Objective To investigate the effects of Pinella's ingredients on the viability of cells, morphology, microstructure, cell cycle and apoptosis in human cervical cancer cell lines. Methods The β-sitosterol and/or total protein of Pinella were incubated at different concentrations with cervical cancer cell line SiHa. The effects of β-sitosterol and/or total protein of Pinella on the viability of cells were tested by MTT assay. The effects of β-sitosterol on morphology, microstructure, cell cycle and apoptosis were studied by phase-contrast microscope, electron microscope and flowcytometry, respectively. Results β-sitosterol could obviously inhibit the viability of SiHa cells in a time-and dose-dependent manner. The total protein of Pinella had no effects on SiHa cells' viability. 20 μmol/L β-sitosterol induced the accumulation of SiHa cells in S phase in the cell cycle. And the percents of apoptosis and necrosis increased. The morphology and microstructure of SiHa cells changed significantly after treated with 20 μmol/L β-sitosterol. Conclusions The total protein of Pinella had little influence on the viability of cervical cancer cells SiHa. The viability of SiHa cells could be suppressed by β-sitosterol. β-sitosterol could induce the accumulation of cells in S phase and the percent of apoptosis and necrosis. The morphology and microstructure changed significantly after treated with β-sitosterol. Therefore, β-sitosterol might be a prospect safe and low toxicity anti-cervical cancer drug.

Objective To explore the early clinical characteristics and influencing factors in children with autism spectrum disorders(ASD).Methods From January 2005 to December 2014,193 children with ASD were collec-ted by continuous grouping method from Children's Rehabilitation Training Center in Xi'an.According to the 1∶1 matched case-control study requirements,and the other 193 children from kindergartens and primary schools in the urban areas of Xi'an were collected as healthy control group from March 1 to July 1,2016.The age of children in the case group was(40.78±14.86)months and the age of the healthy control group was(40.61±14.40)months.There were 167 boys and 26 girls in 2 groups and the ratio of boys to girls was 6.42∶1.00.The general status questionnaires,medical history questionnaire,diagnostic chart,Autism Behavior Checklist(ABC)and Family Environment Scale of Chinese version(FES-CV)were completed by parents between 2 groups.Childhood Autism Rating Scale(CARS)was completed by doctors in the case group.By using Excel software,the original questionnaires were completed in 2 entries by 2 persons to set up the database.All data were analyzed by SPSS 17.0 statistical software and conditional Logistic regression was used for multivariate analysis.Results Seventy point eight percent(137/193 cases)of children with ASD had been found abnormal under 2 years old or at 2 years old,and 54.9％(106/193 cases)had been diagnosed under 3 years old or at 3 years old.The average delay from the discovery to the diagnosis was 17 months.The initial abnormalities appea-rances were mainly manifested as no response to calling in 153 cases(79.3％),very little active contact with others in 141 cases(73.1％),silent or less use of oral language in 137 cases(71％),avoiding contact with the eyes of others or lack of facial expressions in 121 cases(62.7％).Their signs were easy to be misdiagnosed as mental retardation and language retardation.Children in the case group began to walk alone at the age of 8 months to 3 years old,and only 62.2％(120/193 cases)of them could walk alone at the age of 18 months or before.The age of conscious speech was at 8 months to 4 years and 4 months,and only 39.4％(76/193 cases)of the ASD children could speak at the age of 18 months or before.The total scores of the ABC scale of the case group were(56.520±22.140)scores and the sub-scales and total scores were significantly higher than those of the healthy control group,and the difference was statistically