Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Objective:To investigate the difference of condylar bone density,volume and surface area in adult female patients with different vertical skeletal features of skeletal Class Ⅱ.Methods:This study was a retrospective case-control study.150 female patients aged 18-30 years were included as the subjects.Lateral cephalic radiographs were measured and the cases with high angle,average an-gle and low angle of skeletal Class Ⅱ were included and respectively grouped(n=50),the CBCT images were collected,the condyles were reconstructed by Mimics Reseach 20.0 software,the bone density,volume and surface area of the condyles were measured.Univa-riate analysis of variance(ANOVA)was used to compare the difference of condyle measurements among the 3 groups.Results:The overall difference of condylar bone density,volume and surface area among the 3 groups were statistically significant(P＜0.05).Pair-wise comparison showed that the condyle bone mineral density in high angle group was lower than that in average angle group(P＜0.05),in average angle group was lower than that in low angle group(P＜0.05),in high angle group was lower than that in low angle group(P＜0.001).Condyle volume and surface area in high angle group were lower than those in low angle group(P＜0.05),in aver-age angle group was lower than those in low angle group(P＜0.001),in high angle group was lower than those in average angle group(P＞0.05).Conclusion:The condyle bone density,volume and surface area of the different vertical skeletal features of skeletal class Ⅱ in adult female patients are different.

Objective To investigate the role of miRNAs in maternal amniotic fluid exosomes in development of isolated ventriculomegaly(VM)in fetuses.Methods Amniotic fluid samples were collected from 9 cases of moderate isolated VM and 8 normal control cases to extract exosomal miRNA,and miRNA sequencing technique was used to identify differentially expressed miRNAs between the two groups.Three miRNAs with significant differential expression between the two groups,whose high expression was associated with VM,were selected for verification with RT-qPCR.Dual luciferase reporter assays were used to verify the regulatory effect of miR-122-5p on its predicted target genes AKT3 and CCDC88C.Gene ontology(GO)and KEGG pathway analyses were performed to explore the possible roles of the top 40 significant differential miRNAs in the pathophysiology of VM.Results We identified a total of 272 differentially expressed miRNAs in VM cases,including 43 up-regulated and 229 down-regulated miRNAs.The target genes of these differential miRNAs were associated with DNA and transcription factor binding,transmembrane transporter and nucleic acid binding transcription factor activity,and cell developmental process.These miRNAs were mostly enriched in the MAPK,cGMP-PKG and Wnt signaling pathways.Verification with RT-qPCR showed that miR-122-5p expression level was significantly lower in VM group than in the control group(P<0.05),which was consistent with miRNA sequencing results;let-7b-5p expression level was significantly lower in VM group,which was contrary to miRNA sequencing result.Dual luciferase reporter assays showed that miR-122-5p was not capable of regulating AKT3 or CCDC88C expressions.Conclusions The highly abundant differentially expressed miRNAs in maternal amniotic fluid exosomes play important roles in the occurrence of fetal VM possibly by regulating the MAPK,PI3K-Akt,Wnt and cGMP-PKG signaling pathways.

Objective To study the population genetic structure and phylogenetic relationships by combining Y-STR haplotype genetic information from the Han population in Dalian with 32 domestic and foreign groups.Methods Blood samples of 958 Han male volunteers from Dalian were collected.Genetic typing of 42 genetic loci was completed using Y-STR fluorescent reagent kits and capillary electrophoresis.Related forensic parameters were calculated.Nei's standard genetic distances among 33 populations based on 17 Y-STR loci were computed,in order to create a principal coordinate analysis as well as construct a phylogenetic tree.Results The analysis of genetic polymorphisms at 42 Y-STR loci revealed 30 unconventional alleles at 10 loci.Genetic analysis of the population based on 17 Y-STR loci confirmed that Dalian's Han population had the closest genetic distance to the Anshan's Han population,followed by populations from Henan,Heilongjiang,Jilin,Shandong,and Chongqing.Furthermore,the genetic distances between the Han population in Dalian and the Qiang population in Beichuan or the Miao population in Guizhou were relatively closer than that to the Manchu population living in Liaoning.Conclusion The genetic distance between the Han population in Dalian and other groups is not entirely proportional to ethnicities and geographical proximity.Both population migration and ethnic assimilation or isolation may have influence on it.

