Objective To investigate the molecular epidemiology of hypervirulent Klebsiella pneumoniae(HvKp)and its independent risk factors for infection,and to provide research basis for anti-infection treat-ment.Methods A total of 519 Klebsiella pneumoniae strains were collected from Inner Mongolia Forestry General Hospital from January 2020 to December 2022.String test was used to distinguish hypermyxoid strains(HMV-Kp)and non-HMV-KP.The rmpA,rmpA2,and iutA genes were detected by common PCR agarose gel electrophoresis to screen the HvKp strains.Multilocus sequence typing analysis was performed on 60 HvKp strains and the minimum spanning tree was drawn.Multivariate Logistic regression was used to ana-lyze the risk factors of HvKp infection.Results The positive rate of HMV-Kp was 39.69％,the positive rate of HvKp was 37.19％,and HMV-Kp accounted for 76.68％of HvKp.The detection rate of HvKp in general surgery department was the highest,and the detection rate of HvKp in pus specimens was the highest.By ST typing comparison,a total of 18 types of 60 HvKp strains were detected,ST23 type was the most common type(50.00％),followed by ST86 type(8.33％).Multivariate Logistic regression analysis showed that male,liver abscess,infection or suppuration of other tissues and organs,and use of macrolide antibiotics in the past 3 months were independent risk factors for HvKp infection(P＜0.05).Conclusion There is a strong associa-tion between HvKp and HMV-Kp strains,and ST23 type is the dominant type in this study.Male,liver ab-scess,infection or suppuration of other tissues and organs,and use of macrolide antibiotics in the past 3 months are independent risk factors for HvKp infection.

Objective:To evaluate the effect of emodin on liver injury in a mouse model of intestinal ischemia-reperfusion (I/R) and the role of heme oxygenase-1-mediated autophagy.Methods:Twenty-four SPF-grade healthy male C57BL/6 mice, aged 6-8 weeks, weighing 18-22 g, were divided into 4 groups ( n=6 each) using a random number table method: sham operation group (Sham group), I/R group, emodin group (E group) and emodin plus HO-1 inhibitor Zinc Protoporphyrin Ⅸ (ZnPP) group (ES group). The intestinal I/R injury model was established by clamping the superior mesenteric artery for 45 min followed by 120 min of reperfusion. Emodin 40 mg/kg dissolved in 5% methylcellulose sodium was given by gastric gavage once a day for 5 days before ischemia in E group. Emodin 40 mg/kg dissolved in 5% methylcellulose sodium was given by gastric gavage once a day for 5 days before intestinal I/R, and ZnPP 7.5 mg/kg was injected via the tail vein at 12 h before ischemia in ES group. Orbital venous blood samples were collected at the end of reperfusion for determination of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) concentrations. Then the mice were sacrificed, and liver tissues were obtained for microscopic examination of the pathological changes (after HE staining) and for determination of the activity of superoxide dismutase (SOD), content of malondialdehyde (MDA), expression of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) mRNA (by fluorescent quantitative polymerase chain reaction), the expression of HO-1, autophagy-related protein Beclin1 and microtubule-associated protein 1 light chain 3 (LC3) (by Western blot). The LC3-Ⅱ/Ⅰ ratio was calculated. Results:Compared with Sham group, the activity of SOD was significantly decreased, the content of MDA and serum ALT and AST concentrations were increased, the expression of IL-6 and TNF-α mRNA and HO-1 was up-regulated, the expression of Beclin1 was down-regulated, the LC3-Ⅱ/Ⅰ ratio was decreased ( P<0.05), and the pathological changes of liver tissues were found in I/R group. Compared with I/R group, the activity of SOD was significantly increased, the content of MDA and serum ALT and AST concentrations were decreased, the expression of IL-6 and TNF-α mRNA was down-regulated, the expression of HO-1 and Beclin1 was up-regulated, the LC3-Ⅱ/Ⅰ ratio was increased ( P<0.05), and the pathological changes of liver tissues were significantly attenuated in E group ( P<0.05). Compared with E group, the activity of SOD was significantly decreased, the content of MDA and serum ALT and AST concentrations were increased, the expression of IL-6 and TNF-α mRNA was up-regulated, the expression of HO-1 and Beclin1 was down-regulated, the LC3-Ⅱ/Ⅰ ratio was decreased ( P<0.05), and the pathological changes of liver tissues were aggravated in ES group. Conclusions:Emodin can alleviate liver injury induced by intestinal I/R in mice, and the mechanism may be related to the activation of HO-1-mediated autophagy.

