OBJECTIVE To explore the value of bronchoalveolar lavage fluid(BALF)metagenomic next generation sequencing(mNGS)technique in diagnosis of the pulmonary cryptococcosis patients with different clinical features so as to raise the clinicians' awareness of the diagnosis of pulmonary cryptococcosis with BALF-mNGS.METHODS Totally 73 patients with pulmonary cryptococcosis and 73 non-pulmonary cryptococcosis patients who were treated in Wenzhou People's Hospital from Jan.2021 to May 2024 were recruited as the research subjects.The clinical data including baseline data,complications,clinical manifestations,clinical laboratory test data,ima-ging findings and serological test data were collected from the patients.The sensitivity and specificity of BALF-mNGS in diagnosis of the pulmonary cryptococcosis were observed,and the difference in the sensitivity in diagno-sis of the patients with various clinical features was analyzed.RESULTS A total of 73 patients with pulmonary cryp-tococcosis were enrolled in the study,44 of whom were male,and 29 were female;62 were positive for BALF-mNGS,11 were negative;all of the 73 non-pulmonary cryptococcosis patients were tested negative for BALF-mNGS,the sensi-tivity of the BALF-mNGS test was 84.93％,with the specificity 100.00％,the positive predictive value 100.00％,the negative predictive value 86.95％.Totally 45 patients had the symptoms of cough and expectoration,42 of whom were positive for BALF-mNGS,the test sensitivity of the patients with the symptoms of cough and expectoration was 93.33％,higher than that of the patients without the symptoms of cough and expectoration(P＜0.05).Totally 31 pa-tients had the lesion site of right lung,30 of whom were positive for BALF-mNGS,and the test sensitivity of the patients with lesion site of right lung was 96.77％,higher than that of the patients with the lesion site of left lung or both lungs(P＜0.05).Totally 44 patients had multiple lesions,41 of whom were positive for BALF-mNGS,the test sensitivity of the patients with multiple lesions was 93.18％,higher than that of the patients with single lesion(P＜0.05).CONCLUSIONS The sensitivity of the BALF-mNGS is high in diagnosis of the patients with pulmonary cryptococ-cosis,especially for the patients with symptoms of cough and expectoration,lesion site of right lung and multiple lesions.It can be used for the early diagnosis of pulmonary cryptococcosis.

Background and Aims:Obesity is often accompanied by symptoms of disordered eating.Although bariatric metabolic surgery can alleviate these symptoms,there are significant individual differences in postoperative outcomes,and effective predictive indicators are lacking.Liver and kidney function,along with lipid profiles,are closely related to metabolic status and may serve as useful markers for preoperative risk stratification and prognosis prediction.This study was conducted to explore the relationship between preoperative metabolic indicators and symptoms of disordered eating,thereby identifying postoperative recovery patterns among obese patients to support individualized management strategies.Methods:A total of 41 obese patients who underwent sleeve gastrectomy at Shanghai Sixth People's Hospital between September 2020 and June 2023 were enrolled,along with 36 healthy volunteers recruited during the same period.Participants completed the Eating Disorder Inventory-2(EDI-2)questionnaire,and serum samples were collected to assess liver function,kidney function,and lipid levels prior to surgery.The Mantel test was used to analyze correlations between metabolic indicators and EDI-2 scores.Latent profile analysis(LPA)was conducted using the indicators significantly correlated with EDI-2 scores to identify subgroups within the obese cohort.Linear mixed models were then applied to examine the trajectories of postoperative symptom changes across subgroups.Results:Levels of cystatin C,cholinesterase,gamma-glutamyl transferase,triglycerides,and apolipoprotein E were significantly higher in the obese group compared to the healthy group(all P＜0.05),and EDI-2 total score was also significantly elevated(P＜0.05);the prealbumin level in the healthy group was significantly higher than that in the obese group(P＜0.05).These six indicators were positively correlated with EDI-2 score(all r＞0.20,P＜0.05).Based on these markers,the LPA classified the obese group into two subgroups,with subgroup 2 exhibiting higher levels of most metabolic indicators than subgroup 1.During the 18-month postoperative follow-up,both subgroups showed reductions in EDI-2 score,but symptom improvement in subgroup 2 occurred later(month 6)compared to subgroup 1(month 4).Conclusion:Preoperative levels of cholinesterase,gamma-glutamyl transferase,prealbumin,triglycerides,and apolipoprotein E may serve as predictive indicators for improvement in disordered eating symptoms.Recovery patterns after bariatric surgery vary among obese patients with different metabolic profiles,highlighting the need for tailored intervention strategies.

