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Objective To analyze the frequency of interleukin (IL)-22+CD161+CD4+ T cells in the peripheral blood mononuclear cells (PBMCs) in rheumatoid arthritis (RA) patients compared with healthy control subjects and investigate the relationship of IL-22+CD4+CD161+ T lymphocyte frequency changes with RA disease activity.In addition to explore the pathogenesis of RA,and to look for new treatment targets for RA.Methods Twenty-one RA cases were included in the Department of Rheumatology of Tangshan Gongren Hospital from 2017 to 2018.Fourteen patients were female and 7 were male with the age ranged from 36 to 74 years old.The average age of this group of patients was (55±10) years,the average disease course was (60±50) months.All patients fulfilled the classification criteria of American College of Rheumatology [American College of Rheumatology (ACR)].Twenty-one subjects were enrolled as the control group,all of them came to Tangshan Gongren Hospital for regular health check-up.Fifteen subjects in the control group were female and 6 were male.Their age ranged between 40-78 years old with the average age of (55±9) years.IL-22+CD4+CD161+ T cells in PBMCs were detected by flow cytometry.The frequency variation of different CD4+CD161 + T was compared between case and control groups.The correlation was studied between the frequency and RA disease activity score (DAS28),tender joints number,swollen joints number,red blood cell sedimentation rate,high sensitive C reactive protein and white blood cell counts,red blood cell counts,platelet counts,IgG,IgA,IgM,complement C3 level,complement C4 level.T-test or Mann-Whitney U test were used for single-factor analysis,Pearson's test was used for correlation analysis.Results The percentage of RA group secreted CD4+ T cells (0.33± 0.20)％ of INF-γand IL-22,CD4+ T cells (0.51±0.29)％ of IL-22,and CD4+CD161+ T cells of IL-22 simultaneously.The number (0.55 ±0.28)％ was.significantly higher than that of the healtby control group [(0.22±0.14)％,(0.25±0.18)％,(0.36±0.24)％],and the differences were statistically significant [P=0.002,P=-0.0.45,P=0.026].Conclusion The percentage of IL-22+CD4+CD161+ T lymphocytes in the peripheral blood monocytes in RA patients is significantly higher than that in the healthy controls.The results of this study suggest that IL-22+CD4+CD161+ T lymphocytes in RA patients maybe related to RA disease activity and joint lesions.
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Objective@#To compare the predictive validity of writhing movements assessments of neurological outcome between high-risk preterm and full-term infants.@*Methods@#High-risk preterm and full-term infants who accepted the general movements(GMs) assessments from July 2013 to April 2014 and took part in follow-up regularly for 1 year in the Newborn Pediatrics Department of the Fifth Central Hospital of Tianjin were confirmed as the participants.GMs recordings during writhing movements period (at least once) and fidgety movements period (at least once) were collected and assessed.Neurological outcome was confirmed by Peabody Developmental Motor Scale-2(PDMS-2) when the patients were 1 year old.Then the predictive validities of writhing movements assessments of neurological outcome between high-risk preterm and full-term infants were calculated and compared.@*Results@#There was no significant difference in the detection rate of writhing movements between preterm and full-term infant groups(χ2=1.592, P=0.207). There was no significant difference in the detection rate of fidgety movements between preterm and full-term infant groups(χ2=1.605, P=0.205). The sensitivity was 92.9%, the specificity was 90.0%, and the negative predictive value was 97.8% in the stage of writhing movement to the motor development outcome in preterm infant group; the sensitivity was 85.7%, the specificity was 94.0%, and the negative predictive value was 95.9% in the stage of fidgety movement to the motor development outcome in preterm infant group; there was a good consistency between the assessment of writhing movement and neurological outcome confirmed by PDMS-2(Kappa=0.703, P<0.01). The specificity was 71.0%, the positive predictive value was 55.6% in the stage of writhing movement to the motor development outcome in full-term infant group; there was a worse consistency between the assessment of writhing movement and neurological outcome confirmed by PDMS-2(Kappa=0.555, P<0.01). Both the sensitivity and the specificity were 75.0% in the stage of writhing movement to the cerebral palsy in preterm infant group; there was a poor consistency between the assessment of writhing movement and neurological outcome confirmed by PDMS-2(Kappa=0.311, P<0.05). The specificity was 85.4%, the positive predictive value was 22.2% in the stage of writhing movement to the cerebral palsy in full-term infant group; there was still a poor consistency between the assessment of writhing movement and neurological outcome confirmed by PDMS-2(Kappa=0.319, P<0.05). Both the sensitivity and the negative predictive value were 100.0% in the stage of fidgety movement to the cerebral palsy in both preterm and full-term infant groups.@*Conclusions@#The predictive validity of writhing movements assessments to the motor development outcome in preterm infant group is higher than in full-term infant group, and it can be used as a tool for early and accurate prediction of neural development outcome of brain injured premature infants.The predictive validity of writhing movements assessments of the cerebral palsy is poor.Both the sensitivity and the negative predictive value were high in the stage of fidgety movement to the cerebral palsy in both preterm and full-term infant groups, and it may be used to predict the cerebral palsy earlier.
