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Objective:To investigate the characteristics of implicit memory and its related factors in schizophrenic patients with negative and positive symptoms.Methods:Ninety-three schizophrenic patients (including 52 cases of negative symptoms group and 41 cases of positive symptoms group) and 30 normal controls (normal group) were tested with the method of Chinese character word completion method. The reaction time and correct rate were recorded and compared with analysis of variance and t-test, and the correlation with demographic factors was analyzed with Pearson correlation analysis. Results:Accuracy of implicit memory test in schizophrenia group: there were statistically significant differences in the accuracy of patients with different course of disease (≤5 years: (38±5)%, 5-15 years: (34±8)%, ≥15 years: (34±7)%, P<0.05).The differences were statistically significant in the accuracy of patients with different education levels(primary school: (35±6)%, junior and senior high school: (34±7)%, secondary college education and above: (39±5)%, P<0.05).The accuracy of patients with education years < 10 years ((34±7)%) was significantly lower than those with education years ≥10 years ((37±6)%, P<0.05).Reaction time results of implicit memory test in schizophrenia group : there were significant differences in reaction time of patients with different disease course(≤5 years: (3 248±971)ms, 5-15 years: (3 515±672)ms, ≥15 years: (3 925±842)ms, P<0.05).The differences were statistically significant in the reaction time of workers ((3 495±712)ms), farmers ((3 870±878)ms) and soldiers ((3 024±924)ms, P<0.05).The reaction time of patients with clozapine ((3 869±871)ms) was significantly higher than that of patients with olanzapine ((3 393±626)ms, P<0.05).Intergroup results of accuracy: the accuracy of normal control group ((40±5)%) was significantly higher than that of negative symptom group ((33±7)%, P<0.01).The accuracy of negative symptom group was significantly lower than that of positive symptom group ((37±6)%, P<0.01).Intergroup reaction time results: the reaction time of normal control group ((2 660±667)ms) was significantly lower than that of negative symptom group ((3 678±951)ms, P<0.01) and positive symptom group ((3 072±865)ms, P<0.05).The reaction time of negative symptom group ((3 678±951)ms)was significantly higher than that of positive symptom group( P<0.01).There was significant negative correlation between the accuracy of implicit memory and the course of disease in schizophrenia ( r=-0.22, P<0.05). Conclusion:The implicit memory of schizophrenic patients is related to the course of disease, taking drugs, and occupation.There may have differences in implicit memory between patients with negative and positive symptoms of schizophrenia.
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@#The purpose of this experiment is to achieve the soluble expression of immunotoxins in Escherichia coli and avoid the complicated operation caused by the formation of inclusion bodies. An anti-mesothelin monoclonal antibody SS1 and it′s derivative SS1P which composing of SS1 and a truncated pseudomonas exotoxin PE38KDEL were used as the passenger protein. We took advantages of solubility promoting fusion tags and the self-cleaving split intein in recombinant antibody fragment and immunotoxin expression and purification. We constructed solubilizing tags-NpuCD118G fusion tags, and recombinant SS1/SS1P were fused at the C-terminal of the fusion tags. The constructs were expressed in E. coli(SHuffle T7)cytoplasm in soluble form at low temperature. Dextrin Beads 6FF column and Nickel column was used to purify the fusion protein. The self-cleavage of the fusion protein was achieved by adding the NpuN fragment. The released solubilizing tag and unreacted precursor were removed by Nickel column and the cleaved antibody fragment and immunotoxinwere further captured by Capto L. The dissociation constant of the obtained Fv and immunotoxin were determined by Fortebio. In summary, the current method could enhance the solubility of antibody fragment and immunotoxin in E. coli, and could improve the purification process. This method provides a reference for the development of a method for soluble expression and purification of immunotoxin-type pharmaceutical recombinant proteins based on the Escherichia coli expression system.
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@#Naturally split Npu DnaE intein can mediate rapid trans-splicing and C-cleavage, which is of great use in many aspects of protein engineering. However, the degradation of NpuC during expression and purification reduces the yield and purity of recombinant protein. N2C, an extended NpuN2-containing N-terminal NpuC fragment, was constructed to improve NpuC stability. N2C was expressed in BL21(DE3) and purified by affinity chromatography. The degradation ratio was calculated by ImageJ, and the factors affecting the C-terminal cleavage reaction of intein, such as temperature, DTT concentration and N/C ratio, were also investigated. The results showed that N2C lowered the proportion of degradation to 2.7%-7.2% and the yield of C-terminal cleavage reached 90% in 30 min at 37 °C with an N/C ratio of 5∶1 catalyzed by 1 mmol/L DTT. N2C can not only improve the stability of NpuC in Escherichia coli expression system, but also retain the activity of C-terminal cleavage reaction, which is of great significance for its application in protein purification.