Objective To establish an individualized nomogram model and evaluate its efficacy to provide a possible evaluation basis for the prognosis of lower third and abdominal part of oesophageal adenocarcinoma (EAC). Methods Lower third and abdominal part of EAC patients from 2010 to 2015 were chosen from the SEER Research Plus Database (17 Regs, 2022nov sub). The patients were randomly allocated to the training cohort and the internal validation cohort with a ratio of 7∶3 using bootstrap resampling. The Cox proportional hazards regression analysis was used to determine significant contributors to overall survival (OS) in EAC patients, which would be elected to construct the nomogram prediction model. C-index, calibration curve and receiver operating characteristic (ROC) curve were performed to evaluate its efficacy. Finally, the efficacy to evaluate the OS of EAC patients was compared between the nomogram prediction model and TNM staging system. Results In total, 3945 patients with lower third and abdominal part of EAC were enrolled, including 3475 males and 470 females with a median age of 65 (57-72) years. The 2761 patients were allocated to the training cohort and the remaining 1184 patients to the internal validation cohort. In the training and the internal validation cohorts, the C-index of the nomogram model was 0.705 and 0.713, respectively. Meanwhile, the calibration curve also suggested that the nomogram model had a strong capability of predicting 1-, 3-, and 5-year OS rates of EAC patients. The nomogram also had a higher efficacy than the TNM staging system in predicting 1-, 3-, and 5-year OS rates of EAC patients. Conclusion This nomogram prediction model has a high efficiency for predicting OS in the patients with lower third and abdominal part of EAC, which is higher than that of the current TNM staging system.

Objective To observe the therapeutic efficacy of Jisheng Shenqi Pills combined with endocrine therapy in alleviating lower urinary tract obstruction symptoms of patients with advanced prostatecancer(PCa).Methods A total of 68 patients with advanced PCa complicated by lower uri-nary tract obstruction symptoms were randomly divided into observation group and control group,with 34 cases in each group.The control group received endocrine therapy alone,while the observation group received Jisheng Shenqi Pills on the basis of the control group.Both groups underwent a course of 3-month treatment.The total therapeutic effectiveness rate,traditional Chinese medicine(TCM)syndrome scores,maximum urinary flow rate(Qmax),residual urinary volume(RUV),International Prostate Symptom Score(IPSS),prostate-specific antigen(PSA)levels,and Quality of Life(QOL)Scale scores were compared between the two groups.Results The total therapeutic effectiveness rate in the observation group was 94.12％,which was significantly higher than 82.35％in the control group(P＜0.05).After treatment,TCM syndrome scores decreased in both groups,with the obser-vation group had significantly lower scores than the control group(P＜0.05).Both groups exhibited an increase in Qmax and a decrease in RUV after treatment,with the observation group had significant-ly higher Qmax and lower RUV compared to the control group(P＜0.05).PSA level decreased signif-icantly in both groups after treatment(P＜0.05),but no significant difference was observed between the two groups(P＞0.05).IPSS score was decreased while QOL score was increased in both groups after treatment,with the observation group had significantly lower IPSS score and higher QOL score compared to the control group(P＜0.05).Conclusion Jisheng Shenqi Pills combined with endo-crine therapy effectively alleviates urinary obstruction symptoms,improves TCM syndromes,and en-hances quality of life in patients with advanced PCa.

