Objective:To highly express the secretory gene engineering anti CD20F(ab') 2 in the E.coli,simplify working processes and enhance the bioactivity of the antibody fragment.Methods:Using the single factor analysis to optimize the ferment parameters.A culture method which most suit for E.coli secreting anti CD20F(ab') 2 was selected.The anti CD20F(ab') 2 was extracted from periplasm then purified it using the strptococal protein G affinity colume and the S200 HR size exclusion chromatography.The activity of anti CD20F(ab') 2 inhibiting the growth of cell Daudi in vitro was eamined using MTT.Results:Using the optimized culture method,the yield of anti CD20F(ab') 2 was distinctly enhanced from 1.9～2.2 mg/L to 3.7～4.3 mg/L and the proportion of anti CD20F(ab') 2 in all the extract also was improved from 9.7～13.2% to 38.1%～46.8%.After the S200 HR size exclusion chromatography,the purity of anti CD20F(ab') 2 could exceed 85%.The outcome of MTT exmiination showed the IC 50 of anti CD20F(ab') 2 and Fab' were 14.6 ?g/ml and 39.5 ?g/ml,respectively.Conclusion:The gene engineering anti CD20F(ab') 2 could highly express in the E.coli by using the optimized culture method.The anti CD20F(ab') 2 inhibited the growth of Daudi cells in vitro more strong than anti CD20Fab'.

Objectives To compare prospective memory (PM) deficits with retrospective memory (RM) deficits and to explore the correlation between PM and RM in chronic schizophrenia. Methods Fifty chronic schizophrenia pa?tients and fifty healthy controls were recruited. The PM performance [event-based PM (EBPM) and time-based PM (TB?PM)] were evaluated by the Chinese version of the Cambridge Prospective Memory Test (C-CAMPROMPT); working memory (WM) was evaluated by the digital span subtest (DS);immediate auditory logical memory (IALM), delayed audito?ry logical memory (DALM), immediate visual reproduction memory (IVRM) and delayed visual reproduction memory (DVRM) were evaluated by the logical memory and visual reproduction subtest. The score of each test was transformed to comparable standard score. Results Patients performed significantly worse on EBPM [(7.9 ± 3.4) vs. (13.7 ± 2.9)], TBPM [(6.9±3.6) vs. (13.0±3.2)], DS [sequence:(5.8±2.0) vs. (7.5±2.2);backward:(6.5±1.9) vs. (8.2±2.8)], IALM [(8.3±3.1) vs. (11.9 ± 2.5)], DALM [(7.4 ± 3.7) vs. (11.8 ± 2.6)], IVRM [(8.0 ± 2.7) vs. (11.2 ± 3.8)], and DVRM [(7.7 ± 3.5) vs. (10.8 ± 2.7)] scores than controls (P<0.05). The extent of deficits of EBPM and TBPM were greater than those of DS (sequence and backward), IALM, DALM, IVRM and DVRM (P<0.05), but not DALM (P>0.05). The performance of PM in chronic schizophrenia was significantly related to DS (sequence and backward), IALM, DALM and DVRM (P<0.05), but not IVRM (P=0.155). Conclusion:There are greater prospective memory deficits than retrospective memory deficits in chron?ic schizophrenia and the prospective memory deficits are correlated with the retrospective memory deficits in chronic schizophrenia.

ObjectiveTo investigate the influence of dual positivity of HBsAg and anti-HBs on the development of hepatocellular carcinoma (HCC) in patients with hepatitis B virus (HBV) infection. MethodsPubMed, Embase, Cochrane Library, CNKI, and Wanfang Data were searched for articles on the influence of dual positivity of HBsAg and anti-HBs on the risk of HCC published from July 1, 1975 to March 27, 2019. RevMan5.3 and Stata11.2 were used for statistical analysis of data. A heterogeneity analysis was performed for the studies included; a random effects model was used in case of significant heterogeneity, and a fixed effects model was used in case of non-significant heterogeneity. Odds ratio (OR) and corresponding 95% confidence interval (CI) were used to investigate the association of dual positivity of HBsAg and anti-HBs with the development of HCC. Begg funnel plots were used to investigate publication bias. By removing one article each time, the sensitivity analysis was used to assess the quality and reliability of the Meta-analysis. ResultsA total of 4 articles were included, with 2 studies in the Korean population and 2 in the Chinese population, and there were 3042 patients in total. The meta-analysis showed that there was no significant association between dual positivity of HBsAg and anti-HBs and the development of HCC (OR=1.46, 95%CI: 0.76-2.80, P=0.25). A country-based subgroup analysis showed significant association between dual positivity of HBsAg and anti-HBs and the development of HCC in the Korean population (OR=2.67, 95%CI: 1.61-4.43, P=0.000 1), while no significant association was found in the Chinese population (OR=0.89, 95%CI: 0.48-1.64, P=0.70). ConclusionThere is no significant association between dual positivity of HBsAg and anti-HBs and the development of HCC, and further studies are needed in future.

