Objective To study the effects of scutellarin on endometrial carcinoma Ishikawa cells,and analyze the correlation between the effects and phosphatidyl inositol 3 kinase(PI3 K)/protein kinase B(Akt)signaling pathway.Methods Ishikawa cells were divided into blank group and experimental-L,-M,-H groups,each group was treated with complete medium containing 0,5,10 and 20 μmol·L-1 scutellarin,respectively.Cell viability,clonal formation ability,metastatic ability,invasion,apoptosis and protein expression were detected by cell counting kit-8(CCK-8),plate cloning,scratch,Transwell,flow cytometry and Western blot assay,respectively.Results The cell viability of blank group and experimental-L,-M,-H groups at 48 h were(100.00±0.00)％,(78.51±7.54)％,(52.93±4.91)％and(41.62±5.33)％;the clone formation rate were(100.00±0.00)％,(56.59±6.34)％,(35.23±4.62)％and(10.66±1.91)％;the scratch healing rate were(53.70±6.19)％,(40.59±4.75)％,(34.25±4.40)％and(15.78±2.14)％;the number of invasive cells were 189.70±14.06,106.82±12.67,84.37±8.13 and 53.74±6.78;the relative expression levels of matrixmetallo proteinase-2 were 0.96±0.10,0.73±0.06,0.68±0.08 and 0.42±0.05;tissue inhibitor of MMP-1(TIMP-1)were 0.35±0.04,0.51±0.05,0.74±0.08 and 1.20±0.14;the apoptosis rates were(4.21±0.53)％,(15.83±2.42)％,(22.72±3.85)％and(34.41±4.67)％;the relative expression levels of B cell lymphoma-2(Bcl-2)were 1.38±0.15,0.90±0.10,0.56±0.06 and 0.24±0.03;Bcl-2 associated X protein(Bax)were 0.31±0.02,0.44±0.04,0.93±0.11 and 1.26±0.14;the relative expression levels of PI3Kp85 were 0.67±0.05,0.42±0.04,0.36±0.02 and 0.28±0.03;phosphorylated Akt(p-Akt)were 0.78±0.06,0.53±0.04,0.46±0.05 and 0.42±0.03.Compared with the blank group,the above indexes in the experimental-L,-M,-H groups were statistically significant(P＜0.05 or P＜0.01).Conclusion Scutellarin can inhibit the proliferation,metastatic ability and invasion of endometrial carcinoma Ishikawa cells and promote apoptosis by regulating PI3K/Akt signaling pathway.

To explore the mechanism of spleen- were obtained for the treatment of acute-on-chronic livstrengthening and moisture-nourishing liver prescription er failure, and 244 intersecting target genes and 7 core (JPLSYGF) in the treatment of acute-on-chronic liver target genes were screened. Molecular docking showed failure using network pharmacology and the molecular that the core target genes AKT1, SRC, VEGFA, docking. Methods Relying on TCMSP and Gene- STAT3 , EGFR, MAPK3 , HRAS had good affinity with Cards and other databases, the relevant targets of JPL- quercetin, the main active component in the JPLSYGF in the treatment of acute-on-chronic liver failure SYGF, and had strong binding activity. In addition, in were obtained. String and Cytoscape were used to con- vivo tests verified that the JPLSYGF could reduce the struct PPI networks of targets, core targets were expression of HRAS, EGFR, STAT3 , SRC, and VEGscreened out, and DAVID was used for GO function FA, to delay the progression of acute-on-chronic liver annotation and KEGG pathway enrichment analysis. failure. Conclusions JPLSYGF may act on core tar- The main active ingredients of the traditional Chinese gets such as HRAS, EGFR, STAT3, SRC, VEGFA medicine compound formula for JPLSYGF were select- and so on, to achieve the effect of treating acute-oned with a bioavailability OB value of =Э 30% and a chronic liver failure. drug-like DL^O. 18 as the screening conditions, and.