Objective:To investigate the efficacy of radiotherapy in patients with advanced esophageal cancer receiving first-line chemoimmunotherapy.Methods:A retrospective analysis was conducted on the data of 137 patients with Stage Ⅳ esophageal squamous cell carcinoma (ESCC) treated at our hospital from January 2018 to May 2023. These patients were divided into two groups: a group treated with first-line chemoimmunotherapy combined with radiotherapy (chemoimmunotherapy + radiotherapy group, n = 43) and a group treated with only chemoimmunotherapy ( n = 94). Inverse probability of treatment weighting (IPTW) was applied to balance baseline characteristics between the groups. With overall survival (OS) and progression-free survival (PFS) as study endpoints, the survival data were analyzed using the Kaplan-Meier method, the log-rank test, and the Cox regression method. Results:Before calibration, the chemoimmunotherapy + radiotherapy group significantly outperformed the sole chemoimmunotherapy group in median PFS (13.6 months vs. 7.0 months; HR: 0.501, 95% CI: 0.309-0.811, P = 0.005). After calibration using the COX proportional-hazards model for age, gender, Eastern Cooperative Oncology Group (ECOG) performance status, smoking history, T/N/M stage, and tumor location, the chemoimmunotherapy + radiotherapy group still had significant advantages in PFS (14.7 months vs. 7.0 months; HR: 0.441, 95% CI: 0.261-0.745, P = 0.002). IPTW analysis further confirmed this trend (13.9 months vs. 7.0 months; HR: 0.492, 95% CI: 0.304-0.795, P < 0.001). Specifically, the median OS of the chemoimmunotherapy + radiotherapy group demonstrated significant improvement in all analyses: pre-calibration (29.5 months vs. 18.0 months; HR: 0.507, 95% CI: 0.297-0.867, P = 0.013), after calibration using the Cox model (27.5 months vs. 16.7 months; HR: 0.470, 95% CI: 0.266-0.830, P = 0.009), and after calibration using IPTW (29.5 months vs. 16.9 months; HR: 0.448, 95% CI: 0.262-0.764, P < 0.001). Conclusions:The combination of radiotherapy and first-line chemoimmunotherapy can significantly improve survival outcomes of patients with advanced ESCC, suggesting its potential as a standard treatment strategy.

Objective:To construct a sizeable naive human Fab phage display antibody library to screen high-affinity specific antibodies in vitro. Methods:Total RNA was extracted from peripheral blood mononuclear cells (PBMCs) of 126 healthy individuals, subsequently reverse-transcribed into cDNA, and used as a template. PCR amplification was performed to obtain the V H from IgG, IgM and light chain κ, λ, separately, with the initial PCR products serving as templates for a second round of PCR. Overlap extension PCR was employed to generate fragments of the κ and λ light chains. These fragments were ligated with the phage vector pNC3, which harbors the variable region 1 of the heavy chain, to construct a recombinant phage plasmid. This plasmid was then electroporated into competent Escherichia Coli TG1 cells to establish a naive human Fab phage display antibody library. One hundred clones were randomly selected for identification and sequencing, and antibody gene polymorphisms were analyzed using the IMGT database and MAFFT software. Recombinant α-hemolysin from Staphylococcus aureus was utilized to screen Fab antibody fragments through biopanning of the antibody library, followed by random selection of phage ELISA-identified clones. The positive clones (antigen A450∶blank control A450≥2.1) were sequenced. Results:Two large naive Fab phage display antibody libraries were successfully constructed, in which the capacity of κ and λ chain antibody libraries were 1.25×10 11 and 1.54×10 11, respectively. The titers for two antibody libraries were 6.04×10 13 CFU/ml and 3.50×10 13 CFU/ml. The positive transformation insertion rates for κ and λ chain antibody libraries were 96% (96/100) and 100% (100/100), respectively. Sequence analysis revealed that all antibody sequences were unique. The amino acid sequences in the skeletal region were relatively conserved. In contrast, significant variations in the length of the complementarity determining region (CDR) were found, and the diversity of amino acid sequence of the complementary determining region was high, especially the CDR3. Analysis using the IMGT database indicated that the sequences exhibited a broad distribution across variable-diversity-joining gene families. After six rounds of panning, specific phage antibodies enrichment targeting α-hemolysin were achieved. A total of 142 monoclonal antibodies were sequenced, yielding 8 distinct Fab antibody sequences. Conclusion:This study successfully constructed two naive human Fab phage display antibody libraries with large capacity and good diversity, which can be used for screening human antibodies for serum epidemiology.