Objective Gallstone disease(GSD)is one of the common digestive tract diseases with a high worldwide prevalence.The effects of GSD on patients include but are not limited to the symptoms of nausea,vomiting,and biliary colic directly caused by GSD.In addition,there is mounting evidence from cohort studies connecting GSD to other conditions,such as cardiovascular diseases,biliary tract cancer,and colorectal cancer.Early identification of patients at a high risk of GSD may help improve the prevention and control of the disease.A series of studies have attempted to establish prediction models for GSD,but these models could not be fully applied in the general population due to incomplete prediction factors,small sample sizes,and limitations in external validation.It is crucial to design a universally applicable GSD risk prediction model for the general population and to take individualized intervention measures to prevent the occurrence of GSD.This study aims to conduct a multicenter investigation involving more than 90 000 people to construct and validate a complete and simplified GSD risk prediction model.Methods A total of 123 634 participants were included in the study between January 2015 and December 2020,of whom 43 929 were from the First Affiliated Hospital of Chongqing Medical University(Chongqing,China),11 907 were from the First People's Hospital of Jining City(Shandong,China),1 538 were from the Tianjin Medical University Cancer Institute and Hospital(Tianjin,China),and 66 260 were from the People's Hospital of Kaizhou District(Chongqing,China).After excluding patients with incomplete clinical medical data,35 976 patients from the First Affiliated Hospital of Chongqing Medical University were divided into a training data set(n=28 781,80% )and a validation data set(n=7 195,20% ).Logistic regression analyses were performed to investigate the relevant risk factors of GSD,and a complete risk prediction model was constructed.Factors with high scores,mainly according to the nomograms of the complete model,were retained to simplify the model.In the validation data set,the diagnostic accuracy and clinical performance of these models were validated using the calibration curve,area under the curve(AUC)of the receiver operating characteristic curve,and decision curve analysis(DCA).Moreover,the diagnostic accuracy of these two models was validated in three other hospitals.Finally,we established an online website for using the prediction model(The complete model is accessible at https://wenqianyu.shinyapps.io/Completemodel/,while the simplified model is accessible at https://wenqianyu.shinyapps.io/Simplified/).Results After excluding patients with incomplete clinical medical data,a total of 96 426 participants were finally included in this study(35 876 from the First Affiliated Hospital of the Chongqing Medical University,9 289 from the First People's Hospital of Jining City,1 522 from the Tianjin Medical University Cancer Institute,and 49 639 from the People's Hospital of Kaizhou District).Female sex,advanced age,higher body mass index,fasting plasma glucose,uric acid,total bilirubin,gamma-glutamyl transpeptidase,and fatty liver disease were positively associated with risks for GSD.Furthermore,gallbladder polyps,total cholesterol,high-density lipoprotein cholesterol,low-density lipoprotein cholesterol,and aspartate aminotransferase were negatively correlated to risks for GSD.According to the nomograms of the complete model,a simplified model including sex,age,body mass index,gallbladder polyps,and fatty liver disease was constructed.All the calibration curves exhibited good consistency between the predicted and observed probabilities.In addition,DCA indicated that both the complete model and the simplified model showed better net benefits than treat-all and treat-none.Based on the calibration plots,DCA,and AUCs of the complete model(AUC in the internal validation data set=74.1%[95% CI:72.9%-75.3%],AUC in Shandong=71.7%[95% CI:70.6%-72.8%],AUC in Tianjin=75.3%[95% CI:72.7%-77.9%],and AUC in Kaizhou=72.9%[95% CI:72.5%-73.3%])and the simplified model(AUC in the internal validation data set=73.7%[95% CI:72.5%-75.0%],AUC in Shandong=71.5%[95% CI:70.4%-72.5%],AUC in Tianjin=75.4%[95% CI:72.9%-78.0%],and AUC in Kaizhou=72.4%[95% CI:72.0%-72.8%]),we concluded that the complete and simplified risk prediction models for GSD exhibited excellent performance.Moreover,we detected no significant differences between the performance of the two models(P>0.05).We also established two online websites based on the results of this study for GSD risk prediction.Conclusions This study innovatively used the data from 96 426 patients from four hospitals to establish a GSD risk prediction model and to perform risk prediction analyses of internal and external validation data sets in four cohorts.A simplified model of GSD risk prediction,which included the variables of sex,age,body mass index,gallbladder polyps,and fatty liver disease,also exhibited good discrimination and clinical performance.Nonetheless,further studies are needed to explore the role of low-density lipoprotein cholesterol and aspartate aminotransferase in gallstone formation.Although the validation results of the complete model were better than those of the simplified model to a certain extent,the difference was not significant even in large samples.Compared with the complete model,the simplified model uses fewer variables and yields similar prediction and clinical impact.Hence,we recommend the application of the simplified model to improve the efficiency of screening high-risk groups in practice.The use of the simplified model is conducive to enhancing the self-awareness of prevention and control in the general population and early intervention for GSD.

