Objective·To construct a pH-responsive manganese-based nanoprobe and explore the therapeutic efficacy of chemotherapy/ferroptosis synergistic treatment in breast cancer and the effect of pH-responsive magnetic resonance-activated imaging.Methods·BSA-MnO2@CPT(BMC)nanoprobes were prepared by biomineralization,and their physicochemical properties were characterized by transmission electron microscope(TEM)and dynamic light scattering.The magnetic resonance imaging(MRI)was used to evaluate the pH-responsive MRI T1 activation and time-dependent activation efficacy at the cellular level,with quantitative analysis of MRI T1 signal intensity.The reactive oxygen species(ROS)generation and glutathione(GSH)depletion by BMC nanoprobes were respectively detected by methylene blue(MB)and DTNB in vitro.The synergistic efficacy of chemotherapy and ferroptosis mediated by the nanoprobes in 4T1 breast cancer cells was evaluated using the Thiazolyl Blue Tetrazolium Bromide(MTT)assay.After co-incubation 4T1 cells with BMC,intracellular ROS levels were determined through the staining of ROS fluorescence indicator 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)and the level of lipid peroxide(LPO)expression was detected by using BODIPY581/591 C11 probe.A subcutaneous xenograft tumor model of 4T1 breast cancer was established in mice,with four experimental groups:Control group(PBS group),CPT group,BSA-MnO2(BM)group,and BMC group.The pH-responsive T1 activation effect of the BMC nanoprobes was dynamically monitored in vivo,while the ferroptosis-based antitumor efficacy was evaluated by measuring tumor volume and ferroptosis biomarkers(LPO and ROS).Results·TEM revealed that the prepared BMC nanoprobes exhibited a spherical morphology with an average diameter of approximately 150 nm.The MRI results demonstrated that the nanoprobes were pH-activable,exhibiting progressively enhanced T1 signal intensity under acidic conditions,and displaying pH-dependent r1 relaxivity enhancement.These findings validated their dual pH/time-responsive activation efficacy at the cellular level.In vitro solution-level MB and DTNB assays demonstrated that the BMC nanoprobes effectively enhanced the generation of ROS and the consumption of GSH.Fluorescence staining with DCFH-DA and BODIPY581/591 C11 demonstrated that the combination of ferroptosis effect and chemotherapy significantly enhanced intracellular generation of ROS and LPO accumulation.The MTT assay demonstrated that the survival rate of tumor cells significantly decreased to 17%(P=0.003).In vivo MRI demonstrated that the T1 signal was significantly enhanced and reached its peak at 4 h after tail vein injection of the BMC nanoprobes.Furthermore,in vivo antitumor therapy showed that the BMC group exhibited upregulated levels of LPO and ROS in tumor tissues,accompanied by marked tumor suppression(P=0.009).Conclusion·The pH-responsive theranostic BMC nanoprobes enhances antitumor efficacy via the synergistic interaction of chemotherapy and ferroptosis,while enabling tumor microenvironment-activated MRI.

Objective·To construct a pH-responsive manganese-based nanoprobe and explore the therapeutic efficacy of chemotherapy/ferroptosis synergistic treatment in breast cancer and the effect of pH-responsive magnetic resonance-activated imaging.Methods·BSA-MnO2@CPT(BMC)nanoprobes were prepared by biomineralization,and their physicochemical properties were characterized by transmission electron microscope(TEM)and dynamic light scattering.The magnetic resonance imaging(MRI)was used to evaluate the pH-responsive MRI T1 activation and time-dependent activation efficacy at the cellular level,with quantitative analysis of MRI T1 signal intensity.The reactive oxygen species(ROS)generation and glutathione(GSH)depletion by BMC nanoprobes were respectively detected by methylene blue(MB)and DTNB in vitro.The synergistic efficacy of chemotherapy and ferroptosis mediated by the nanoprobes in 4T1 breast cancer cells was evaluated using the Thiazolyl Blue Tetrazolium Bromide(MTT)assay.After co-incubation 4T1 cells with BMC,intracellular ROS levels were determined through the staining of ROS fluorescence indicator 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)and the level of lipid peroxide(LPO)expression was detected by using BODIPY581/591 C11 probe.A subcutaneous xenograft tumor model of 4T1 breast cancer was established in mice,with four experimental groups:Control group(PBS group),CPT group,BSA-MnO2(BM)group,and BMC group.The pH-responsive T1 activation effect of the BMC nanoprobes was dynamically monitored in vivo,while the ferroptosis-based antitumor efficacy was evaluated by measuring tumor volume and ferroptosis biomarkers(LPO and ROS).Results·TEM revealed that the prepared BMC nanoprobes exhibited a spherical morphology with an average diameter of approximately 150 nm.The MRI results demonstrated that the nanoprobes were pH-activable,exhibiting progressively enhanced T1 signal intensity under acidic conditions,and displaying pH-dependent r1 relaxivity enhancement.These findings validated their dual pH/time-responsive activation efficacy at the cellular level.In vitro solution-level MB and DTNB assays demonstrated that the BMC nanoprobes effectively enhanced the generation of ROS and the consumption of GSH.Fluorescence staining with DCFH-DA and BODIPY581/591 C11 demonstrated that the combination of ferroptosis effect and chemotherapy significantly enhanced intracellular generation of ROS and LPO accumulation.The MTT assay demonstrated that the survival rate of tumor cells significantly decreased to 17%(P=0.003).In vivo MRI demonstrated that the T1 signal was significantly enhanced and reached its peak at 4 h after tail vein injection of the BMC nanoprobes.Furthermore,in vivo antitumor therapy showed that the BMC group exhibited upregulated levels of LPO and ROS in tumor tissues,accompanied by marked tumor suppression(P=0.009).Conclusion·The pH-responsive theranostic BMC nanoprobes enhances antitumor efficacy via the synergistic interaction of chemotherapy and ferroptosis,while enabling tumor microenvironment-activated MRI.

