Objective:To investigate impact of farrerol(Far)on LPS-induced microglial inflammatory injury by regulating cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING)signal pathway.Methods:Mice BV2 microglial cell lines were grouped into control group(normal culture),LPS group(1 μg/ml),Far low,medium and high doses groups(1,5,10 μmol/L),activator group(10 μmol/L Far+75 μg/ml cGAS-STING signal pathway activator DMX-AA);proliferation and apoptosis of BV2 cells were detected by MTT,plate cloning assay and flow cytometry;qRT-PCR and ELISA were applied to detect levels of IL-6,IL-1β and TNF-α in cells and supernatants;NO content in cell supernatant was detected by nitrate reductase method;Western blot was applied to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti-apoptotic factor B cell lymphoma protein-2(Bcl-2)and cGAS-STING pathway protein in BV2 cells.Results:Compared with control group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in LPS group were decreased,apoptosis rate,mRNA expressions of IL-6,IL-1β,TNF-α,contents of IL-6,IL-1β,TNF-α,NO,and protein expres-sions of Bax,cGAS and STING were increased(P<0.05);compared with LPS group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in Far low,medium and high dose groups were increased,apoptosis rate,mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were decreased (P<0.05); compared with Far high-dose group, A490 of BV2 cells, number of cloned cells, expressions of PCNA and Bcl-2 proteins in activator group were decreased, apoptosis rate, mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were increased (P<0.05).Conclusion:Far may inhibit apoptosis and inflammatory injury of BV2 cells and promote proliferation of BV2 cells by inhibiting cGAS-STING pathway, and thus alleviate inflammatory injury of BV2 cells induced by LPS.

Objective:To investigate impact of farrerol(Far)on LPS-induced microglial inflammatory injury by regulating cyclic guanosine monophosphate-adenosine monophosphate synthase(cGAS)-stimulator of interferon genes(STING)signal pathway.Methods:Mice BV2 microglial cell lines were grouped into control group(normal culture),LPS group(1 μg/ml),Far low,medium and high doses groups(1,5,10 μmol/L),activator group(10 μmol/L Far+75 μg/ml cGAS-STING signal pathway activator DMX-AA);proliferation and apoptosis of BV2 cells were detected by MTT,plate cloning assay and flow cytometry;qRT-PCR and ELISA were applied to detect levels of IL-6,IL-1β and TNF-α in cells and supernatants;NO content in cell supernatant was detected by nitrate reductase method;Western blot was applied to detect expressions of proliferating cell nuclear antigen(PCNA),Bcl-2 associated X protein(Bax),anti-apoptotic factor B cell lymphoma protein-2(Bcl-2)and cGAS-STING pathway protein in BV2 cells.Results:Compared with control group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in LPS group were decreased,apoptosis rate,mRNA expressions of IL-6,IL-1β,TNF-α,contents of IL-6,IL-1β,TNF-α,NO,and protein expres-sions of Bax,cGAS and STING were increased(P<0.05);compared with LPS group,A490 of BV2 cells,number of cloned cells,expressions of PCNA and Bcl-2 proteins in Far low,medium and high dose groups were increased,apoptosis rate,mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were decreased (P<0.05); compared with Far high-dose group, A490 of BV2 cells, number of cloned cells, expressions of PCNA and Bcl-2 proteins in activator group were decreased, apoptosis rate, mRNA expressions of IL-6, IL-1β, TNF-α, contents of IL-6, IL-1β, TNF-α, NO, and protein expressions of Bax, cGAS and STING were increased (P<0.05).Conclusion:Far may inhibit apoptosis and inflammatory injury of BV2 cells and promote proliferation of BV2 cells by inhibiting cGAS-STING pathway, and thus alleviate inflammatory injury of BV2 cells induced by LPS.

