ObjectiveTo understand the epidemiological distribution and gene evolutionary variation of influenza A (H3N2) viruses in Fengxian District, Shanghai, in the surveillance year of 2023, and to provide a reference basis for influenza prevention and control. MethodsThe prevalence of influenza virus in Fengxian District in the 2023 influenza surveillance year (April 2023‒March 2024) was analyzed. The hemagglutinin (HA) gene, neuraminidase (NA) gene, and amino acid sequences of 75 strains of H3N2 influenza viruses were compared with the vaccine reference strain for similarity matching and phylogenetic evolutionary analysis, in addition to an analysis of gene characterization and variation. ResultsIn Fengxian District, there was a mixed epidemic of H3N2 and H1N1 in the spring of 2023, with H3N2 being the predominant subtype in the second half of the year, and Victoria B becoming the predominant subtype in the spring of 2024. A total of 75 influenza strains of H3N2 with HA and NA genes were distributed in the 3C.2a1b.2a.2a.2a.3a.1 and B.4 branches, with overall similarity to the reference strain of the 2024 vaccine higher than that of the reference strain of the 2022 and 2023 vaccine. Compared with the 2023 vaccine reference strain, three antigenic sites and one receptor binding site were changed in HA, with three glycosylation sites reduced and two glycosylation sites added; where as in NA seven antigenic sites and the 222nd resistance site changed with two glycosylation sites reduced. ConclusionThe risk of antigenic variation and drug resistance of H3N2 in this region is high, and it is necessary to strengthen the publicity and education on the 2024 influenza vaccine and long-term monitoring of influenza virus prevalence and variation levels.

ObjectiveTo investigate the pathogenic spectrum and epidemiological characteristics of diarrheal disease in Fengxian District of Shanghai, and to provide scientific basis for the prevention and control of diarrheal diseases. MethodsBasic information of the initial adult cases visited diarrheal disease surveillance sentinel hospital in Fengxian District, Shanghai, was collected from August 2013 to 2023, and fecal samples were collected at 1∶5 sampling intervals to isolate and identify 5 kinds of diarrheagenic Escherichia coli (DEC), Salmonella (SAL), Vibrio parahaemolyticus, Campylobacter, Vibrio cholerae, Shigella and Yersinia enterocolitica (YE). Simultaneously, nucleic acid detection was performed for 3 kinds of rotavirus, 2 kinds of norovirus, intestinal adenovirus, astrovirus and sapovirus. ResultsA total of 1 861 cases of newly diagnosed diarrheal disease were reported, with the peak in July to August. Additionally, 704 surveillance samples were detected, with a total positive detection rate of 50.57%. The detection rates of bacterial, viral and mixed infection were 25.14%, 21.02% and 4.40%, respectively. Among the pathogens detected, DEC accounted for the highest (17.61%, 124/704), followed by norovirus (16.48%, 116/704), rotavirus (6.39%, 45/704), SAL (5.97%, 42/704) and Campylobacter (3.84%, 27/704). DEC detected were mainly enteroaggregative Escherichia coli and enterotoxigenic Escherichia coli, with no detection of Vibrio cholerae, Shigella and YE. The highest total pathogen detection rate was observed from June to September, and the detection peaks of norovirus were from March to June and from October to December, whereas that of DEC was from June to October. The detection rate of rotavirus peaked from January to February, but which was not detected between 2020‒2023. The SAL positive rate peak was in September, whereas that of Campylobacter was from July to September. ConclusionThe main pathogens detected in Fengxian District from 2013‒2019 are DEC, norovirus, rotavirus, SAL and Campylobacter. Different pathogens have different detection peaks, with bacteria predominating in summer and viruses in winter and spring. Prevention and control measures should be carried out according to the epidemiological characteristics of different seasons.

This study aimed to comprehensively examine the association of gallstones, cholecystectomy, and cancer risk. Multivariable logistic regressions were performed to estimate the observational associations of gallstones and cholecystectomy with cancer risk, using data from a nationwide cohort involving 239 799 participants. General and gender-specific two-sample Mendelian randomization (MR) analysis was further conducted to assess the causalities of the observed associations. Observationally, a history of gallstones without cholecystectomy was associated with a high risk of stomach cancer (adjusted odds ratio (aOR)=2.54, 95% confidence interval (CI) 1.50-4.28), liver and bile duct cancer (aOR=2.46, 95% CI 1.17-5.16), kidney cancer (aOR=2.04, 95% CI 1.05-3.94), and bladder cancer (aOR=2.23, 95% CI 1.01-5.13) in the general population, as well as cervical cancer (aOR=1.69, 95% CI 1.12-2.56) in women. Moreover, cholecystectomy was associated with high odds of stomach cancer (aOR=2.41, 95% CI 1.29-4.49), colorectal cancer (aOR=1.83, 95% CI 1.18-2.85), and cancer of liver and bile duct (aOR=2.58, 95% CI 1.11-6.02). MR analysis only supported the causal effect of gallstones on stomach, liver and bile duct, kidney, and bladder cancer. This study added evidence to the causal effect of gallstones on stomach, liver and bile duct, kidney, and bladder cancer, highlighting the importance of cancer screening in individuals with gallstones.
Humans ; Mendelian Randomization Analysis ; Gallstones/complications* ; Female ; Male ; Cholecystectomy/statistics & numerical data* ; Middle Aged ; Risk Factors ; Aged ; Adult ; Neoplasms/etiology* ; Stomach Neoplasms/epidemiology*

