ObjectiveTo identify the pathogen species responsible for the wilt disease of Tetradium ruticarpum in Chongqing， investigate there biological characteristics， and screen effective fungicides， so as to provide a theoretical basis for disease control in production. MethodsThe pathogen was isolated via the tissue culture method. Pathogenicity was verified according to Koch's postulates. The pathogen was identified based on morphological characteristics and multi-gene phylogenetic analysis. The mycelial growth rate method was used for biological characterization of the pathogen and fungicide screening. ResultsThe pathogen colonies were nearly circular with irregular edges， white， short， velvety aerial hyphae， and pale purple undersides. Macroconidia were colorless， sickle-shaped， with 3-5 septa， while microconidia were transparent， elliptical， aseptate or with 1-2 septa. Multi-gene phylogenetic analysis showed that the pathogen clustered in the same clade as Fusarium fujikuroi with 100% support， which， combined with morphological characteristics， identified the pathogen causing wilt of T. ruticarpum in Chongqing as F. fujikuroi. The optimal conditions for the mycelial growth of F. fujikuroi were mung bean agar （MBA） with glucose as the carbon source， beef extract and yeast powder as nitrogen sources， 28 ℃， pH 7.0， and alternating light/dark conditions. The optimal conditions for sporulation were potato dextrose agar （PDA） with glucose as the carbon source， beef extract as the nitrogen source， 28 ℃， pH 7.0， and complete darkness. Among chemical fungicides， phenazine-1-carboxylic acid exhibited the strongest inhibitory effect on F. fujikuroi. Shenqinmycin and tetramycin were the most effective bio-fungicides. ConclusionThis study is the first to report F. fujikuroi as the causal agent of wilt disease in T. rutaecarpa. The chemical fungicide phenazine-1-carboxylic acid and the bio-fungicides shenqinmycin and tetramycin showed strong inhibitory effects against F. fujikuroi.

Objective: To explore the association role of anxiety and depressive symptoms with Internet gaming disorder (IGD) and emotional-behavioral problems among junior high school students, so as to provide references for preventing the occurrence and progression of emotional-behavioral problems in middle school students. Methods: From September to October 2024, a random cluster sampling method was adopted to select 38 069 students from 28 junior high schools in 14 counties (cities and districts) and functional areas of Jining City, Shandong Province. Surveys were conducted by Nine item Internet Gaming Disorder Scale-Short Form, Generalized Anxiety Disorder-7, Patient Health Questionnaire-9, and Strengths and Difficulties Questionnaire. The correlation of emotional and behavioral issues, online game addiction, anxiety and depressive symptoms among junior high school students was analyzed by Speaman correlation analysis. Mediation analysis was used to explore the role of anxiety and depressive symptoms between IGD and emotional-behavioral problems among junior high school students. Results: The detection rates of emotional-behavioral problems, anxiety symptoms, depressive symptoms, and IGD among junior high school students were 19.24 %, 9.48%, 16.69%, and 19.68%, respectively. The scores of emotional-behavioral problems, anxiety symptoms, depressive symptoms and IGD were 10.00 (6.00, 15.00), 3.00 (0.00, 7.00), and 4.00 (1.00, 8.00), 13.00 (10.00, 19.00), respectively. There were positive correlations between each pair of emotional-behavioral problem, IGD, anxiety symptoms, and depressive symptoms scores ( r =0.51-0.81, all P <0.01). Anxiety and depressive symptoms played mediating roles between IGD and emotional-behavioral problems among junior high school students, with mediating effect values of 0.16 (95% CI =0.15-0.17) and 0.12 (95% CI =0.11-0.14), respectively (both P <0.01). Conclusions Anxiety and depressive symptoms mediate the relationship between IGD and emotional-behavioral problems among junior high school students. It is necessary to identify early signs of IGD among junior high school students, strengthen intervention for anxiety and depressive symptoms, to prevent the occurrence and development of emotional and behavioral issues.

