Objective To explore the role of resilience and stress in childhood maltreatment and adolescent depression and anxiety symptoms, and to provide a basis for adolescent psychological intervention. Methods From September to October 2022, a total of 11 217 students from four middle schools in Changsha were selected by multistage stratified sampling, and the Childhood Trauma Questionnaire, Depression–Anxiety–Stress Scales and the Resilience Scale for Chinese Adolescents were used to carry out online questionnaire survey. Results The detection rate of depressive symptoms and anxiety symptoms in adolescent was 19.43% (2 179) and 28.7% (3 224). The direct effect of childhood maltreatment on depressive symptoms was significant (β =0.09), Childhood maltreatment is a negative prediction of resilience (β = -0.57) and a positive prediction of pressure (β =0.06); Resilience can negatively predict stress (β = -0.61) and depressive symptoms (β = -0.25) (all P values <0.001); The direct effect of childhood maltreatment on anxiety symptoms was significant (β =0.03), resilience negatively predicts anxiety symptoms (β = -0.08) (all P values <0.01). Resilience and stress have a partially mediating role between childhood maltreatment and depressive symptoms, the mediation effect value was 0.39, accounting for 81.25% of the total effect. The effect values of the three pathways accounted for 29.17%, 8.33% and 43.75% of the total effect, respectively. Resilience and stress have a partial mediating effect between childhood abuse and anxiety symptoms, and the mediating effect was 0.36, accounting for 92.31% of the total effect, and the effect values of the three pathways accounted for 12.82%, 10.26% and 69.23% of the total effect, respectively. Conclusion Childhood maltreatment could affect adolescents’ depression and anxiety through the chain mediating effect of resilience and stress.

Objective To analyze the epidemiological characteristics of mumps in Guangxi from 2012 to 2024, and to provide a scientific basis for formulating prevention and control strategies. Methods Descriptive epidemiological methods were used to analyze the incidence data of mumps in Guangxi from 2012 to 2024. Results A total of 159 873 mumps cases were reported from 2012 to 2024 in Guangxi, with an average annual reported incidence of 25.41/100 000, and no death. Mumps occurred every month, with the peak incidence mainly concentrated in April to July and October to January of the next year. There were 96,118 male cases (29.43 /100 000), and 63 755 female cases (21.07 /100 000). The male to female ratio was 1.40:1, and the difference between male and female was significant (χ²=4 321.276，P＜0.05). The annual incidence of mumps showed a certain periodic change, with the incidence peak and trough alternating every 4 - 5 years. The majority of patients were under 15 years old, accounting for 85.32% of the total number of cases. The patients mainly included students, preschool children and scattered children. The highest average incidence was in Nanning City with 40 231 cases (42.08/100 000), and the lowest was in Qinzhou City with 3 466 cases (8.16/100 000). From 2012 to 2024, a total of 210 mumps outbreaks with 4 483 cases were reported in Guangxi. Conclusion The incidence of mumps in Guangxi from 2012 to 2024 shows a periodic change and obvious seasonality. People under 15 years old are the key group at risk of mumps. The prevention and control of the epidemic of mumps in schools and kindergartens should be strengthened. It is suggested to carry out long-term monitoring of mumps as well as immune effect research, and continue to maintain a high vaccination rate of 2 doses of mumps-containing vaccines.

