Loss-of-function variants of low-density lipoprotein receptor-related protein 5 (LRP5) can lead to reduced bone formation, culminating in diminished bone mass. Our previous study reported transcription factor osterix (SP7)‍-binding sites on the LRP5 promoter and its pivotal role in upregulating LRP5 expression during implant osseointegration. However, the potential role of SP7 in ameliorating LRP5-dependent osteoporosis remained unknown. In this study, we used mice with a conditional knockout (cKO) of LRP5 in mature osteoblasts, which presented decreased osteogenesis. The in vitro experimental results showed that SP7 could promote LRP5 expression, thereby upregulating the osteogenic markers such as alkaline phosphatase (ALP), Runt-related transcription factor 2 (Runx2), and β‍-catenin (P<0.05). For the in vivo experiment, the SP7 overexpression virus was injected into a bone defect model of LRP5 cKO mice, resulting in increased bone mineral density (BMD) (P<0.001) and volumetric density (bone volume (BV)/total volume (TV)) (P<0.001), and decreased trabecular separation (Tb.‍Sp) (P<0.05). These data suggested that SP7 could ameliorate bone defect healing in LRP5 cKO mice. Our study provides new insights into potential therapeutic opportunities for ameliorating LRP5-dependent osteoporosis.
Animals ; Low Density Lipoprotein Receptor-Related Protein-5/metabolism* ; Osteoporosis/genetics* ; Mice ; Mice, Knockout ; Sp7 Transcription Factor/physiology* ; Osteogenesis ; Bone Density ; Osteoblasts/metabolism* ; Core Binding Factor Alpha 1 Subunit/metabolism* ; Mice, Inbred C57BL ; beta Catenin/metabolism*

Addressing the enduring challenge of evaluating traditional Chinese medicines (TCMs), the integrated evidence chain-based effectiveness evaluation of TCMs (Eff-iEC) has emerged. This paper explored its capacity through a demonstration study that evaluated the effectiveness evidence of six commonly used anti-hepatic fibrosis Chinese patent medicines (CPMs), including Biejiajian Pill (BP), Dahuang Zhechong Pill (DZP), Biejia Ruangan Compound (BRC), Fuzheng Huayu Capsule (FHC), Anluo Huaxian Pill (AHP), and Heluo Shugan Capsule (HSC), using both Eff-iEC and the Grading of Recommendations, Assessment, Development, and Evaluation (GRADE) system. The recognition of these CPMs within the TCM academic community was also assessed through their inclusion in relevant medical documents. Results showed that the evidence of BRC and FHC received higher assessments in both Eff-iEC and GRADE system, while the assessments for others varied. Analysis of community recognition revealed that Eff-iEC more accurately reflects the clinical value of these CPMs, exhibiting superior evaluative capabilities. By breaking through the conventional pattern of TCMs effectiveness evaluation, Eff-iEC offers a novel epistemology that better aligns with the clinical realities and reasoning of TCMs, providing a coherent methodology for clinical decision-making, new drug evaluations, and health policy formulation.