Objective To analyze the innate circuit qi endowment of patients with seasonal allergic rhinitis(SAR)in Renyin year(Ren-Yin stem-branch year of Chinese lunar calendar).Methods A retrospective study was conducted in 8 918 SAR patients who were treated in the Department of Otorhinolaryngology of Xiyuan Hospital,China Academy of Chinese Medical Sciences in pollen seasons of the spring and autumn(i.e.,the period of March 5 to May 21,and the period of August 7 to September 23)in 2022(Renyin year).The characteristics of circuit qi at birth date of the SAR patient were analyzed.Results The occurrence of SAR in Renyin year was related to the suiyun(yearly circuit phase),sitian(the first half of the alterable yearly circuit qi),dominant qi,and circuit qi combination.The two-factor model analysis showed that the occurrence of SAR in Renyin year was closely related with suiyun-sitian and dominant qi-guest qi.High morbidity of SAR was shown in the patients born in the year of heavenly-stem Ren and earthly-branch Xu,and with the circuit qi features of five circuits being wood circuit,Suiyun being excessive wood circuit,sitian of yangming dryness-gold,zaiquan(the second half of the alterable yearly circuit qi)of shaoyin monarch-fire,dominant qi being yangming dryness-gold,and circuit qi combination being disharmony.The two-factor model analysis showed that the morbidity of SAR was increased in the year with the circuit qi features of suiyun-sitian being excessive wood circuit and taiyang cold-water,and the combination of dominant qi and guest qi being yangming dryness-gold and taiyang cold-water.Conclusion Among the innate circuit qi factors,suiyun,sitian,dominant qi,and circuit qi combination have an impact on the occurrence of SAR.The interaction between innate circuit qi and circuit qi at onset may be one of the important mechanisms leading to the occurrence of SAR.

Objective To investigate the dynamic expression with the time change of N6-methyladenosine(m6A)methylation-related factors in the repair process of skeletal muscle injury and its mechanism in the inflammatory response of macrophage in the injure process.Methods In vivo mice models of BaCl2 injury in the gastrocnemius were established.Four mice per group in the control group and injury group.Gastrocnemius tissues were harvested at day 1,3,5,7,and 9 after injury for experiments.Primary gastrocnemius muscle tissue cells,muscle satellite cells,muscle cells,and cell line C2C12 cells were treated with dexamethasone(DEX,50 μmol/L)to mimic injury.Lipopolysaccharide(LPS,100 μg/L)induced RAW264.7 cell lines to mimic the inflammatory response after skeletal muscle injury,and STM2457(30 μmol/L)was added to inhibit the effect of methyltransferase 3(Mettl3)before LPS treatment.The expression of m6A methylation-related factors(Writers,Erasers,Readers)and inflammation factors were detected by Real-time PCR and Western blotting.Results The muscle fibers were dissolved and then gradually repaired with the extension of injury time,the number of monocytes/macrophages increased first and then decreased,and the Pax7 mRNA level increased first and then decreased with the change of injury time.Compared with the control group,the mRNA and protein levels of m6A methylation-related factors in gastrocnemius did not change significantly on the injury-1 day.However,they were significantly increased on the injury-3 days compared with the control group(P＜0.05),and then obviously decreased on the injury-5 days group compared with the injury-3 days group(P＜0.05).Compared with the control group,they were no significant differences on the injury-7 days group and-9 days group.In vitro DEX decreased the mRNA levels of m6A methyltransferase factors in primary muscle satellite cells and C2C12 cells and increased the mRNA expression level of methylation-recognition enzyme factors(P＜0.05).The mRNA levels of m6A methylation-related factors increased significantly in skeletal muscle tissue cells and myocytes after DEX treatment(P＜0.05).After LPS treatment,the mRNA and protein expression levels of m6A methylation-related factors and the mRNA expression levels of inflammatory factors interleukin(IL)-6 and IL-1β in macrophages increased significantly(P＜0.05),while the levels of IL-6 and IL-1β mRNA in macrophages decreased significantly when the Mettl3 was inhibited(P＜0.05).Conclusion m6A methylation-related factors primarily is activated in the damaged muscle cells and inflammation response of macrophages.Inhibition of m6A methyltransferase can reduce the inflammatory response of macrophages.

This study aims to investigate the function of lmo2363 gene in stress resistance of Liste-ria monocytogenes strain LM83-1.In this study,the lmo2363 gene deletion strain and complement-ation strain of Listeria monocytogenes were constructed using overlapping extended PCR and ho-mologous recombination techniques,and the growth ability,stress survival rate and biofilm forma-tion ability of wild,deletion strain and complementation strain were compared under different stress environments.lmo2363 gene deletion strain and complementation strain of Listeria monocy-togenes were successfully constructed in this experiment.The growth curves showed that the growth capacity of the deletion strain was weaker than the wild strain LM83-1 under 4 ℃,7％NaCl,10％NaCl,3.5％ethanol,4.0％ethanol and pH5 stress(P＜0.001).The results of stress survival test showed that the survival rate of the deletion strain was significantly lower than the wild strain after 1 h treatment with pH3 and 10 mmol/L H2 O2 stress(P＜0.010).The biofilm forming ability of the deletion strain was decreased compared with that of the wild strain(P＜0.050).This study confirmed that lmo2363 gene mediated the adaptation of LM to low temperature,high osmotic pressure,ethanol and acid stress environment and affected the formation of LM bio-film.This study laid a foundation for further exploring the function of lmo2363 gene in the stress resistance process of Listeria monocytogenes.

