Objective To evaluate the diagnostic efficacy of metagenomic next-generation sequencing(mNGS)in detecting pathogens causing secondary infections in maintenance hemodialysis(MHD)patients,providing evidence for clinical implementation.Methods Sixty-five MHD patients with suspected secondary infections admitted to the blood purification treatment center of Tianjin University Tianjin Hospital from October 2022 to March 2024 were selected as the research subjects.All enrolled patients underwent routine laboratory examinations and imaging tests,the peripheral blood,sputum,bronchoalveolar lavage fluid(BALF),urine,feces,pleural effusion,and peritoneal effusion were collected for mNGS and traditional isolation and culture respectively.The infection status of MHD patients was analyzed.Compare the sensitivity,specificity,diagnostic accuracy,positive predictive value,negative predictive value,pathogen detection rate,and detection cycle across the two etiological diagnostic methods.Results Among the enrolled patients,11 were excluded from infection after comprehensive analysis and diagnostic treatment,and 54 were definitively diagnosed with MHD-associated secondary infections,including 20 cases of respiratory system infection(accounting for 37.04%),18 cases of bloodstream infection(33.33%),13 cases of urinary system infection(24.07%),2 cases of pleurisy(3.71%),and 1 case of peritonitis(1.85%).Among these infected cases,there were 12 cases of single bacterial infection,6 cases of single fungal infection,3 cases of atypical pathogen infection,and 33 cases of overlapping infections detected.The most common pathogens in respiratory infections,bloodstream infections,and urinary tract infections were Klebsiella pneumoniae,Staphylococcus aureus,and Escherichia coli respectively.Through mNGS,6 additional cases of Pneumocystispneumonia,3 cases of pulmonary Aspergillosis,3 cases of pulmonary tuberculosis,and 1 case of fungal pleurisy were successfully detected.A total of 25 patients had their medication adjusted based on the mNGS results,among whom 18 were discharged after improvement with treatment.The most common pattern of mixed infection was bacteria+fungi(16 cases),followed by bacteria+bacteria(9 cases),bacteria+fungi+atypical pathogens(4 cases),bacteria+atypical pathogens(3 cases),and fungi+fungi(1 case)in overlapping infections.The positive detection rate of overlapping infections by mNGS was significantly higher than that by traditional isolation and culture[78.79%(26/33)vs.12.12%(4/33),P<0.05].The sensitivity of mNGS in diagnosing pathogenic infections was significantly higher than that of traditional isolation and culture(overall diagnostic efficacy:96.30%vs.59.26%,precise diagnostic efficacy:95.35%vs.35.29%,both P<0.05).However,the specificity of the overall diagnostic efficacy of traditional isolation and culture was significantly higher than that of mNGS(100.00%vs.81.82%,P<0.05),the diagnostic coincidence rate,accuracy rate of single pathogen detection,and accuracy rate of overlapping pathogen detection of mNGS were all significantly higher than those of traditional isolation and culture(diagnostic coincidence rate of overall diagnostic efficacy:93.85%vs.66.15%,diagnostic coincidence rate of precise diagnostic efficacy:76.92%vs.35.38%,accuracy rate of single pathogen detection:80.95%vs.52.38%,accuracy rate of overlapping pathogen detection:66.67%vs.0,all P<0.05).The detection cycle of mNGS was significantly shorter than that of traditional isolation and culture[hours:27(26,29)vs.70(28,114),P<0.05].Conclusion mNGS can provide accurate and rapid etiological diagnosis for secondary infections in MHD patients,bringing benefits to precise diagnosis and targeted drug therapy.

