Objective To compare the humoral immune response of BALB/c mice immunized by recombinant plasmids PeDNA3.1-M-NS1 and pcDNA3.1-N-NS1.Methods Dengue type 2 virus(DENV2)NS1 gene were constructed two partial sequences(1-413 bp)of the pcDNA3.1 eukaryotic plasmids and pET28a(+)plasmid for prokaryotic expression,identification,purification and quantification.The BALB/c mice were immunized by pcDNA3.1-M-NS1,pcDNA3.1-N-NS1 recombinant plasmids with adjuvant.Each animal received a primary inoculation and two boosts at 1-week intervals.Then the blood samples of BALB/c mice were collected from different experiment groups at day 7,14,28 and 56,respectively after first immunization.The specific IgM/IgG antibodies for NS1 protein in serum were confirmed by indirect ELISA.And then the activities of the specific protective antibody were determined by cytopathic effect inhibition(CPEI).Results Construction of the pET28a(+)-NS1 m/pET28a(+)-NS1n prokaryotic expression plasmid,SDS-PAGE analysis showed that,NS1 gene partial sequence was expressed,both the relative molecular weight of about 22.3×103:Western blot showed that the protein can bind anti-His tag monoclonal antibody;byNi affinity chromatographywith apurity of 92% protein,on the C6/36 cell toxicity,and can be used ELASA detection.The results showed that the levels of specific IgM/IgG antibody and neutralizing antibody activities were increased in pcDNA3.1-M-NS1 booster immunization group than other groups.The result had been observed longer duration of antibody level in peDNA3.1-M-NS1 booster immunization group.Conclusion Humoral immune response were significantly different between pcDNA3.1-M-NS1 and pcDNA3.1-N-NS1 recombinant plasmid immunized mice groups.

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 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 establish the models with human rheumatoid arthritis (RA) synovial fibroblasts (RASFs)-cartilage-severe combined immune deficient (SCID) mice for the study of the RASFs in the pathogenesis of RA.Methods The 4th passage RASFs were marked with 5-bromodexyuridine (5-Brdu)and injected into a cavity of inert sterile gel sponge,then with the normal human cartilage co-implanted in the back subcutaneously of SCID mice to set up a novel model of RA.Osteoarthritis synovial fibroblasts (OASFs)were injected as control group.Thirty days after the surgery,the mice were killed,the grafts and the knee joints were proceed with histological study and immunohistochemistry to detect the expression of 5-Brdu and Vimentin in synoviocytes.The serum level of interleukin (IL)-6 and matrix metalloproteinase (MMP)-3 were detected with enzyme-linked immunosorbent assay (ELISA).Results Twenty-three mice survived except for one mouse in the RASFs group died of anesthesia.① Only in one case in the RASFs group,the IL-6 was detected,the others were unable to be detected.The MMP-3 in the OASFs group was (40±17) pg/ml,but in RASFs group only one case was detected.② There were 4 and 3 implanted cartilages loss in the RASFs group and OASFs group respectively.The histological scores of cartilage invasion by synoviocytes and cartilage degradation in grafts were higher in RASFs groups than in OASFs groups (0.6±0.7 vs 0.3±0.5,2.3±0.8 vs 1.7±1.0 respectively).③ The histological scores of synovial hyperplasia and cartilage invasion in the knee joints was significantly higher in the RASFs group than in the OASFs group (3.1±0.8 vs 1.7±1.0,P＜0.01,1.6±1.7 vs 0.6±1.4,P＜0.05 respectively).④ During the grafts,a lot of 5-Brdu and Vimentin markers positive synovio-cytes were found in the mice subcutaneous tissue,but manipulus positive synoviocytes were found on the area of cartilage invasion in both groups.In knee joints,single positive synoviocytes could be detected in bone marrow and hyperplasic area of the synovial tissue in both groups.Conclusion The isolated RASFs can survival and have the ability to invade and degrade the cartilage in vivo without the limitation of immunity induced inflammations,and can also migrate to the synovial joints in distance and induce arthritis.

Objective To explore the gender difference in clinical features and outcomes of elderly patients with coronary true bifurcation lesions after percutaneous coronary intervention (PCI).Methods A total of 169 consecutive patients were included in the study, who were diagnosed coronary true bifurcation lesions by coronary angiography (CAG)and received PCI in our hospital from December 2013 to December 2015.All patients were divided into 2 groups according to their gender (71 femals and 98 males).Angiographic characteristics of the coronary lesions and clinical data were analyzed in both groups.Clinical outcomes during follow up were recorded and studied.Results The levels of total cholesterol (TC), triglyceride (TG),high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) in the female group were significantly higher than in the male group (all P<0.05).The levels of BUN,Cr,UA,CyC and TBIL in the female group were significantly lower than in the male group (all P<0.05).Comorbidity rates of hypertension and diabetes were higher in female patients without significant differences (P>0.05).The median follow-up time was 17 months and the incidence rate of overall adverse events in women was higher than that of men (25.4% vs.11.2%, P<0.05).Conclusions The clinical outcomes of elderly patients with coronary true bifurcation lesions after PCI had gender differences.The rates of adverse events in female patients was found significantly higher than male patients.