Objective:To observe the clinical efficacy and safety of hypomethylating agent therapy in chronic myelomonocytic leukemia(CMML).Methods:From February 2014 to June 2021,the clinical data,efficacy,survival time and safety of CMML patients diagnosed in the Second Hospital of Hebei Medical University and treated with hypomethylating agent therapy were retrospectively analyzed.Results:A total of 25 CMML patients received hypomethylating agent therapy,including 18 cases treated with decitabine(DEC)and 7 cases treated with azacytidine(AZA)as the basic treatment.Among them,20 patients responded,and 7 patients got complete remission(CR).All patients with CR were treated with DEC as the basic treatment.Five cases of CR occurred in the first 4 courses of treatment.After a median follow-up of 16.4(9.4-20.5)months,4 patients with CR progressed to acute myeloid leukemia(AML).The median overall survival(OS)time of 25 CMML patients was 17.4 months(95％CI:12.437-22.363).According to MD Anderson prognostic scoring system(MDAPS),CMML-specific prognostic scoring system(CPSS),CPSS molecular(CPSS-mol),Mayo molecular model(MMM),risk stratification of patients was performed,and the difference only between different risk stratification of MDAPS and survival time was statistically significant.Common adverse reactions of hypomethylating agent therapy in CMML patients included infection,gastrointestinal reaction,hematological toxicity,skin allergy and liver function damage.All patients'symptoms were improved after corresponding treatment.Conclusion:Hypomethylating agent therapy is effective and safe for CMML patients.CR mostly occurs in the first 4 courses of treatment,and hypomethylating agent therapy combined with low-dose chemotherapy can be used for patients who do not respond.Hypomethylating agent therapy can delay the disease,but can't prevent progression.

Objective:To investigate the psychological status of the outpatients in the andrology clinic and the effect of risk warning combined with multidisciplinary team(MDT)intervention on their satisfaction.Methods:Using convenience sampling,we enrolled 600 outpatients seeking medical attention in the Department of Andrology of our hospital from July to October 2022.We ran-domized the patients into a control(n=300)and an observation group(n=300),obtained their basic information,evaluated their psychological status with the Hospital Anxiety and Depression Scale(HADS),and assessed their satisfaction with the Xijing Hospital Outpatients'Satisfaction Questionnaire(HOSQ).The controls followed the routine procedure of treatment,while the patients in the ob-servation group received early warning before intervention based on their HADS scores.We provided normal medical care for those with HADS scores≤7,employed empathetic communication for those with HADS scores of 8-10,and conducted MDT intervention for those with HADS scores≥1l,followed by comparison of the patients'satisfaction with the outpatient service between the two groups.Results:There were no statistically significant differences in general conditions between the groups of patients(P＞0.05).The mean prevalence rate of anxiety and depression was 47.83％among the male subjects,lower in the control than in the observation group(47.00％vs 48.67％,P＞0.05),but higher in the patients with the education of junior high school or below(60.99％)than in those with that of senior high school(22.34％)and university or above(16.67％),and also higher in those aged 18-40 years(67.38％)than in those aged 41-60 years(51.82％)and over 60 years old(38.33％).A significantly higher rate of satisfaction with the outpatient service was found in the observation group than in the controls(97.18％vs 90.39％,P＜0.05).Conclusion:Anxie-ty and depression are prevalent among the outpatients in the andrology clinic,with a higher prevalence rate in those with lower educa-tion and at a younger age.Early risk warning combined with MDT intervention can improve the satisfaction of the patients.

AIM To study the chemical constituents from n-butanol fraction of Corydalis impatiens(Pall.)Fisch.and their antioxidant activities.METHODS The n-butanol fraction was isolated and purified by silica gel,MCI,ODS,Sephadex LH-20 and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The antioxidant activities were determined by DPPH method and tyrosinase method.RESULTS Fourteen compounds were isolated and identified as nicotinamide(1),methyl L-pyroglutamate(2),bungeanoline F(3),monomethyl fumarate(4),5-hydroxymethylfurfural(5),4-hydroxybenzoic acid(6),hydroxybenzoate(7),methyl 3,4-dihydroxybenzoate(8),methyl ferulate(9),dimethylcaffeic acid(10),dimethyl feruloyl malate(11),(-)-4-O-feruloylquinic acid(12),syringaresinol(13)and(-)-loliolide(14).Compounds 1,8,11 and 13 showed strong antioxidant activites on DPPH free radicals,with IC50 values ranging from 54.47 to 97.4 μmol/L.Compound 13 had potential inhibitory effect on tyrosinase.CONCLUSION Compounds 4-14 are first isolated from Corydalis genus,and 3 is isolated from this plant for the first time.Compounds 1,8,11 and 13 have strong antioxidant activities.