significant(t=16.845,27.390,16.527,26.320,23.371,32.206,all P<0.001).The positive consistent rate of ABC and clinical diagnosis was 56.5％.The total scores of CARS in the case group was(36.4±8.6)scores,and the positive consistent rate of CARS and clinical diagnosis was 78.8％.There was a statistical significance between the 2 groups in parental education,mother's occupation,family history(x2=29.670,44.593,15.439,6.095,all P<0.05),and there were statistical significance in the main caregivers,family harmony and family income(x2=19.006,7.129,109.027,all P<0.05).There was no statistical significance between the 2 dimensions of independence and achievement orientation between the 2 groups(t=-1.559,-0.139,P=0.120,0.890).The case group in the family cohesion,expressiveness,intellectual-cultural orientation,active-recreational orientation,moral-religious emphasis,organization and control of the 7 dimension scores were significantly lower than those in the healthy control group,and the differences were statistically significant(t=-7.683,-5.734,-8.762,-14.109,-2.026,-4.530,-2.464,all P<0.05).In the case group,the scores of the conflict dimension were higher than those of the healthy control group,and the difference was statistically significant(t=4.925,P<0.001).There was a statistical significance between the 2 groups in gestational age and birth hypoxia(x2=6.898,27.180,all P<0.05).According to multivariate analysis of Logistic regression,people other than parents serving as the primary support,anoxia of newborn,mother of non professional and technical personnel and lower scores of family active-recreational orientation might be the risk factors of ASD,family per capita income of 3 000 Yuan RMB or more monthly,mother education level of high school and above,and lower scores of family conflict might be the protective factors for ASD.Conclusions Clinical features of most ASD children can be easily identified under 2 years old,but if the diagnosis is delayed,the related intervention is late,so importance should be attached to early diagnosis.Mother's occupation and education level,family economic status,family environment,their supervisors,and anoxia of newborn may be the effective entry points in the prevention and treatment of ASD.

Objective This study was aimed to characterize the swine leukocyte antigen( SLA) class I genes of GGTA1 -/ - Wuzhishan minipigs and compare their similarity to human leukocyte antigen( HLA) . It has important implica?tions for understanding the cellular rejection in xenotransplantation. Methods Specimens of ear tissue from six founding GGTA1 -/ - Wuzhishan minipigs were collected, and the SLA class I genes (SLA?1, SLA?3, SLA?2) were amplified by RT?PCR. Purified products were cloned into pEASY?T1 vectors and sequenced, followed by BLAST alignment and using bioin? formatc analysis to characterize the SLA class I genes and compare with the similarity to HLA. Results A total of six al?leles were detected, among them alleles were previously reported (SLA?1?0703,SLA?2?1102, SLA?3?0401, SLA?3?0403), and the other were novel (SLA?1?0401wz01, SLA?2?11wz01). The homology between alleles of SLA class I genes in Wuzhishan minipigs and HLA was from 70?5% to 72?1%. The homology analysis of critical amino acid residues on HLA binding with human CD8 + molecules showed that SLA?1?0401wz01, SLA?1?0703, SLA?2?11wz01, SLA?2?1102 and SLA?3?0401 occurred mutant at amino acid positions 225 and 228 ( T→S,T→M) , whereas the other loci were highly conserved. There was a high homology at amino acid level between SLA?2?11wz01, SLA?2?1102 and HLA class I genes which are NK cell KIRs binding sites. Conclusions The amino acid sequences of SLA class I genes of GGTA1 -/ -Wuzhishan minipigs have a high homology to HLA. From the point of view of cell?mediated xenograft rejection, the amino acid sequences of SLA class I genes of GGTA1 -/ - Wuzhishan minipigs have a high homology to HLA, therefore, Wzhishan minipigs may become a good potential donor for pig?human xenotransplantation.

