patients’ age, peak body temperature, WBC count and CRP level were the influencing factors of surgical drainage (P<0.05). Regression analysis showed that CRP (P<0.001), age (P=0.003) and WBC count (P=0.014) were independent risk factors for surgical drainage. The area under the ROC curve (AUC) of CRP, age, and WBC count were 0.923, 0.601, and 0.796, respectively. The cutoff value of CRP was 29.87 mg/L (sensitivity 79.4%, specificity 90.0%). Logistic regression model showed CRP was a significant clinical predictor. 【Conclusion】 Ureteral calculi with upper urinary tract infection need to be diagnosed and treated in time. Positive anti-infection should be performed during emergency treatment, and surgical drainage could be selected according to the value of CRP.

Purpose: The unique chromosomal rearrangements of endometrial stromal sarcoma (ESS) make it possible to distinguish high-grade ESS (HGESS) and low-grade ESS (LGESS) from the molecular perspective. Analysis of ESS at the genomic and transcriptomic levels can help us achieve accurate diagnosis of ESS and provide potential therapy options for ESS patients. Materials and Methods: A total of 36 ESS patients who conducted DNA- and/or RNA-based next-generation sequencing were retrospectively enrolled in this study. The molecular characteristics of ESS at genomic and transcriptomic levels, including mutational spectrum, fusion profiles, gene expression and pathway enrichment analysis and features about immune microenvironment were comprehensively explored. Results: TP53 and DNMT3A mutations were the most frequent mutations. The classical fusions frequently found in HGESS (ZC3H7B-BCOR and NUTM2B-YWHAE) and LGESS (JAZF1-SUZ12) were detected in our cohort. CCND1 was significantly up-regulated in HGESS, while the expression of GPER1 and PGR encoding estrogen receptor (ER) and progesterone receptor (PR) did not differ significantly between HGESS and LGESS. Actionable mutations enriched in homologous recombination repair, cell cycle, and phosphoinositide 3-kinase/AKT/mammalian target of rapamycin pathways were detected in 60% of HGESS patients. Genes with up-regulated expression in HGESS were significantly enriched in five immune-related pathways. Most HGESS patients (85.7%) had positive predictors of immunotherapy efficacy. Moreover, immune microenvironment analysis showed that HGESS had relatively high immune infiltration. The degree of immune infiltration in HGESS patients with ZC3H7B-BCOR fusion was relatively higher than that of those with NUTM2B-YWHAE fusion. Conclusion This study investigated the molecular characteristics of ESS patients at the genomic and transcriptomic levels and revealed the potentially high sensitivity of targeted therapy and immunotherapy in a subset of HGESS with specific molecular features, providing a basis for guiding decision-making of treatment and the design of future clinical trials on precision therapy.

Objective : To analyze the genome structure and genetic characteristics of IncpGRT1 plasmids from Pseud⁃ omonas , and elucidate its potential transmission risk . Methods : The genomic DNA of the clinical isolate 15420352 was extracted after purification and preservation of the strain , and then the whole genome was sequenced , and then the type of the plasmid was identified . Sequence annotation and comparison of the backbone region and the accessory modules were performed on all five same type sequenced plasmids , including one plasmid p420352 - strA in this study and four from GenBank . The plasmids were annotated by RAST , Plasmidfinder , Blast , ResFinder , and ISfinder. The ORFs of the plasmid were annotated and drug resistance genes were found . Results : All five plasmids were classified as new IncpGRT1 type plasmids . The IncpGRT1 backbone genes or gene loci were in all five plasmids , and they contained an auxiliary replicon besides the primary IncpGRT1 replicon . Five IncpGRT1 plasmids carried at least three different accessory modules , including the srp region , the msr region , and a Tn5053 family transposon . Three resistance genes strA , strB , and mer were obtained in these plasmids , which were involved in resistance to two categories of antibiotics and heavy metals . We also found that these plasmids carried at least one virulence gene msr and five key transporters srp , emrE , mod , phn , and lpt , which could improve the environmental adaptability of the strains . Conclusion The IncpGRT1 plasmids have become the important vector for the accumulation and spread of some drug resistance genes and virulence genes in Pseudomonas , and have improved the environmental adaptability of the strain.

Objective:To study the clinicopathological features of metaplastic breast cancer (MBC) , and its prognostic factors.Methods:Data of 49 MBC cases and 30 cases of invasive ductal carcinoma (IDC) during the same period as matched control were collected. The immunohistochemistry staining of CK5/6, CK, P63, ER, HER2, Ki67 was performed in all MBC samples.Results:In MBC cases, the median age was 55 years. Median tumor size was 2.5 cm (range, 0.6-19 cm). Fifteen cases were classified as metaplastic carcinoma with heterologous mesenchymal differentiation (8 as matrix-producing carcinoma), 12 as spindle cell carcinoma, 7 as squamous cell carcinoma, 2 as low-grade adenosquamous carcinoma, 2 as fibromatosis-like metaplastic carcinoma, and 11 as mixed metaplastic carcinoma. The 5-, 10-year overall survival rate was 50%, 41%, respectively, lower than those of IDC (76%,63%) (all P<0.05). Lymph node metastasis rate, and expression of ER, PR, HER2 in MBC were lower than those in IDC (all P<0.05). Triple-negative cases in MBC were more than those of IDC ( χ2=26.244, P=0.000). The proliferative index of Ki67 was statistically different between the two groups ( t=2.624, P=0.011). Conclusions:MBC is a rare and heterogenous breast cancer. Compared to IDC, MBCs are usually larger, lower in lymph nodes metastasis, higher in proliferative index of Ki67, more triple-negative, hence with a poorer prognosis.

Histones are important structural proteins of chromatin in the nucleus, which can regulate gene transcription, and can be released from the nucleus to the outside of the cell under injury and inflammatory stimulations, thereby causing cytotoxicity and immune stimulation, and aggravating tissue damage. Extracellular histones are involved in the occurrence and development of many diseases, including sepsis, autoimmune diseases, liver injury, and acute lung injury. Therefore, its application not only can be used as a body’s biomarker of inflammation, but also it is expected to become a molecular target for the treatment of diseases. This article reviews the role of extracellular histones in the inflammatory process of liver injury.