AIM:To compare the formation and release of neutrophil extracellular traps(NETs)af-ter exposure to the high-altitude hypoxic environ-ment for different periods,and their relationship with the dynamic changes of microcirculatory disor-ders associated with pulmonary edema.METH-ODS:SD rats were raised under normoxic condi-tions at an altitude of 400 m and under hypoxic conditions at an altitude of 4 200 m.The rats raised under hypoxic conditions at an altitude of 4 200 m for 7 days were returned to normoxic con-ditions at 400 m to observe the changes in physio-logical and pathological indicators of the rats.The dynamic changes of neutrophil extracellular traps(NETs)release and microcirculatory disorders relat-ed to pulmonary edema after high-altitude hypoxia exposure and after returning to the plain were ex-plored by blood routine determination,rat arterial blood gas analysis,ELISA experiment,lung water content determination,H&E staining,and immuno-histochemical staining.RESULTS:In the hypoxic en-vironment at 4 200 m,rats exhibited significant re-ductions in arterial oxygen saturation(SaO2)and partial pressure of oxygen(PO2)(P＜0.01),accompa-nied by a marked increase in lung-tissue water con-tent(P＜0.01).The complete blood count revealed elevated levels of red blood cells,white blood cells,lymphocytes,and neutrophils(P＜0.01).In addition,the formation and release of NETs in neutrophils in-creased,accompanied by an aggravation of the in-flammatory response.After returning to the low-al-titude normoxic area at 400 m,the above indica-tors gradually returned to normal levels on the 7th day.Pathological changes such as alveolar epitheli-al cell shedding and inflammatory cell infiltration were observed in the lung tissues of rats in the high-altitude area,and the pathological changes were restored after returning to the low-altitude normoxic environment.CONCLUSION:The release of NETs from neutrophils is closely related to the re-covery of pulmonary edema and pulmonary ede-ma-related microcirculatory disorders after high-al-titude hypoxia exposure.

Objective: To analyze the disease burden of chronic obstructive pulmonary disease (COPD) and changes in its risk factors among residents in Zhejiang Province from 1990 to 2021, so as to identify key priorities for COPD prevention and control. Methods: Data on COPD mortality and disability-adjusted life years (DALY) for residents in Zhejiang Province from 1990 to 2021 were collected from the Global Burden of Disease (GBD) 2021 database. Standardized mortality and standardized DALY rate were calculated using the GBD 2021 world population standard structure. Premature mortality was computed via the life table method. The average annual percent change (AAPC) was applied to analyze trends in COPD mortality, DALY rate, and premature mortality. Changes in deaths of COPD risk factors were evaluated using population attributable fraction (PAF). Results: From 1990 to 2021, the standardized COPD mortality in Zhejiang Province decreased from 272.40/100 000 to 70.56/100 000 (AAPC=-4.395%), and the standardized DALY rate declined from 4 167.37/100 000 to 1 071.89/100 000 (AAPC=-4.396%). Similar downward trends were observed in both males (AAPC=-3.933%, -4.173%) and females (AAPC=-4.785%, -4.480%), all P<0.05. Crude mortality and DALY rates increased with age, and the crude mortality and DALY rates of various age groups in Zhejiang Province showed decreasing trends from 1990 to 2021 (all P<0.05). The premature mortality declined from 4.37% to 0.60% from 1990 to 2021 (AAPC=- 6.206%), with consistent trends across males and females (AAPC=- 6.144%, - 6.379%, all P<0.05). From 1990 to 2021, particulate matter pollution showed the largest reduction in PAF (- 56.76%), while ambient ozone pollution had the largest increase (103.07%) in Zhejiang Province. By 2021, smoking became the leading risk factor for deaths of COPD (PAF=43.32%). Conclusions The standardized mortality, standardized DALY rate, and premature mortality for COPD show consistent declining trends in Zhejiang Province from 1990 to 2021. However, risk factors such as smoking and ambient ozone pollution require intensified focus to further reduce disease burden of COPD.