Objective:To explore the responsibilities of respiratory nurses in the management of chronic obstructive pulmonary disease (COPD) patient and the difficulties in disease management, so as to provide reference basis for formulating COPD management training strategies for respiratory nurses.Methods:Using phenomenological methodology, 14 nurses from the Department of Respiratory and Critical Care Medicine in the General Hospital of Ningxia Medical University and the First People′s Hospital of Yinchuan were interviewed in a personal semi-structured way from May to August 2021, and the data were sorted and analyzed according to Colaizzi 7-step analysis method.Results:Nurses′understanding of the responsibilities of COPD patient management could be summarized into five themes: dynamic monitoring and management of patients′ health, nursing decision-making of sudden changes in patients′ condition, implementation of patients′ out of hospital follow-up, promotion of improvement of patients′ self-management behavior, and cooperation among multidisciplinary team members. The difficult experience of nurses in the management of COPD patients abstracted five themes: lack of professional knowledge of COPD management, lack of clinical nursing decision-making authority, lack of human and financial support for follow-up, lack of communication skills with patients, and lack of multidisciplinary team formation in the hospital.Conclusions:Respiratory nurses have a clear understanding of the responsibilities of COPD patient management, but there are multiple difficulties in performing their responsibilities. We should pay attention to the responsibility positioning and difficulty support of nurses′ COPD management, and formulate targeted training strategies to promote the improvement of COPD nursing quality.

Objective:To observe the effects of salvianolic acid B on migration and tube formation of the retinal vascular endothelial cell (RVEC) in high glucose, and explore its mechanism with network pharmacology.Methods:The cells were divided into normal group, model group and 1.0, 0.5, 0.1 μg/ml salvianolic acid B group according to the random number table method. The cells of each group were added with 5.5 mmol/L glucose for intervention, and the salvianolic acid B group was added with 1.0, 0.5, and 0.1 μg/ml salvianolic acid B for intervention. After 72 h, the cell viability of each group was detected by the CCK-8 method. The cells were divided into normal group, model group and low-, medium-, and high-dose salvianolic acid B group according to the random number table method. Then the cells of the normal group were added with 5.5 mmol/L glucose; the model group was added with 25 mmol/L glucose; the low-, medium-, and high-dose salvianolic acid B group was added with 25 mmol/L glucose and 0.062 5, 0.1250, 0.250 0 μg/ml salvianolic acid B. Then by taking Transwell test to detect the number of cell migration, and Matrigel test to analyze the total length of cells tubes. The active targets of Salvianolic acid B were screened by SuperTarget and Swiss TargetPrediction. Then, the targets of diabetic retinopathy were obtained by searching the GAD database, pharmGkb database, TTD database, DiGSeE database and OMIM database. The effective targets of drug-disease interaction were screened, and the component-target-disease interaction network was constructed by Cytoscape. Finally, the effective targets were analyzed by DAVID for GO analysis and KEGG pathway enrichment analysis. Molecular docking was performed by using Accelrys Discovery Studio Client 2.5 software.Results:The CCK-8 method showed that the cell absorbance values of 0.5 and 0.1 μg/ml salvianolic acid B group were not significantly different from those of the normal group ( P>0.05). The results of Transwell experiment and Matrigel experiment showed that compared with the model group, the relative number of migrating cells and the total length of tubule formation in each dose group of salvianolic acid B decreased ( P<0.05 or P<0.01). The interaction network revealed that salvianolic acid B acted on 46 targets and 8 signaling pathways. Conclusions:Salvianolic acid B could inhibit the migrating and tube forming ability of RVEC cultivated by high glucose. The results suggest that salvianolic acid B may play roles in preventing diabetic retinopathy.