Antibody-drug conjugate (ADC) has the characteristics of low toxicity and high efficiency, and plays an important role in cancer treatment. However, due to the complexity of its structure, it brings difficulties in pharmacokinetic (PK) bioanalysis. This study established an analytical method for the detection of ADC (RC108) in cynomolgus monkey plasma by ligand-binding assay (LBA) and liquid chromatography tandem mass spectrometry (LC-MS/MS), which was used to analyze and quantify the total antibody, bound antibody and free drug in cynomolgus monkey plasma. Based on the LBA method, rabbit anti-RC108 Fab and mouse anti-MMAE (monomethyl auristatin E) mAb were pre-coated in 96-well plates as the total antibody and antibody binding reagents, respectively. The samples to be tested were added, and then the detection reagents were added in turn. Goat anti-human IgG (H+L)-HRP, chromogenic solution tetramethylbenzidine (TMB), H₂SO₄ terminate the reaction, read data at 450 nm/630 nm wavelength of microplate reader; LC-MS/MS analysis method quantifies MMAE concentration, and refer to relevant regulations for methodological validation. The analytical method for quantifying total antibody, bound antibody and free drug of RC108 drug obtained good accuracy and precision, and the selectivity, dilution linearity, hook effect, parallelism and stability were verified. Meet the requirements of biological analysis. Finally, a bioanalytical method for the determination of the concentration of the test substance RC108 (total antibody, conjugated antibody, free MMAE) in cynomolgus monkey plasma with high sensitivity and high throughput was established by LBA and LC-MS/MS method. Subsequent non-clinical research on PK research in cynomolgus monkeys will provide technical support.

OBJECTIVE: To examine the anti-inflammatory effect of grape seed extract (GSE) in animal and cellular models and explore its mechanism of action. METHODS: This study determined the inhibitory effect of GSE on macrophage inflammation and Th1 and Th17 polarization in vitro. Based on the in vitro results, the effects and mechanisms of GSE on multiple sclerosis (MS)-experimental autoimmune encephalomyelitis (EAE) mice model were further explored. The C57BL/6 mice were intragastrically administered with 50 mg/kg of GSE once a day from the 3rd day to the 27th day after immunization. The activation of microglia, the polarization of Th1 and Th17 and the inflammatory factors such as tumor necrosis factor- α (TNF- α), interleukin-1 β (IL-1 β), IL-6, IL-12, IL-17 and interferon-γ (IFN-γ) secreted by them were detected in vitro and in vivo by flow cytometry, enzyme linked immunosorbent assay (ELISA), immunofluorescence staining and Western blot, respectively. RESULTS: GSE reduced the secretion of TNF-α, IL-1 β and IL-6 in bone marrow-derived macrophages stimulated by lipopolysaccharide (P<0.01), inhibited the secretion of TNF-α, IL-1 β, IL-6, IL-12, IL-17 and IFN-γ in spleen cells of EAE mice immunized for 9 days (P<0.05 or P<0.01), and reduced the differentiation of Th1 and Th17 mediated by CD3 and CD28 factors (P<0.01). GSE significantly improved the clinical symptoms of EAE mice, and inhibited spinal cord demyelination and inflammatory cell infiltration. Peripherally, GSE downregulated the expression of toll-like-receptor 4 (TLR4) and Rho-associated kinase (ROCKII, P<0.05 or P<0.01), and inhibited the secretion of inflammatory factors (P<0.01 or P<0.05). In the central nervous system, GSE inhibited the infiltration of CD45⁺CD11b⁺ and CD45⁺CD4⁺ cells, and weakened the differentiation of Th1 and Th17 (P<0.05). Moreover, it reduced the secretion of inflammatory factors (P<0.01), and prevented the activation of microglia (P<0.05). CONCLUSION GSE had a beneficial effect on the pathogenesis and progression of EAE by inhibiting inflammatory response as a potential drug and strategy for the treatment of MS.
Mice ; Animals ; Encephalomyelitis, Autoimmune, Experimental/pathology* ; Grape Seed Extract/therapeutic use* ; Interleukin-17 ; Interleukin-1beta ; Tumor Necrosis Factor-alpha/metabolism* ; Interleukin-6/metabolism* ; Th1 Cells ; Mice, Inbred C57BL ; Interferon-gamma/therapeutic use* ; Th17 Cells/metabolism* ; Interleukin-12/therapeutic use* ; Cytokines/metabolism*