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Objective@#To investigate the association between expression of phosphoglycerate kinase 1 (PGK1) in liver cancer tissue and prognosis, as well as its influence on metastasis and invasion of hepatocellular carcinoma (HCC) cells.@*Methods@#Overexpression and downregulated expression of PGK1 in HCC cells were mediated by lentivirus to establish hepatoma cell lines with different expression levels of PGK1. The Transwell chamber invasion assay, wound healing assay, and colony-forming assay were used to investigate the influence of PGK1 on metastasis, invasion, and proliferation of HCC cells. Immunohistochemistry was used to measure the expression of PGK1 in liver cancer tissue samples from 116 patients with hepatocellular carcinoma who underwent radical surgery, and the Kaplan-Meier method and the log-rank test were used to determine the association between PGK1 expression and prognosis of patients with liver cancer.@*Results@#HCCLM3 and MHCC97H HCC cells with high metastatic potential had significantly higher expression of PGK1 than Hep3B and Huh7 HCC cells with low metastatic potential. Downregulation of PGK1 expression significantly inhibited the migration (31.2% ± 2.4% vs 12.0% ± 1.3%, t = 21.57, P < 0.01), invasion (58 ± 11 vs 21 ± 8, t = 4.687, P < 0.05), and colony-forming ability (168.6 ± 15.1 vs 118.4 ± 8.1, t = 6.650, P < 0.05) of MHCC97H cells, while overexpression of PGK1 enhanced the migration (62.8% ± 4.4% vs 83.6% ± 6.1%, t = 20.56, P < 0.01), invasion (80 ± 12 vs 121 ± 15, t = 4.603, P < 0.05), and colony-forming ability (52.3 ± 8.6 vs 84.7 ± 9.0, t = 27.18, P < 0.01) of Hep3B cells. The high PGK1 expression group had significantly shorter median disease-free survival time and mean survival time than the low PGK1 expression group (22.00 ± 8.51 vs not reached, P < 0.05; 46.00 ± 16.87 vs not reached, P < 0.01).@*Conclusions@#PGK1 is involved in the regulation of metastasis and invasion of HCC cells and can promote the migration and invasion of HCC cells. Therefore, PGK1 may be an important predictor of prognosis and postoperative recurrence in patients with liver cancer.
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Objective To investigate the value of tissue motion mitral annular displacement (TMAD) in assessment of left ventricular systolic synchrony in patients with dilated cardiomyopathy (DCM).Methods Thirty-eight DCM patients and 30 healthy subjects were enrolled in this study.The left ventricular end diastolic volume (LVEDV),left ventricular end systolic volume (LVESV),and left ventricular ejection fraction (LVEF) in two groups were investigated using conventional echocardiographyexamination.The time of systolic mitral annular peak displacement (TP) in two groups were measured by the technique of TMAD at six sites (anteroseptal,posteroseptal,anterior,lateral,posterior and inferior).The standard deviation of TP (TP-SD) and the maximal difference of TP (TP-DIF) were calculated.And the correlation between TP-SD and LVEF as well as TP-DIF and LVEF in the DCM group were analyzed.The standard deviation and maximal difference of the time to point with minimal systolic volume of 16 segments (Tmsv16-SD,Tmsv16-DIF) of the DCM group were measured by the technique of real-time three-dimensional echocardiography (RT-3D).And the correlation between Tmsv-16SD and TP-SD as well as Tmsv16-DIF and TP-DIF were analyzed.Results Compared with the healthy subjects,LVESV and LVEDV increased (t=14.023,16.643,P ＜ 0.01) and the LVEF decreased significantly (t=26.112,P ＜ 0.01) in the DCM group.TP,TP-SD and TP-DIF in the DCM group obviously increased (t=7.628,6.869,7.507,6.616,5.631,4.738,7.368,5.777,6.781,P ＜ 0.01).TP-SD and TP-DIF were negatively correlated with LVEF (r=-0.645,-0.513,P ＜ 0.05),while TP-SD and Tmsv1 6-SD,TP-Dif and Tmsv1 6-Dif were positively correlated(r=0.643,P ＜ 0.01;r=0.563,P ＜ 0.05).Conclusions In this study,the TMAD technique was used to measure the time of peak displacement of mitral annulus to evaluate the synchrony of left ventricle movement.It is expected to be a new method in evaluation of the synchrony of left ventricle,which is simple,feasible approach without angle dependence.