Objective To establish an index system for assessing the quality of hospital accreditation data and determine the hierarchical weights of indices at different levels,providing a reference for enhancing the governance of such data.Methods A quality assessment system for hospital accreditation data was developed using literature analysis method and Delphi method.The analytic hierarchy process was employed to create a pairwise comparison judgment matrix to determine the weights of indices.Results An evaluation system was constructed with the following levels:"Indicator Screening Quality-Data Reporting Quality-Data Quality Control Quality-Data On-Site Verification Quality."The system includes 4 first-level indicators and 18 second-level indicators.The indicator with the highest weight among the first-level indicators was Data Reporting Quality(0.496 5),followed by Data Quality Control Quality(0.313 2).Among the second-level indicators,Timeliness(0.428 6)and Cooperation(0.428 6)had the highest weights.Conclusion The establishment of the quality evaluation system of hospital accreditation da-ta is a strategy to optimize the control of hospital accreditation data quality and is beneficial for continuous improvement of hospital accreditation data quality.

The density and composition of lymphocytes infiltrating colon tumors serve as predictive factors for the clinical outcome of colon cancer.Our previous studies highlighted the potent anti-cancer properties of the principal compounds found in Garcinia yunnanensis(YTE-17),attributing these effects to the regu-lation of multiple signaling pathways.However,knowledge regarding the mechanism and effect of YTE-17 in the prevention of colorectal cancer is limited.In this study,we conducted isobaric tags for relative and absolute quantification(iTRAQ)analysis on intestinal epithelial cells(IECs)exposed YTE-17,both in vitro and in vivo,revealing a significant inhibition of the Wnt family member 5a(Wnt5a)/c-Jun N-terminal kinase(JNK)signaling pathway.Subsequently,we elucidated the influence and mechanism of YTE-17 on the tumor microenvironment(TME),specifically focusing on macrophage-mediated T helper 17(Th17)cell induction in a colitis-associated cancer(CAC)model with Wnt5a deletion.Additionally,we performed the single-cell RNA sequencing(scRNA-seq)on the colonic tissue from the Wnt5a-deleted CAC model to characterize the composition,lineage,and functional status of immune mesenchymal cells during different stages of colorectal cancer(CRC)progression.Remarkably,our findings demon-strate a significant reduction in M2 macrophage polarization and Th17 cell phenotype upon treatment with YTE-17,leading to the restoration of regulatory T(Treg)/Th17 cell balance in azoxymethane(AOM)/dextran sodium sulfate(DSS)model.Furthermore,we also confirmed that YTE-17 effectively inhibited the glycolysis of Th17 cells in both direct and indirect co-culture systems with M2 macrophages.Notably,our study shed light on potential mechanisms linking the non-canonical Wnt5a/JNK signaling pathway and well-established canonical β-catenin oncogenic pathway in vivo.Specifically,we proposed that Wnt5a/JNK signaling activity in IECs promotes the development of cancer stem cells with β-catenin activity within the TME,involving macrophages and T cells.In summary,our study undergoes the po-tential of YTE-17 as a preventive strategy against CRC development by addressing the imbalance with the immune microenvironment,thereby mitigating the risk of malignancies.

Objective To study the causes of two Chinese families with Branchio-oto syndrome.Methods The clinical data of two families were collected,and the pathogenic genes and variants of Branchio-oto syndrome were screened and verified by whole exome sequencing and Sanger sequencing.Results Two proband patients were diagnosed with Branchio-oto syndrome.Proband 1 presented with preauricular and anterior cervical fistulas,as well as congenital severe sensorineural hearing loss.On the other hand,proband 2 displayed a preauricular fistula and an anterior cervical cyst.At the age of 5,progressive deterioration of binaural hearing was observed,leadingtothe cur-rent diagnosis of severe mixed deafness.Genetic analysis showed that proband 1 and 2 carried nonsense variants of EYA1 gene:NM_000503.6:c.1408G＞T(p.Glu470Ter),and c.889C＞T(p.Arg297Ter).According to the guide-lines of the American College of Medical Genetics and Genomics(ACMG),the above variants were rated as patho-genic variants.After reviewing the literature,the c.1408G＞T variant had not been previously reported,and the c.889C＞T is a known variant.Conclusion The variants c.1408G＞T(p.Glu470Ter)and c.889C＞T(p.Arg297Ter)of EYA1 gene are the cause of these two families with Branchio-oto syndrome.The first report of c.1408G＞T broadens the mutational spectrum of EYA1 gene and provids a clinical reference for the diagnosis of Branchio-oto syndrome.