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@#Intein is a functional protein that mediates self-cleavage from precursor protein and simultaneously connects the exteins on both sides of the intein via peptide bonds. Among all kinds of inteins, split intein has a wide range of applications in the field of antibody ligation. However, since the naturally split intein specifically recognizes the first three amino acid sequences “cysteine, phenylalanine, and asparagine”(CFN)of the extein, exogenous amino acids are inevitably introduced after the protein splicing reaction. In this study, the amino acid sequence “cysteine, aspartic acid, and lysine”(CDK)of the antibody hinge region was substituted for “CFN” as the recognition site for the intein and the split intein Npu DnaE was mutated into Npu*GEP DnaE. The results showed that the mutant could recognize “CDK” and the intein splicing reaction could successfully take place. The factors affecting the intein splicing reaction platform, such as pH, temperature and the concentration of NaCl and DTT were investigated in this study. The results showed that the splicing reaction of the mutant performed well, which indicated its potential usefulness in bispecific antibody assembly. In conclusion, the problem of introducing foreign amino acids was alleviated, the broadness of intein substrate was further expanded, and further technical support for the application of intein to the antibody assembly was provided.
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Objective To investigate the treatment options and prognostic factors of limited-stage small cell cancer of the esophagus.Methods A retrospective analysis of 58 limited-stage cases admitted to Shandong Cancer Hospital Affiliated to Shandong University from June 2009 to July 2017 was performed.Kaplan-Meier and log-rank methods were used for survival analysis.Cox regression model was used for prognostic factors analysis.Results The median overall survival (OS) of the whole group was 21.3 months (5.3-97.2 months).The 6-month,1-year,2-year,3-year and 5-year survival rates were 93％,84％,44％,28％ and 11％ respectively.Univariate analysis suggested that treatment and stage affected patient survival.The median OS of the chemotherapy,chemotherapy + radiotherapy,surgery + chemotherapy and surgery + chemotherapy + radiotherapy groups were 14.5,18.0,23.8 and 46.5 months respectively,with a significant difference (x2 =11.148,P =0.011).The combination therapy was better than chemotherapy alone (all P ＜ 0.05),but there was no significant difference between the different combinations of treatments (all P ＞ 0.05).The median OS of the stage Ⅱ,Ⅲ,patients were 27.0,17.8 and 9.9 months respectively,with a significant difference (x2 =48.114,P ＜ 0.001).The prognosis of patients with stage Ⅱ and Ⅲ was better than that of patients with stage Ⅳ (both P ＜0.001),but there was no significant difference in OS between stage Ⅱ and stage Ⅲ patients (P ＞0.05).Multivariate analysis found that treatment (HR =0.567,95％ CI:0.387-0.830,P =0.004) and stage (HR =3.009,95 ％ CI:1.811-4.999,P ＜ 0.00 1) were independent prognostic factors for OS.The stratified analysis found no significant difference in the prognosis between the surgical and non-surgical patients (median OS:28.6 and 16.9 months;x2 =3.938,P =0.052).Preoperative neoadjuvant therapy did not improve the prognosis of the patients (17.8 months vs.43.4 months;x2 =0.571,P =0.450).The analysis showed that there was no statistical difference in OS between patients with Ki-67 index ≤ 80％ and ＞ 80％(median OS:16.9 and 24.5 months;x2 =3.341,P =0.068).Conclusion The treatment and stage are independent prognostic factors for patients with limited-stage small cell cancer of the esophagus.The effect of chemotherapy alone is poor for patients with limited-stage small cell cancer of the esophagus.Multimodality therapy can benefit patients.
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Objective To investigate the differences of the working memory tasks' performance between the patients with no neurological syphilis disease and the health controls (HC).Methods 2-back test of N-back task paradigm were applied to measure the performances of working memory.A total of 30 patients with syphilis disease were compared with 35 gender-,age-,years of education-matched healthy persons,then compared the accurate rate and reaction time on the tasks between the two groups.Results Participants with anxiety or depression had to be excluded.The results showed that patients impaired on working memory task.The accurate rate and reaction time of the working memory of the patients was worse than that of the healthy persons ((65.31 ± 8.78) ％ vs (71.00 ±9.64)％ ;(1521.14 ±290.57)ms vs (1296.08 ±253.20)ms respectively) (t=-2.472,P=0.016;t=3.337,P =0.001 respectively).The study indicated the reaction time of the working memory for the patients was negative correlation to the age of final diagnosis (r =-0.362,P =0.049).Conclusion The patients with no neurological syphilis disease show impairment on working memory.This probably because treponema pallidum injures the central nervous system.