Objective:To investigate the effect of oxidized-low density lipoprotein (ox-LDL) on contrast-induced renal tubular epithelial cell apoptosis and its mechanism.Methods:Human renal proximal tubular epithelial HK-2 cells were cultured in vitro and divided into four groups using the random number table method, including a control group, an ox-LDL group (50 μg/ml ox-LDL), an iohexol group (100 mg I/ml iohexol) and an ox-LDL+iohexol group (50 μg/ml ox-LDL+100 mg I/ml iohexol). Apoptosis rates were detected using the in situ terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling assay. Western blotting was used to detect the level of apoptosis by evaluating the presence of cleaved cysteine aspartic acid specific protease-3 (cleaved Caspase-3) protein. Mitochondrial membrane potential was detected using the JC-1 method. Mitochondrial/cytoplasmic cytochrome C (Cyt C) levels were detected by Western blotting to assess mitochondrial damage. Western blotting was used to detect the levels of the mitochondrial fission-related dynamin-related protein 1 (Drp1) and the mitochondrial fusion-related optic atrophy protein 1 (OPA1) to evaluate mitochondrial fusion/fission. Independent sample t test and one-way analysis of variance were used for data analysis. Results:The apoptosis rate [(30.12±3.19)%] and the relative expression of cleaved Caspase-3 protein (0.34±0.07) in the iohexol group were higher than those in the control group [(4.77±1.68)%, 0.23±0.05] ( P<0.05, 0.01). The apoptosis rate [(48.81±4.96)%] and the relative expression of cleaved Caspase-3 protein (0.53±0.05) in the ox-LDL+iohexol group were higher than those in the iohexol group (both P<0.01). The mitochondrial membrane potential (1.61±0.15) and mitochondrial/cytoplasmic Cyt C ratio (1.09±0.14) in the iohexol group were lower than those in the control group (6.15±0.52 and 3.09±0.45) (both P<0.01). The mitochondrial membrane potential (0.27±0.01) and the mitochondrial/cytoplasmic Cyt C ratio (0.24±0.04) in the ox-LDL+iohexol group were lower than those in the iohexol group (both P<0.01). The relative expression of Drp1 protein in the iohexol group (0.49±0.05) was higher than that in the control group (0.29±0.03), the relative expression of long OPA1 protein (0.24±0.03) was lower than that of the control group (0.40±0.03) (both P<0.01). The relative expression of Drp1 protein in the ox-LDL+iohexol group (0.64±0.02) was higher than that in the iohexol group, the relative expression of long OPA1 protein (0.07±0.02) was lower than that in the iohexol group (both P<0.01). Conclusions:ox-LDL can exacerbate contrast-induced renal tubular epithelial cell apoptosis, and its mechanism may be due to an imbalance in mitochondrial fusion and division caused by the contrast agent, leading to the increase mitochondrial damage.

Objective To evaluate the diagnostic efficacy of metagenomic next-generation sequencing(mNGS)in detecting pathogens causing secondary infections in maintenance hemodialysis(MHD)patients,providing evidence for clinical implementation.Methods Sixty-five MHD patients with suspected secondary infections admitted to the blood purification treatment center of Tianjin University Tianjin Hospital from October 2022 to March 2024 were selected as the research subjects.All enrolled patients underwent routine laboratory examinations and imaging tests,the peripheral blood,sputum,bronchoalveolar lavage fluid(BALF),urine,feces,pleural effusion,and peritoneal effusion were collected for mNGS and traditional isolation and culture respectively.The infection status of MHD patients was analyzed.Compare the sensitivity,specificity,diagnostic accuracy,positive predictive value,negative predictive value,pathogen detection rate,and detection cycle across the two etiological diagnostic methods.Results Among the enrolled patients,11 were excluded from infection after comprehensive analysis and diagnostic treatment,and 54 were definitively diagnosed with MHD-associated secondary infections,including 20 cases of respiratory system infection(accounting for 37.04%),18 cases of bloodstream infection(33.33%),13 cases of urinary system infection(24.07%),2 cases of pleurisy(3.71%),and 1 case of peritonitis(1.85%).Among these infected cases,there were 12 cases of single bacterial infection,6 cases of single fungal infection,3 cases of atypical pathogen infection,and 33 cases of overlapping infections detected.The most common pathogens in respiratory infections,bloodstream infections,and urinary tract infections were Klebsiella pneumoniae,Staphylococcus aureus,and Escherichia coli respectively.Through mNGS,6 additional cases of Pneumocystispneumonia,3 cases of pulmonary Aspergillosis,3 cases of pulmonary tuberculosis,and 1 case of fungal pleurisy were successfully detected.A total of 25 