Objective To investigate the effects of substrate stiffness on the proliferation of human umbilical vein endothelial cells ( HUVEC) during dengue virus infection.Methods Polyacrylamide gels were prepared for cell culture [(0±4) kPa].The proliferation of HUVEC cultured on substrates with differ-ent stiffness was determined by using 3-(4,5-diethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfo-phenyl)-2H-etrazolium,inner salt ( MTS) assay.The cycle and apoptosis of HUVEC were determined by flow cytometry analysis.Dengue virus serotype 2 (DENV-2) strains were propagated and identified by con-ventional assays.The HUVEC were infected with DENV-2 strains at a MOI of 10 and cultured on traditional plastic and hydrogel substrates, respectively.The levels of nitric oxide (NO) and endothelin-1 (ET-1) were detected by nitric acid reductase assay and double antibody sandwich ELISA.Results Young′s modulus E value of the hydrogels was (3030 ±0.44) Pa.The proliferation of HUVEC and the expression of NO and ET-1 were enhanced along the increased substrate stiffness.However, no significant differences with the cell cycle and apoptosis were observed between cells cultured on different substrates.Conclusion The stiffness of substrates affected not only the proliferation of HUVEC, but also the release of cytokines during DENV-2 infection.The development of dengue fever was associated with the decreased secretion of vascular active substances as a result of blood vessel injury.The establishment of hydrogel substrates as the model of vascu-lar basement membranes might provide a new way for the in vitro investigation of the pathogenesis of DENV infection.

Objective To analyze the effects of dengue virus 2 ( DENV-2 ) infection on the ex-pression of IL-29 in primary human umbilical vein endothelial cells ( HUVECs) cultured on hydrogel sub-strates .Methods Primary HUVECs were isolated and cultured on hydrogel substrates .DENV-2 stains were used to infect the primary HUVECs at a multiplicity of infection( MOI) of 10.Flow cytometry analysis was performed to detect the apoptosis and infection rate of HUVECs after 48 hours of culturing .The gene chip profiling was performed to analyze mRNA expression .The expression of IL-29 at mRNA and protein levels were measured by real-time fluorescent quantitative PCR analysis and double antibody sandwich ELISA as -say, respectively.Results Compared with 96.36%of baby hamster kidney (BHK) cells that were infected with DENV-2 stains, only 4.71%primary HUVECs cultured on hydrogel substrates were infected .The pri-mary HUVECs cultured on hydrogel substrates with or without DENV-2 infection showed no significant differ-ences with the rates of cell apoptosis and infection (P>0.05).A significant difference was observed with the expression of IL-29 at mRNA level between primary HUVECs cultured on hydrogel substrates and the cells cultured in plastic bottles (P<0.05).The results of the real-time quantitative PCR analysis and ELISA as-say showed that IL-29 was highly expressed in DENV-2 infected primary HUVECs cultured on hydrogel sub-strates as compared with those in control groups (P<0.05).Conclusion The expression of IL-29 was de-tected in DENV-2 infected primary HUVECs cultured on hydrogel substrates , which was significantly differ-ent from that in DENV-2 infected primary HUVECs cultured in plastic bottles .The successful establishment of hydrogel substrates as the model of vascular basement membranes might provide a new way for the investi -gation of the pathogenesis of DENV infection .

Objective To observe the efficacy and safety of Shenqiwuweizikeli for treating chronic insomnia.Methods One hundred and ninety-six cases of subjects were randomly divided into Shenqiwuweizikeli group (n =98) and Estazolam tablets group (n =98).The pittsburg sleep quality index (PSQI) was adopted to evaluate the clinical effects and records of adverse reactions during the study period.Also the lab routine inspection(blood routine,urine routine,liver and kidney function, electrocardiogram were conducted to evaluate safety.Results The Shenqiwuweizikeli and Estazolam tablets all had significant effects for chronic insomnia.The total effective rate of Shenqiwuweizikeli group was 92.86％ (91/98), of Estazolam tablets group was 93.88％(92/98) ,and there was no significant difference (P＞0.05).There were no abnormalities in terms of each routine inspection index.After stopping take the medicine, The adverse reactions including bounce sex insomnia(60 cases), daytime sleepiness/drowsiness (55 cases), dizziness with lacking of power and light headedness(23 cases) in Estazolam tablets group were all more than Shenqiwuweizikeli group with significant difference(P＜0.01).Conclusion The Shenqiwuweizikeli has definite efficacy and safety for treated with chronic insomnia without withdrawal of recoil and dependence.