Objective To explore the effects of Living with Hope Program(LWHP) on family caregivers of patients with advanced lung cancer.Methods Totally 60 family caregivers of patients with advanced lung cancer were randomly divided into the experimental group and the control group.The experimental group received LWHP intervention and routine care knowledge education,the control group only received routine care knowledge education.Hope,self-efficacy,anxiety and depression,and quality of life were evaluated at the first week,the second week,the first month,and the third month.Differences between two groups were compared using variance analysis of repeated measurements.Results The self-evaluation of the intervention was 76.6％,and the main effects of hope level,selfefficacy,anxiety and depression,and mental health were statistically significant (P＜0.05);the time effect on hope level and self-efficacy were statistically significant (P＜0.05);there were interaction effects between intervention and time on hope level,self-efficacy,anxiety and depression,and mental health (P＜0.05).Conclusion LWHP can effectively improve hope level,self-efficacy and mental health status,and alleviate anxiety and depression of family caregivers of patients with advanced lung cancer.

Objective To evaluate the efficacy and safety of domestic human recombinant FSH (rhFSH) in women with anovulation of WHO groupⅡ. Methods A randomized, blind, parallel-controlled, non-inferiority and multicenter study was performed. A total of 534 admitted to 13 hospitals from May 2008 to August 2009. There were 531 women with ovulatory disorder was included in the statistical analysis, were randomly divided into test group (domestic rhFSH, n=352) and control group (imported rhFSH, n=179). Percentage of cycle with mature follicle, ovulation rate, clinical pregnancy rate, multiple pregnancy rate, ovarian hyperstimulation syndrome (OHSS) and adverse events were observed. Results No statistical significant differences (P>0.05) were observed between the two groups in terms of the efficiency on mature follicle [91.8%(323/352) versus 88.8%(159/179)], ovulation rate [91.3%(295/323) verus 90.6%(144/159)], clinical pregnancy rate [19.2%(62/323) verus 18.2%(29/159)], the number of the follicles<14 mm, the level of serum LH and progesterone, the thickness of endometrium on the day of hCG administration. The number of follicle≥18 mm and 14 mm≤follicle<18 mm and the level of serum estradiol on the day of hCG in the test group were significantly higher than those in the control group (P<0.05). The number of days of rhFSH administration in the test group was significantly less than that in the control group [(9.8±2.2) versus (11.4± 0.6) days, P<0.05], the dosage of rhFSH was significantly lower than that in the control group [(879 ± 419) versus (1 043 ± 663) U, P<0.05]. The multiple pregnancy rate in the test group was significantly higher than that in the control group [21% (13/62) versu 10% (3/29), P<0.05]. The incidence of OHSS and adverse events were similar between the two groups (P>0.05), and no other adverse events were observed in test group during treatment. Conclusion Ovarian stimulation with domestic rhFSH is effective, safe and economical in women with anovulation of WHO groupⅡ.

Aim To investigate the effect of curcumin against high-fat-diet induced C57BL/6J mice bone changes and the correlation between the expression of cathepsin K and curcumin.Methods Curcumin treated C57BL/6J mice had been on high fat diet for 12 weeks.The HE, Alizarin red S staining and Safranin O/fast green staining of femur were employed to evaluate bone microstructure, bone metabolism and bone development.The expressions of cathepsin K were assessed by Western blot and immunohistochemical staining.Results Histopathological results showed that curcumin could improve the destruction of trabecular bone structure, cartilage development and bone calcification.Biomechanical results proved that curcumin could improve the bone strength of the type 2 diabetic mice induced by high fat.The results of immunohistochemistry and Western blot assay indicated that curcumin could significantly inhibit the expression of cathepsin K in bone tissues of mice.Conclusion Curcumin can increase bone strength, improve bone microstructure, and enhance the degree of bone calcification, which may be achieved by inhibiting the expression of cathepsin K.

Abstract The possible mechanisms of developmental dyslexia mainly include the hypothesis of language framework and the hypothesis of non-verbal framework. The language framework assumes that people with developmental dyslexia may exhibit defects in phonetic awareness, rapid naming, phonetic memory, and orthographic processing. Studies of developmental dyslexia in Chinese have found that deficiencies in orthography may be an important cause of dyslexia, but there are diverse views and opinions regarding orthography processing. This article sorts out the research progress in behavioral and neuroimaging aspects of orthography studies, and provides references for further development of processing test materials and methods in the research of processing mechanism of developmental dyslexia orthography.

Objective: To investigate the characteristics of visual motor integration in children with developmental dyslexia and ADHD, and to provide a reference for the diagnosis and treatment of these children. Methods: During July to November of 2020，students from grade 3 to grade 5 of 7 primary schools in Xinjiang were selected by using random cluster sampling method. A total of 56 dyslexia group (group DD), attention deficit hyperactivity disorder group (group ADHD), comorbidity group and normal control group were selected and compared the differences of their visual motor integration ability and related factors ability. Multiple linear regression analysis was used to explore the predictive effects of related factors on visual motor integration ability. Results: The scores of visual integration ability and visual perception in comorbidity group (95.05±14.01, 12.71±7.40) were lower than those in DD group (104.77±17.19，23.04±11.48), ADHD group (104.00±14.11，25.70±10.74) and normal control group (129.80±12.91, 44.05±16.56) ( F/Z =58.24,110.49, P <0.05). The visual working memory score of the comorbidity group ( 73.64 ±5.36) was lower than the normal control group (78.96±4.68) ( P <0.05)，and there was no significant difference between the DD group (74.48±7.06) and the ADHD group (75.98±7.36) ( P >0.05). The results of multiple regression showed that visual perception, age, IQ and visual working memory were associated with visual and motor integration ability of dyslexia children with ADHD ( R 2=0.32，0.17，0.11， 0.04 ， P <0.05). Conclusion Visual motor integration and visual perception among children with DD combined ADHD are more severely impaired than those with ADHD and DD alone. Visual perception, age, IQ and visual working memory could help predict the development of visual and motor integration ability in children with DD combined ADHD.