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Objective To investigate the effects of alprostadil injection on IL-6, IL-8, TNF-αand insulin resistance in diabetic patients.Methods 90 patients with diabetes mellitus in our hospital from January 2014 to May 2016 were selected,and randomly divided into observation group and control group,45 cases in each groups.The control group received conventional western medicine , the observation group was treated with alprostadil injection.The levels of serum IL-6, IL-8, TNF-α, insulin and insulin resistance index were compared before and after treatment in both two groups. Results After treatment,the total effective rate of treatment group was 91.11%,which was significantly higher than the control group 71.11%( P<0.05).Before treatment,the levels of IL-6,IL-8,TNF-α,insulin and insulin resistance index of two groups were had no statistical significance,after treatment,the levels of IL-6,IL-8,TNF-α,insulin and insulin resistance index were decreased in both groups,and the observation group was significantly lower than the control group,the difference was statistically significant (P<0.05).Conclusion The alprostadil injection can significantly reduce the levels of IL-6,IL-8,TNF-α,and control the level of blood glucose,reduce the effect of insulin resistance.
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<p><b>OBJECTIVE</b>To observe the effect of sodium bicarbonate combined with ulinastatin on cholinesterase activity for patients with acute phoxim pesticide poisoning.</p><p><b>METHODS</b>A total of 67 eligible patients with acute phoxim pesticide poisoning, Who were admitted to the emeryency department of hospital from March 2011 to February 2014, Acording to different treatments au patients were randomly divided into the conventional treatment group (n=34) and the sodium bicarbonate+ulinastatin group (n=35) . The conventional treatment group were given thorough gastric lavage with water, the sodium bicarbonate + ulinastatin group were given gastric lavage with 2% sodium bicarbonate solution. Both groups were given such treatments as catharsis, administration of oxygen, fluid infusion, diuresis, and antidotes such as atropine and pralidoxime methylchloride. On the basis of comprehensive treatment, people in the sodium bicarbonate+ulinastatin group were given 5% sodium bicarbonate injection and ulinastatin. The clinical effect of the two groups were compared.</p><p><b>RESULTS</b>The serum cholinesterase activity of the sodium bicarbonate+ulinastatin group was significantly higher than the conventional treatment group from the 5th day, and the difference was statistically significant (P<0.05) . The total atropine dosage, total pralidoxime methylchloride dosage and hospitalization days were better than the conventional treatment group, and the differences were statistically significant (P<0.05) . The difference in the time of atropinization between the two groups was not statistically significant (P>0.05) . The results of arterial blood pH, HCO3- of the sodium bicarbonate + ulinastatin group were higher than the conventional treatment group, and the difference of HCO3- at the 10th day was statistically significant (P<0.05) .</p><p><b>CONCLUSIONS</b>Sodium bicarbonate combined with ulinastatin can improve the therapeutic effect and reduce complications in the treatment of acute phoxim pesticide poisoning, and have beneficial effects on the recovery of cholinesterase activity.</p>
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Humans , Atropine , Therapeutic Uses , Cholinesterases , Metabolism , Glycoproteins , Therapeutic Uses , Organophosphate Poisoning , Drug Therapy , Organothiophosphorus Compounds , Poisoning , Pesticides , Poisoning , Pralidoxime Compounds , Therapeutic Uses , Sodium Bicarbonate , Therapeutic UsesABSTRACT
Objective To study the role of autophagy in the replication process of Toxoplasma proliferation. Meth-ods As the experimental groups, these cells were infected by Toxoplasma gondii tachyzoites at given MOI (2 ∶ 1, 4 ∶ 1 ,8 ∶ 1 ,16 ∶ 1 ) . Host cell autophagy was detected through acridine orange staining and MDC fluorescence stai-ning at different time points (1, 2, 4, 8, 24, 48, 96 hs post infection). Detect the condition of HEF cells autoph-agy with acridine orange fluorescence staining and MDC fluorescence staining, and detect the replication kinetics of Toxoplasma gondii infection at different time points using Giemsa staining. Results The results of acridine orange and MDC fluorescence staining showed that autophagy inhibitors and inducers could inhibit and promote the autoph-agy of HEF cells respectively. From the results of Giemsa staining, it was found that the proliferation of Toxoplasma gondii in HEF cells could be promoted with autophagy inducers and be inhibited with autophagy inhibitors. Conclu-sion The regulation on autophagy of host cell could regulate the proliferation and replication of Toxoplasma gondii.