Objective To investigate the mechanism of circadian clock protein Bmal1(Bmal1)on renal injury with chronic periodontitis,we established an experimental rat periodontitis model.Methods Twelve male Wistar rats were randomly divided into control and periodontitis groups(n=6,each group).The first maxillary molars on both sides of the upper jaw of rats with periodontitis were ligated by using orthodontic ligature wires,whereas the control group re-ceived no intervention measures.After 8 weeks,clinical periodontal parameters,including probing depth,bleeding index,and tooth mobility,were evaluated in both groups.Micro-CT scanning and three-dimensional image recon-struction were performed on the maxillary bones of the rats for the assessment of alveolar bone resorption.Histopatholo-gical observations of periodontal and renal tissues were conducted using hematoxylin-eosin(HE)and periodic acid-Schiff(PAS)staining.Renal function indicators,such as creatinine,albumin,and blood urea nitrogen levels,and oxida-tive stress markers,including superoxide dismutase,glutathione,and malondialdehyde levels,were measured using bio-chemical assay kits.MitoSOX red staining was used to detect reactive oxygen species(ROS)content in the kidneys.The gene and protein expression levels of Bmal1,nuclear factor erythroid 2-related factor 2(Nrf2),and heme oxygenase-1(HO-1)in rat renal tissues were assessed using real-time quantitative polymerase chain reaction(RT-qPCR)and immuno-histochemical staining.Results Micro-CT and HE staining results showed significant bone resorption and attachment loss in the maxillary first molar region of the periodontitis group.Histological examination through HE and PAS staining revealed substantial histopathological damage to the renal tissues of the rats in the periodontitis group.The findings of the assessment of renal function and oxidative stress markers indicated that the periodontitis group exhibited abnormal levels of oxidative stress,whereas the renal function levels showed abnormalities without statistical significance.Mito-SOX Red staining results showed that the content of ROS in the renal tissue of the periodontitis group was significantly higher than that of the control group,and RT-qPCR and immunohistochemistry results showed that the expression levels of Bmal1,Nrf2,and HO-1 in the renal tissues of the rats in the periodontitis group showed a decreasing trend.Conclu-sion Circadian clock protein Bmal1 plays an important role in the oxidative damage process involved in the renal of rats with periodontitis.

Objective This study aims to explore changes in uncoupling protein 2(UCP2)in experimental periodonti-tis-associated renal injury induced by ligation and investigate the effect of UCP2 on renal injury induced by periodontitis.Methods Twelve Wistar male rats were randomly divided into two groups:control and periodontitis groups.A periodon-tal model was built by ligating the maxillary first molars area with 0.2 mm orthodontic ligature wire.After 8 weeks,the in-traoral condition of the rats was observed and periodontal clinical indices such as gingival bleeding index(BI),periodontal probing depth(PD),and tooth mobility(TM)were detected.The maxillary bone was scanned by Micro CT to observe the alveolar bone resorption.The tissue mineral density(TMD),bone mineral density(BMD),bone volume fraction(BV/TV),trabecular thickness(Tb.Th),trabecular bone separation(Tb.Sp)were recorded,and the distance from the enamel bone boundary to the alveolar crest(CEJ-ABC)of the maxillary first molar was measured.The oxidative stress indexes such as malondialdehyde,glutathione(GSH),and superoxide dismutase(SOD)were detected using frozen rat kidney tissue.The gene expression of UCP2,nuclear factor erythroid 2-related factor 2(Nrf2),and peroxisome proliferator-activated receptor gamma coactivator-1α(PGC-1α)was observed by quantitative real-time polymerase chain reaction(qRT-PCR)test.The gingival tissue of the rats was used for immunohistochemical staining to observe the expression of the UCP2 protein.The fixed rat kidney tissue was used for hematoxylin-eosin(HE),periodic acid-schiff(PAS),MitoSOX Red,JC-1,and immu-nohistochemical staining to observe the renal histopathology,the level of reactive oxygen species(ROS),the level of mito-chondrial membrane potential,and the expression of UCP2,Nrf2,and PGC-1α protein.Rat serum was collected to detect renal function indices,namely,blood urea nitrogen(BUN),creatinine(Cre),and albumin(Alb).Results Compared with the control group,the periodontitis group showed red,swollen,and