Objective·To prepare self-assembled drug-loaded nanoprobes with activatable chemical exchange saturation transfer(CEST)imaging capability,and evaluate their imaging performance and therapeutic potential for photodynamic-sensitized pyroptosis in breast cancer in vivo and in vitro.Methods·GC nanoprobes co-loaded with gemcitabine(Gem)and chlorin e6(Ce6)were constructed by using a self-assembly strategy.The physicochemical properties of the GC nanoprobes were characterized by scanning electron microscopy(SEM)and dynamic light scattering(DLS).The pH-/time-dependent CEST activation and drug release profiles were investigated.The 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe was used to detect the generation of reactive oxygen species(ROS),and enzyme-linked immunosorbent assay(ELISA)was used to detect the release of inflammatory factors such as interleukin-1β(IL-1β)and IL-18 in mouse breast cancer 4T1 cells after treatment with GC nanoprobes with synergistic laser irradiation.Immunofluorescence was performed to detect immunogenic cell death(ICD)markers,including calreticulin(CRT)and high mobility group box 1 protein(HMGB1).The 4T1 breast cancer mouse models were established to validate tumor-specific CEST activation and evaluate anti-tumor efficacy by measuring tumor volume and detecting inflammatory factors and ICD markers.Results·SEM and DLS confirmed the uniform spherical morphology of the GC nanoprobes.The CEST imaging results showed that the nanoprobes had excellent pH-concentration and time-dependent activation imaging effects both in the simulated acidic microenvironment and at the cellular level in vitro.The drug release from this drug-loaded nanoprobe was 80％at pH 5.0,which was significantly higher than at pH 7.4(P=0.003).DCFH-DA fluorescence staining demonstrated that GC-mediated photodynamic therapy induced a significant generation of ROS.Analysis of pyroptosis-related factors revealed a marked increase in the release levels of IL-1β and IL-18(both P＜0.05),along with elevated fluorescence expression of CRT and HMGB1.The in vivo CEST imaging results showed that the CEST signal at the tumor site was significantly enhanced,peaking at 4 h with tail vein injection of GC.The GC nanoprobes with synergistic laser irradiation group showed markedly elevated inflammatory factors(IL-1β,IL-18),changed ICD biomarkers(HMGB1 and CRT),and significant tumor suppression,compared to the PBS control group(all P＜0.05).Conclusion·The GC nanoprobes enables specific CEST imaging-guided photodynamic therapy,effectively inducing pyroptosis and precise ablation of breast cancer.