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This study was aimed to reveal the roles of biological factors T3, T4 and TSH in the pathogenesis of liver-qi invasion syndrome and liver-qi depression syndrome of premenstrual syndrome (PMS) in the body. Thus, we may expound the connotation of conceptpotential stagnation ofqi and blood. Female workers of Jinan and Qingdao were selected as target groups by epidemiological cross-sectional survey. And specially trained investigators were responsible for screening patients referred to criterion on the international diagnostic standards and syndrome diagnosis of PMS. Patients' blood was taken as samples during the follicular phase (6 to 14 days), luteal phase (15 to 23 days), premenstrual phase (24 to 28 days) and the menstrual phase (1 to 5 days), respectively. Blood samples were disposed according to requirements of radioimmunoassay. Single factor analysis of variance was used in the comparison with the level of T3, T4 and TSH in menstrual blood for different time intervals of the case group and the normal group by SPSS 10.0 statistical software. The results showed that T3 levels of patients with either syndrome were significantly reduced during the premenstrual phase and the menstrual phase (P < 0.05). There were no significant changes on the level of serum T4 and TSH before and after the occurrence of the disease. Both of them were significantly increased during each phase of the menstrual cycle. It was concluded that changes of T3, T4 and TSH were one of the mechanisms in disease with liver-qi invasion syndrome and liver-qi depression syndrome. The micro change was the essence ofpotential stagnation of qi and blood, which was the body condition of this disease.
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This study was aimed to explore brain regions which were closely related to the disease onset of premenstrual syndrome (PMS) with liver-qi depression. The BOLD-functional magnetic resonance imaging (fMRI) was used in the study. The processing of imaging data was based on the SPM 8 software and the REST software of the matlab platform. Each cluster was more than 389 continuous voxel. The brain region with single voxel of P < 0.05 (corrected) was defined as region with statistical significance. The 2 Sample T-Test was applied in the case group and the control group. The results showed that compared with the normal control group, the frontal lobe, occipital lobe, insula, limbic lobe, basal nuclei, and cingulate gyrus were activated in the PMS with liver-qi depression cases. It was concluded that the disease onset of PMS with liver-qi depression cases was related to brain regions such as frontal lobe, occipital lobe, insula, limbic lobe, basal nuclei, and cingulate gyrus.
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Objective The purpose of this study is to study the expression of miR -221 in cervical cancer tissues and its relationship with HPV infection .Methods HR-HPV infection was detected by HC2,and 30 cases of HR-HPV negative and 5 cases of HR-HPV positive cervical cancer tissues were collected .Mean-while,30 cases of normal cervical tissues in patients with benign disease were collected as control group .The ex-pression of miR -221was detected by RT -PCR,preliminarily investigating the relationship between miR -221 expression and the occurrence of cervical cancer and HPV infection .Through transfection of miR-221 and anti-miR-221 into HPV16-positive cervical carcinoma cell line Caski and HPV 16-negative cervical carcinoma cell line C33a,we observed the role of miR -221 on the migration and invasion of Caski cells and C 33a cells.Results Compared with normal cervical tissues , the expression of miR -221 in cervical cancer was significantly in-creased,the difference was statistically significant( P<0.01);and the expression of miR-221 was closely correl-ative to the patients with or without lymph node metastasis ,pathological grade and clinical stage ( P<0 .01 );the expression of miR-221 in HR-HPV positive cervical cancer tissues was higher than in HR -HPV negative cer-vical cancer tissues(P<0.01);transfection of miR -221 and anti-miR-221 could promote or downregulate C33 a and Caski cells migration and invasion ,and the changes between two groups had statistical significance ( P<0.05).Conclusion The increased expression of miR -221 in cervical cancer tissues is closely related to the oc-currence and development of cervical cancer and HPV infection .