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Objective To observing the effects of health education on modified lifestyle and behavior patterns among migrant workers. Methods Individuals from two factories were randomly assigned to the intervention group ( n = 262 ; men 129, women 133 ; mean age 31.8) and the control group ( n = 147 ;men 75, women 72; mean age 31.8). The intervention group received 4-months' health education before changes of awareness of health knowledge, cigarette smoking, alcohol consumption, physical activities,mental health and medical consultation. Results In the intervention group, the awareness of the risk of cigarette smoking and alcohol consumption and normal levels of blood pressure and obesity was significantly improved at 4 months(χ2 =59.65, 47.69, 19.50, and 30. 17; all P＜0. 01 ). However, no significant improvement in awareness of AIDS was found in the intervention group. Of the intervention group, some unfavorable lifestyles and behavior patterns were modified ( cigarette smoking χ2 = 4.50 ; altitude toward clinic visit χ2 = 7.09 ; both P＜0. 05 ). Conclusion Health education could help to improve the awareness of health knowledge and modify lifestyle and behavior patterns of migrant workers.
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Aim: To clone and over-express the gene encoding aminoglycoside(AG)phosphotransferase(APH)from clinical MRSA isolates in E.coli and to develop an assay method for the recombinant APH.Methods: The susceptibility of clinical MRSA isolates to AGs was tested by disk diffusion.A nucleic acid sequence encoding APH was amplified from the genomic DNA of an isolate and ligated to expression vector pET-28a,and then trans-formed into E.coli BL21(DE3).After purification of the recombinant protein by affinity chromatography,the phosphorylation activity of the enzyme was determined by ESI-MS and disk diffusion.Flesults: All 6 clinical MRSA isolates were unsusceptible to AGs.After cloning and expression,the recombinant APH was purified to90%.The in vitro activity assay indicated that the recombinant protein could inactivate kanamycin B in the assay mixture within 2 h.Conclusion: The recombinant APH showed excellent enzymatic activity.The assay method was simple and convenient,which may provide the basis of developing a screening model for APH inhibitors.
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[Objective]To assess the effect of a new invented spondylolisthesis reduction system:elevating-pull reduction system for surgical treatment of lumbar spinal spondylolisthesis associated with isthmic spondylolysis.[Method]Sixty-two patients with lumbar spinal spondylolisthesis were treated with elevating-pull reduction system combined with SRS(spondylolisthesis reduction system),after surgery the patients were evaluated for symptom improvement,intervertebral bone graft fusion,and internal fixation.[Result]The patients were followed up from 1 to 6 years(average 2 years and 5 months) in 62 patients.According to the criterion,excellent results were in 43 patients,good in 16,moderate in 3.No patient showed worse.No internal fixation loosening and breakage were found.[Conclusion]The new invented spondylolisthesis elevating-pull reduction system combined with SRS are more effective and reliable,especially for those osteoporosis patients and serious cases,the reduction and fixation are more effectively.In this procedure the pedicle entrance mouth can be seen straightly and the pedicle screw is inserted more easily.It can prevent the nerve injury,save the operation time,reduce the blood lossing,and excellent outcome can be obtained.
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[Objective]To observe the clinical therapeutic effect of Jing Kang Yin decoction on nerve-root cervical spondylosis.[Method]152 patients with Cervical Spondylosis neuiodynia were selected and divided into two groups;treatment group with Jing kang Yin decoction(87 cases) and control group with Jingfukang(65 cases).Pain degree(VAS methods),hemorheological indexes,serum cytokine IL-10 and adhesion molecule were observed before and after treatment and the clinical therapeutic effect of two groups were compared.[Result]Pain scores of two groups were lower after treatment than treatment before(P
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Objective To investigate the role of activated T lymphocytes in the pathogenesis and effect of CD95 and bcl 2 expression on the development of alopecia areata. Methods The lymphocytes were isolated from the peripheral blood of the patients. Then CD4 +, CD8 +,and/or CD95 and bcl 2 positive lymphocytes were sorted out by flow cytometry . Results In the active stage of the disease, CD4 + lymphocytes decreased while CD8 + lymphocytes increased significantly. At the same time, the expression of bcl 2 was much more frequently detected in CD8 + lymphocytes than in CD4 +lymphocytes. CD95 expression was also increased markedly in T lymphocytes. Conclusions Down regulation of bcl 2 leads to the decrease in number of CD4 + lymphocytes. Meanwhile, CD8 + lymphocytes increased duo to the over expression of bcl 2. As a result , the ratio of CD4 + to CD8 +lymphocytes has dropped markedly and the cytotoxic effect of CD8 +lymphocytes on the hair follicles is enhanced. On the other hand, over expression of CD95 in T lymphocytes triggers cell apoptosis in the hair follicles. Both effects promote the development of alopecia areata . [