Bacterial therapy has attracted increasing attention due to its special mechanism and abundant applications. With the flourishing development of synthetic biology, therapeutic genes have been introduced into engineering bacteria to improve their antitumor efficacy. However, it is difficult to spatiotemporally control the expression of these therapeutic genes at the tumor site in vivo, thereby considerably limiting the application of engineered bacteria in tumor treatment. To resolve this problem, we constructed a temperature-responsive bacterial strain capable of triggering the expression of exogenous genes in a laser-controllable way. Noxa, a pro-apoptotic protein, is chosen to test the expression of exogenous protein and its anti-tumor effect in engineered bacteria upon laser irradiation. Firstly, Noxa was fused to the C-terminus of the bacterial outer membrane protein cytolysin A (ClyA), and then the recombinant gene fragment ClyA-Noxa was inserted into the temperature-sensitive plasmid pBV220 and the recombinant plasmid was transformed into non-pathogenic Escherichia coli MG1655. Thus, we constructed the engineering strain (TRB@Noxa) that could express Noxa on the bacterial surface. TRB@Noxa could target and colonize the tumor tissue without causing notable host toxicity. The bacterial infection triggered thrombosis in the tumor tissue, resulting in the darkness of tumor sites. In a xenograft mouse tumor model, our strategy demonstrated precise tumor targeting and strong tumor inhibition. In conclusion, we successfully constructed a new engineering bacterial strain TRB@Noxa. TRB@Noxa combined with photothermal therapy could arrest tumor growth in the absence of photosensitizers, which represents an appealing method for antitumor therapy in the future.
Escherichia coli/radiation effects* ; Animals ; Humans ; Lasers ; Mice ; Proto-Oncogene Proteins c-bcl-2/biosynthesis* ; Neoplasms/therapy* ; Genetic Engineering ; Cell Line, Tumor ; Escherichia coli Proteins/genetics*

Purpose To investigate the effect of MYD88 gene overexpression on the proliferation and apoptosis of human diffuse large B cell lymphoma(DLBCL)cells,and to prelimi-narily explore the mechanism of MYD88 gene action.Methods PEGFP-C2-MYD88 overexpressing MYD88 L265P gene was transfected into DLBCL cells by plasmid transfection.The exper-iment was divided into blank control group,negative control group and MYD88 L265P overexpression group.The fluores-cence expression of MYD88 L265P after overexpression was ob-served under inverted fluorescence microscope.RT-PCR and Western blot were used to detect the mRNA and protein expres-sion of MYD88 L265P,IRAK4,NF-κB and BCL2 in DLBCL cells before and after overexpression of MYD88 L265.CCK8 method was used to detect DLBCL cells proliferation and Ho-echst staining was used to detect DLBCL cells apoptosis.Re-sults After overexpression of MYD88 L265P,compared with the blank control group(0.670 4±0.017 5)and the negative control group(0.715 3±0.019 6),the MYD88L265P overex-pression group(1.157 2±0.010 2)increased significantly,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(0.69 ±0.04)and the negative control group(0.81±0.07),the MYD88L265P overexpression group(0.48±0.05)was signifi-cantly decreased,with statistical significance(all P＜0.05).After overexpression of MYD88 L265P,compared with the blank control group(mRNA:1.0158±0.0115,0.987 3±0.010 2,1.007 6±0.015 3,protein:0.183 4±0.058 9,0.096 8± 0.015 7,0.147 5±0.0418)and negative control group(mR-NA:0.9132±0.0098,1.0032±0.0156,0.9327± 0.011 2,protein:0.187 9±0.042 3,0.088 9±0.0513,0.134 8±0.050 1),the mRNA(3.243 2±0.013 6,2.976 6 ±0.0213,1.585 9±0.019 8)and protein expressions(0.452 7±0.052 4,0.218 9±0.047 5,0.301 4±0.059 8)of IRAK4,NF-κB and anti-apoptosis protein BCL2 in MYD88L265P overexpression group were significantly increased,which was statistically significant(all P＜0.05).Conclusion After overexpression of MYD88 L265P,the apoptosis rate of DLBCL cells decreased and the cell proliferation rate increased.The mechanism may be related to the mutation of MYD88 L265P gene,activation and amplification of NF-κB pathway,and pro-motion of the overexpression of antiapoptotic protein BCL2.