patients had their medication adjusted based on the mNGS results,among whom 18 were discharged after improvement with treatment.The most common pattern of mixed infection was bacteria+fungi(16 cases),followed by bacteria+bacteria(9 cases),bacteria+fungi+atypical pathogens(4 cases),bacteria+atypical pathogens(3 cases),and fungi+fungi(1 case)in overlapping infections.The positive detection rate of overlapping infections by mNGS was significantly higher than that by traditional isolation and culture[78.79%(26/33)vs.12.12%(4/33),P<0.05].The sensitivity of mNGS in diagnosing pathogenic infections was significantly higher than that of traditional isolation and culture(overall diagnostic efficacy:96.30%vs.59.26%,precise diagnostic efficacy:95.35%vs.35.29%,both P<0.05).However,the specificity of the overall diagnostic efficacy of traditional isolation and culture was significantly higher than that of mNGS(100.00%vs.81.82%,P<0.05),the diagnostic coincidence rate,accuracy rate of single pathogen detection,and accuracy rate of overlapping pathogen detection of mNGS were all significantly higher than those of traditional isolation and culture(diagnostic coincidence rate of overall diagnostic efficacy:93.85%vs.66.15%,diagnostic coincidence rate of precise diagnostic efficacy:76.92%vs.35.38%,accuracy rate of single pathogen detection:80.95%vs.52.38%,accuracy rate of overlapping pathogen detection:66.67%vs.0,all P<0.05).The detection cycle of mNGS was significantly shorter than that of traditional isolation and culture[hours:27(26,29)vs.70(28,114),P<0.05].Conclusion mNGS can provide accurate and rapid etiological diagnosis for secondary infections in MHD patients,bringing benefits to precise diagnosis and targeted drug therapy.

Background and purpose:Human epidermal growth factor receptor 2(HER2)is closely associated with drug efficacy and prognosis in urothelial carcinoma(UC).HER2 is a significant biomarker and therapeutic target in various tumors.In recent years,anti-HER2 antibody-drug conjugates have shown significant clinical efficacy in UC patients with HER2 overexpression.Therefore,an in-depth understanding of HER2 expression and its characteristics in Chinese UC patients is crucial to guide treatment decision-making,optimize treatment strategies and achieve personalized therapy.This study aimed to thoroughly investigate correlation of HER2 expression and clinicopathological characteristics in Chinese patients with UC.Methods:This study was a multicenter study that retrospectively included UC patients from urology departments of 8 tertiary hospitals in 5 geographical regions of China(North China,East China,South China,Central China and Northwest)whose tissue samples were collected from January 2023 to March 2024.Inclusion criteria:① age above 18 years;② UC diagnosed by histopathological or cytological examination;③ complete results of HER2 expression detection using immunohistochemistry(IHC)in the primary tumor site were required.Exclusion criteria:① diagnosed patients with tumors in other parts of the body;② physicians evaluated other situations that were not suitable for inclusion in this study.IHC results for HER2 expression and clinicopathological data were collected.HER2 expression was determined according to the criteria outlined in"Clinical pathological expert consensus on HER2 testing in urothelial carcinoma in China",with HER2 2+and 3+defined as HER2 overexpression.The HER2 expression and clinicopathological features were analyzed.This study was approved by the medical ethics committee of Fudan University Shanghai Cancer Center(ethical number:2301268-12)and was registered at China Clinical Trial Registry(registration number:ChiCTR2300069746).Results:A total of 1054 patients with UC were included.Most of the tumors were bladder UC(n=807,76.6%).The mean age of patients was(66.8±10.5)years,and the majority were male(78.5%).The HER2 overexpression rate was 58.4%(n=616),with an additional 23%of patients having HER2 1+expression(n=242),and a small proportion exhibiting negative HER2 expression(n=196,18.6%).HER2 expression was significantly associated with various clinical and pathological characteristics such as Eastern Cooperative Oncology Group(ECOG)performance status,history of cardiovascular disease,history of metabolic disorders,smoking,UC disease location,differentiation grade,pathological type,and tumor stage.Conclusion:Retrospective analysis of multi-center data shows that HER2 expression is frequently observed in Chinese UC patients,with an overexpression rate of up to 58.4%.Furthermore,HER2 expression is closely associated with various clinical and pathological features of UC patients.This study underscores the critical importance of accurately assessing HER2 expression in UC patient to guide personalized therapies.

