Objective To investigate the effects of ATP-binding cassette subfamily B transporter 6 subfamily B(ABCB6)on cognitive dysfunction in Alzheimer's disease(AD)rats and its possible potential regulatory molecular mechanisms.Methods Amyloid β-protein(Aβ)was injected to construct the AD mouse model in vivo.Water maze test and Y maze test were used to evaluate the learning and memory ability and space exploration ability of rats.An in vitro AD cell model was constructed by HT22 cells and Aβ.The binding relationship between YTH domain containing 1(YTHDC1)and ABCB6 was analyzed by RNA immuniprecipitation(RIP).Quantitative real time polymerase chain reaction(qRT-PCR)was used to detect overexpression and knockdown transfection efficiency.Western blot analysis was performed to detect the expression levels of YTHDC1 and ABCB6 proteins,as well as ferroptosis related proteins[Solute Carrier Family 7 Member 11(SLC7A11),Glutathione peroxidase 4(GPX4)].Cell viability was detected with CCK-8.Malondialdehyde(MDA),Glutathione(GSH),Reactive oxygen species(ROS)levels and Fe2+content were analyzed by the assay kit.Results The ABCB6 mRNA(3.51±0.17 vs 1.02±0.01,3.45±0.21 vs 1.02±0.01)and protein(3.25±0.14 vs 1.01±0.01,3.14±0.16 vs 1.01±0.01)levels in the hippocampus of AD mice and Aβ-induced HT22 cells were up-regulated,and the differences were statistically significant(t=-46.238,-20.349;-50.468,-23.013,all P<0.001).Knocking down ABCB6 decreased the time and distance of AD mice reaching the platform,and increased the ratio of spontaneous exchange rate to the number of times of entered the new arm(t=27.007,11.264,24.414,19.901,all P<0.001).Knockdown ABCB6 promoted HT22 cell proliferation,decreased levels of MDA and Fe2+,increased GSH levels,reduced ROS generation,and promoted expression of SLC7A11 and GPX4 proteins(t=2.883～26.122,all P<0.05).YTHDC1 protein promoted its stability by binding to ABCB6 mRNA and up-regulated the expression of ABCB6 protein.Knockdown of YTHDC1 decreased ABCB6 protein level(t=18.504,P<0.001),promoted the proliferation of HT22 cells,increased GSH content,SLC7A11 and GPX4 protein levels,decreased MDA and Fe2+content,and inhibited ROS production(t=4.404～14.486,all P<0.05).Knocking down YTHDC1 could improve the learning and memory ability and spatial exploration ability of AD mice.Over-expression of ABCB6 reversed the effects of YTHDC1 knockdown on ferroptosis in HT22 cells and cognitive dysfunction in AD mice.Conclusion YTHDC1 may induce ferroptosis of neuronal cells by mediating the up-regulation of ABCB6,thus promoting cognitive dysfunction in AD mice.

OBJECTIVE: To analyze the differentially phosphorylated proteins in DENV-2-infected human umbilical venous endothelial cells (HUVECs) and explore the possible pathogenic mechanism of DENV-2 infection. METHODS: The total proteins were extracted from DENV-2-infected HUVECs and blank control HUVEC using SDT lysis method. The phosphorylated proteins were qualitatively and quantitatively analyzed using tandem mass spectrometry (TMT). The identified differentially phosphorylated proteins were analyzed by bioinformatics analyses such as subcellular localization analysis, GO enrichment analysis, KEGG pathway analysis and protein-protein interaction (PPI) analysis. Western blotting was used to detect the expressions of phosphorylated Jun, map2k2 and AKT1 proteins in DENV-2-infected HUVECs. RESULTS: A total of 2918 modified peptides on 1385 different proteins were detected, and among them 1346 were significantly upregulated (FC > 1.2, P < 0.05) and 1572 were significantly downregulated (FC < 0.83, P < 0.05). A total of 49 phosphorylated conserved motifs were obtained by amino acid conservative motif analysis. The most abundant differentially phosphorylated peptides in protein domain analysis included RNA recognition motif, protein kinase domain and PH domain. Subcellular localization analysis showed that the differentially modified peptides were mainly localized in the nucleus and cytoplasm. GO enrichment and KEGG pathway analysis showed that the differential peptides were mainly enriched in the regulation of stimulation response, biosynthesis of small molecules containing nuclear bases, and migration of phagosomes and leukocytes across the endothelium. PPI and KEGG joint analysis showed that the up-regulated and down-regulated differentially phosphorylated proteins were enriched in 15 pathways. In DENV-2-infected HUVECs, Western blotting detected differential expressions of phosphorylated proteins related with the autophagy pathway, namely JUN, MAP2K2 and AKT1, and among them p-JUN was significantly down-regulated and p-AKT1 and p-MAP2K2 were significantly upregulated (P < 0.01). CONCLUSION DENV-2 infected HUVECs show numerous differentially expressed proteins. The downregulation of p-JUN and upregulation of p-MAP2K2 and p-AKT1 suggest their potential roles in regulating autophagy, which is probably involved in the mechanism of DENV-2 infection.
Humans ; Autophagy ; Cell Death ; Cell Nucleus ; Human Umbilical Vein Endothelial Cells/virology* ; Dengue ; Proteome

