[摘 要] 目的：探究组蛋白去乙酰化酶（HDAC）抑制剂帕比司他对黑色素瘤生长和免疫性的影响及其机制。方法：常规培养黑色素瘤细胞B16F0，用不同浓度的帕比司他处理细胞，WB法检测帕比司他对B16F0细胞中HDAC表达的影响，CCK-8法、划痕愈合实验、Transwell实验和流式细胞术分别检测帕比司他对B16F0细胞增殖、迁移和侵袭能力，以及细胞凋亡和周期的影响。转录组学检测帕比司他对B16F0细胞基因表达的影响，用qPCR法加以验证。流式细胞术检测帕比司他对B16F0细胞表面MHC Ⅰ/Ⅱ类分子表达的影响，B16F0与骨髓来源树突状细胞（BMDC）共培养检测帕比司他对BMDC细胞表达CD11c、CD80和CD86的影响，B16F0细胞移植瘤实验检测帕比司他对移植瘤生长和裸鼠免疫功能的影响。结果：帕比司他促进B16F0细胞中组蛋白3（H3）和α-微管蛋白（α-TUB）蛋白乙酰化（P < 0.01或P < 0.001或P < 0.000 1），抑制B16F0细胞增殖、迁移和侵袭能力，促进其凋亡，并使细胞周期阻滞于G1期（P < 0.05或P < 0.001或P < 0.000 1），促进B16F0细胞表面表达MHC Ⅰ/Ⅱ类分子表达并促进共培养BMDC成熟（均P < 0.01）。转录组学检测结果显示，帕比司他促进B16F0细胞中E-cadherin和抗原提呈相关基因的表达，抑制N-cadherin、vimentin、c-Myc和CDK1的表达，qPCR法验证了这些结果。帕比司他抑制裸鼠移植瘤的生长并增强荷瘤裸鼠的免疫功能（P < 0.05, P < 0.000 1）。结论：帕比司他可抑制B16F0细胞的恶性生物学行为，促进其凋亡，调控其免疫性，增强荷瘤裸鼠的免疫功能。

This study aimed to explore the molecular mechanism of rubioncolin C(RuC) in inhibiting gastric cancer(GC). AGS and MGC803 cell lines were selected as cellular models. After treating the cells with RuC at different concentrations, the effects of RuC on the proliferation ability of GC cells were assessed using the CCK-8 method, real-time cellular analysis(RTCA), and colony formation assays. Transmission electron microscopy was used to observe subcellular structural changes. Immunofluorescence was applied to detect LC3 fluorescent foci. Acridine orange staining was used to evaluate the state of intracellular lysosomes. Western blot was employed to detect the expression of autophagy-related proteins LC3Ⅱ, P62, and lysosomal cathepsin D(CTSD). The SuperPred online tool was used to predict the target proteins that bound to RuC, and molecular docking analysis was conducted to identify the interaction sites between RuC and CTSD. The drug affinity responsive target stability(DARTS) assay was performed to detect the direct binding interaction between RuC and CTSD. The results showed that RuC significantly inhibited the proliferation and colony formation of GC cells at low concentrations, with 24-hour half-maximal inhibitory concentrations(IC_(50)) of 3.422 and 2.697 μmol·L~(-1) for AGS and MGC803 cells, respectively. After 24 hours of treatment with RuC at concentrations of 1, 2, and 3 μmol·L~(-1), the colony formation rates for AGS cells were 61.0%±1.5%, 28.0%±0.5%, and 18.2%±0.5%, respectively, while the rates for MGC803 cells were 56.0%±0.5%, 23.3%±1.0%, and 11.8%±1.0%, all of which were significantly reduced. Transmission electron microscopy revealed that RuC promoted an increase in autophagosome formation in GC cells. Immunofluorescence detection showed that LC3 fluorescent foci of GC cells increased with the increase in RuC dose. RuC up-regulated the expression of autophagy-related proteins LC3Ⅱ and P62 in GC cells. Acridine orange staining indicated that RuC altered the acidic environment of lysosomes. SuperPred online prediction identified CTSD as a potential target protein of RuC. Western blot analysis revealed that RuC induced the up-regulation of the inactive precursor of CTSD in GC cells. CTSD activity assays indicated that RuC reduced the activity of CTSD. Molecular docking simulations found that RuC bound to the substrate-binding region of CTSD, forming hydrogen bonds with the Tyr205 and Asp231 residues. Microscale thermophoresis and DARTS assays further confirmed that RuC directly bound to CTSD. In summary, RuC inhibits lysosomal activity by targeting and down-regulating the expression of CTSD, thereby inducing autophagosome accumulation in GC cells.
Humans ; Stomach Neoplasms/enzymology* ; Cathepsin D/chemistry* ; Cell Line, Tumor ; Molecular Docking Simulation ; Cell Proliferation/drug effects* ; Autophagosomes/metabolism* ; Autophagy/drug effects*