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Around the core issue how to evaluate and test hypothesis, starting from the concept of hypothesis and scientific hypothesis, learning form recent progress and understanding at home and abroad, we have analyzed the type of scientific hypothesis, test methods and pre-mortem and post-mortem evaluation criteria in depth, established scientific vision and discipline ruler, explored the awareness level of the Chinese medicine hypothesis, pointed out the problems and gaps. Whereby, we have analyzed and demonstrated brain regulating five internal organs concept of wholism scientific hypothesis, and proven it belongs to the scientific hypothesis. We were confident that these works mentioned above would lead the theoretical reform changing Chinese medicine hypothesis from traditional description to the nature clarification.
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Start to explore the theory conditions around the theme why to build the modern basic theory of tradi-tional chinese medicine and what kind of theory we want to establish . On the basis of making clear the related concepts such as theory, knowledge, scientific theory and scientific knowledge, discuss the theoretical development pathways and evaluation criteria, compare the development mode of Chinese and Western medicine, providing a refer-ence for evaluation and future development of the modern basic theory of traditional chinese medicine. According to Carl G. Hempel's bridge principle, summarize the five bridge principles and a logic protection principle of the research and foundation of the modern basic theory of traditional chinese medicine, and explain its role and signifi-cance. Put forward a theoretical framework of the modern basic theory of traditional chinese medicine Three levels and five grades like a tree structure, demonstrate the scientific connotation of 11 principles and concepts under the frame. Analyze and demonstrate its scientific rationality according to the role and evaluation criteria of scientific theory mentioned above, clearing direction for future development.
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Objective To investigate the effect of early rehabilitation treatment on functional improvement after total knee arthroplasty. Methods 48 patients (53knees) of rehabilitation in our hospital from 2010 to 2013 were divided into 2 groups, 22 patients (25knees) in control group were given common rehabilitation training, 26 patients (28 knees) in observation group were given early rehabilitation treatment. Results All the ROM, knee score and function score of 2 groups were improved, and observation group were better than that of control group (P<0.05),at the last follow-up, the PF, BP and VT of observation group were better than control group (P<0.05) . Conclusion Early rehabilitation treatment can significantly promote the knee function recovery after surgery and improve the life quality of patients, so it deserves to be generalized.
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To evaluate the reagent 2-methoxy-4,5-dihydro-1H-imidazole used for isotopic labeling in quantitative proteomics, we synthesized 2-methoxy-4,5-dihydro-1H-imidazole and its tetradeuterated analog in three steps. Prior to tryptic cleavage, bovine serum albumin (BSA) was reduced and alkylated. Tryptic peptides were derivatized with an equal volume of either DO or D4 and D4-derivatized peptides were mixed with at variable ratio (from 10:1 to 1:5) prior to MS and MS/MS analysis. We used matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS) and Electro spray ionization-mass spectrometry (ESI-MS) to evaluate the quantitative capability of labeling. The specificity of the reagent is excellent: only lysine side chains were modified among tryptic peptides. MALDI and ESI ionization modes not only could achieve the quantification of differentially expressed proteins but also facilitate the de novo sequencing. This side-chain modification can be used for quantitative analysis with proteomic strategies involving liquid chromatography. Reverse phase liquid chromatography (RPLC) kept a good resolution, and the introduction of D atoms did not introduce a variation of retention time between heavy and light peptides in RPLC.
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Imidazoles , Chemistry , Isotope Labeling , Methods , Lysine , Chemistry , Peptides , Chemistry , Proteomics , Methods , Serum Albumin, Bovine , Chemistry , Spectrometry, Mass, Electrospray Ionization , Methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , MethodsABSTRACT
To study the relevant correlation of anger-out (expression of anger toward other persons or objects in the environment) or anger-in (holding in or suppression of angry feelings) with life events, so as to gain a better understanding of present-day social influences and assess which life events trigger anger. This study proposes the new hypothesis that "life events constitute originating factors of anger-triggered emotional action".