Objective To investigate whether long non-coding RNA(lncRNA)myocardial infarct-associat-ed transcription factor(MIAT)promotes sepsis-induced acute kidney injury(AKI)by regulating microRNA-338-3p(miR-338-3p)/platelet thromboplastin-1(THBS1)axis.Methods Septic AKI model was established by cecal ligation and puncture.The rats were divided into control group and sepsis AKI group,with 10 rats in each group.The expression levels of lncRNA MIAT,miR-338-3p and THBS1gene in renal tissue were detec-ted by quantitative real-time PCR(qPCR).The levels of serum urea nitrogen(BUN)and creatinine(Cre)were detected by automatic biochemical analyzer.Renal tubular epithelial NRK-52E cells were induced by li-popolysaccharide(LPS)to establish a cell model of sepsis induced AKI.NRK-52E cells were divided into CK group,LPS group,LPS+si-NC group,LPS+si-lncRNA MIAT group,LPS+si-lncRNA MIAT+inhibitor NC group,LPS+si-lncRNA MIAT+miR-338-3p inhibitor group,LPS+si-lncRNA MIAT+oe-NC group,LPS+si-lncRNA MIAT+oe-THBS1 group.qPCR was used to detect the expression levels of lncRNA MIAT,miR-338-3p and THBS1 gene in each group.Cell counting kit 8 was used to detect cell proliferation in each group.Flow cytometry was used to detect the apoptosis rate of each group.Enzyme-linked immunosorbent assay was used to detect the levels of lactate dehydrogenase(LDH),tumor necrosis factor-α(TNF-α),interleukin(IL)-6,IL-10,malondialdehyde(MDA)and the activities of superoxide dismutase(SOD)and catalase(CAT)in each group.The expression of NOD-like receptor pyrin domain-containing protein 3(NLRP3),Caspase-1,cleaved caspase-3 and THBS1 protein were detected by Western blot.The targeting relationship between miR-338-3p and lncRNA MIAT and THBS1 was verified.Results The expression levels of BUN,Cre,lncRNA MI-AT and THBS1 gene were increased(P<0.05),and the expression level of miR-338-3p was decreased in sep-sis AKI group(P<0.05).Compared with CK group,the expression of lncRNA MIAT,THBS1 gene,apopto-sis rate and the levels of IL-6,LDH,TNF-αand MDA were significantly increased(P<0.05),the protein ex-pression levels of NLRP3,Caspase-1,cleaved caspase-3 and THBS1 were significantly increased(P<0.05),while the expression of miR-338-3p,A450(24,48 h)value and IL-10 level were significantly decreased(P<0.05),and the activities of CAT and SOD were significantly decreased in LPS group and LPS+si-NC group(P<0.05).Compared with LPS+si-NC group,the expression of lncRNA MIAT,THBS1 gene,apoptosis rate and the levels of IL-6,LDH,TNF-α and MDA were significantly decreased(P<0.05),the protein expression levels of NLRP3,Caspase-1,cleaved caspase-3 and THBS1 were significantly decreased(P<0.05),while the expression of miR-338-3p,A450(24,48 h)value and IL-10 level were significantly increased(P<0.05),and the CAT and SOD activities were significantly increased in LPS+si-lncRNA MIAT group(P<0.05).Silen-cing miR-338-3p expression or up-regulation of THBS1 gene expression could attenuate the improvement effect of lncRNA MIAT on sepsis AKI(P<0.05).Conclusion lncRNA MIAT promotes sepsis-induced AKI through regulating miR-338-3p/THBS1 axis.

Objective To investigate the effects of open reading frame 1 protein(ORF1p),encoded by long interspersed nuclear element-1(LINE-1),on the proliferation,migration,and invasion of esophageal squamous cell carcinoma(ESCC)cells,and explore the underlying molecular mechanism.Methods① Western blotting was performed to compare the expression of ORF1p between normal esophageal squamous epithelial cells and ESCC cells.② Immunohistochemistry(IHC)assay was used to examine ORF1p expression in ESCC tissues and paired normal tissues adjacent to tumor.③ The effects of ORF1p knockdown and overexpression on malignant behaviors in ESCC cells were determined through functional assays.④ Xenograft tumor model in nude mice was established to evaluate the impact of ORF1p on tumor growth in vivo.⑤ Transcriptome sequencing combined with cell functional rescue experiments were conducted to identify downstream targets regulated by ORF1p.Results ① Western blot analysis demonstrated the expression of ORF1p was significantly higher in the ESCC cell lines than the normal esophageal squamous epithelial cells(P<0.05).② IHC confirmed remarkable up-regulation of ORF1p in ESCC tissues than paired adjacent normal tissues(P<0.000 1).③ Functional assays and experiments on xenograft tumor models revealed that ORF1p substantially enhanced the proliferation,migration,and invasion of ESCC cells,as well as tumorigenic potential in vivo(P<0.05).④ Functional rescue experiments showed that ORF1p facilitated the proliferation,migration,and invasion of ESCC cells by modulating AJUBA expression(P<0.05).Conclusion ORF1p is significantly up-regulated in ESCC and promotes the proliferation,migration,and invasion of ESCC cells by regulating AJUBA expression.