Hydroxyectoine, a vital compatible solute, is widely utilized in cosmetics, food, pharmaceutical industries, and biologics. However, the current microbial fermentation methods for hydroxyectoine production face challenges including insufficient precursor supply and low yields. Therefore, developing engineering microbial strains capable of efficiently synthesizing hydroxyectoine is of great significance. In this study, we first constructed a high-yield ectoine-producing strain ECT04 by multi-copy integration of the ectoine synthesis genes ectABC into the pseudogene loci of Escherichia coli MG1655(DE3), achieving an ectoine titer of 6.03 g/L. Subsequently, we employed plasmids with varying copy numbers to express ectD from Chromohalobacter salexigens to enable the conversion for hydroxyectoine production. We further investigated the effects of promoter, co-substrate ɑ-ketoglutarate, Fe²⁺ concentration, and dissolved oxygen on hydroxyectoine synthesis. Through fed-batch fermentation in a 7-L bioreactor, we significantly enhanced the hydroxyectoine production efficiency, attaining a final titer of 8.58 g/L and a productivity of 0.24 g/(L·h). This work successfully achieved the de novo synthesis of hydroxyectoine in E. coli, laying a foundation for the efficient bioproduction of this compound.
Escherichia coli/genetics* ; Fermentation ; Amino Acids, Diamino/biosynthesis* ; Bioreactors/microbiology* ; Metabolic Engineering/methods* ; Chromohalobacter/genetics* ; Plasmids/genetics*

With the extended survival period of breast cancer patients and the increasing health demands, the concomitant diseases of breast cancer have gradually attracted the attention of both doctors and patients, and it is imperative to conduct comprehensive management of these diseases, in which the general practitioners, as the more comprehensive and complex medical talents, have not yet played their due roles. In this article, we report two cases of comprehensive management of concomitant diseases of breast cancer through collaboration of general practitioners and specialists (integrated general and specialist care). The role and function of general practitioners in this process were deeply analyzed, and the establishment of a consultation-liaison general practice model to further promote the role of integrated general and specialist care in integrated oncology care was advocated.

Objective:To retrospectively evaluate the clinical efficacy and safety of total 3D laparo-scopic ileal ureters replacement for bilateral ureters combined with bladder augmentation in patients with post-radiotherapy long-segment bilateral ureteral strictures and contracted bladder.Methods:Clinical data of two patients(aged 72 and 54 years)with radiation-induced long-segment bilateral ureteral stric-tures and reduced bladder capacity,treated at the Sixth Affiliated Hospital of Jinan University from Octo-ber 2023 to June 2024,were analyzed.Both presented with bilateral flank pain,recurrent chills/fever,urinary frequency,and urgency.Preoperative ureteral stricture lengths were measured as follows:left 10.4 cm and right 8.7 cm in the first case;left 10.6 cm and right 11.7 cm in the second case.Bladder capacity assessed by nephrostomy-assisted antegrade urography was 90 mL and 130 mL respectively.Both underwent single-position,one-stage totally 3D laparoscopic bilateral ileal ureteral replacement and blad-der augmentation based on membrane anatomy principles,with regular postoperative follow-up.Results:Procedures were completed by the same experienced urologist.Operative times were 420 min and 355 min,with intraoperative blood loss of 50 mL(no transfusion required).Postoperative bowel function re-sumed at the end of 4.5 and 3 days.No major perioperative complications occurred.Ureteral stents were removed at 2 months postoperatively,with imaging showing improved hydronephrosis,unobstructed ure-teral drainage,symmetrical bladder morphology,and smooth walls.Postoperative bladder capacities were 230 mL and 250 mL.Follow-up durations were 10 and 8 months.Both patients experienced significant relief of flank pain and lower urinary tract symptoms.No complications(enteric fistula,urinary fistula,or metabolic acidosis)were observed.At the final follow-up,one patient had mildly elevated serum cre-atinine,while the other showed reduced levels compared with preoperative values;both remained stable.Conclusion:Membrane anatomy-based dissection facilitates safe mobilization of fibrotic ureters with mini-mal bleeding and collateral damage.Total intracorporeal 3D laparoscopic ileal ureters replacement for bi-lateral ureters combined with bladder augmentation effectively addresses long-segment ureteral obstruction and improves bladder capacity.This approach is technically safe and feasible,though further validation with larger clinical cohorts is warranted.