soft gingival tissue,with gingival probing bleeding,periodontal PD increased,tooth loosening,alveolar bone resorption,decreased TMD,BMD,BV/TV,and Tb.Th indices,and increased Tb.Sp index,CEJ-ABC,and gingival UCP2 protein expression.Compared with the control group,the levels of MDA and ROS in the kidney tissue of periodontitis rats and the gene and protein expression of UCP2 increased,and the levels of MMP,GSH,and SOD and the gene and protein expression of Nrf2 and PGC-1α decreased.Renal functional indi-ces,namely,BUN,Cre,and Alb,were not significantly different between the two groups.Conclusion UCP2 may play a role in renal injury induced by periodontitis through oxidative stress.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Objective To investigate the mechanism mediating the regulatory effect of lncRNA MAGI2-AS3 on cisplatin(DDP)resistance in non-small cell lung cancer(NSCLC).Methods MAGI2-AS3 and miR-1269a expression levels were detected by qRT-PCR in DDP-sensitive lung cancer cell lines(A549 and H1299)and their resistant counterparts(A549/DDP and H1299/DDP).In A549 and H1299 cells with MAGI2-AS3 silencing and A549/DDP and H1299/DDP cells overexpressing MAGI2-AS3,the effects of 20 μmol/L DDP on cell viability and apoptosis were examined with CCK-8 assay,colony formation assay,flow cytometry and Western blotting,and the changes in epithelial-mesenchymal transition(EMT)were assessed with wound healing and Transwell assays.The interaction between MAGI2-AS3,miR-1269a and PTEN was predicted using GEPIA,StarBase and miRDB and verified with luciferase reporter gene assay and radioimmunoprecipitation(RIP)assay.A miR-1269a mimic and pcDNA3.1-PTEN plasmid were used to perform the rescue assay.Results MAGI2-AS3 expression was significantly downregulated in lung cancer tissues(P<0.05)in association with a poor prognosis(P<0.05).In the two DDP-resistant lung cancer cell lines,MAGI2-AS3 expression was significantly lowered as compared with the sensitive cells.Silencing MAGI2-AS3 significantly enhanced cell viability and promoted EMT of A549 and H1299 cells irrespective of DDP treatment,and also decreased DDP-induced apoptosis of the cells.In A549/DDP and H1299/DDP cells,MAGI2-AS3 overexpression strongly repressed cell viability and EMT irrespective of DDP treatment and promoted DDP-induced cell apoptosis.Luciferase reporter gene and RIP assays confirmed the binding of MAGI2-AS3 with miR-1269a and the binding of miR-1269a with 3'-UTR domain of PTEN.The rescue assay demonstrated that MAGI2-AS3 acted as a sponge for miR-1269a to promote PTEN expression and downregulate AKT phosphorylation,thus inhibiting EMT and promoting DDP-induced apoptosis of A549/DDP cells.Conclusion MAGI2-AS3 enhances DDP sensitivity of NSCLC by targeted regulation of the miR-1269a/PTEN/AKT signaling axis.

Objective To investigate the molecular mechanism through which calenduloside E inhibits hepatocellular carcinoma(HCC)cell proliferation and migration.Methods HCC cell lines HepG2 and Huh7 treated with calenduloside E were examined for changes in cell viability using CCK-8 assay and expressions of GPX4,SLC7A11,LC3,P62 and phosphorylation of Akt/mTOR using Western blotting.The effects LY294002 and Rapamycin(the inhibitor and activator of autophagy,respectively)on proliferation and migration of calenduloside E-treated HCC cells were evaluated using EdU and Transwell assays.The TCGA database was used to explore the expression levels of GPX4 and SLC7A11 in HCC and normal liver tissues and their correlation with the patients'survival outcomes.GPX4 and SLC7A11 expressions were also detected in HCC cells and normal hepatocytes using RT-qPCR and Western blotting.Results Calenduloside E obviously inhibited the viability of HCC cells.GPX4 and SLC7A11 were highly expressed in HCC tissues and cell lines,and their expression levels were negatively correlated with the patients'survival.In HCC cell lines,calenduloside E significantly inhibited the expressions of GPX4 and SLC7A11 proteins,activated the Akt-mTOR pathway,and enhanced the expression of LC3 II.The inhibitory effect of calenduloside E on GPX4 and SLC7A11 expressions was significantly enhanced by rapamycin but attenuated by LY294002.Inhibiting the autophagy pathway obviously diminished the inhibitory effect of calenduloside E on proliferation and migration of HCC cells,while activating this pathway produced the opposite effect.Conclusion Calenduside E inhibits the proliferation and migration of HCC cells by down-regulating GPX4 and SLC7A11 expression via the autophagy pathway.