Objective To explore the protective effects of electro-acupuncture (EA) serum onβ-amyloid protein (Aβ) induced primary rat hippcampal neurons. Methods The rat models of Alzheimer's disease were established by intracerebral injection of Aβ1-40. After treated them with EA, the serum was harvested. Primary cultured hippocampal neurons were treated with Aβ25-35 to establish neuronal damage model in vitro, and divided into normal group, model group and EA serum group. The proliferation of neurons was detected by MTT test. Neuronal apoptosis was examined by TUNEL staining, and expression of cysteine aspartic acid proteases-3 (Caspase-3) was detected by immunocytochemical staining. Results MTT test showed that the cell viability was significantly decreased after treated with Aβ. While compared with the model group, cell proliferation of EA serum group was significantly enhanced (P<0.01). TUNEL staining showed that the number of apoptotic cells in EA serum group decreased significantly compared with the model group (P<0.01). After 48 h of Aβ treatment, Caspase-3 expression levels were significantly elevated. However, compared with the model group, the number of Caspase-3 positive cells in EA serum group was significantly reduced (P<0.01). Conclusion The EA serum could promote the proliferation of hippocampal neurons, reduce the expression of Caspase-3, counteract the neurotoxicity of β-amyloid protein, and reduce neuronal apoptosis.

Objeetive To develop decision-making training system in fundamentals of nursing for nursing students and evaluate its application effect.Methods The training contents and level requirements were determined first.Then based on the five steps of clinical decision-making theory (put forward questions,determine goals,make decisions,implement and evaluate),the simulated training cases were designed and integrated into software of the training system.And 28 nursing students used the training system.Results Most of the nursing students liked this learning method.They thought it was very interesting,challenging,and useful.It could help increase the enthusiasm learning and the participation of students.Using it the students could integrate the theoretical knowledge into the practice.So it was helpful to train the capacity of solving clinical problems and clinical decision-making of students.The results of the open-ended question showed that the comprehensive training system could help students grasp the knowledge,test and consolidate the knowledge.It contributed to the combination of theory and practice,the cultivation the capacities of basic nursing skills,teamwork,self-learning,and solving clinical problems.Conclusions The comprehensive training system could help nursing students develop the ability to solve clinical problems and improve the clinical decision-making ability.It can be applied in the related fields.

Objective To identify the clinical profile and neuro-imaging findings of the right temporal lobe variant of semantic dementia (RTLV),and to understand the relationship between anatomic and function by comparison to the left temporal lobe atrophy.Methods Of 70 patients with diagnosis of frontotemporal dementia in our memory clinic,three patients with right temporal lobe atrophy were identified based on the MRI scans.We obtained the profile of cognitive function,behavior and personality changes in these 3 patients and compared them with those in 5 semantic dementia (SD) patients with predominant leftsided temporal lobe atrophy.We also underwent 18F-fluoro-2-deoxy-D-glucose positron emission tomography (18F-FDG-PET) with statistical parametric mapping analysis in these 3 patients.Results In RTLV patients,the most prominent cognitive deficits were impairment of prosopagnosia,and getting lost.Variety behavioral symptoms including apathy,social disinhibition,stereotypy,compulsive behaviors were more prevalent in early course for patients with RTLV,which were different from SD patients with semantic loss began with anomia or single-word comprehension.18F-FDG-PET scan showed decreased metabolism mainly in right temporal lobe and in other brain regions with varying degrees.Conclusions The RTLV is clinical heterogeneous.Prosopagnosia,behavioral changes and getting lost are the main features in the early course.It therefore,might be proposed a separate clinical entity.

Objective:To explore the relationship between the polymorphisms in gene FGFR1, FGF10, FGFI8 and the nonsyndromic cleft lip with or without cleft palate (NS CLP) in Chinese population. Methods: Genomic DNA was isolated from peripheral lymphocytes of 75 patients with NS CLP and their parents and 75 unimpaired healthy children. The polymorphisms in FGFRI gene rs13317, p. E467K, p. M3691 and p. S393S, FGF10 gene rs1448037 and FGFI8 gene rs4043716 were detected by applying three-dimensional (3-D) polyacrylamide gel microarray technology. The data were performed using statis-tical analysis : the genotype frequenc+ y and allele frequency between patients with NSCL/P and control subjects were performed. Haplotype relative risk (HRR) , family based association test (FBAT) , and transmission disequilibrium test (TDT) in nuclear family were performed. Results: There were no poly-morphism in FGFR1 gene p. E467K, p. M369I and p. $393S site, the corresponding base was all G. The polymorphisms of rs13317 and rs1448037 were detected and their genotype frequency and allele frequen-cy showed no significant difference between 75 patients with NSCL/P and 75 normal children. TDT, HRR and FBAT were also no significant differences. The genotype frequency of gene FGF18 rs4043716 showed significant difference, but allele frequency were no significant difference. TDT, HRR and FBAT were also no significant difference. Conclusion: Our studies suggest an association between gene FGF18 rs4043716 and the NS CLP in Chinese population, and no association among gene FGFR1 rs13317, p. FA67K, p. M3691, p. S393S and gene FGF10 rs1448037.