Atypical teratoid/rhabdoid tumor(AT/RT)is a rare and highly malignant embryonal tumor of the central nervous system in children,characterized by diverse histological morphology,high malignancy,rapid clinical progression,and poor prognosis for affected children.The pathogenesis of AT/RT involves mutations in chromosomes and genes,particularly the loss of function of the SMARCB1 gene.The diagnosis of AT/RT primarily relies on histological and immunohistochemical analysis.Currently,there is no standardized treatment protocol for AT/RT.The main treatment modalities include surgery,chemotherapy,radiotherapy,as well as emerging targeted therapy and immunotherapy.Despite progress in research and clinical trials on AT/RT in recent years,the prognosis for affected children remains poor,necessitating further research to develop more effective treatment strategies to improve patient outcomes.

OBJECTIVE To establis h the fingerprint of Cynanchum auriculatum，to conduct its chemical pattern recognition analysis，and to determine the contents of four components at the same time. METHODS High performance liquid chromatography （HPLC）method was adopted. The determination was performed on ACE UExcel C 18 column with mobile phase consisted of acetonitrile-0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1.2 mL/min. The determination wavelength was set at 210 nm，and the column temperature was 40 ℃. The sample size wa s 10 μL. Taking qingyang shengenin as the reference ，HPLC fingerprints of 16 batches of C. auriculatum medicinal materials were drawn and similarity was evaluated by using the Similarity Evaluation of Chromatographic Fingerprints of Traditional Chinese Medicine （2012 edition）， and the common peaks were determined. SPSS 26.0 software andSIMCA 14.0 software were used for cluster analysis ，principal component analysis and orthogonal partial least squares- discriminant analysis. The differential components affecting the quality of C. auriculatum were screened by taking the value of variable importance in projection （VIP）greater than 1 as the standard ；same HPLC method was used to determine the contents of syringic acid ，acyl asclepiadelenin ，baishouwubenzophenone and qingyang shengenin. RESULTS There were 29 common peaks in 16 batches of C. auriculatum ，with a similarity of 0.723-0.998. Four common peaks were identified ，namely syringic acid （peak 7），acyl asclepiadoidin （peak 9），baishouwubenzophenone（peak 13）and qingyang shengenin（peak 15）. The results of cluster analysis showed that 16 batches of C. auriculatum could be clustered into three categories ，among which S 1 were grouped into one category，S3 were grouped into one c ategory，S2，and S 4-S16 were grouped into one category. The results of principal component analysis showed that the cumulative variance contribution rate of the five principal components was 88.706％，and the classification results were consistent with the results of cluster analysis. The results of orthogonal partial least squares-discriminant analysis showed that the common peaks （from large to small ）with VIP value greater than 1 were peak 20，peak 10，peak 25，peak 12， peak 15（qingyangshengenin），peak 21，peak 14，peak 16，peak 26，peak 22 and peak 17. The linear ranges of syringic acid，acyl asclepterin，baishouwubenzophenone and qingyangshengenin were 0.715 3-45.778 0，2.379 4-152.281 0，0.642 0- 41.085 0，14.541 6- 930.662 0 µg/mL respectively （all R 2＞0.999）. The quantitative limits were 0.357 7，0.475 9，0.642 0 and 2.423 6 μg/mL；the detective limits were 0.146 0，0.164 1，0.248 8 and 0.833 3 μg/mL，respectively. RSDs of precision ，repeatability and stability （24 h）tests were less than 3％；the average recoveries were 99.11％（RSD＝2.00％，n＝9），98.54％（RSD＝2.21％，n＝9）， 96.33％（RSD＝2.54％，n＝9）and 95.96％（RSD＝2.93％，n＝9）；the contents were 17.12-147.80，95.23-583.10（S8 below the quantitative limit ），16.91-210.88 and 211.68-3 587.15（S1 below the quantitative limit ）μg/g，respectively. CONCLUSIONS Established HPLC fingerprint and the method of content determination are stable ，reliable，accurate and reproducible. Combined with analysis of chemical pattern recognition ，it can be used for the quality control of C. auriculatum .