Objective·To prepare self-assembled drug-loaded nanoprobes with activatable chemical exchange saturation transfer(CEST)imaging capability,and evaluate their imaging performance and therapeutic potential for photodynamic-sensitized pyroptosis in breast cancer in vivo and in vitro.Methods·GC nanoprobes co-loaded with gemcitabine(Gem)and chlorin e6(Ce6)were constructed by using a self-assembly strategy.The physicochemical properties of the GC nanoprobes were characterized by scanning electron microscopy(SEM)and dynamic light scattering(DLS).The pH-/time-dependent CEST activation and drug release profiles were investigated.The 2',7'-dichlorodihydrofluorescein diacetate(DCFH-DA)probe was used to detect the generation of reactive oxygen species(ROS),and enzyme-linked immunosorbent assay(ELISA)was used to detect the release of inflammatory factors such as interleukin-1β(IL-1β)and IL-18 in mouse breast cancer 4T1 cells after treatment with GC nanoprobes with synergistic laser irradiation.Immunofluorescence was performed to detect immunogenic cell death(ICD)markers,including calreticulin(CRT)and high mobility group box 1 protein(HMGB1).The 4T1 breast cancer mouse models were established to validate tumor-specific CEST activation and evaluate anti-tumor efficacy by measuring tumor volume and detecting inflammatory factors and ICD markers.Results·SEM and DLS confirmed the uniform spherical morphology of the GC nanoprobes.The CEST imaging results showed that the nanoprobes had excellent pH-concentration and time-dependent activation imaging effects both in the simulated acidic microenvironment and at the cellular level in vitro.The drug release from this drug-loaded nanoprobe was 80％at pH 5.0,which was significantly higher than at pH 7.4(P=0.003).DCFH-DA fluorescence staining demonstrated that GC-mediated photodynamic therapy induced a significant generation of ROS.Analysis of pyroptosis-related factors revealed a marked increase in the release levels of IL-1β and IL-18(both P＜0.05),along with elevated fluorescence expression of CRT and HMGB1.The in vivo CEST imaging results showed that the CEST signal at the tumor site was significantly enhanced,peaking at 4 h with tail vein injection of GC.The GC nanoprobes with synergistic laser irradiation group showed markedly elevated inflammatory factors(IL-1β,IL-18),changed ICD biomarkers(HMGB1 and CRT),and significant tumor suppression,compared to the PBS control group(all P＜0.05).Conclusion·The GC nanoprobes enables specific CEST imaging-guided photodynamic therapy,effectively inducing pyroptosis and precise ablation of breast cancer.

Objective:To explore the optimization of the standardized assessment tool for clinical diagnosis of Chinese developmental dyslexia (DD).Methods:A cross-sectional study was conducted from May to December 2023, in which 130 primary school children in grades 1 to 3 with clinical signs of literacy lag and positive screening results on the screening scales were recruited from the outpatient clinic of Child Health Care Medical Division, Shanghai Children′s Hospital, Shanghai Jiao Tong University School of Medicine. Chinese dyslexia screening behavior checklist for primary students (CDSBC) was used as the screening scales, and supplemented by dyslexia checklist for Chinese children. Referring to the standard procedure of the"expert advice on diagnosis and intervention of Chinese developmental dyslexia", the developmental dyslexia scale for standard mandarin (DDSSM) was used to evaluate the children′s literacy-related cognitive abilities and conduct the diagnostic assessment, and divided the children into learning backward group and the DD group. The t-test and χ2 test were used to compare the differences in the distribution of intelligence, literacy and attention deficit hyperactivity disorder between the two groups. Spearman′s correlation was used to analyze the correlation between the scores for each cognitive ability in the DDSSM and the CDSBC. Results:Of the 130 children, 90 were male, aged (8.3±1.0) years; 40 were female, aged (8.1±0.9) years. A final diagnosis of DD was made in 59 cases, of which 41 were males. There was no statistically significant difference in operational intelligence quotient (101±15 vs.100±15, t=0.53, P>0.05) and statistically significant difference in literacy of DDSSM (32±5 vs.21±4, t=11.56, P<0.001) between the learning backward group and the DD group. Eighteen cases (25.4%) of the learning backward group were children with attention deficit subtype attention deficit hyperactivity disorder (ADHD-I), and 16 cases (27.1%) in DD group, the difference in incidence between the two groups was not statistically significant ( χ2=0.05, P>0.05). There were correlations between the DDSSM (for oral vocabulary, morphological awareness and orthographic awareness) and the CDSBC total score ( r=-0.42, -0.32, -0.35, all P<0.01), but the correlations for visuospatial perception and rapid automatized naming with CDSBC total score were not statistically significant ( r=-0.09 and -0.20,both P>0.05). Conclusion:For literacy-related cognitive abilities, screening scales CDSBC are not sufficiently useful for assessment, so the introduction of standardized assessment tools DDSSM is an optimization of the clinical diagnosis of Chinese DD, which is crucial for achieving accurate diagnosis and intervention.