Jiuwei Xifeng Granules have become a Chinese patent medicine in the market. Because the formula contains Os Draconis, a top-level protected fossil of ancient organisms, the formula was to be improved by replacing Os Draconis with Ostreae Concha. To evaluate whether the improved formula has the same effectiveness and safety as the original formula, a randomized, double-blind, parallel-controlled, equivalence clinical trial was conducted. This study enrolled 288 tic disorder(TD) of children and assigned them into two groups in 1∶1. The treatment group and control group took the modified formula and original formula, respectively. The treatment lasted for 6 weeks, and follow-up visits were conducted at weeks 2, 4, and 6. The primary efficacy endpoint was the difference in Yale global tic severity scale(YGTSS)-total tic severity(TTS) score from baseline after 6 weeks of treatment. The results showed that after 6 weeks of treatment, the declines in YGTSS-TSS score showed no statistically significant difference between the two groups. The difference in YGTSS-TSS score(treatment group-control group) and the 95%CI of the full analysis set(FAS) were-0.17[-1.42, 1.08] and those of per-protocol set(PPS) were 0.29[-0.97, 1.56], which were within the equivalence boundary [-3, 3]. The equivalence test was therefore concluded. The two groups showed no significant differences in the secondary efficacy endpoints of effective rate for TD, total score and factor scores of YGTSS, clinical global impressions-severity(CGI-S) score, traditional Chinese medicine(TCM) response rate, or symptom disappearance rate, and thus a complete evidence chain with the primary outcome was formed. A total of 6 adverse reactions were reported, including 4(2.82%) cases in the treatment group and 2(1.41%) cases in the control group, which showed no statistically significant difference between the two groups. No serious suspected unexpected adverse reactions were reported, and no laboratory test results indicated serious clinically significant abnormalities. The results support the replacement of Os Draconis by Ostreae Concha in the original formula, and the efficacy and safety of the modified formula are consistent with those of the original formula.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Male ; Double-Blind Method ; Drugs, Chinese Herbal/therapeutic use* ; Tic Disorders/drug therapy* ; Treatment Outcome

OBJECTIVE: The study aim was to investigate the effects of exposure to multiple environmental organic pollutants on cardiopulmonary health with a focus on the potential mediating role of oxidative stress. METHODS: A repeated-measures randomized crossover study involving healthy college students in Beijing was conducted. Biological samples, including morning urine and venous blood, were collected to measure concentrations of 29 typical organic pollutants, including hydroxy polycyclic aromatic hydrocarbons (OH-PAHs), bisphenol A and its substitutes, phthalates and their metabolites, parabens, and five biomarkers of oxidative stress. Health assessments included blood pressure measurements and lung function indicators. RESULTS: Urinary concentrations of 2-hydroxyphenanthrene (2-OH-PHE) ( β = 4.35% [95% confidence interval ( CI): 0.85%, 7.97%]), 3-hydroxyphenanthrene ( β = 3.44% [95% CI: 0.19%, 6.79%]), and 4-hydroxyphenanthrene (4-OH-PHE) ( β = 5.78% [95% CI: 1.27%, 10.5%]) were significantly and positively associated with systolic blood pressure. Exposures to 1-hydroxypyrene (1-OH-PYR) ( β = 3.05% [95% CI: -4.66%, -1.41%]), 2-OH-PHE ( β = 2.68% [95% CI: -4%, -1.34%]), and 4-OH-PHE ( β = 3% [95% CI: -4.68%, -1.29%]) were negatively associated with the ratio of forced expiratory volume in the first second to forced vital capacity. These findings highlight the adverse effects of exposure to multiple pollutants on cardiopulmonary health. Biomarkers of oxidative stress, including 8-hydroxy-2'-deoxyguanosine and extracellular superoxide dismutase, mediated the effects of multiple OH-PAHs on blood pressure and lung function. CONCLUSION Exposure to multiple organic pollutants can adversely affect cardiopulmonary health. Oxidative stress is a key mediator of the effects of OH-PAHs on blood pressure and lung function.
Humans ; Oxidative Stress/drug effects* ; Male ; Cross-Over Studies ; Female ; Young Adult ; Environmental Pollutants/toxicity* ; Environmental Exposure/adverse effects* ; Biomarkers/blood* ; Adult ; Blood Pressure/drug effects* ; Polycyclic Aromatic Hydrocarbons/urine* ; Beijing