Objective To investigate the relationship between serum microRNA-183-5p(miR-183-5p)and microRNA-339-3p(miR-339-3p)levels and the severity and prognosis of patients with acute pancreatitis(AP).Methods A total of 175 AP patients admitted to the hospital from July 2021 to July 2023 were collect-ed as AP group.According to the acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ)score,the AP patients were divided into mild group(108 cases)and severe group(67 cases).According to the progno-sis,the AP patients were divided into good prognosis group(114 cases)and poor prognosis group(61 cases).A total of 160 healthy people who underwent physical examination in the hospital during the same period were selected as the control group.The expression levels of serum miR-183-5p and miR-339-3p were detected by re-al-time fluorescent quantitative PCR.The levels of tumor necrosis factor(TNF)-α and C-reactive protein(CRP)were determined by enzyme-linked immunosorbent assay.Pearson correlation analysis was used to ana-lyze the correlation of serum miR-183-5p and miR-339-3p levels with TNF-α,CRP and APACHE Ⅱ score in AP patients.Multivariate Logistic regression was used to analyze the influencing factors of poor prognosis in AP patients.Receiver operating characteristic(ROC)curve was used to analyze the predictive value of serum miR-183-5p and miR-339-3p levels for poor prognosis.Results Compared with the control group,the level of miR-183-5p in the AP group was increased,and the level of miR-339-3p was decreased,the difference was sta-tistically significant(t=17.497,19.113,all P＜0.05).Compared with the mild group,the serum levels of miR-183-5p,TNF-α,CRP and APACHE Ⅱ score in the severe group were increased,and the level of miR-339-3p was decreased(t=10.582,5.972,11.991,13.864,2.224,all P＜0.05).The level of serum miR-183-5p in AP patients was positively correlated with TNF-α,CRP and APACHE Ⅱ score(r=0.623,0.570,0.679,all P＜0.05).The level of miR-339-3p was negatively correlated with TNF-α,CRP and APACHE Ⅱ score(r=-0.655,-0.600,-0.756,all P＜0.05).The level of serum miR-183-5p in the poor prognosis group was higher than that in the good prognosis group,and the level of miR-339-3p was lower than that in the good prognosis group,the difference was statistically significant(t=5.324,5.436,all P＜0.05).TNF-α,CRP,A-PACHEⅡ score,and miR-183-5p were independent risk factors for poor prognosis in AP patients,and miR-339-3p was a protective factor for poor prognosis(all P＜0.05).The area under the curve of miR-183-5p,miR-339-3p levels and their combination to predict poor prognosis of AP patients was 0.760,0.731 and 0.836,respectively,and the predictive value of combined miR-183-5p and miR-339-3p was higher than that of single prediction(Z=2.952,2.892,all P＜0.05).Conclusion The serum level of miR-183-5p is increased and miR-339-3p is decreased in AP patients,which are closely related to the severity and prognosis of AP pa-tients and may be used as markers for the evaluation of the condition and prognosis of AP.

Objective:To investigate whether quercetin reduces neuroinflammation in epileptic rats by regulating high mobility group protein B1(HMGB1),receptor for advanced glycation end products(RAGE)and nuclear factor kappa-B(NF-κB),and to explore the possibility of HMGB1/RAGE/NF-κB pathway as a new target of quercetin.Methods:Twelve SD rats were randomly selected from 60 SD rats as healthy group,and the remaining rats were used to construct experimental models of epilepsy rats.Rats that were successfully modeled were divided into model group,high-dose quercetin group,low-dose quercetin group,quercetin+pathway activator group,with 12 rats in each group.Pathological changes of hippocampal tissue of rats were observed;neurobehavioral function of rats was evaluated;levels of tumor necrosis factor-α(TNF-α),IL-6 and IL-1β,and mRNA and protein expression levels of HMGB1,RAGE,NF-κB in hippocampal tissue of rats were detected.Results:Structure of hippocampus of rats in healthy group was complete;compared with healthy group,structure of hippocampal tissue of rats in model group was scattered,the number of surviving neurons was observably reduced,apoptotic index was observably increased,the Racine grade was observably increased,TNF-α,IL-1β,IL-6 contents and mRNA and protein expression levels of HMGB1,RAGE,NF-κB in hippocampal tissue were significantly increased;com-pared with model group,structure of hippocampal tissue of rats in low-and high-dose quercetin groups was relatively complete,the number of surviving neurons was observably increased,apoptotic index was observably decreased,the Racine grade was significantly decreased,TNF-α,IL-1β,IL-6 contents and mRNA and protein expression levels of HMGB1,RAGE,NF-κB were significantly re-duced,and the improvement effect of high-dose quercetin group was better;compared with high-dose quercetin group,the number of surviving neurons in quercetin+pathway activator group was significantly reduced,apoptotic index was significantly increased,the Ra-cine grade was significantly increased,TNF-α,IL-1β,IL-6 contents and mRNA and protein expression levels of HMGB1,RAGE,NF-κB in hippocampal tissue were significantly increased.Conclusion:Quercetin can effectively reduce neuroinflammation in epilep-tic rats by inhibiting the HMGB1/RAGE/NF-κB pathway and reducing the mRNA and protein expressions of related genes.