PURPOSE: To evaluate the efficacy of closed reduction on the humeroradial joint in the treatment of Bado type Ⅰ, Ⅱ and Ⅲ fresh Monteggia fractures in children and investigate the effect of clinical factors, including Bado classification, age and time of treatment on the success rate of closed reduction. METHODS: We retrospectively studied the data of children ≤10 years old with fresh Monteggia fractures (injury within two weeks) treated by manual reduction with plaster immobilization from January 2014 to April 2019. All patients were followed up in the outpatient department every two weeks for 4-6 weeks until plaster removal and then 3, 6 and 12 months. Online or telephone interview was provided for some inconvenient patients after 6 months. Mackay criteria were used to evaluate the clinical effect. Radiographic data were collected and reviewed to assess the reduction of the humeroradial joint. Function of the elbow joint and forearm was evaluated and risk factors related to the failure of reduction were assessed. The successful manual reduction was analyzed from three aspects, respectively Bado fracture type (Ⅰ, Ⅱ, Ⅲ), patient age (<3 year, 3-6 years, >6 years) and time interval from injury to treatment (group A, <1 day; group B, 1-3 days; group C, >3 days). RESULTS: Altogether 88 patients were employed in this study, including 58 males (65.9%) and 30 females (34.1%) aged from 1 to 10 years. There were 29 cases (33.0%) of Bado type Ⅰ Monteggia fractures, 16 (18.2%) type Ⅱ and 43 (48.7%) type Ⅲ. Successful manual reduction was achieved in 79 children (89.8%) at the last follow-up. The failed 9 patients received open surgery. Mackay criteria showed 100% good-excellent rate for all the patients. The success rate of manual reduction was 89.7%, 87.5% and 90.7% in Bado type Ⅰ, Ⅱ and Ⅲ cases, respectively, revealing no significant differences among different Bado types (χ = 0.131, p = 0.937). Successful closed reduction was achieved in 13 toddlers (13/13, 100%), 38 preschool children (28/42, 90.5%) and 28 school-age children (28/33, 84.8%), suggesting no significant difference either (χ = 2.375, p = 0.305). However time interval from injury to treatment showed that patients treated within 3 days had a much higher rate of successful manual reduction: 67 cases (67/71, 94.4%) in group A, 10 cases (10/11, 90.9%) in group B, and 2 cases (2/6, 33.3%) in group C (χ = 22.464, p < 0.001). Fisher's test further showed significant differences between groups A and C (p = 0.001) and groups B and C (p = 0.028). CONCLUSION Closed reduction is a safe and effective method for treating fresh Monteggia fractures in children. The reduction should be conducted as soon as possible once the diagnosis has been made.
Child ; Child, Preschool ; Closed Fracture Reduction ; methods ; Female ; Follow-Up Studies ; Humans ; Infant ; Male ; Monteggia's Fracture ; classification ; surgery ; therapy ; Retrospective Studies ; Time Factors ; Treatment Outcome

Objective To investigate the changes of metabolites in serum of paroxysmal patients with atrial fibrillation based on metabonomics.Methods The metabolites of serum in 15 patients with atrial fibrillation (case group) and 20 healthy subjects (control group) were measured by gas chromatography-mass spectrometry (GC-MS) group.The metabolic profile difference in the two groups was analyzed and compared by orthogonal design method (OPLS).Results The metabolic profile of the control and case groups was significantly distinguished,and 29 statistically significant differences metabolites were identified using commercially available metabolite libraries (such as the Wiley and NIST mass pools) and the standard metabolite library established in our laboratory,The areas under the ROC curve of 29 metabolites were calculated.The area under the curve of glycerol was 0.937,the area under the serine curve was 0.853,the area under the curve of aspartic acid was 0.933,the area of threonine was 0.823,the area of tryptophan was 1.Conclusion There exist differences in serum of sma] l molecular metabolites between patients with atrial fibrillation and normal healthy people.Glycerol,serine,aspartic acid,threonine and tryptophan of serum in patients with atrial fibrillation may be biomarkers of diagnosis of atrial fibrillation.