OBJECTIVE To explore the value of bronchoalveolar lavage fluid(BALF)metagenomic next generation sequencing(mNGS)technique in diagnosis of the pulmonary cryptococcosis patients with different clinical features so as to raise the clinicians' awareness of the diagnosis of pulmonary cryptococcosis with BALF-mNGS.METHODS Totally 73 patients with pulmonary cryptococcosis and 73 non-pulmonary cryptococcosis patients who were treated in Wenzhou People's Hospital from Jan.2021 to May 2024 were recruited as the research subjects.The clinical data including baseline data,complications,clinical manifestations,clinical laboratory test data,ima-ging findings and serological test data were collected from the patients.The sensitivity and specificity of BALF-mNGS in diagnosis of the pulmonary cryptococcosis were observed,and the difference in the sensitivity in diagno-sis of the patients with various clinical features was analyzed.RESULTS A total of 73 patients with pulmonary cryp-tococcosis were enrolled in the study,44 of whom were male,and 29 were female;62 were positive for BALF-mNGS,11 were negative;all of the 73 non-pulmonary cryptococcosis patients were tested negative for BALF-mNGS,the sensi-tivity of the BALF-mNGS test was 84.93％,with the specificity 100.00％,the positive predictive value 100.00％,the negative predictive value 86.95％.Totally 45 patients had the symptoms of cough and expectoration,42 of whom were positive for BALF-mNGS,the test sensitivity of the patients with the symptoms of cough and expectoration was 93.33％,higher than that of the patients without the symptoms of cough and expectoration(P＜0.05).Totally 31 pa-tients had the lesion site of right lung,30 of whom were positive for BALF-mNGS,and the test sensitivity of the patients with lesion site of right lung was 96.77％,higher than that of the patients with the lesion site of left lung or both lungs(P＜0.05).Totally 44 patients had multiple lesions,41 of whom were positive for BALF-mNGS,the test sensitivity of the patients with multiple lesions was 93.18％,higher than that of the patients with single lesion(P＜0.05).CONCLUSIONS The sensitivity of the BALF-mNGS is high in diagnosis of the patients with pulmonary cryptococ-cosis,especially for the patients with symptoms of cough and expectoration,lesion site of right lung and multiple lesions.It can be used for the early diagnosis of pulmonary cryptococcosis.

Methods: This retrospective study analyzed 32 consecutive IDS patients who underwent UEDD surgery. Clinical features, laboratory data (erythrocyte sedimentation rate and C-reactive protein), and treatment outcomes were analyzed. Results: Definite microorganisms were identified in 27 patients (84.3%), with 24 (88.9%) meeting cure criteria. The cure rate was significantly higher in the detected pathogen group compared to the undetected pathogen group (88.9% vs. 80%; χ²=19.36, p<0.0001). Metagenomic next generation sequencing (mNGS) provided faster diagnosis (41.72±6.81 hours) compared to tissue culture (95.74±35.47 hours, p<0.05). The predominant causative pathogen was Mycobacterium tuberculosis, followed by Staphylococcus aureus. Significant improvements were observed in Visual Analog Scale pain scores, from a mean of 7.9 preoperatively to 1.06 at 1 year postoperatively. The Oswestry Disability Index revealed a similar trend, showing significant improvement (p<0.05). Conclusions UEDD is a viable alternative to traditional open surgery for managing IDS in high-risk patients. UEDD offers a dual therapeutic-diagnostic advantage during the initial admission phase, enabling simultaneous debridement, neurological decompression, and targeted biopsy in a single intervention. Compared with traditional tissue culture, mNGS enables rapid microbiological diagnosis and extensive pathogen coverage.