AIM:To explore the efficacy of lenvatinib(Len)in enhancing the therapeutic effects of immune checkpoint inhibitor for hepatocellular carcinoma(HCC)and to delve into its immunomodulatory mechanisms within the tumor microenvironment.METHODS:The effects of various concentrations of Len on the migration of human umbilical vein endothelial cells(HUVECs)and the secretion of CXC chemokine ligand 10(CXCL10)were investigated,and the mechanism by which Len modulates CXCL10 secretion was validated.An orthotopic HCC model was established,and the mice bearing tumors were randomly allocated into 4 groups:PBS group,BMS-202(PD-1/PD-L1 inhibitor)group,Len group,and Len/BMS-202 group.The progression of the orthotopic liver tumors was monitored with small animal in vivo im-aging techniques.On the 13th day after the treatment,mice were sacrificed and tumor tissues were harvested for analysis.Immunofluorescence was employed to identify apoptosis,vascular architecture,and hypoxic status within the tumor tis-sue.The expression levels of proliferation marker Ki67,transforming growth factor-β(TGF-β),and the infiltration de-grees of CD4+T cells and CD8+T cells in the tumor tissue were monitored with immunohistochemistry.The secretion of im-mune factors interferon-γ(IFN-γ),CXCL10 and TGF-α in the mouse serum was quantified with ELISA.Above all data were followed by statistical analysis.RESULTS:(1)Len could facilitate endothelial cell migration within a specific range and potentiated the response of tumor cells to IFN-γ by blocking fibroblast growth factor receptor(FGFR),thereby increasing the secretion of CXCL10 from the tumor cells.(2)Compared with PBS group,tumor growth was slower in all treatment groups,with Len/BMS-202 group showing the most significant inhibition of tumor growth in tumor-bearing mice(P<0.05).(3)Compared with PBS group and monotherapy groups,Len/BMS-202 significantly promoted tumor tissue apoptosis and inhibited tumor cell proliferation(P<0.05).(4)Compared with PBS group and BMS-202 group,both Len group and Len/BMS-202 group manifested a substantial enhancement in pericytes coverage rate(P<0.01),concomitantly showing a marked improvement in hypoxic conditions(P<0.01).(5)Compared with PBS group and monotherapy groups,Len/BMS-202 group showed a significant increase in the infiltration of CD4+T cells and CD8+T cells within the tumor(P<0.01),along with a marked decrease in the expression of TGF-β(P<0.01).(6)Compared with PBS group,all treatment groups collectively induced varying degrees of secretion of IFN-γ,CXCL10 and TGF-α in mouse serum(P<0.05),with Len/BMS-202 group demonstrating the most pronounced effects(P<0.01).CONCLUSION:Lenvatinib may augment the therapeutic efficacy of BMS-202 in HCC by facilitating tumor vascular normalization,alleviating hypoxic conditions,and enhancing the secretion of CXCL10,thereby synergistically activating the tumor immune microenvironment.

ObjectiveTo evaluate the effect of transcranial direct current stimulation (tDCS) on the cognitive function and quality of life in patients with Parkinson's disease. MethodsRandomized controlled trials (RCTs) on tDCS for Parkinson's disease were searched in PubMed, Web of Science, Embase, Cochrane Library, CNKI, CBM, VIP and Wanfang Data from the inception to September, 2023. Control group was administered standard Parkinson's medications or placebo, physical therapy, and cognitive rehabilitation, while treatment group received tDCS additionally. The quality of the researches was evaluated using the Cochrane Risk of Bias Tool. Data synthesis and analysis were performed using RevMan 5.4 and Stata 17.0, with heterogeneity and sensitivity analyses. ResultsEight articles were included. tDCS significantly improved the scores of Montreal Cognitive Assessment (MD = 2.00, 95%CI 1.13 to 2.87, P < 0.001). However, there was no significant difference in the scores of Parkinson's Disease Questionnaire (MD = 0.73, 95%CI -5.78 to 7.23, P = 0.830), Beck Depression Inventory-Ⅱ(MD = -0.77, 95%CI -7.14 to 5.60, P = 0.810), and Unified Parkinson Disease Rating Scale-Ⅲ (MD = 1.60, 95%CI -0.77 to 3.97, P = 0.190). ConclusiontDCS may improve cognitive function of patients with Parkinson's disease.

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

OBJECTIVE To observe the clinical efficacy of Kanglaite injection assisted with camrelizumab combined with chemotherapy in the treatment of advanced non-small cell lung cancer （NSCLC）. METHODS A total of 192 patients with advanced NSCLC and hospitalized in the TCM oncology department of our hospital from January 1st， 2018 to December 1st， 2022 were retrospectively selected as the study objects， and were divided into observation group （additional use， n＝104） and control group （without additional use， n＝88） according to whether the patients additionally received Kanglaite injection based on camrelizumab combined with chemotherapy （carboplatin+pemetrexed）. The short-term therapeutic effects of 2，4 and 6 cycles were compared between the two groups. The levels of peripheral blood immune function indexes and serum tumor markers were compared before treatment， after 3 cycles of treatment and after treatment. The long-term therapeutic effects as well as the occurrence of adverse drug reaction（ADR） during hospitalization were compared between the two groups. RESULTS After 3 treatment cycles and at the end of treatment， the CD4+ T lymphocyte ratio and CD4+/CD8+ in the observation group were notably greater than the control group （P＜0.05）； the levels of serum carcinoembryonic antigen and cytokeratin 19 fragment antigen 21-1 were significantly lower than those in the control group （P＜0.05）. The overall survival of the observation group was significantly longer than that of the control group （P＜0.05）， and the median overall survival was （185.27±38.21） d and （132.11±34.23） d， respectively. There were no significant differences in the whole ADR and grade ≥3 ADR between the two groups during hospitalization（P＞0.05）. CONCLUSIONS Based on camrelizumab combined with chemotherapy， the addition of Kanglaite injection can enhance immunological response and prolong overall survival in advanced NSCLC patients.