Objective:To report the nationwide surveillance results of pathogenic profiles and antimicrobial resistance patterns of Gram-positive bloodstream infections in China in 2023.Methods:The clinical isolates of Gram-posttive bacteria from blood cultures were collected in member hospitals of National Bloodstream Infection Bacterial Resistant Investigation Collaborative System（BRICS）during January to December 2023. Antimicrobial susceptibility testing was performed using the dilution method recommended by the Clinical and Laboratory Standards Institute（CLSI）. Statistical analyses were conducted using WHONET 5.6 and SPSS 25.0 software.Results:A total of 4 385 Gram-positive bacterial isolates were obtained from 60 participating center. The top five pathogens were Staphylococcus aureus（ n=1 544，35.2%），coagulase-negative Staphylococci（ n=1 441，32.9%）， Enterococcus faecium（ n=574，13.1%）， Enterococcus faecalis（ n=385，8.8%），and α-hemolytic Streptococci（ n=187，4.3%）. The prevalence of methicillin-resistant Staphylococcus aureus（MRSA）and methicillin-resistant coagulase-negative Staphylococci（MRCNS）was 26.2%（405/1 544）and 69.8%（1 006/1 441），respectively. Notably，all Staphylococci remained susceptible to glycopeptide or daptomycin. Staphylococcus aureus demonstrated excellent susceptibility（>97.0%）to cephalobiol，rifampicin，trimethoprim-sulfamethoxazole，linezolid，minocycline，tigecycline，and eravacycline. No Enterococcus exhibiting resistance to linezolid were detected. Glycopeptide resistance was uncommon but more frequent in Enterococcus faecium（resistance to vancomycin and teicoplanin：both 1.7%）compared to Enterococcus faecalis（both 0.3%）. The detection rates of MRSA and MRCNS exhibited significant regional variations across the country（ χ2=17.674 and 148.650，respectively，both P<0.001）. No vancomycin-resistant Enterococci were detected in central China. Institutional comparison demonstrated higher prevalence of MRSA（ χ2=14.111， P<0.001）and MRCNS（ χ2=4.828， P=0.028）in provincial hospitals than that in municipal hospitals. Socioeconomic analysis identified elevated detection rates of both MRSA（ χ2=18.986， P<0.001）and MRCNS（ χ2=4.477， P=0.034）in less developed regions（per capita GDP

Objective:To report the results of bacterial resistant investigation collaborative system（BRICS）on the distribution and antimicrobial resistance profile of clinical Gram-negative bacteria isolates from bloodstream infections in China in 2023，and provide reference for clinical tretment of bloodstream infections and prevention and control of bacterial resistance.Methods:The clinical isolates of Gram-negative bacteria from blood cultures in member hospitals of BRICS were collected during January 2023 to December 2023. Antibiotic susceptibility tests were conducted by agar dilution or broth dilution methods recommended by Clinical and Laboratory Standards Institute（CLSI）. WHONET 5.6 and SPSS 25.0 were used to analyze the data.Results:During the study period，11 492 strains of Gram-negative bacteria were collected from 60 hospitals，of which 10 098（87.9%）were Enterobacterales and 1 394（12.1%）were non-fermentative bacteria. The top 5 bacterial species were Escherichia coli（50.0%）， Klebsiella pneumoniae（26.1%）， Pseudomonas aeruginosa（5.1%）， Acinetobacter baumannii complex（5.0%）and Enterobacter cloacae complex（4.1%）. The ESBL-producing rates in Escherichia coli， Klebsiella pneumoniae and Proteus mirablilis were 46.8%（2 685/5 741），18.3%（549/2 999）and 44.0%（77/175），respectively. The prevalence of carbapenem-resistant Escherichia coli（CREC）and carbapenem-resistant Klebsiella pneumoniae（CRKP）were 1.3%（76/5 741）and 15.0%（450/2 999）；32.9%（25/76）and 78.0%（351/450）of CREC and CRKP were sensitive to ceftazidime/avibactam combination，respectively. 