Objective:To screen the candidate genes in a patient with Kabuki syndrome (KS), providing basis for genetic counseling, prenatal screening, prenatal diagnosis and facilitating early treatment.Methods:This study included a 16-year-old female KS patient born of non-consanguineous Chinese parents who presented to Department of Orthognathic & Cleft Lip and Palate Plastic Surgery, School and Hospital of Stomatology, Wuhan University. Genomic DNA was extracted from the peripheral blood of the subjects and analyzed by whole-exome sequencing (WES). Sanger sequencing was performed to validate the mutation in the candidate gene. The conformational and physicochemical changes of the mutant were analyzed by Alphafold2, Antheprot and DOG.2.0.1, respectively. Distribution of KMT2D mutations in patients with KS was analyzed based on the Human Gene Mutation DatabaseResults:The proband manifested a typical KS facial gestalt, congenital cleft palate, fifth finger deformity, hypodontia, renal hypoplasia and hydronephrosis. Two de novo mutations c.[1166A>C; 1167dupC] (NM_003482) in cis on the same allele in the KMT2D gene were identified by WES and confirmed by allele-specific PCR. Bioinformatics analysis showed that three more α-helixes were added, and a (β-) turn and a (β-) sheet were reduced in KMT2D p. Y389S, p.V390Rfs*26 compared with the wild type. Meanwhile, the interceptive mutant-KMT2D protein p.V390Rfs*26 lost all four domains (FYRN domain, FYRC domain, SET domain, and PostSET domain), which may cause functional disabilities. Conclusions:Our study is the first to identify two novel and de novo KMT2D mutations in cis on the same allele in a KS patient and extends the KMT2D mutation spectrum of KS, providing evidence for genetic susceptibility counseling, prenatal screening and diagnosis, and early treatment of KS.

AIM: To evaluate the bioequivalence of the test and reference formulations of mycophenolate mofetil capsule in Chinese healthy male subjects under fasting and fed conditions. METHODS: This was a 2-treatment, 2-sequence, 4-period, fully replicated crossover study that included 80 Chinese healthy male subjects (40 subjects in the fasting group and 40 subjects in the fed group, respectively). Subjects were assigned to receive a single oral administration of the test or reference formulation at a dose of 0.25 g in each period. The plasma concentration of mycophenolate mofetil (MMF) and metabolite mycophenolic acid (MPA) were analyhed by LC-MS/MS. The major pharmacokinetic parameters of MMF and MPA were calculated using non-compartmental analysis by WinNonlin 8.0. The statistical analysis was performed by SAS 9.4. Average bioequivalence (ABE) analysis was applied where it has been demonstrated that the within-subject standard deviation of the reference formulation (S

Development of thoracolumbar vertebra (TLV) and rib primordium (RP) is a common evolutionary feature across vertebrates, although whole-organism analysis of the expression dynamics of TLV- and RP-related genes has been lacking. Here, we investigated the single-cell transcriptome landscape of thoracic vertebra (TV), lumbar vertebra (LV), and RP cells from a pig embryo at 27 days post-fertilization (dpf) and identified six cell types with distinct gene expression signatures. In-depth dissection of the gene expression dynamics and RNA velocity revealed a coupled process of osteogenesis and angiogenesis during TLV and RP development. Further analysis of cell type-specific and strand-specific expression uncovered the extremely high level of HOXA10 3'-UTR sequence specific to osteoblasts of LV cells, which may function as anti-HOXA10-antisense by counteracting the HOXA10-antisense effect to determine TLV transition. Thus, this work provides a valuable resource for understanding embryonic osteogenesis and angiogenesis underlying vertebrate TLV and RP development at the cell type-specific resolution, which serves as a comprehensive view on the transcriptional profile of animal embryo development.

Objective To investigate the changes of serum 8-hydroxy-2'-deoxyguanosine(8-OHDG) in Kawasaki disease (KD) children and to explore the importance of 8-OHDG in predicting the severity of coronary artery lesions in Kawasaki Disease.Methods The serum 8-OHDG was measured in KD patients group (n =60),fever patients group (n =12) and health control group (n =12) by ELISA method.Among the KD patients,30 KD patients were in acute stage (10 cases had coronary artery lesions,20 coronary were normal) and 30 patients were in recovery stage.The serum 8-OHDG and brain natriuretic peptide (BNP) were also compared between patients with coronary artery lesions (CALs) and patients with no coronary artery lesions NCALs).ROC curve analysis was used to test the 8-OHDG and BNP predictive values in KD with coronary artery lesions.Results The serum level of 8-OHDG was higher in acute KD patients than recovery KD patients,fever patients and healthy children (P ＜ 0.05) The serum 8-OHDG was higer in CALs patients than the NCALs patients (P ＜ 0.05).The serum BNP was higer in CALs patients than the NCALs patients (P ＜ 0.01).Analysis of the ROC curve showed that serum 8-OHDG had a positive predictive value of 64.3％ and a negative predictive value of 93.8％for distinguishing KD with CALs from KD NCALs when cutoff value was 57.02pg/ml.The area under the curve was 0.820.The serum BNP had a a positive predictive value of 47.6％ and a negative predictive value of 100％ for distinguishing KD with CALs from KD NCALs when cutoff value was 815pg/ml.The area under the curve was 0.745.Conclusion The serum 8-OHDG may have better predictive value than BNP in diagnose KD children with coronary artery lesions.