Objective:To evaluate the role of docosahexaenoic acid (DHA) in long-term cognitive impairment after multiple sevoflurane anesthesia in newborn mice.Methods:Clean-grade healthy male C57BL/6 mice, aged 6 days, were used in this study. Ten mice were divided into 2 groups ( n=5 each) by the random number table method: control group (group C) and sevoflurane group (group S). The animals inhaled 3% sevoflurane for 2 h at 6, 7 and 8 days after birth. The DHA content was detected by ultra-high performance liquid chromatography-mass spectrometry at 9 days of age. Fifty-two mice were selected and divided into 4 groups ( n=13 each) by a random number table method: control+ normal saline group (group C+ S), sevoflurane anesthesia + normal saline group (group S+ S), control+ DHA group (group C+ D), and sevoflurane anesthesia+ DHA group (group S+ D). The sevoflurane anesthesia method was the same as the one mentioned above. DHA 50 mg/kg was administered by intragastric gavage from postnatal days 6-19 (at 6, 7 and 8 days after birth, 2 h before anesthesia) in C+ D and S+ D groups. The equal volume of normal saline was given instead in C+ S group and S+ S group. The novel object recognition test was conducted at 37 days of age, and the Morris water maze test was performed at 42 days of age. The corpus callosum and hippocampal tissues were isolated at 47 days of age for examination of the ultrastructure of myelin (with a transmission electron microscope) and for determination of the expression of myelin basic protein (MBP) in hippocampal tissues (by Western blot). The G-ratio was calculated. Results:Compared with group C, the content of DHA in hippocampal tissues was significantly decreased in group S ( P<0.05). Compared with group C+ S, the discrimination index was significantly decreased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were decreased, the expression of MBP was down-regulated, and the G-ratio in the original platform and hippocampus was increased in S+ S group ( P<0.05). Compared with group S+ S, the discrimination index was significantly increased, the percentage of duration of staying at the target platform quadrant and the number of crossing the original platform were increased, the expression of MBP was up-regulated, and the G-ratio in the original platform and hippocampus was decreased in S+ D group ( P<0.05). Conclusions:The mechanism of long-term cognitive impairment following multiple sevoflurane anesthesia may be related to a decrease in the content of DHA, which subsequently leads to myelin structural damage in neonatal mice.

Objective To explore the clinical characteristics of children with bloodstream infections caused by Klebsiella pneumoniae(KP),analyze the high-risk factors for developing carbapenem-resistant Klebsiella pneumoniae(CRKP)bloodstream infection and poor prognosis.We hope to provide clinical evidence for reducing carbapenem-resistant strains and improving prog-nosis.Methods Clinical data of children with blood cultures confirmed KP infection admitted to the Department of Pediatrics,Pediatric Surgery,PICU,and NICU of Shanghai Jiao Tong University School of Medicine Affiliated Xinhua Hospital from 2016 to 2022 were collected.Based on antibiotics sensitivity results and prognosis,the patients were divided into CRKP and non-CRKP infection groups,good prognosis(recovered or improved)and bad prognosis(death or withdrawing treatment)groups.Clinical characteristics,laboratory tests,and risk factors were compared between these groups.Results Among 177 blood culture samples positive for KP,97 strains(54.8％)were found to be CRKP.Logistic regression analysis suggests a histo-ry of surgery during hospitalization(OR=2.678,95％CI:1.248-5.746,P=0.011),mechanical assisted ventilation(OR=2.774,95％CI:1.235-6.229,P=0.017),hospital stay≥25 days(OR=3.467,95％CI:1.431-8.401,P=0.006),platelet count≥237 × 109/L(OR=3.005,95％CI:1.268-7.124,P=0.012),and mean platelet volume(MPV)≥12fL(OR=3.011,95％CI:1.140-7.955,P=0.026)were possible risk factors for CRKP bloodstream infection.Mechanical assisted venti-lation(OR=2.819,95％CI:1.138-6.985,P=0.025),platelet count＜149 × 109/L(OR=0.238,95％CI:0.079-0.717,P=0.011)were possible risk factors for poor prognosis in pediatric patients.Conclusion The antibiotic resistance rate of KP blood-stream infection is high and shows an increasing trend.In KP blood stream infection children,hospital stay longer than 25 days,surgery history,mechanical assisted ventilation,platelet count≥237 × 109/L and MPV≥12 fL are at high risk for CRKP in-fection.CRKP infection is not a risk factor for bad prognosis,however,KP blood stream infection children with platelet count＜149 × 109/L and mechanical assisted ventilation high-risk factors for CRKP bloodstream infection are at risk for bad progno-sis.Special attention and timely intervention should be paid to these patients.