94.7%（72/76）and 90.2%（406/450）of CREC and CRKP were sensitive to aztreonam/avibactam combination. Furthermore，57.9%（44/76）and 79.1%（356/450）were sensitive to imipenem/relebactam combination. The prevalence of carbapenem-resistant Acinetobacter baumannii（CRAB）complex was 64.6%（370/573），while more than 80.0% of CRAB complex was sensitive to tigecycline，eravacycline and polymyxin B. The prevalence of carbapenem-resistant Pseudomonas aeruginosa（CRPA）was 17.0%（99/581）. There were differences in the composition ratio of Gram-negative bacteria in bloodstream infections and the prevalence of important Gram-negative bacteria resistance among different regions in China，with statistically significant differences in the prevalence of CREC，CRKP，CRPA and CRAB complex（ χ2=10.6，28.6，10.8 and 19.3， P<0.05）. The prevalence of ESBL-producing Escherichia coli， CREC，CRAB complex and CRKP were higher in provincial hospitals than those in municipal hospitals（ χ2=12.5，9.8，12.7 and 57.8，all P<0.01）. Conclusions:Gram-negative bacteria are the main pathogens causing bloodstream infections in China，and Escherichia coli is ranked in the top，while the trend of Klebsiella pneumoniae increases continuously with time. CRKP infection shows a slow upward trend，CREC infecton maintains a low prevalence level，and CRAB complex infection continues to exhibit a high prevalence rate. The composition and resistance patterns of pathogens causing bloodstream infections vary to some extent across different regions and levels of hospitals in China.

Objective:To investigate the short-term efficacy and safety of rituximab (RTX) in children with calcineurin inhibitor (CNI) resistant steroid resistant nephrotic syndrome (SRNS).Methods:A retrospective case analysis was conducted. Thirteen children with CNI resistant SRNS who were regularly treated with RTX (375 mg/m 2 per dose (maximum dose 500 mg), 1 dose per week, a total of 4 doses) in Department of Nephrology, Children′s Hospital of Fudan University from January 2016 to December 2023 were enrolled. The general data, disease related information, urinary protein/creatinine, serum albumin, blood creatinine before RTX treatment, immunosuppressants, adverse events, and monthly urinary protein/creatinine, serum albumin, and blood creatinine indexes within 6 months after RTX treatment were collected. The changes of urinary protein/creatinine, serum albumin and estimated glomerular filtration rate (eGFR) before and after RTX at 3 and 6 months were analyzed by using paired sample t test and Wilcoxon signed-rank test. Results:Among the 13 patients, 8 were male and 5 were female. The age of disease onset was 4.0 (2.9, 6.8) years and the age of RTX treatment was 9.8 (5.9, 13.6) years. There were 8 cases of focal segmental glomerulosclerosis, 3 cases of minimal change disease and 2 cases of mesangial proliferative glomerulonephritis. No clinically significant gene variation was detected in 12 cases and the other one did not receive gene test. Before RTX treatment, 11 cases were in chronic kidney disease stage G1, and 1 case each was in stage G2 and stage G3. Ten children completed 4 doses of RTX treatment, 1 patient completed 3 doses, and 2 patients completed 2 doses. Urinary protein/creatinine in 13 children at 3 and 6 months after RTX treatment was significantly lower than baseline (0.60 (0.13, 2.04), 0.49 (0.28, 1.10) vs. 1.44 (0.76, 4.11) mg/mg, Z=-2.34, -2.34, both P<0.05), and serum albumin was significantly higher than baseline ((35±8), (34±7) vs. (30±6) g/L, t=2.30, 2.60, both P<0.05). The eGFR at 6 months after RTX treatment was not significantly different from the baseline ((110±32) vs. (113±35) ml/(min·1.73 m 2), t=-0.76, P>0.05)). No serious adverse reactions occurred in this study. Conclusion:RTX could reduce urinary protein and increase serum albumin in short-term treatment in children with CNI resistant SRNS without significant side effects.