Objective To clarify the understanding of types of laboratories and manufacturers for GB 9706.1-2020 Medical electrical equipment-Part 1:General requirements for basic safety and essential performance by laboratory proficiency testing for creepage distance and electrical clearance test.Methods An operation guide was formed according to the testing program in GB 9706.1-2020,and the homogeneity and stability of the samples were evaluated according to CNAS-GL003:2018 Guidance on Evaluating the Homogenneity and Stability of Samples Used for Proficiency Testing.Robust statistic methods were used to assess the quantitative parameters of the test results of the participating laboratories according to the requirements in GB/T 28043-2019 Statistical methods for use in proficiency testing by interlaboratory comparison;the results reported by the expert laboratories were used as the specified values of the qualitative parameters.SPSS 25.0 statistical software was used for data analysis.Results All the results of the crreepage distance and electrical clearance tests met the requirements for homogeneity and stability.Of the 46 laboratories involved in,37 ones did have comprehensive satisfactory determinations while the remained 9 ones not.Conclusion Some laboratories don't behave well in understanding the standard,which have to be reformed accordingly to enhance their proficiencies.[Chinese Medical Equipment Journal,2024,45(10):54-59]

Objective To investigate the distribution of pathogens and change of homology of Acinetobacter bau-mannii(AB)in intensive care units(ICUs)of partial medical institutions in Shanghai before and after the pandemic of coronavirus disease 2019(COVID-19).Methods Pathogens were isolated and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry(MALDI-TOF MS),the whole genome of AB was sequenced using Illumina Miseq sequencing platform,and its genetic relationship was explored based on multilocus sequence typing(MLST)and core genome multilocus sequence typing(cgMLST).Results The total detection rate of patho-gens in ICU environment after the pandemic was lower than before the pandemic([6.10％,101/1 656]vs[10.77％,181/1 680];P＜0.05).Before and after the pandemic,the proportion of AB detected on the surface of bedding in the ICU environment remained at the highest level.After the pandemic,the ST types showed a diverse distribution.MLST_Pasteur results showed that 162 strains of AB were divided into 20 ST types,with ST2(80.25％,n=130)being the main type.MLST_Oxford results showed that there were 19 ST types among 162 strains,with ST208(37.04％,n=60)being the main type.The clustering analysis based on cgMLST showed that ST208_Oxford had closer genetic relationship with ST540_Oxford and ST369_Oxford after the pandemic.ST164_Pasteur clone changed from ST234_Oxford before the pandemic to ST1418_Oxford after the pandemic,and 2 new types of ST_Pasteurr and 11 new types of ST_Oxford were discovered.Conclusion The detection rate of pathogens from ICU environment after the pandemic is lower than before the pandemic,and the distribution of ST types is slightly different at the same detection sites before and after the pandemic.ST2_Pasteur/ST208_Oxford is still the dominant epidemic clone before and after the pandemic,but some alleles have changed.cgMLST is more accurate than MLST_Oxford and MLST_Pasteur in homology analysis,evolution,spread,and outbreak analysis.

Objective To verify the therapeutic effect of platelet lysate(PL)on spinal cord injury in rats.Methods Thirty male SD rats were randomly divided into the sham operation group,the injury group and the treatment group,with 10 rats in each group.Rats in the sham operation group did not smash the spinal cord after exposure;rats in the injury group were established the rat spinal cord injury model by smashing the spinal cord with a 20 g of self-made smashing machine weights from a height of 5 cm for 5 seconds after exposure;rats in the treatment group were given intraperitoneal injection of 2 mL of PL once a day for 19 days after established the spinal cord injury model.The hind limb function was evaluated by Basso Beattie Bresnahan(BBB)score and slope experiment;six rats were randomly selected from each group,and the spinal cords of the rats were taken on the 7th day after surgery for HE staining and electron microscopy scanning to observe their pathological changes.Results There were statistically significant differences in BBB scores on the 3rd,7th and 14th days after surgery among the three groups(P<0.05),and the BBB scores of the treatment group were higher than those of the injury group(P<0.05).There were statistically significant differences in the results of slope experiment on the 7th and 14th days after surgery between the treatment group and the injury group(P<0.01).HE staining showed that the neuronal degeneration and necrosis decreased,the cell swelling improved and the microglial phagocytosis decreased in the treatment group compared with those in the injury group.The results of electron microscopy showed that the astrocyte cytoplasmic swelling and local cell membrane damage were alleviated,and the mitochondrial swelling and degeneration in the treatment group were relieved compared with those in the injury group.Conclusion PL can improve the hind limb function and reduce the pathological changes after spinal cord injury in rats,which has therapeutic effect on spinal cord injury in rats.