Objective To investigate the effects of pretreatment with Xuebijing injection on renal tubular injury in rats with contrast-induced acute kidney injury(CI-AKI).Methods Twenty-four Sprague-Dawley(SD)rats were selected and divided into normal group,model group,control group,and treatment group according to the random number table method,with 6 rats in each group.The animal model of CI-AKI was prepared by adopting iohexol,and the normal group was not subjected to any treatment.The rats in the treatment group were injected with Xuebijing injection via the tail vein 15 hours before modeling until 24 hours after modeling.The injection volume was 10 mL/kg for every 6 hours.The control group was injected with normal saline at the same time point.After 24 hours of modeling,the urine of rats in each group was collected to determine the levels of blood urea nitrogen(BUN)and urine N-acetyl-β-D-gluco-aminidase(uNAG),and the blood was collected to determine the levels of serum creatinine(SCr).Then the rats were killed and the kidney tissues were extracted,and then stained with hematoxylin-eosin(HE),and the pathological changes of the kidney tissues were observed under the light microscope.Results BUN,SCr and uNAG were significantly higher in the model group than those in the normal group[BUN(μmol/L):37.29±6.18 vs.6.37±1.19,SCr(mmol/L):30.43±4.44 vs.14.90±1.61,uNAG(U/L):47.77±4.71 vs.11.32±3.62,all P<0.01];BUN,SCr and uNAG levels were obviously decreased in the treatment group compared to the model group[BUN(μmol/L):9.45±3.04 vs.37.29±6.18,SCr(mmol/L):19.83±2.16 vs.30.43±4.44,uNAG(U/L):21.70±6.21 vs.47.77±4.71,all P<0.05],however,BUN and uNAG in the treatment group were still significantly higher than those in the normal group[BUN(μmol/L):9.45±3.04 vs.6.37±1.19,uNAG(U/L):21.70±6.21 vs.11.32±3.62,P<0.05 or P<0.01];SCr in the treatment group was not statistically significant compared to the normal group(μmol/L:19.83±2.16 vs.14.90±1.61,P>0.05).Under the light microscope,the renal tubular epithelial cells at the junction of cortex and dermatomedulla in the kidneys of the model group were obviously vacuolated,accompanied by cell detachment and necrosis,and the tubules were dilated,with no obvious lesions in the glomeruli.The degree of damage in the control group and the treatment group was reduced compared with that in the model group.The degree of renal tubular damage in the model group was higher than that in the normal group;while the degree of renal tubular damage in the control group was significantly lower than that in the model group;and the degree of renal tubular damage in the treatment group was lower than that in the model group.There was no statistically significant difference in the degree of renal tubular damage between the treatment group and the control group.Conclusion Xuebijing injection may exert a protective effect on renal function in rats with CI-AKI by attenuating renal tubular injury.

Objective　To explore the application of time series models based on meteorological factors in the population density of Aedes albopictus in Hainan Province, and to provide a scientific basis for the prevention and control of dengue fever in Hainan Province. Methods　The density of Aedes albopictus in different habitats in 18 cities and counties of Hainan Province from 2017 to 2022 was monitored monthly using the double-mosquito net trapping index and the Breteau index. Mann-Kendall trend test was used to analyze the temporal trends of the two density indices; Spearman's correlation analysis was employed to assess the correlation strength between each meteorological factor and the two indexes, eliminating unrelated variables, and further selecting the final variables through the full-subset regression method. Three time-series models were constructed for the two density indices, with root mean square error (RMSE), mean absolute error (MAE), and other accuracy metrics used to determine the optimal model; predictions for the density indices for 2023-2024 were made. All statistical analyses were performed in R (4.3.1). Results　The net trapping and Breteau indices showed an overall decrease over the years (Z-values of Mann-Kendall trend test were -6.15 and -4.03, respectively, and P<0.05). The meteorological factor most strongly associated with the trap index was the monthly average minimum temperature; monthly mean minimum temperature and monthly mean relative humidity were strongly correlated with the Breteau index. Based on various evaluation indicators, the multivariate time series model demonstrated the highest accuracy. The study predicts one to two peaks in both the trap index and Breteau Index for the years 2023 and 2024, with peak periods occurring between June to September and May to September, respectively. The predicted value for 2023 aligns with the measured value, demonstrating outstanding predictive accuracy. Conclusions　This study has introduced meteorological factors into the seasonal time series model, allowing for more accurate predictions of the density of Aedes albopictus in Hainan Province from 2023 to 2024, providing a model framework for the prevention and control of dengue fever in Hainan Province.