Background and purpose:Adenosine triphosphate-binding cassette superfamily G member 2 (ABCG2), which has been found over-expressed in a variety of cancer cells, takes part in the drug resistance of cancer through effux of anticancer drugs. The purpose of this study was to investigate the mechanisms of human glioblastoma cells sensitivity to pyropheophorbide-a methyl ester (MPPa)-mediated photodynamic therapy (PDT) eradicating tumour cells and its relationship to ABCG2.Methods:U87 and A172 glioma cell lines in the logarithmic growth phase were selected and exposed to the treatment of MPPa-PDT and MPPa-PDT+fumitremorgin C (FTC) respectively. The cell viability was measured with the use of CCK-8 assay. The expression of ABCG2 was detected by Western blot. The intracellular contents of MPPa in each group without illumination were tested by lfow cytometry. Flow cytometry with AnnexinⅤ-FITC/PI double staining was used to detect the cell apoptotic rate. DCFH-DA staining was used to assess the generation of intracellular reactive oxygen species (ROS).Results:The MPPa-mediated PDT could eradicate A172 and U87 cancer cells in an energy-dependent manner. The light energy density in A172 was 8 times of that in U87 when the cell viability reached median lethal dose after MPPa-mediated PDT. The high expression of ABCG2 in A172 cells affected the accumulation of intracellular MPPa. Inhibition of ABCG2, not only could enhance the eradicating effect of MPPa-PDT on A172 cells, but also could increase the yield of ROS triggered by MPPa-PDT and the accumulation of intracellular MPPa.Conclusion:The human glioblastoma cell line A172 is insensitive to MPPa-mediated PDT. The mechanism may relate to ABCG2, which decreases the MPPa content in cancer cells through effux of MPPa, resulting in decline of cytotoxicity.

Objective To investigate the suppression of mrp1 and MRP1 induced by small interfering RNA and the restoration of sensitivity to chemotherapeutic drugs in the multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. Methods mrp1-targeted small interfering RNA duplexes were designed and composed and introduced into multidrug-resistant hepatocellular carcinoma cell lines HepG2/mrp1. The suppression of mrp1 mRNA and its gene product MRP1 was examined by RT-PCR and flow cytometry (FCM), respectively. MTT assay was performed to measure the reverse effect of small interfering RNA based on the results of ICs0. Results The overexpression of mrp1 mRNA and MRP1 was effectively suppressed by small interfering RNAs. The level of mrp1 mRNA in the transfected HepG2/mrp1 cells was reduced to (86.36±2.76)% and MRP1 to (89.38±3.76)%compared with those of the controls. The resistance to ADR was reversed five-fold, which indicated the restoration of sensitivity to drugs. Conclusion Small interfering RNA can inhibit mrp1 expression effectively and reverse the multidrug resistance mediated by MRP1.

Objective To evaluate the efficacy and safety of ligation-assisted endoscopic dissection (ED-L) technique for the removal of gastric tumors originating from muscularis propria.Methods A total of 33 patients with gastric tumors originating from muscularis propria less than 10 mm were treated with ED-L procedures.The tumor was ligated by elastic bands.Endoscopic dissection was performed until the tumor was partially or completely dissected from muscularis propria by using Hook knife and/or IT-knife.The wound was closed with metallic clips and medical adhesive.The patients were followed up 1 week,1 month,3 months,6 months and 12 months thereafter with endoscopy,respectively.Results Of the 33 gastric tumors,there were 25 partial dissections and 8 complete dissections.All of the tumors sloughed completely.Pathological diagnoses of all the patients were acquired.No complications like perforation occurred except for one self-limiting and non-life-threatening hemorrhage.There was no recurrent case during the 3-18 months of follow-up period.Conclusion ED-L is a safe,effective and relatively simple technique for excision of small gastrointestinal tumors originating from muscularis propria,providing a histopathological diagnosis as well.

OBJECTIVETo explore whether the vitamin D receptor gene (VDR) polymorphisms are associated with the outcomes of hepatitis B virus (HBV) infection in Chinese Han population.

METHODSPolymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to detect the polymorphisms of Fok I locus in exon 2 and Taq I locus in exon 9 of VDR gene. One hundred and eighty-four chronic hepatitis B patients and 205 asymptomatic HBV carriers were recruited to make the comparison of frequencies of genotype and haplotype of the VDR gene between the patients and the carriers.

RESULTSThe univariate analysis showed a significant difference in Fok I polymorphism between chronic hepatitis B patients group and asymptomatic HBV carriers group. The FF genotype frequency in chronic hepatitis B patients group was 44.6%,higher than 31.7% in asymptomatic HBV carriers group (P<0.05). After adjusting the confounders by multiple logistic regression analysis, the result still showed a significant difference in Fok I site polymorphism between chronic hepatitis B patients group and asymptomatic HBV carriers group (OR=1.95, P<0.05). The FT haplotype frequency in chronic hepatitis B patients group was higher than that in asymptomatic HBV carriers group (OR=1.45, P<0.05). The fT haplotype frequency in chronic hepatitis B patients group was lower than that in asymptomatic HBV carriers group (OR=0.72, P<0.05).

CONCLUSIONVDR gene polymorphism may be an influence factor of genetic susceptibility to HBV infection.

Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Haplotypes ; Hepatitis B ; genetics ; Humans ; Polymerase Chain Reaction ; Polymorphism, Genetic ; genetics ; Polymorphism, Restriction Fragment Length ; Receptors, Calcitriol ; genetics