Objective:To explore the current status and research hotspots of structural equation modeling in nursing from 2012 to 2021.Methods:We searched for literature on the structural equation modeling in nursing based on Web of Science core collection database. CiteSpace and Excel were used to analyze the time, space, and keywords included in 742 articles.Results:From 2012 to 2021, the number of domestic and international publications showed a rapid growth trend with the growth of the year. China was the country with the highest number of publications. The Western University in Canada was the institution with the highest number of publications. The frequency of keywords such as structural equation modeling, quality of life, job satisfaction, health, social support and stress was high. The clustering effect of keywords was significant ( Q=0.72, S=0.87) , and research hotspots mainly included physical and mental health care and nursing work management. Conclusions:The structural equation modeling in nursing is showing a rapid development trend. In the future, numerous studies should be conducted on the construction and validation of nursing structural relationship models to promote the development of nursing.

Objective To clarify the expression of leptin and leptin receptor in normal brain tissues and gliomas and investigate the effect of exogenous leptin on the proliferation,migration and invasion of human glioma U251 cell line.Methods Immunohistochemical staining was used to detect the expression of leptin and leptin receptor in 50 cases of different grades of glioma tissues and 20 cases of normal brain tissues.The effects of exogenous leptin on proliferation,migration and invasion of U251 cells were detected by MTT assay,cell scratch assay and Transwell invasion assay.Results (1) The positive expression rates of leptin and leptin receptors in glioma tissues were 50.0％ and 92.0％,respectively.(2)Proliferation activity:leptin concentrations of 0 ng/ml,10 ng/ml,and 50 ng/ml had no significant difference in the proliferation of U251 cells (absorbance:0.263±0.015,0.273±0.017 and 0.277±0.006,respectively),and the leptin concentration of 100 ng/ml had a significant effect on the proliferation of U251 cells (absorbance:0.315±0.005,P＜0.05).(3)Migration ability:the migration rate of U251 cells treated with different concentrations of leptin increased significantly with the passage of time,and the migration rate was most significant at the concentration of 100 ng/ml ((93.313±3.080) ％),and the difference was statistically significant (P＜0.05).(4)Invasive ability:with the increase of leptin concentration and the prolongation of the action time,the invasive ability of U251 cells was enhanced.When leptin was used at a concentration of 100 ng/ml,the number of penetrating cells were the biggest(135±2).Conclusion Leptin and leptin receptors are involved in the occurrence of gliomas;and exogenous leptin promotes the proliferation of U251 cells and has time and dose dependability on the migration and invasion of U251 cells.

OBJECTIVE: To explore the pathogenesis of two siblings (including a fetus) from a pedigree affected with Joubert syndrome. METHODS: Peripheral blood samples of the proband and his parents as well as amniotic fluid and abortion tissues of the fetus were collected. Part of the samples were used for the extraction of DNA, and whole exome sequencing (WES) was carried out to screen potential variants in the proband and his parents. Suspected variants were subjected to bioinformatics analysis with consideration of the clinical phenotype, and were verified by Sanger sequencing of the proband, fetus and their parents.The remainders were used for the extraction of RNA, and the mechanism of splicing variant was validated by reverse transcription-PCR (RT-PCR). RESULTS: WES showed that both patients have carried c.175C>T (p.R59X) and c.553+1G>A compound heterozygous variants of the TMEM237 gene. Among these, c.175C>T was a nonsense mutation inherited from the asymptomatic mother, while c.553+1G>A was an alternative splicing mutation inherited from the asymptomatic father. RT-PCR showed that this variant has resulted in aberrant splicing by exon skipping. CONCLUSION The compound heterozygous variants of the TMEM237 gene probably underlay the etiology of Joubert syndrome in this pedigree. Above finding has enriched the phenotype and variant spectrum of the TMEM237 gene, and facilitated genetic counseling and prenatal diagnosis for the family.
Abnormalities, Multiple/genetics* ; Cerebellum/abnormalities* ; Eye Abnormalities ; Female ; Genotype ; Humans ; Kidney Diseases, Cystic ; Mutation ; Pedigree ; Phenotype ; Pregnancy ; Retina/abnormalities*