Objective:To explore the white matter structural characteristics in patients with bipolar disorder(BD)using diffusion tensor imaging(DTI)and investigate their relationship with cognitive function.Methods:A total of 15 patients with BD type Ⅰ and 26 patients with BD type Ⅱ who met the Diagnostic and Statistical Manual of Mental Disorders,Fourth Edition(DSM-Ⅳ)diagnostic criteria and 37 normal controls were included.Cognitive function was assessed with the Trail Making Test(TMT)and Repeatable Battery for the Assessment of Neuropsy-chological Status(RBANS).The tract-based spatial statistics(TBSS)method was used to explore the differences in fractional anisotropy(FA)and mean diffusivity(MD)among the three groups and perform correlation analyses with cognitive function.Results:Patients with BD Ⅰ and BD Ⅱ had lower scores in attention(P＜0.001),delayed memory(P＜0.01),and total scores(P＜0.001)on the RBANS compared to the normal control group.They also exhibited lower FA values in the corpus callosum and right superior corona radiata compared to the normal control group(P＜0.05).In the BD Ⅰ group,there was a positive correlation between FA values in the genu of corpus cal-losum and visuospatial/constructional scores(r=0.74,P＜0.05),while in the BD Ⅱ group,a positive correlation was found between FA values in the same region and language function scores(r=0.55,P＜0.05).Conclusion:It suggests that patients with bipolar disorder may have impaired white matter integrity in the corpus callosum and right superior corona radiata,which may be associated with cognitive impairment.

Aim To investigate the effect of Dahuang Tangluo pills(DHTL)on NOD-like receptor protein 3(NLRP3)/cysteine aspartate proteolytic enzyme-1(caspase-1)/apodermic D(GSDMD)pathway-media-ted pyroptosis in db/db mice with diabetic kidney dis-ease(DKD)and the underlying mechanism.Methods Eight db/m mice were selected as control group,and forty db/db mice were randomly divided into mod-el group,low dose group,medium dose group,high dose group and dapagliflozin group,with eight mice in each group.The control group and model group were given equal volume normal saline intragastric adminis-tration,the low,medium and high dose groups were given DHTL solution of 0.9,1.8 and 3.6 mg·kg-1,respectively,and the dapagliflozin group was given dapagliflozin tablet solution of 1.5 mg·kg-1,and the six groups were given intragastric administration once a day for 10 weeks.The body weight of mice was meas-ured daily and the dose was adjusted during adminis-tration.Fasting blood glucose(FBG)and body weight were measured after administration.The levels of 24-hour urinary total protein(24h-UTP),blood creatinine(Scr)and urea nitrogen(BUN)were measured by au-tomatic biochemical analyzer.The levels of interleukin-1 β(IL-1β),interleukin-6(IL-6),interleukin-18(IL-18)and tumor necrosis factor-α(TNF-α)in re-nal tissue of mice were determined by enzyme-linked immunosorbent assay(ELISA).The pathological changes of renal tissue were observed by hematoxylin-eosin(HE)staining.The DNA damage in mouse kid-ney tissue was observed using in situ end labeling(TUNEL)staining.The mRNA and protein expres-sions of NLRP3,caspase-1 and GSDMD in mouse kid-ney tissues were detected by Real-time quantitative PCR and Western blot.Results Compared with the control group,FBG,body weight,IL-1β,IL-6,IL-18 and TNF-α in the model group significantly increased(P＜0.01).The mRNA and protein expressions of NLRP3,caspase-1 and GSDMD in mouse kidney tis-sues significantly increased(P＜0.01).Compared with the model group,the levels of FBG,body weight,IL-1β,IL-6,IL-18 and TNF-α in each administration group significantly decreased(P＜0.05).The patho-logical morphology of renal tissue was improved in dif-ferent degrees,and the number of positive cells in re-nal tissue was significantly reduced(P＜0.05).The mRNA and protein expressions of NLRP3,caspase-1 and GSDMD in renal tissue of mice in high and medi-um dose of DHTL and dapagliflozin group significantly decreased(P＜0.05).Conclusions DHTL can im-prove the renal injury of DKD,and its mechanism may be through the regulation of NLRP3/caspase-1/GSD-MD pathway to inhibit pyroptosis and relieve the in-flammatory response of DKD mice.