Objective:To explore the choice willingness and related factors of peritoneal dialysis patients regarding automated peritoneal dialysis (APD), in order to provide a basis for future technological improvements and enhancing the application rate of APD.Methods:This was a single-center cross-sectional study. Patients newly initiated on peritoneal dialysis due to end-stage renal disease in the Renal Division, Peking University First Hospital between January 1, 2021 and May 30, 2023 were enrolled. Demographic characteristics, clinical information, and laboratory data were collected. After completing initial training on APD and providing hands-on experience, a self- designed semi-quantitative questionnaire survey was administered to assess patients' willingness to adopt APD and to evaluate potential barriers to its implementation. Factors associated with patients' choice of APD were further analyzed.Results:A total of 203 newly enrolled peritoneal dialysis patients were included in the study, with an age of (53.40±15.86) years, and 57.1% (116 patients) of the participants were male. Among them, 71 patients (35.0%) chose APD, while 132 (65.0%) did not. There were no significant differences in the clinical characteristics between the APD group and the non-APD group (all P>0.05). Compared with the non-APD group, the APD group had a higher proportion of individuals who frequently traveled for business ( χ2=11.442, P<0.001) and a higher proportion with smaller peritoneal cavity volumes ( χ2=5.639, P=0.018). Among patients in the two group, significant differences were observed in self-rated difficulty of APD operation ( χ2=22.291, P<0.001), availability of space for APD setup ( χ2=21.773, P<0.001), 8-hour nocturnal APD treatment ( χ2=17.540, P<0.001), learning difficulty of machine operation ( P<0.001), learning difficulty of machine setting ( P<0.001), impact of APD on sleep ( χ2=6.826, P=0.033), impact of APD on nocturnal urination ( χ2=19.428, P<0.001), and annual cost of APD equipment ( χ2=39.066, P<0.001). Multivariate logistic regression analysis indicated that reimbursement for APD supplies ( OR=3.736, 95% CI 1.655-8.435, P=0.002), with smaller abdominal cavity volume ( OR=58.610, 95% CI 5.000-687.007, P=0.001), the acceptability of 8-hour nocturnal APD treatment ( OR=5.312, 95% CI 1.256-22.461, P=0.023), the low degree of difficulty in learning machine setting ( OR=29.299, 95% CI 2.025-423.812, P=0.013) and the annual cost of accessible APD equipment ( OR=3.643, 95% CI 1.348-9.842, P=0.011) were independent factors associated with patients' choice of APD treatment. Conclusion:Reimbursement for APD supplies, smaller abdominal cavity volume, the acceptability of 8-hour nocturnal APD treatment, the low degree of difficulty in learning machine setting, and the annual cost of accessible APD equipment are the main factors related patients' decision to choose APD treatment.

A 57-year-old female patient with small cell lung cancer was treated with serplulimab (100 mg by intravenous infusion on the first day, 21 days as a cycle). After the 10th cycle of treatment, the patient suddenly presented drowsiness, shortness of breath, dry mouth, nausea, vomiting, fatigue and other symptoms. Laboratory tests showed fasting blood glucose 38.9 mmol/L, glycosylated hemoglobin 7.9%, serum C peptide 0.2 μg/L, pH value 7.05, anion gap 31 mmol/L, anti-islet cell antibody 36 kU/L, anti-glutamic acid decarboxylase antibody 131 kU/L, urine ketone body (+++). The patient was diagnosed with fulminant type 1 diabetic ketoacidosis, which was considered to be related to serplulimab. The drug was stopped, and insulin, rehydration, correction of acid-base imbalance, and other treatments were given. After 3 days, the patient′s consciousness returned to normal. After 12 days, her breathing was stable, dry mouth, nausea, vomiting and other symptoms were relieved. Laboratory tests showed random blood glucose 13.6 mmol/L, pH value 7.44, anion gap 6 mmol/L, and urine ketone body negative. After 26 days, her random blood glucose was controlled at 10.0-12.0 mmol/L.

Objective To compare the efficacy,acceptability and safety of a low-volume magnisium sodicum potassium sulfate oral sulfate solution(OSS)with polyethylene glycol(PEG)electrolytes powder in bowel preparation for colonoscopy.Methods A prospective,single-blinded and single-center cohort study was conducted.The ambulatory and hospitalized 1 037 patients who underwent colonoscopy from April 2023 to January 2024 were enrolled.Participants were divided into OSS group(639 cases)and PEG group(398 cases),according to the bowel cleansing drugs taken orally.After propensity score matching(PSM),each group included 385 cases.The success rate of bowel preparation,scores of Boston bowel preparation scale(BBPS),medication taste,patients'satisfaction and the occurrence of adverse events were compared.Results The success rate of bowel preparation in the OSS group was 96.4％(371/385),higher than the 91.7％(353/385)in the PEG group,and the difference was statistically significant(P＜0.05).The total and segmented BBPS scores of the OSS group were higher than those of the PEG group,the differences were statistically significant(P＜0.05).The medication taste and patients satisfaction of the OSS group were significantly better than those of the PEG group,the differences were statistically significant(P＜0.05).There was no statistically significant difference in incidence of adverse reactions between the two groups(P=0.800).Conclusion Compared to PEG,OSS has a better intestinal cleaning effect,medication taste,and patients satisfaction.In addition,OSS has security that is not inferior to PEG.