BACKGROUND: LY01005 (Goserelin acetate sustained-release microsphere injection) is a modified gonadotropin-releasing hormone (GnRH) agonist injected monthly. This phase III trial study aimed to evaluated the efficacy and safety of LY01005 in Chinese patients with prostate cancer. METHODS: We conducted a randomized controlled, open-label, non-inferiority trial across 49 sites in China. This study included 290 patients with prostate cancer who received either LY01005 or goserelin implants every 28 days for three injections. The primary efficacy endpoints were the percentage of patients with testosterone suppression ≤50 ng/dL at day 29 and the cumulative probability of testosterone ≤50 ng/dL from day 29 to 85. Non-inferiority was prespecified at a margin of -10%. Secondary endpoints included significant castration (≤20 ng/dL), testosterone surge within 72 h following repeated dosing, and changes in luteinizing hormone, follicle-stimulating hormone, and prostate specific antigen levels. RESULTS: On day 29, in the LY01005 and goserelin implant groups, testosterone concentrations fell below medical-castration levels in 99.3% (142/143) and 100% (140/140) of patients, respectively, with a difference of -0.7% (95% confidence interval [CI], -3.9% to 2.0%) between the two groups. The cumulative probabilities of maintaining castration from days 29 to 85 were 99.3% and 97.8%, respectively, with a between-group difference of 1.5% (95% CI, -1.3% to 4.4%). Both results met the criterion for non-inferiority. Secondary endpoints were similar between groups. Both treatments were well-tolerated. LY01005 was associated with fewer injection-site reactions than the goserelin implant (0% vs . 1.4% [2/145]). CONCLUSION: LY01005 is as effective as goserelin implants in reducing testosterone to castration levels, with a similar safety profile. TRIAL REGISTRATION ClinicalTrials.gov, NCT04563936.
Humans ; Male ; Antineoplastic Agents, Hormonal/therapeutic use* ; East Asian People ; Gonadotropin-Releasing Hormone/agonists* ; Goserelin/therapeutic use* ; Prostate-Specific Antigen ; Prostatic Neoplasms/drug therapy* ; Testosterone

Objective: To explore the predictive value of circulating tumor cells (CTCs) characterization for time to castration resistance of newly diagnosed high volume metastatic castration sensitive prostate cancer (mCSPC) patients. Methods: Newly diagnosed high volume mCSPC patients were prospectively enrolled in this study from September 2015 to February 2017. The inclusion criteria include that the patients' age should be between 18 to 85 years old. The Prostate cancer should be diagnosed by biopsy or cytopathology. No endocrinological therapy, radiative therapy or chemotherapy was used before the study. High-volume metastatic lesion was confirmed by imaging. Those patients who accepted previous endocrinological therapy, radiative therapy or chemotherapy were excluded in this study. Those patients combined with concomitant tumor were also excluded. The health males were enrolled in the control group. All patients received androgen deprivation therapy (ADT) with goserelin plus bicalutamide (goserelin 3.6 mg subcutaneous injection, once a month plus bicalutamide 50mg orally, once a day). CanPatrol system was used to count CTCs in peripheral blood of patients and characterize CTCs based on expressions of epithelial markers(EpCAM and CK8/18/19) and mesenchymal markers(vimentin and twist). Primary endpoint was time to castration resistance. Survival analysis was conducted using Kaplan-Meier method and log-rank test was used to assess the difference of survival between groups, and univariate and multivariate analyses of prognostic factors were conducted using the Cox proportional hazards model. Results: A total of 108 newly diagnosed high volume mCSPC patients were enrolled in this study. The median age of enrolled patients was 68 years old (ranging 51-85 years old), and median PSA was 196.2 ng/ml(ranging 5.8-5 011.9 ng/ml). The median level of hemoglobin was 32 g/L(ranging 9-172 g/L). The median level of LDH was 179 U/L(ranging 49-630 U/L). The ECOG scores was 0-1 score in 94 cases(87.0%), 2 scores in 14 cases (13.0%). The Gleason scores was 6-7 in 20 cases (18.5%) and more than 8 in 88 