BACKGROUND:The development of steroid-induced osteonecrosis of the femoral head is a complex process involving multiple mechanisms.There is still no standard therapeutic drug for early intervention of this disease.Current studies have shown that baicalein has various pharmacological activities such as regulating lipid metabolism,bone metabolism,apoptosis and anti-oxidative stress,which provides an idea for the prevention and treatment of steroid-induced osteonecrosis of the femoral head. OBJECTIVE:To observe the preventive effect of baicalein against steroid-induced osteonecrosis of the femoral head and to investigate its possible mechanism. METHODS:Thirty-six 10-week-old male Sprague-Dawley rats were randomly divided into three groups(n=12 per group):blank control group,model group,and baicalein intervention group.In the model group and baicalein intervention group,intraperitoneal lipopolysaccharide and intramuscular injection of methylprednisolone sodium succinate were performed for modeling,while normal saline was used as a substitute for the modeling drug in the blank control group.Baicalein 300 mg/kg was administered by gavage(once a day for 6 weeks)at the time of initial intramuscular glucocorticoid injection in the baicalein intervention group,and baicalein was replaced by normal saline in the other two groups.The serum level of malondialdehyde in rats was detected at 2 weeks of the experiment.Blood lipid indicators and bone formation metabolic markers were detected at 6 weeks of the experiment,the histomorphometric changes of the femoral head were analyzed by hematoxylin-eosin staining,anti-tartaric acid phosphatase staining and TUNEL staining,and the femoral head was subjected to Micro-CT scanning and three-dimensional reconstruction of the bone in order to analyze the alterations of bone tissue structure and parameters. RESULTS AND CONCLUSION:The serum levels of malondialdehyde,triglyceride,β-collagen type Ⅰ carboxy-terminal peptide were increased and the serum levels of bone specific alkaline phosphatase and pre-collagen type Ⅰ amino-terminal peptide were decreased in the model group compared with the blank control group(P＜0.05).The serum level of malondialdehyde decreased in the baicalein intervention group compared with the model group(P＜0.05),but there was no significant difference between the baicalein intervention group and blank control group(P＞0.05).The serum level of triglyceride was higher in the baicalein intervention group than the blank control group(P＜0.05),but had no significant difference between the baicalein intervention group and model group(P＞0.05).There were also no significant differences in the levels of bone specific alkaline phosphatase and β-collagen type Ⅰ carboxy-terminal peptide between the baicalein intervention group and the other two groups(P＞0.05).The serum level of the baicalein intervention group was lower in the baicalein intervention group than the blank control group(P＜0.05)but had no significant difference between the baicalein intervention group and model group(P＞0.05).Histomorphological analysis of the femoral head showed that the rate of bone empty lacuna,osteoclast counting and cell apoptosis rate in the femoral head of model group rats were significantly higher than those of the other two groups(P＜0.05).There was a significant increase in the number of adipocytes in the bone marrow cavity of the femoral head,bone trabeculae were thinned and sparsely arranged with more disruptions in the continuity.The incidence of osteonecrosis was higher in the model group(75%)than in the baicalein intervention group(25%;bilateral and unilateral exact significance results were both 0.05).There was also an increase in the number of adipocytes in the bone marrow cavity of the femoral head in the baicalein intervention group,and the trabecular changes were roughly similar to those in the model group.Micro-CT results showed that bone volume fraction,trabecular thickness,trabecular number,and bone mineral density decreased and trabecular separation increased in the model group compared with the blank control group(P＜0.05).Overall significant bone mass loss was observed in the model group.Bone tissue parameters in the baicalein intervention group were significantly improved than those in the model group,which were reflected in bone volume fraction,trabecular thickness and trabecular separation(P＜0.05),and trabecular number and bone mineral density had no significant difference between the baicalein intervention group and blank control group(P＞0.05).Although baicalein failed to significantly ameliorate dyslipidemia and promote bone formation in rats with steroid-induced osteonecrosis of the femoral head,it could reduce the incidence of steroid-induced osteonecrosis of the femoral head by reducing oxidative stress damage,decreasing cell apoptosis,inhibiting osteoclasts,suggesting its effectiveness in the early prevention of steroid-induced osteonecrosis of the femoral head.