Objective To explore the value of analysis on the incidence of healthcare-associated infection(HAI)based on disease diagnosis-related grouping(DRG),case mix index(CMI),and relative weight(RW).Methods All discharged cases,DRG and HAI status in a tertiary first-class general hospital from January 1 to December 31,2023 were analyzed retrospectively.Incidences of HAI in different departments were adjusted and compared by CMI.Incidences of HAI in different DRG groups were adjusted by RW.Results Among the 47 695 cases included in the analysis,757 were HAI cases,including 225 DRG groups.The department of critical care medicine had the highest incidence of HAI(11.98％).After CMI adjustment,departments with higher incidence of HAI were main-ly the department of respiratory and critical care medicine(3.96％),department of critical care medicine(3.04％),and department of neurology(2.85％),et al.DRG groups with the top five high incidence of HAI were AH11(tracheotomy and with ventilator support ≥96 hours or extracorporeal membrane oxygenation[ECMO],accompa-nied by major complications and comorbidity[MCC],50.00％),BC29(ventricular shunt and revision surgery,31.43％),BB21(craniotomy other than trauma,accompanied by MCC,27.56％),BB11(craniotomy of brain trauma,accompanied by MCC,26.32％),and GB1A(major surgery of esophagus,stomach,and duodenum,accompanied by major or moderate complications and comorbidity,16.00％).After RW adjustment,the DRG groups with the top five high incidence of HAI were ES21(respiratory system infection/inflammation,accompanied by MCC,5.89％),BR21(cerebral ischemic disease,accompanied by MCC,5.17％),FR11(heart failure,shock,accompanied by MCC,4.80％),BC29(4.57％)and AH11(3.57％).Conclusion Analyzing the incidence of HAI based on CMI and RW can help to identify key departments and disease groups for infection prevention and control,and provide reference for precise prevention and control of HAI in the new era.

Objective To investigate the preventive effect of ipratropium bromide on sufentanil-induced cough during general anesthesia induction.Methods A total of 201 patients undergoing elective surgery under general anesthesia from June 2022 to September 2024 were selected and divided into the ipratropium bromide group(group P,n=100)and the normal saline group(group C,n=101).Thirty minutes before anesthesia in-duction,group P and group C received oral administration of ipratropium bromide inhalation aerosol(2 puffs,equivalent to 42 μg C20H30BrNO3·H2O)and 0.9%normal saline spray(2 puffs),respectively.Anesthesiolo-gists intravenously administered sufentanil(0.5 μg/kg)within 5 seconds,followed by propofol(2 mg/kg)and rocuronium(0.6 mg/kg)after 3 minutes.Endotracheal intubation was performed using a videolaryngo-scope after achieving satisfactory muscle relaxation.The occurrence of cough within 1 minute after sufentanil administration was recorded.Heart rate(HR),mean arterial pressure(MAP),and pulse oxygen saturation(SpO2)were measured at different time points.Results Within 1 min after sufentanil administration,the in-cidence of severe cough and total cough in group P was significantly lower than that in group C(P<0.05).There was no significant difference in HR,MAP,or SpO2 between the two groups at different times(P>0.05).No adverse reactions such as nausea,vomiting,or arrhythmia occurred in either group.Conclusion In-haling ipratropium bromide 30 min before anesthesia induction could effectively reduce sufentanil-induced cough and decrease the incidence of severe cough.

Somatostatin receptor 1 (SSTR1) is a crucial therapeutic target for various neuroendocrine and oncological disorders. Current SSTR1-targeted treatments, including the first-generation somatostatin analog lanreotide (Lan) and the second-generation analog pasireotide (Pas), show promise but encounter challenges related to selectivity and efficacy. This study presents high-resolution cryo-electron microscopy structures of SSTR1 complexed with Lan or Pas, revealing the distinct mechanisms of ligand-binding and activation. These structures illustrate unique conformational changes in the SSTR1 orthosteric pocket induced by each ligand, which are critical for receptor activation and ligand selectivity. Combined with the biochemical assays and molecular dynamics simulations, our results provide a comparative analysis of binding characteristics within the SSTR family, highlighting subtle differences in SSTR1 activation by Lan and Pas. These insights pave the way for designing next-generation therapies with enhanced efficacy and reduced side effects through improved receptor subtype selectivity.