Objective To explore the reliability and validity of the computerized Chinese version of Cambridge Prospective Memory Test (C-CAMPROMPT) for assessment of prospective memory (PM) in chronic schizophrenia patients. Methods 50 patients and 50 healthy controls formed the study sample. PM performance was measured with computerized C-CAMPROMPT, while the Wechsler Adult Memory Scale-Forth Edition (WMS-IV), Wisconsin Card Sorting Test (WCST) and Category Fluency Test (CFT) were administered to assess logical memory (LM), visual representation (VR), executive function and processing speed. Results The test- retest reliability (0.981, P<0.001), split half reliability (0.627, P<0.001) and internal consistency reliability (0.742) of C-CAMPROMPT were satisfied. The scores of C-CAMPROMPT and its subtest in schizophrenia were lower than that in healthy control (P<0.001). The performance of PM in patients with schizophrenia closely related to the scores of LM, VR, WCST-CC and CFT (r=0.34~0.89, P<0.05). The sensitivity (86%) and specificity (92%) of the scale were satisfied. Factor analysis extracted 2 factors. Conclusion The computerized C-CAMPROMPT shows a good reliability and validity for assessment of PM function in chronic schizophrenia, and is a sensitive, adaptable, stable instrument.

Objective To develop Prospective Memory Test and explore its reliability and validity in normal and schizophrenic population. Methods According to the structure of prospective memory, Prospective Memory Test was developed based on the previous researches. 40 inpatients with schizophrenia and 30 healthy subjects were assessed with Prospective Memory Test and Wechsler Memory Scale-Fourth Edition, Chinese (WMS-Ⅳ). Results Factor analysis extracted 2 main factors, named time-based prospective memory (TBPM) and event-based prospective memory (EBPM), including 10 items. The average score of Prospective Memory Test was lower in the patients than in the healthy controls by 1~2 standard deviations (P<0.001). Discriminant analysis showed that the specificity, sensitivity, and diagnostic consistency were 93.3%, 72.5% and 81.4%, respectively. Prospective Memory Test scores and two subtest scores positively correlated with total score of WMS-Ⅳ (r=0.44~0.53, P<0.001). The Cronbach's α among all the items was 0.76, and was 0.68 in the TBPM and 0.59 in the EBPM. Split-half reliability of the scale was 0.65 (P<0.001). Conclusion The reliability and validity of Prospective Memory Test are acceptable for schizophrenics.

AIM: To study the effects of fibroblast growth factor 8 (FGF8) on directional differentiation of human dental pulp stem cells (hDPSCs) into odontoblasts and pulp tissue.METHODS: hDPSCs were isolated and cultured, and identified with flow cytometry by detecting cell surface markers of hDPSCs.FGF8 at concentration of 50 μg/L was added into the mineralization fluid to induce the differentiation of the hDPSCs.The mRNA expression of dentin sialophosphoprotein (DSPP), alkaline phosphatase (ALP), bone sialoprotein (BSP) and core-binding factor alpha 1 (Cbfa-1) in differentiated cells was detected by real-time PCR.FGF8 and mouse E11.5 dental epithelium formed restructuring cell group with hDPSCs, and then the restructuring cell group was transplanted under renal capsule membrane in nude mice for tissue culture.DNA in situ hybridization was used to identify the sources of odontoblasts and pulp cells.RESULTS: The surface markers of CD29 and CD90 showed positive in isolated hDPSCs.FGF8 induced hDPSCs to form a distinct mineralization nodule, and the expression of dentin-specific proteins, DSPP, BSP and Cbfa-1, was increased.hDPSCs were induced to differentiate into odontoblasts and pulp cells by E11.5 dental epithelium and FGF8.CONCLUSION: FGF8 can assist dental epithelium to induce directional differetiation of hDPSCs into odontoblasts and pulp cells, and formation of dentin and dental pulp cavity structure.

Background: Feline calicivirus (FCV) is a common pathogen of felids, and FCV vaccination is regularly practiced. The genetic variability and antigenic diversity of FCV hinder the effective control and prevention of infection by vaccination. Improved knowledge of the epidemiological characteristics of FCV should assist in the development of more effective vaccines. Objectives: This study aims to determine the prevalence of FCV in a population of cats with FCV-suspected clinical signs in Hangzhou and to demonstrate the antigenic and genetic relationships between vaccine status and representative isolated FCV strains. Methods: Cats (n = 516) from Hangzhou were investigated between 2018 and 2020. The association between risk factors and FCV infection was assessed. Phylogenetic analyses based on a capsid coding sequence were performed to identify the genetic relationships between strains. In vitro virus neutralization tests were used to assess antibody levels against isolated FCV strains in client-owned cats. Results: The FCV-positive rate of the examined cats was 43.0%. Risk factors significantly associated with FCV infection were vaccination status and oral symptoms. Phylogenetic analysis revealed a radial phylogeny with no evidence of temporal or countrywide clusters. There was a significant difference in the distribution of serum antibody titers between vaccinated and unvaccinated cats. Conclusions This study revealed a high prevalence and genetic diversity of FCV in Hangzhou. The results indicate that the efficacy of FCV vaccination is unsatisfactory. More comprehensive and refined vaccination protocols are an urgent and unmet need.