Objective To obtain tree shrew Vasorin(VASN)recombinant protein through prokaryotic expression and purification,prepare monoclonal antibody against tree shrew VASN by immunizing mice with this protein,and preliminarily evaluate its application value.Methods Reverse transcription-polymerase chain reaction(RT-PCR)was used to amplify the full-length sequence of tree shrew VASN gene in vitro.The tree shrew VASN gene fragment was inserted into pET-30a vector to construct pET-30a-VASN recombinant plasmid.The recombinant plasmid was subjected to double digestion with BamH Ⅰ and Sal Ⅰfor identification,and its correctness was further verified by sequencing.The recombinant plasmid with correct sequencing was transformed into BL21(DE3)competent cells,and isopropyl β-D-thiogalactoside(IPTG)was used to induce expression of VASN recombinant protein.Proteins were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE),and the VASN recombinant protein was purified by KCI.Purified recombinant protein was used to immunize BALB/c mice for four times,and serum antibody titer was detected by enzyme-linked immunosorbent assay(ELISA).Splenocytes from mice with serum antibody titer above 1:10 000 were used for cell fusion with myeloma cells.Hypoxanthine-aminopterin-thymidine(HAT)culture medium was first used to screen hybridoma cells.ELISA was used to screen positive hybridoma cell lines that could secrete specific antibodies,and monoclonal hybridoma cell lines were obtained by limiting dilution method.VASN monoclonal antibodies were prepared in large quantities by ascites induction method,purified using rProtein G,and the affinity and in vitro reaction specificity of the monoclonal antibodies were detected by ELISA and Western blotting.Results The full-length sequence of the tree shrew VASN gene was successfully amplified and the recombinant plasmid vector of tree shrew pET-30a-VASN was constructed.The sequence obtained by sequencing of the recombinant plasmid vector was identical to the tree shrew VASN target gene sequence.Recombinant protein VASN mainly existed in the form of inclusion bodies,and the purity after purification reached 90％,meeting the requirements of subsequent immunization experiments.After four immunizations with recombinant protein VASN,mouse serum antibody titer reached 1:729 000.Monoclonal positive hybridoma cell lines were obtained through ascites induction and purification,and the constant affinity value of monoclonal antibodies measured by ELISA reached 2.59x107 L/mol.Western blotting results showed that the tree shrew VASN monoclonal antibody could bind to tree shrew VASN recombinant protein,but it showed no binding reaction with porcine retinol-binding protein 4 recombinant protein,human VASN-leucine rich repeat recombinant protein,or bovine serum albumin.Anti-tree shrew VASN monoclonal antibody could specifically recognize VASN protein in tree shrew heart,liver,spleen,lung,kidney and muscle,with clear bands and clean background.Immunohistochemical detection results showed that this monoclonal antibody could recognize VASN protein in tree shrew spleen,lung,and tree shrew immortalized fibroblasts with high VASN mRNA expression levels,and the detection results were positive.Conclusion Monoclonal antibody against tree shrew VASN is successfully prepared.This antibody can be used for immunohistochemical detection of tree shrew immortalized fibroblasts,spleen tissue,and lung tissue,providing an important tool for further research on the function of VASN in tree shrew models.