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Objective To prepare the infected Oncomelania hupensis by artificial method for the research on the activity, vaccine, and genetic variation of Schistosoma Japonicum (S. Japonicum).Methods The mature eggs of S. Japonicum were collected by Nylon silk method and the miracidia were incubated under appropriate conditions. Negative snails were infected with miracidia in different proportion by means of individual or collective infection to seek the best method and proportion of infection between miracidia and snails. Infected snails were divided into 12 groups in total. Ⅰ-Ⅵ groups were for individual infection and Ⅶ-Ⅻ groups were for collective infection. There were 200 snails in each group. The infection ratios between snails and miracidia in Group Ⅰ-Ⅵ or screened, numbered, and reared singly. The amount of cercariae was calculated once every 10 days until the infected snails died. Then cercariae shedding quantity, infection quantity, and mortality of infected snails in every group were compared to find the best infection method and the best infection proportion between miracidia and snails. The cercariae were collected from the first generation of infected snails and were used to infect experimental animals. The mature eggs of S. Japonicum were saved from the infected experimental animals and incubated to get miracidia. The snails were artificially infected by miracidium to get the second generation of infected snails. The developmental rates of adult worms, the egg density in fecal and liver were compared between artificially and naturally infected snails. Results In individual infection GroupⅠ-Ⅵ,the average infection value of snails were 0±0,22.7±4.2,31.7±4.5,53.0±5.3,39.3±5.9,32.7±4.7,the average fatality of snails were 21.7±3.1,25.0±3.6,31.3±4.9,44.7±6.5,78.3±9.5,89.7±13.6, and the average value of cercariae shedding from infected snails were 0.0±0.0,308.0±96.6,428.1±146.2,527.0±171.1,571.4±148.9,602.9±356.3, respectively. In collective infection Group Ⅶ-Ⅻ,the average infection value of snails were 0±0,12.3±2.5,18.7±4.7,28.3±4.2,33.3±4.7,29.3±5.5,and the average fatality of snails were 22.7±3.8,23.7±4.5,28.3±5.5,47.0±9.5,75.7±8.5,86.3±12.2, and the average value of cercariae shedding from infected snails were 0±0,244.5±57.3,292.3±74.8,347.1±100.8,477.2±142.1,447.3±161.4, respectively. The second generation of artificially infected snails was obtained successfully. The average infection rate and fatality rate for the second generation of artificially infected snails were 24.65% and 24.50%, both of which were not obviously different from that of the first generation of artificially infected snails (P>0.05). In the animal experiment, the worm growth rate for the naturally infected snails, the first or second generation of artificially infected snails were 68.50%,73.50% or 71.00%. There was no obvious difference among them (P>0.05). The fecal (or liver) eggs per gram for the naturally infected snails, the first or the second generation of artificially infected snails were 1 503±269,1 683±233, or 1 541±117 (or 6 641±1 819,6 272±1 419, or 7 263±1 643). There was no significant difference among the 3 groups (P>0.05). Conclusion Infected snails can be obtained through the artificial method by using S. Japonicum miracidia to infect snails. Individual infection has the advantage over collective infection. The optimal proportion of infection between first and the second generation of artificially infected snails in the average of cercariae shedding, infection, and fatality average of snails. There was no significant difference between artificially and naturally infected snails in the developmental rate of adult worms, fecal and liver eggs per gram.
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To determine the possibility whether DNA vaccines pcDNA3/SjSDISP of Schistosoma japonicum to induce autoimmunity and immune tolerance in the vaccinated mice, the titer of the specific antibodies against SjSDISP and the production of the autoimmune antibodies, such as presence of anti-nuclear and anti-dsDNA antibodies in the vaccinated mice were detected by ELISA assay and the toxicity of the plasmid DNA was studied through the observation of the change in body weight and the pathological examination of the major organs in mice. It was found that the titer of the specific antibody against SjSDISP was 1∶400 as determined by ELISA, but no autoimmune antibody could detected. The difference of the body weight of mice between the experimental and the control groups was not significant. No abnormal results of histopathological examination were obtained in both groups. From these observations, it is clear that there is no evidence to show that DNA vaccine pcDNA3/SjSDISP can induce auto-immunity and immune tolerance in mice.