Objective Although principal components analysis profiles greatly facilitate the visualization and interpretation of the multivariate data,the quantitative concepts in both scores plot and loading plot are rather obscure.This article introduced three profiles that assisted the better understanding of metabolomic data.Methods The discriminatory profile,heat map,and statistic profile were developed to visualize the multivariate data obtained from high-throughput GC-TOF-MS analysis.Results The discriminatory profile and heat map obviously showed the discriminatory metabolites between the two groups,while the statistic profile showed the potential markers of statistic significance.Conclusion The three types of profiles greatly facilitate our understanding of the metabolomic data and the identification of the potential markers.

Background: Feline calicivirus (FCV) is a common pathogen of felids, and FCV vaccination is regularly practiced. The genetic variability and antigenic diversity of FCV hinder the effective control and prevention of infection by vaccination. Improved knowledge of the epidemiological characteristics of FCV should assist in the development of more effective vaccines. Objectives: This study aims to determine the prevalence of FCV in a population of cats with FCV-suspected clinical signs in Hangzhou and to demonstrate the antigenic and genetic relationships between vaccine status and representative isolated FCV strains. Methods: Cats (n = 516) from Hangzhou were investigated between 2018 and 2020. The association between risk factors and FCV infection was assessed. Phylogenetic analyses based on a capsid coding sequence were performed to identify the genetic relationships between strains. In vitro virus neutralization tests were used to assess antibody levels against isolated FCV strains in client-owned cats. Results: The FCV-positive rate of the examined cats was 43.0%. Risk factors significantly associated with FCV infection were vaccination status and oral symptoms. Phylogenetic analysis revealed a radial phylogeny with no evidence of temporal or countrywide clusters. There was a significant difference in the distribution of serum antibody titers between vaccinated and unvaccinated cats. Conclusions This study revealed a high prevalence and genetic diversity of FCV in Hangzhou. The results indicate that the efficacy of FCV vaccination is unsatisfactory. More comprehensive and refined vaccination protocols are an urgent and unmet need.

Objective:To investigate the effectiveness of mindfulness training on the emotional regulation and resilience of college students.Methods:Eighty one college students were recruited and randomly divided into mindfulness training group and control group, of which 74 completed the study (36 in mindfulness training group and 38 in control group). Participants in mindfulness training group received an 8-week mindfulness training, while participants in control group received no intervention. Participants were assessed with the Five Facet Mindfulness Questionnaire (FFMQ, measuring mindfulness level), Profile of Mood State (POMS, measuring emotion) and the Resilience Scale for Chinese Adolescent(RSCA, measuring resilience) before and after intervention (4 and 8 weeks).Results:Before and after the intervention, the interaction effects of total score of five factors mindfulness, scores of observing, actaware, non-judging, total score of resilience, scores of goal planning, help-seeking and the fatigue score were significant(all P<0.05). There were significant differences in mindfulness level, mood state and resilience scores in mindfulness training group before and after intervention(all P<0.05). There were significant differences of scores of FFMQ , POMS and RSCA post-training compared with baseline( P<0.05). The total score of FFMQ was improved significantly after 4 weeks of intervention ((114.92±11.70) vs(110.36±11.24), P<0.05), while the scores of observing((26.19±5.21) vs(24.00±5.34)), describing((23.36±4.82) vs(21.19±4.76)), non-judging((23.75±4.20) vs(22.06±4.99)) and non-reacting((19.72±3.16) vs(18.58±3.61)) were improved significantly after 8 weeks of intervention. There were significant differences of the scores of fatigue, depression-dejection, panic, vigor-activity and total mood disturbance before and after the intervention in mindfulness training group (all P<0.05). The scores of fatigue((12.31±4.08) vs(13.39±5.12)), panic((12.08±3.57) vs(13.67±4.35)) and total mood disturbance((132.94±17.71) vs(145.75±22.68)) were reduced significantly after 8 weeks of intervention (all P<0.05), while the scores of vigor-activity were improved significantly after 8 weeks of intervention ((18.31±4.61) vs (15.39±4.99), P<0.01). The participants in mindfulness training group got significantly higher total score, scores of goal planning, help-seeking, and affect control of RSCA after intervention compared with baseline (all P<0.05). Conclusion:Mindfulness training can enhance the mindfulness level and resilience of college students, decrease negative affect such as fatigue, depression, panic, and increase positive affect such as activity.