To develop a method for determining the antibody-dependent cell-mediated phagocytosis (ADCP) activity of human epidermal growth factor receptor 2 (HER2)-targeted antibody drug conjugate (ADC) based on the reporter gene assay, we established an ADCP activity assay with Jurkat/NFAT/FcγRIIa cells as the effector cells and BT474 as the target cells. Then, the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were optimized by the method of design of experiment (DOE). The method showed a significant dose-response relationship, which was complied with the four-parameter equation: y=(A-D)/[1+(x/C)^B]+D. The durability ranges of the target cell density, the ratio of effector to target cells, the target cell adhesion time, the incubation time for drug administration, and the induction time after adding effector cells were (2.5-4.0)×10⁵ cells/mL, 3-5, 1.0-2.0 h, 0 h, and 5.0-6.0 h, respectively. The results of the methodological validation showed that the linear equation was y=1.106 8x-0.011 6, r=0.969 2. The established method showed the relative accuracy ranging from -6.59% to 2.98% and the geometric coefficient of variation less than 11% in the intermediate precision test. Furthermore, the method was target-specific. The method was then applied to the determination of ADCP activity of HER2-targeted ADC, demonstrating the result of (103.5±5.7)%. We developed a reporter gene assay for determining the ADCP activity of HER2-targeted ADC and the assay demonstrated high accuracy and good reproducibility, which proposes a highly efficient and approache for evaluating ADCP effect of this HER2-targeted ADC, and also provides a referable technique for characterizing the Fc effector functions of ADCs with diverse targets.
Humans ; Receptor, ErbB-2/immunology* ; Phagocytosis/drug effects* ; Immunoconjugates/immunology* ; Genes, Reporter ; Antibody-Dependent Cell Cytotoxicity ; Jurkat Cells

Glycosylation modification is an important post-translational modification of proteins, which participates in regulating protein half-life, biological activity, and immunogenicity, thereby affecting their functions. Glycoproteins expressed in Pichia pastoris predominantly carry high-mannose type glycans, primarily composed of mannose residues, which starkly contrasts with the complex-type glycans synthesized by mammalian cells. This study aims to transform the high mannose glycosylation modification of P. pastoris into a hybrid glycosylation modification similar to that of mammalian cells through genetic engineering technology. We introduced the mannosidase Ⅰ gene (MDSⅠ) from Trichoderma viride and the human β-1,2-N-acetylglucosaminyltransferase I gene (GnTⅠ) into a previously constructed P. pastoris strain (∆och1) capable of producing Man₈GlcNAc₂ glycans. To precisely regulate the expression of MDSⅠ and GnTⅠ, we designed various promoter combinations, including the strong inducible AOX promoter and the constitutive GAP promoter. The receptor-binding domain (RBD, residues 377-588) of the spike protein from the Middle East respiratory syndrome coronavirus (MERS-CoV) was selected as the reporter protein for this investigation (MERS-RBD). The N-glycosylation profile of MERS-RBD was systematically analyzed using PNGase F digestion coupled with mass spectrometry. The results showed that after the knockout of och1 and the introduction of MDSⅠ and GnTⅠ genes with different promoter combinations, P. pastoris strains capable of producing GlcNAcMan₅GlcNAc₂ glycans were successfully generated. When the AOX promoter was used to control the MDSⅠ gene and the GAP promoter was used to control the GnTⅠ gene, the engineered strain exhibited the highest proportion of hybrid-type GlcNAcMan₅GlcNAc₂ glycans, which accounted for 68.38% of the total N-glycosylation. In conclusion, we successfully engineered a P. pastoris strain capable of synthesizing hybrid-type GlcNAcMan₅GlcNAc₂ glycans, establishing a foundation for subsequent research on the biosynthesis of complex-type N-glycans in P. pastoris.
Glycosylation ; Glycoproteins/genetics* ; Polysaccharides/metabolism* ; N-Acetylglucosaminyltransferases/metabolism* ; Pichia/metabolism* ; Humans ; Mannosidases/metabolism* ; Genetic Engineering ; Trichoderma/genetics* ; Recombinant Proteins/genetics* ; Saccharomycetales