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Objective To clone, express Tp0319 gene from Treponemapallidum (T. pallidum), and to assess the immunocompetence of recombinant protein. Methods The immuno-dominant region of Tp0319gene was chosen by computer analysis, amplified from T. pallidum complete genome by PCR, subcloned into the expression vector pQE32 to construct a recombinant plasmid, pQE32/Tp0319, which was then expressed in E. coli M15. The recombinant protein was purified with Ni-NTA affinity chromatography, and identified by using sodium dodecyl sulfate polyacrylamide gel electropheresis (SDS-PAGE) and Western blot. New Zealand rabbits were immunized with the recombinant protein, and the titer of anti-Tp0319 antibodies in sera from immunized rabbits were measured with indirect ELISA. Also, indirect ELISA with the recombinant Tp0319 as coating antigen was performed to detect the anti-Tp0319 antibody in sera from 200 normal human controls and 200 patients with syphilis. Results The prokaryotic expression vector pQE32/Tp0319 was constructed successfully, and the recombinant protein Tp0319 with a molecular weight of about 30 000 was attained. Specific humoral response was elicited by the recombinant protein in New Zealand rabbits and the specific antibody titer was more than 1: 10 240 after immunization for 3 times. Western blot proved that the recombinant protein could specifically react with anti-T. pallidum IgG antibody-positive sera. Indirect ELISA was successfully developed with the recombinant Tp0319, and detected antibodies to T. pallidum in control sera with a sensitivity and specificity of 100% (40/40), respectively. Compared with T. pallidum particle agglutination (TPPA) assay, the sensitivity and specificity of the indirect ELISA were 92.6% and 100%, respectively, in the detection of T. pallidum in sera from patients and controls, and the concordance between the indirect ELISA and TPPA was 96%. Conclusions The prepared recombinant protein shows a satisfactory immunocompetence, which may lay a foundation for its further application in the serodiagnosis of syphilis.
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The aim of this research was to study the immunoprotections of Schistosoma japonicum (S. japonicum) DNA vaccines SjRPS4 and SjRPL7 in mice. Fourty Kunming mice were randomly divided into 4 groups (A, B, C and D), and the pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 plasmid DNA vaccines were prepared for experiment. Mice in group A were intramuscularly injected with 100μL normal saline, whereas mice in group B were injected with 100 (g naked plasmid pcDNA3.0 into the quadriceps. Mice in groups C and D were injected with 100μg/100μL eukaryotic recombinant plasmids pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 into the hind leg muscles respectively. The initial injections were followed by two sets of boosters at 2 weeks intervals. In addition, levels of the specific antibodies were detected 2 weeks after the last immunization and all mice were percutaneously infected with 20( 1) S. japonicum cercariae on abdomen. Fourty-two days after the infection, all mice were killed to detect the worm reduction rate and the egg reduction rate. Significant differences of worm burden reduction rates, LEPG reduction rates, IEPG reduction rates and intrauterine eggs reduction rates were observed in both test group (group C and D), comparing with the control groups (group A and B). Results indicated that the DNA vaccines of pcDNA3.0/SjRPS4 and pcDNA3.0/SjRPL7 could induce strong protective immunity against S. japonicum in mice.
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OBJECTIVE@#To develop a rapid and simple immunoassay to detect antibodies in the sera of patients infect Schistosoma japonicum (S. japonicum).@*METHODS@#Soluble egg antigen (SEA) of S. japonicum conjugated with colloidal carbon in advance was used to react with the antibodies in the sera of patients with schistosomiasis. Then the carbon-antigen-antibody complex would be captured by SEA which had been absorbed on the nitrocellulose membrane and a gray band was shown.@*RESULTS@#A total of 137 sera samples from S. japonicum epidemic area were tested, and the consistency, sensitivity, and specificity of colloidal carbon dipstick assay were 98.54%, 98.99%, and 97.37%, respectively, compared with the IHA method. The gray scale of bands on the dipstick was curvilinear to serum titer which revealed that the assay could be used semi-quantitatively in serum analysis.@*CONCLUSION@#Colloidal carbon dipstick assay is not only rapid and simple, but also sensitive and specific for the detection of serum antibodies of schistosomiasis japonica. It will be a practical immunological assay for the diagnosis of schistosomiasis in the field testing.