Objective This study aimed to investigate how β-glucosidase(GBA)knockdown affects malignant progression in cisplatin(DDP)-resistant ovarian cancer(OC)cells and the role of the EGFR signaling pathway.Methods The A2780/DDP cells were categorized into four groups,with one of them serving as blank control(Con)group,si-NC group(transfected with negative control si-NC),si-GBA group(transfected with si-GBA),and NSC 228155 group(transfected with si-GBA and treated with 2 μmol/L NSC 228155).The protein expression levels of GBA,E-cadherin,N-cadherin,Vimentin,EGFR,p38 mitogen-activated protein kinase(p38 MAPK),phospho(p)-p38 MAPK,extracellular regulated protein kinase(ERK)and p-ERK were detected through Western blot.The relative expression of GBA was evaluated through reverse transcription quantitative polymerase chain reaction.The proliferation activity,migration,and invasion potential were evaluated using cell counting kit-8(CCK-8),plate clone formation,cell scratch healing,and Transwell migration assays.Thirty-six nude mice were divided into six groups(six mice per group):blank control(injected with normal saline),blank control+DDP(treated with DDP),negative control(injected with A2780/DDP cell suspension transfected with si-NC),negative control+DDP(injected with A2780/DDP cell suspension transfected with si-NC and treated with DDP),knockdown(injected with A2780/DDP cell suspension transfected with si-GBA),and knockdown+DDP groups(injected with A2780/DDP cell suspension transfected with si-GBA and treated with DDP).The tumor volume and weight of nude mice were evaluated.Results The relative protein and mRNA expression levels of GBA were significantly higher in the A2780/DDP group than in the A2780 group(P＜0.05).Compared with estimates in the Con and si-NC groups,the proliferation activity,number of cloned cells,scratch repair rate,and number of transmembrane cells in the si-GBA group were significantly lower(P＜0.05).The abundance of E-cadherin expression exhibited a notable elevation(P＜0.05),and expression levels of Vimentin,N-cadherin,and EGFR as well as the p-p38 MAPK/p38 MAPK and p-ERK/ERK ratios were significantly decreased(P＜0.05).The proliferation activity,number of cloned cells,scratch repair rate,and the count of transmembrane cells and the expression level of E-cadherin in the NSC 228155 group were markedly higher and lower,respectively,than those in the si-GBA group(P＜0.05),and the expression levels of Vimentin,N-cadherin,and EGFR as well as the ratios of p-p38 MAPK/p38 MAPK and p-ERK/ERK were significantly increased(P＜0.05).In the nude mouse xenograft study,the tumor size and mass in the blank control+DDP group were notably smaller and lighter,respectively,compared to those in the blank control group(P＜0.05).The tumor volume and weight were significantly lower in the negative control+DDP group than in the negative control group(P＜0.05),significantly lower in the knockdown+DDP group than in the knockdown group(P＜0.05),moreover,they were markedly reduced in the knockdown group in comparison to both the blank control and negative control groups(P＜0.05).Compared with those in the blank control+DDP and negative control+DDP groups,the tumor volume and weight in the knockdown+DDP group were significantly reduced(P＜0.05).Conclusions GBA knockdown suppresses the proliferation,migration,and invasion of DDP-resistant OC cells significantly as well as the growth of subcutaneous xenografts derived from A2780/DDP cells in nude mice.These effects may be mediated through the inhibition of the EGFR/MAPK/ERK signaling pathway.