Objective To study the clinical manifestations and antibiotic sensitivity features of early-and late-onset invasive infections caused by group B Streptococcus (GBS). Methods A total of 96 infants with invasive GBS infections were enrolled prospectively from seven tertiary hospitals of GBS Infection Research Cooperative Group in southwest Fujian, such as Xiamen Maternal and Child Care Hospital, etc., from January 2016 to June 2018. According to the onset time of infection after birth, they were divided into early-onset GBS disease (GBS-EOD) group (<7 d, n=67) and the late-onset GBS disease (GBS-LOD) group (7-89 d, n=29). Clinical manifestations, disease spectrum, complications and outcomes of the two groups were compared. Drug sensitivity test was carried out using disk diffusion test. Chi-square or Fisher's exact test, two independent sample t-test or Mann-Whitney U tests were used for statistical analysis. Results (1) The average ages at onset in GBS-EOD and GBS-LOD groups were (15.8±6.7) h (0.5-142.0 h) and (25.0±8.1) d (9-89 d), respectively. The incidence of tachypnea, pallor, fever and convulsion were noted in 68.7% (46/67) vs 44.8% (13/29), 52.2% (35/67) vs 17.2% (5/29), 23.9% (16/67) vs 65.5% (19/29) and 7.5% (5/67) vs 48.3% (14/29) of GBS-EOD and GBS-LOD groups with χ2 values of 6.282, 10.199, 15.146 and 21.237 (all P<0.05). The main clinical manifestations of GBS-EOD were tachypnea and pallor, while most of the patients in the GBS-LOD group developed fever and convulsions. (2) The incidence of pneumonia, sepsis, meningitis, sepsis complicated by septic joints, pneumonia complicated by sepsis, sepsis complicated by meningitis and pneumonia complicated by sepsis and meningitis were noted in 43.3% (29/67) vs 20.7% (6/29), 9.0% (6/67) vs 17.2% (5/29), 0.0% (0/67) vs 3.4% (1/29), 0.0% (0/67) vs 6.9% (2/29), 31.3% (21/67) vs 13.8% (4/29), 6.0% (4/67) vs 31.0% (9/29) and 10.4% (7/67) vs 6.9% (2/29) of GBS-EOD and GBS-LOD groups. There was a statistically significant difference in the disease spectrum between the two groups (Fisher's exact test, all P<0.001). Compared with the GBS-LOD group, the GBS-EOD group had a higher incidence of pneumonia [85.1% (57/67) vs 41.4% (12/29), χ2=19.116, P<0.001] and a lower incidence of meningitis [16.4% (11/67) vs 41.4% (12/29), χ2=6.922, P=0.009]. Complications such as acute respiratory distress syndrome (ARDS), pulmonary hemorrhage, shock and persistent pulmonary hypertension of the newborn (PPHN) occurred much more in the GBS-EOD group than the GBS-LOD group [28.4% (19/67) vs 6.9% (2/29), 13.4% (9/67) vs 0.0% (0/29), 11.9% (8/67) vs 10.3% (3/29), 4.5% (3/67) vs 0.0% (0/29), χ2=13.683, P<0.001]. (3) Among the 96 patients, 23 (24.0%) had meningitis and 73 (76.0%) developed pneumonia and sepsis. Meningitis resulted in a higher fatality rate [17.4% (4/23) vs 4.1% (3/73), χ2=4.564, P=0.035] and longer average hospital stay [(37.2±12.6) vs (14.1±5.3) d, t=7.831, P<0.001] than pneumonia and sepsis. Seven out of the 19 meningitis survivors developed intracranial complications. (4) The overall fatality rate in this study was 7.3% (7/96) and no significant difference was found between GBS-EOD and GBS-LOD group [7.5% (5/67) vs 6.9% (2/29), χ2=0.010, P=0.982]. Among the 67 GBS-EOD infants, 58 (86.6%) occurred within 24 h and five of them died, but no death was reported in the other nine cases occurred after 24 h. (5) Totally 96 strains of GBS were isolated with 100% sensitivity to penicillin, ampicillin, cefazolin and meropenem, and 97% to vancomycin. Around 79.3%-91.0% of GBS isolates were resistant to clindamycin and erythromycin. Conclusions Clinial features vary greatly in GBS-LOD and GBS-EOD cases. Infants with meningitis have poor prognosis. The drug resistance rate of GBS to erythromycin and clindamycin are relatively high.