Objective:To evaluate the relationship between preoperative gut microbiota and post-operative ventilator-associated pneumonia (VAP) in patients undergoing coronary artery bypass grafting.Methods:This was a secondary analysis of a previous research project study. Patients who received invasive mechanical ventilation treatment after elective off-pump coronary artery bypass grafting at the Second Hospital of Hebei Medical University from April to September 2023 were selected and divided into VAP group and non-VAP group based on whether VAP occurred after surgery. Fecal samples were collected from patients before surgery, and 16S rRNA gene sequencing was used to analyze the characteristics of preoperative gut microbiota in the two groups. The differences in the diversity of gut microbiota between the two groups were compared. The linear discriminant analysis was used to identify the gut microbiota with significant differences between groups (differential bacteria), and logistic regression analysis was conducted to assess the association between differential bacteria and VAP. Receiver operating characteristic curves were drawn to analyze the predictive value of the differential bacteria for VAP.Results:A total of 79 patients were finally included, with 25 in VAP group and 54 in non-VAP group. The Beta diversity analysis showed statistically significant differences between VAP group and non-VAP group (pseudo- F=2.00, P=0.002). The linear discriminant analysis indicated that Bifidobacterium, Blautia and Megamonas were enriched in non-VAP group, while Klebsiella was enriched in VAP group. Multivariate logistic regression analysis showed that the relative abundance of Bifidobacterium was a protective factor for postoperative VAP ( OR=0.32, 95% confidence interval [ CI] 0.15-0.71, P=0.005), and the relative abundance of Klebsiella was a risk factor for postoperative VAP ( OR=2.49, 95% CI 1.143-5.43, P=0.022). The area under the receiver operating characteristic curve of the relative abundance of Bifidobacterium for predicting VAP was 0.80 (95% CI 0.69-0.90, P<0.001) and of the relative abundance of Klebsiella was 0.70 (95% CI 0.57-0.83, P=0.005). Conclusions:Bifidobacterium is a protective factor, while Klebsiella is a risk factor for postoperative VAP in patients undergoing coronary artery bypass grafting, and the relative abundance of both bacteria has a certain predictive value for VAP.

Objective To analyze the clinical characteristics of co-infection of Talaromyces marneffei(TM)and non-tuberculous Mycobacterium(NTM)in human immunodeficiency virus(HIV)-negative patients.Methods Clinical data of 8 HIV-negative patients with co-infection of TM and NTM in a hospital from 2019 to 2022 were co-llected.Clinical manifestations,auxiliary examination,treatment and prognosis were retrospectively analyzed.Results Among the 8 patients,5 were females and 3 were males,with an average age of(52.25±12.31)years old.All patients presented TM and NTM disseminated infection.The major involved organs were lung(100％),lymph nodes(87.5％),and skin(75.0％).Clinical symptoms mainly included cough and expectoration(87.5％),fatigue(62.5％),joint and lumbosacral pains(62.5％),fever(50.0％),as well as skin and soft tissue abscess(50.0％),etc.Anti-interferon-γ(INF-γ)autoantibodies were detected in 4 patients and the results were positive.All 8 patients(100％)had pulmonary lesions,with chest CT mainly showing spots,patches,and striped shadows in both lungs.Among them,7 cases(87.5％)had increased and enlarged mediastinal lymph nodes,4 cases(50.0％)had pleural thickening and pleural effusion,2 cases each(25.0％for each)were accompanied by pulmonary mass shadows,bronchial stenosis,as well as increased and enlarged hilar lymph nodes.One case each(12.5％for each)had pulmonary cavity formation,bronchiectasis,and pericardial effusion.Conclusion The co-infection of TM and NTM in non-HIV patients presents disseminated infection,with multiple clinical symptoms.Chest imaging shows a wide variety of pulmonary lesions.It is prone to miss diagnosis in clinic,and the effect is not ideal after treatment for single pathogen infection.