Objective:To observe the regulatory effect of microRNA-10b(miR-10b)on the immune effect of glioma cells and explore its mechanism.Methods:Human glioma cell U251 was cultured to obtain cells in logarithmic growth stage.The cell suspen-sion was prepared according to the concentration of 1.0×105 cells/ml,and the control group,overexpression group,low expression group and blank group were set up,with 6 wells in each group.The negative control,miR-10b mimics and miR-10b inhibitor were transfected by liposome transfection in control group,overexpression group and low expression group,respectively.The blank group was given the same amount of sterile normal saline.Natural killer(NK)cells from peripheral blood of a healthy volunteer was isolated and cultured.The killing activity of NK cells was detected by MTT method.The expression of NK cell activated receptor(NKG2D)on the surface of NK cells in each group were detected by flow cytometry,and the expression of major histocompatibility complex class Ⅰ chain-related gene A(MICA),UL16 binding protein 2(ULBP2)and UL16 binding protein 3(ULBP3)on the surface of U251 hu-man glioma cells in each group were detected.Results:The transfection efficiency of control group,overexpression group and low ex-pression group were(93.55±2.05)％,(95.67±3.14)％,(94.18±3.26)％,respectively.Compared with control group and blank group,the expression of miR-10b increased in overexpression group and decreased in low expression group,and the difference were statisti-cally significant(P<0.05).There was no significant difference in the expression of miR-10b between control group and blank group(P>0.05).Compared with control group and blank group,the killing activity of NK cells with different effect target ratios in overex-pression group decreased,the expression of NKG2D decreased,the killing activity of NK cells with different effect target ratios in low expression group increased,and the expression of NKG2D increased,and the difference were statistically significant(P<0.05).The killing activity of NK cells in each group increased with the increase of effect target ratio,and the difference were statistically signifi-cant(P<0.05),and there was no significant difference in NK cell killing activity and NKG2D expression between control group and blank group(P>0.05).Compared with control group and blank group,the expression of MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251 in overexpression group decreased,and the expression of MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251 in low expression group increased,the difference were statistically significant(P<0.05),and there was no signifi-cant difference in the expression of MICA,ULBP2 and ULBP3 on the surface of U251 glioma cells between control group and blank group(P>0.05).Conclusion:Inhibiting the expression of miR-10b can increase the expression of NKG2D on the surface of NK cells and MICA,ULBP2 and ULBP3 on the surface of human glioma cell U251,and enhance the killing activity of NK cells against human glioma cell U251.

Objective:To investigate the clinical practicability of positive end-expiratory pressure (PEEP) titrated by lung stretch index (SI) in patients with acute respiratory distress syndrome (ARDS).Methods:A parallel randomized controlled trial was conducted. Patients with moderate to severe ARDS who required mechanical ventilation admitted to the department of critical care medicine of General Hospital of the Yangtze River Shipping from August 2022 to February 2023 were enrolled. They were randomly divide into SI guided PEEP titration group (SI group) and pressure-volume curve (P-V curve) inspiratory low inflection point (LIP) guided PEEP titration group (LIP group). All patients were ventilated in a supine position after admission, with the head of the bed raised by 30°. The primary disease was actively treated, prone position ventilation for 12 h/d, and lung protective ventilation strategies such as controlled lung expansion were used for lung recruitment. On this basis, mechanical ventilation parameters were titrated with SI in the SI group; the LIP group titrated mechanical ventilation parameters with P-V curve inspiratory LIP+2 cmH 2O (1 cmH 2O ≈ 0.098 kPa). The oxygenation index (PaO 2/FiO 2), and respiratory mechanics indicators such as lung dynamic compliance (Cdyn), peak airway pressure (Pip) were monitored before recruitment maneuver and after 1, 3, and 5 days of treatment. The therapeutic effect of the two groups was compared. Results:There were 41 patients in the SI group and 40 patients in the LIP group. There was no significant difference in general information such as gender, age, and disease type between the two groups. The mechanical ventilation time and the length of intensive care unit (ICU) stay in the SI group were significantly shorter than those in the LIP group (days: 9.47±3.36 vs. 14.68±5.52, 22.27±4.68 vs. 27.57±9.52, both P < 0.05). Although the 28-day mortality of the SI group was lower than that of the LIP group, the difference was not statistically significant [19.5% (8/41) vs. 35.0% (14/40), P > 0.05]. On the fifth day, the PaO 2/FiO 2 was higher in SI group [mmHg (1 mmHg≈0.133 kPa): 225.57±47.85 vs. 198.32±31.59, P < 0.05], the Cdyn was higher in SI group (mL/cmH 2O: 47.39±6.71 vs. 35.88±5.35, P < 0.01), the Pip was lower in SI group (mmHg: 35.85±5.77 vs. 43.87±6.68, P < 0.05). The Kaplan-Meier survival curve showed no statistically significant difference in the 28 days cumulative survival rate between the two groups (Log-Rank: χ2 = 2.348, P = 0.125). Conclusion:The application of SI titration with PEEP in the treatment of ARDS patients may improve their prognosis.