Objective To explore the effect of exercise preconditioning on inflammatory response in ischemic stroke brain tissue which mediated by miR-146a in extracellular vesicles in rats with middle cerebral artery oc-clusion(MCAO),and its mechanism.Methods Sixty 6-week-old male Sprague-Dawley rats were randomly divid-ed into a non-exercise group and an exercise group.The non-exercise group was further divided into a sham-operation control group(C,n=15)and an MCAO model group(M,n=15),while the exercise group was further di-vided into an exercise only group(E,n=15)and an exercise plus MCAO model group(EM,n=15).Rats in the E and EM groups underwent 8 weeks of treadmill exercise,6 days per week,30 minutes per day.Then rats in the M and EM groups received MCAO to induce ischemic stroke,while the C and E groups underwent a sham surgery.Twenty-four hours after the surgery,neurobehavioral tests were performed.Plasma was collected to ex-tract extracellular vesicles,and brain tissue was collected to measure the volume of cerebral infarction by using the triphenyl tetrazolium chloride(TTC)staining.Moreover,the Nissl staining was conducted to observe neuronal and Nissl body.Mean while,the content of miR-146a in plasma extracellular vesicles and brain tissue was mea-sured by using the quantitative polymerase chain reaction(qPCR),and the expression of TNF receptor associat-ed factor 6(TRAF6),nuclear factor kappa-B(NF-κB)and tumor necrosis factor-α(TNF-α)in brain tissues were determined using Western blotting.The targeting relationship between miR-146a and TRAF6 was detected by using the dual luciferase reporter gene assay.Results(1)The neurological behavioral scores of the EM and M groups were higher than those of the C group(P<0.01 and P<0.01),with that of the EM group lower than the M group(P<0.01).(2)TTC staining showed that the infarct volume of the EM and M groups was larger than that of the other two groups(P<0.01 and P<0.01),with that of the EM group smaller than the M group(P<0.01).(3)Nissl staining results showed that the neuronal arrangement was loose,the number of neurons re-duced,and the Nissl bodies were lightly stained and decreased in the M group compared with the C and E groups.Moreover,compared with the M group,the number of neurons and Nissl bodies increased in the EM group.(4)The qPCR analysis showed that the expression of miR-146a in the plasma-derived exosomes and brain tissues of the EM and M groups decreased compared with the C group(P<0.05 and P<0.01),with that of the EM group higher than the M group(P<0.05).(5)According to Western blotting,compared with the C group,the expression levels of TRAF6,NF-κB,and TNF-α proteins increased significantly(P<0.05 and P<0.01),with that of group EM signfiicantly lower than group M(P<0.05 and P<0.05).(6)Dual-luciferase report-er gene assay showed that miR-146a had a specific binding site with TRAF6.Conclusion Eight weeks of exer-cise preconditioning reduces the infarct area and the extent of brain damage,which may be mediated by miR-146a via exosomes,increasing the expression of miR-146a in brain tissue,targeting TRAF6,negatively regulat-ing TRAF6/NF-κB,and reducing the expression of TNF-α,thus alleviating the inflammatory response in brain tissue and exerting a protective effect on ischemic brain injury.

Enteral nutrition preparation is one of most important tools to perform clinical nutrition support. Food for special medi-cal purposes ( FMSP) as a special category of enteral nutrition preparation was developed from foods gradually. At present, the man-agement of FMSP bases on the evidence at home and its development has been accelerated. A complete understanding and appropriate use of FMSP will become one essential clinical skill for doctors, pharmacists and dietitians in a multidisciplinary medical team. The re-view revealed the current situation of FMSP and addressed the authors' understanding on the differences between FMSP, medications and foods in order to provide reference for the clinical application of FMSP.