Objective Objective To explore the best dog urethra rectum fistula operation method.Methods In June 2016 to October 2016,15 healthy male dogs were selected as research objects,aged 2-3 years (mean 2.5 years),with a body mass of 12-15 kg (mean 13.5 kg).They were randomly divided into three groups,each group of five.5 male dogs were cut through the rectum.5 male dogs were cut through the perineum.5 male dogs were cut through the rectal sphincter of the anus.Postoperative retrograde urethrectomy rectum angiography,the urethral pressure injection of methylene blue white gauze aizen rectum fistula and histologic evaluation of each group were compared 1,2,3 moths after the operation.Results Anal rectum drag cold knife cut group,1,2,3 moths after the operation,postoperative indicated the exist of orificium fistulae.The urethral pressure injection of methylene blue could observe rectum white gauze aizen,and pathological histology suggested the mucosal lamina propria disorder.There were a large number of inflammatory cells,5 dogs models were successfully built.Abdominal perineal pathway group:1 dog was killed by itself.1 dog one month after operation in anus at 2 cm below diameter about 3 cm appeared in a mixture of urine fistula and urethral orifice outflow,no urine outflow.More than 3 dogs after a week in the edge of serious infection,a canine model of severe dysuria,1,2,3 moths after the operation,postoperative indicated the exist of orificium fistulae.The urethral pressure injection of methylene blue could observe rectum white gauze aizen.3 dogs models were built.Through anal sphincter incision group:five dogs of the urethral fistula were successful,and one dog was incontinent with fecal incontinence.Conclusions The rectal rectum could successfully produce the animal model of the urethral rectal fistula and provide the experimental subjects for the treatment of iatrogenic urethral fistula.

Glutathione (GSH) is the most important small-molecule, active oligopeptide in the maintenance of redox balance. GSH contributes to antioxidant and thiol equilibrium, as well as modulates the activities of many signaling molecules and redox-sensi-tive transcription factors by S-glutathionylation. Several studies have shown that the GSH level increased in various tumors. Additional-ly, increased GSH significantly contributes to drug resistance by eliminating ROS, detoxifying drugs, or participating in DNA repair. GSH-related metabolic enzymes are overexpressed in resistant cells, thereby regulating cellular response to chemotherapy drugs. Deple-tion of GSH or downregulation of GSH-related metabolic enzymes may effectively reverse drug resistance and promote resistant cells to restore sensitivity. This potential indicates that the GSH antioxidant system plays an important role in drug resistance. The GSH anti-oxidant system, as a potential target for antitumor therapy and reversal of drug resistance, has recently become an attractive focus in cancer research. This paper presents a review of the role of the GSH antioxidant system in drug resistance and discusses the therapeutic strategies targeting the GSH antioxidant system.

Cancer cells frequently share biological characteristics and energy metabolic processes distinct from normal cells. The specific metabolic phenotype was originally known as the Warburg effect. Researchers later discovered that cancer cells prefer to synthesize fatty acid de novo . Moreover, key enzymes involved in fatty acid synthesis and β-oxidation are overexpressed in tumor tissues, with low or without expression in normal tissues. Abnormal fatty acid metabolism is related to the survival and invasiveness of cancer cells, indicating that abnormal fatty acid metabolism provides the crucial components and energy sources of cancer cells. In recent years, the specific phenotype of abnormal fatty acid metabolism and the exploration of the role of this metabolic alteration in cancer biology and therapeutic strategies targeting the fatty acid metabolic pathways have become attractive focuses in cancer research. The role of active fatty acid metabolism in tumorigenesis and development, as well as the research progress in the development of the specific inhibitors, is reviewed in this paper.

Objective: To evaluate the pharmacokinetics of palonosetron hydrochloride in healthy volunteers. Methods: Thir-ty-one healthy volunteers were grouped into three palonosetron hydrochloride dosage regimens of 0.125, 0.25, and 0.5 mg. The plasma concentrations of palonosetron were determined by ultra high-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). DAS 2.1 software was applied to assess the plasma concentration-time data. Results:After intravenous injection of 0.125, 0.25, and 0.5 mg palonosetron to the subjects, the AUC0-168h values of palonosetron were (7.5±2.5), (15.2±4.0), and (34.8±9.7) μg· h·mL-1. The t1/2 values were (27.2±9.5), ( 27.2±6.5), and (31.4±5.6) h. Palonosetron exposure increased proportionally with the dose range of 0.125 mg to 0.5 mg. The correlation coefficient was 0.998. No grade 3 or grade 4 toxicity was observed during the study. Con-clusion:A rapid, sensitive, and selective UPLC-MS/MS method for palonosetron quantification in human plasma was developed and validated. All the participants indicated high tolerance throughout the study. Our data showed that palonosetron exhibits linear pharma-cokinetics over the the dose range of 0.125 mg to 0.5 mg.