cases (81.5%). All patients had bone metastatic lesions, among which 41 (38.0%) patients had more than 10 metastatic lesions and 6 (5.6%) patients with visceral metastasis, 30(27.8%) patients with limb bone metastasis. The median CTCs count was four, and ranging 0-35. Mesenchymal CTCs positive and negative (negative included CTCs negative, epithelial CTCs positive and biophenotypic CTCs positive) patients were 58(53.7%) and 50, respectively. There was no correlation between CTCs characterization with age, baseline PSA, Gleason score, ALP and other clinical parameters (P>0.05). In control group, the mean age was 26 years old (ranging 20-31 years old). No CTCs were detected among those people. After a median follow-up of 24 months (ranging 18-32 months), 90 patients (83.3%) progressed to castration resistant prostate cancer (CRPC). The median time to CRPC for patients of mesenchymal CTCs positive and negative was (10.5±1.4) and (14.0±3.4) months, respectively(P<0.001). Univariate analysis revealed CTCs characterization(HR=1.647, P=0.003), the number of metastatic lesions (HR=1.624, P=0.025)and limb bone metastasis(HR=1.706, P=0.019) were prognostic factors of time to CRPC; further multivariate analysis showed that only baseline mesenchymal CTCs positive (HR=1.562, P=0.008) was independent prognostic factors of unfavorable time to CRPC. Conclusions CTCs characterization can predict time to CRPC of newly diagnosed high volume mCSPC patients receiving ADT, and patients of baseline mesenchymal CTCs positive are more likely to progress to CRPC.

Acute kidney injury (AKI) is a clinical syndrome characterized by a rapid decline of renal function in a short period of time. In recent years, the incidence of AKI has been increasing gradually. Once the AKI occurs in a patient whose mortality in hospital may be increased significantly, the length of stay in hospital will be prolonged and the hospitalization costs increased in a short term, the long-term consequences include AKI recurrence, development into chronic kidney disease (CKD) or end-stage kidney disease (ESRD), cardiovascular events and death, etc. Ischemia/reperfusion (I/R) injury and contrast agents are common causes of AKI. The nephropathy induced by I/R and contrast agent is associated with intracellular calcium overload caused by Na+-Ca2+ exchanger (NCX). In this article, a systematic review of the relationship between NCX and AKI was conducted, aiming to provide a reference for further recognizing the prevention, treatment and prognosis of AKI.

Objective To evaluate the effects of autophagy on contrast-induced acute kidney injury (CI-AKI) in rat models.Methods Eighteen male rats were divided into control group (Con),CI-AKI group (CI-AKI) and rapamycin-pretreated group (Rapa).In the CI-AKI group,CI-AKI was induced by intraperitoneal injection of iohexol (12.25 g/kg I).In the Rapa group,rapamycin was given by intraperitoneal injection with a dose of 5 mg/(kg ·d) for consecutive 7 days,and then injected with iohexol (12.25 g/kg I).Rats in the Con group were injected by the same dose of saline.The renal function,renal histopathology,and the levels of LC3 Ⅱ / Ⅰ and Beclin-1 as well as catalase (CAT) in the kidneys of rats were evaluated one day after the injection.Results Compared with the Con group,serum creatinine in the CI-AKI group was significantly increased ((239.93±27.00)μmol/L) vs (51.70±10.59) μmol/L,P＜0.05),and the content of CAT was significantly decreased ((14.86 ± 0.32) U/mg vs (18.72±1.46) U/mg,P＜0.05).In the CI-AKI group,renal tubules were severely injured,and the expression of autophagy-related proteins LC3 Ⅲ / Ⅰ and Beclin-1 in renal tissue was increased.Compared with the CI-AKI group,the pretreatment of rapamycin (Rapa group) increased the expression of LC3 Ⅱ / Ⅰ and Beclin-1 as well as the content of CAT in renal tissue ((17.62±1.86) U/mg vs (14.86±0.32) U/mg,P＜0.05),and inhibited the increase of contrast-induced serum creatinine ((187.62± 47.76) μmol/L vs (239.93±27.00) μmol/L,P＜0.05) and renal tubule injury.Conclusions The results showed that contrast administration can induce autophagy activation in kidneys,while enhancing autophagy can attenuate contrast-induced oxidative stress injury and related renal injury.