Objective To observe the effect ofTanreqinginjection combined with routine medicines on the treatment of senile patients with bronchial pneumonia.Methods A total of 146 patients were randomly divided into 2 groups according to the random number table, each group 73 patients. Control group was treated with routine medicines and treatment group was added 20 mlTanreqing once a day. All treatments lasta total of 7 days. The Th17 and Treg cells were detected by cytometry analysis. The expression of IL-17, IL-10, IL-6, and TNF-α were detected by ELISA analysis. The clinical effect and adverse effect of the two groups were compared. Results After treatment, the clinical effective rate of control group was 79.5% (58/73), which was significantly lower than that of 91.8% (67/73) in treatment group (χ2=2.406,P=0.045). After treatment, Th17 cells was significantly lower in the treatment group than that in the control group (5.16% ± 1.24%vs. 8.22% ± 1.84%;t=2.564,P=0.017); but Treg cells was significantly higher in the treatment group than that in the control group (6.32% ± 1.79%vs. 4.32% ± 1.23%;t=2.552,P=0.021). The expression of IL-17 (11.43 ± 2.52 ng/mlvs. 14.15 ± 2.61 ng/ml,t=2.684), IL-6 (12.47 ± 2.16 ng/mlvs. 15.58 ± 3.12 ng/ml,t=2.564), and TNF-α (25.43 ± 4.27 ng/ml vs. 32.55 ± 6.14 ng/ml,t=2.594) was significantly lower in the treatment group than those in the control group. However, the expression of IL-10 (10.07 ± 2.13 ng/mlvs. 7.94 ± 1.83 ng/ml;t=2.673,P=0.023) was significantly higher in the treatment group than that in the control group. The temperature decreased time (4.57 ± 1.24 dvs.
3.25 ± 0.92 d,t=2.628), cough disappeared time (7.53 ± 2.13 dvs.6.14 ± 1.59 d,t=2.416), pulmonary rales disappeared time (6.81 ± 1.82 dvs.5.17 ± 1.06 d,t=2.537) was significantly lower in the treatment group than those in the control group.Conclusions TheTanreqing injection combined with routine western medicine could regulate Th17/Treg cells balance in the senile patients with bronchial pneumonia, and showed the significantly clinical effect.

OBJECTIVE:To establish a method for the dissolution determination of Dronedarone hydrochloride tablet,and eval-uate the quality consistency of its generic and original preparations. METHODS:UV spectrometry was performed on the column of 288 nm,dissolution media of Phosphate buffer solution (pH4.5),0.1 mol/L Hydrochloric acid solution [adding into 0.5% sodium dodecyl sulfate(SDS)],Phosphate buffer solution [pH6.8,adding into 0.5%SDS] and water,volume of dissolution medium was 1 000 ml,rotation speed was 75 r/min,the dissolution of generic and original preparations of Dronedarone hydrochloride tablet was detected,and the similarity of dissolution curve was evaluated by calculating the similarity factor (f2). RESULTS:The linear range of dronedarone hydrochloride was 2.147-25.764 μg/ml;RSDs of precision,stability and reproducibility tests were lower than 2.0%;recoveries 4 dissolution media were 99.53%-101.05%(RSD=0.48%,n=9),98.95%-100.05%(RSD=0.39%,n=9), 99.54%-100.20%(RSD=0.24%,n=9)and 98.54%-100.06%(RSD=0.44%,n=9). In the 4 dissolution media,f2 of the dissolu-tion curve of 3 batches of generic and original preparations of Dronedarone hydrochloride tablet was 56,60,63,68,68,52,59, 67,65,68,76,62,respectively. CONCLUSIONS:The method is suitable for the dissolution determination of Dronedarone hydro-chloride tablet;meanwhile,the in vitro dissolution curves of generic and original preparations of Dronedarone hydrochloride tablet show similarity,so the quality consistency is good.