Enteral nutrition preparation is one of most important tools to perform clinical nutrition support. Food for special medi-cal purposes ( FMSP) as a special category of enteral nutrition preparation was developed from foods gradually. At present, the man-agement of FMSP bases on the evidence at home and its development has been accelerated. A complete understanding and appropriate use of FMSP will become one essential clinical skill for doctors, pharmacists and dietitians in a multidisciplinary medical team. The re-view revealed the current situation of FMSP and addressed the authors' understanding on the differences between FMSP, medications and foods in order to provide reference for the clinical application of FMSP.

Objective To investigate the effects of grape seed proanthocyanidin(GSPE) on mitochondrial injury in hippocampus and learning-memory impairment after obstructive sleep apnea hypoxia in rats.Methods Male SD rats(n=80) were randomly divided into control group,model group,low dose of GSPE treatment group and high dose of GSPE treatment group.Rats in control group were exposed in air,the model group were suffered from intermittent hypoxia conditions (50 ml/L,8-hour-intermittent hypoxia everyday,and the duration of experiment 2 and 6 weeks,respectively).Mitochondrion pathology in hippocampal region was observed using electron microscope;malondialdehyde (MDA) contents and superoxide dismutase activity were detected by colorimetry and apoptotic cells was measured by TUNEL method.The cognitive function of rats in each group was assessed with the Morris water maze (MWM).Results After hypoxia,mitochondrion was significantly injured.The MDA contents were increased(79.86 ± 2.52,88.26 ± 2.86) and SOD level decreased (70.67 ± 6.70,64.26 ± 7.86).The number of neural apoptotic cells was significantly enhanced (9.68 ± 0.79,15.9 ± 2.92).MWM test showed that the escaping latency was prolonged and the frequency of crossing the platform was decreased (P ＜ 0.05).Compared with that in the model group,low dose of GSPE decreased MDA contents (76.38 ± 1.96,82.16 ±2.02),increased SOD level(76.20 ± 6.86,70.58 ± 6.86),and decreased apoptotic cells (6.60 ± 0.69,9.54 ±1.36).MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased in GSPE treatment groups(P ＜ 0.05).Compared with low dose of GSPE,high dose of GSPE decreased MDA contents increased SOD level and decreased apoptotic cells.MWM test showed that the escaping latency was shortened and the frequency of crossing the platform was increased (P＜ 0.05).Conclusion GSPE can attenuate mitochondrial injury and improve learning-memory function after obstructive sleep apnea hypoxia.

Objective To compare the effects of different administration routes and dosages of tropisetron on cisplatin-induced kaolin intake in rats.Methods Ninety-six healthy adult male Wistar SPF rats were randomly divided into 8 groups(n =12 each):intrathecal (IT) control group (group TC) and 3 tropisetron groups receiving IT tropisetron 10,20 and 30 μg,the volume of each group was 30 μl (group T10,T20,T30),intravenous(Ⅳ) control group (group IC) and 3 tropisetron groups receiving Ⅳ tropisetron 0.3,0.5 and 0.7 mg/kg respectively (group I0.3,I0.5,I0.7).In group TC and IC,normal saline 30 μl and 0.5 ml were injected IT and Ⅳ,respectively.All rats received cisplatin 3mg/kg by intraperitoneal injection at the time point of thirty minutes after administration,each rat weight,the daily food and kaolin intakes were detected at the time point of 48 hours after cisplatin administration.Results Compared with group Tc,each rat weight loss,the kaolin intakes were significantly decreased (P ＜ 0.05),and food intake dose was significantly increased in group T20 (P ＜ 0.05).Compared with group IC,each rat weight loss,the kaolin intakes were significantly decreased (P ＜ 0.05),and food intake dose was significantly increased in group I0.5 and I0.7 (P ＜0.05).There was no significant difference between group I0.5,I0.7 and group T20.Conclusions The kaolin intakes and the rat weight loss can be decreased by IT tropisetron,and the food take dose was increased meanwhile,and IT tropisetron 20 μg has equivalent efficacy to IV tropisetron 0.5 or 0.7 mg/kg.IT could be the new administration route of tropisetron.