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Objective:To explore the collaborative development of drug clinical trial institutions and Contract Research Organizations from the perspective of " government-application-industry-academia-research" , and facilitate faster and better conducting of clinical trials.Methods:Based the combination of literature review and the working practice in drug clinical trial management, problems existed during the implementation of clinical trials were summarized, and then the collaborative development of drug clinical trial institutions and Contract Research Organizations were discussed from the perspective of " government-application-industry-academia-research" partnership.Results:Problems identified during the implementation of clinical trials including uneven capacity of CROs, lack of effective supervision department and insufficient cooperation with clinical trial institutions, which resulted difficulties in sharing clinical trial resources and also negatively impacted the quality of clinical trials. Some proposals were offered in this article, including making good use of the " visible hand" of the government to strengthen the supervision of CROs, accelerating the construction of innovation alliance between clinical trial institutions and CROs, establishing the incentive mechanism of collaborative development and the talent team construction, strengthening the personnel professional training.Conclusions:The application of " government-application-industry-academia-research" model in clinical trials would promote the collaboration between drug clinical trial institutions and Contract Research Organizations, which play important roles in the development of clinical trials.
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Objective:To strengthen the quality management of drug clinical trials to ensure that the clinical trial data obtained is true, accurate, complete and standardized.Methods:By review literatures and the data published in NMPA and survey the problems in the drug clinical trials in some hospitals, etc, we summarized the common problems of drug clinical trials in China.At the same time, using PDCA cycle theory, we sorted out the problems in the drug clinical trials in Taizhou People's Hospital, conducted root-cause analysis of the problems, put forward suggestions and measures on how to strengthen the quality management of drug clinical trials.Results:The incidence of quality problems in clinical trials was significantly reduced from 105.6% to 37.4% by formulating relevant incentive policies, strengthening training, strengthening the supervision function of institutions to researchers, and improving the awareness of GCP of researchers.Conclusions:The implementation of PDCA cycle quality management model can effectively improve the quality of drug clinical trials.
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Objective:To investigate the ultrasonographic characteristics and risk factors of breast cancer internal mammary lymph node (IMLN) metastasis.Methods:A retrospective analysis of 296 first diagnosed breast cancer patients in the Fourth Hospital of Hebei Medical University from March 2010 to May 2020. IMLN was divided into metastatic group (236 cases) and non-metastatic group (60 cases) based on pathology. Chi-square test and independent sample t test were used to analyze the ultrasound characteristics of IMLN metastasis and factors related to metastasis. ROC curve analysis of IMLNs were plotted to obtain the diagnostic thresholds and their sensitivity and specificity.Univariate and multivariate Logistic analysis was used to analyze the risk factors of IMLN metastasis. Results:①The appearances of IMLN in ultrasound were normal type, thickened-cortex type, unclear hilus structure type and thickened-nodular soft tissue type. ②In the two groups, the differences in IMLN long diameter, thickness diameter, number, and lymphatic hilum structure type were statistically significant (all P<0.05), and there were no significant differences in IMLN long diameter/thickness diameter and IMLN blood supply (all P>0.05). ③The long diameter threshold of IMLN for diagnosis of metastasis was 10.5 mm, the are under the ROC curve(AUC) was 0.825, with sensitivity of 58.5% and specificity 93.3%; thickness and diameter threshold was 4.5 mm, AUC was 0.790, with sensitivity 66.9% and specificity 75.0%. The sensitivity and specificity of the diagnosis of long-diameter combined structure type were 56.3% and 93.3%, respectively; the sensitivity and specificity of the diagnosis of thick-diameter combined structure type were 64.8% and 81.7%, respectively. The cortical thickness threshold was 1.9 mm, and the diagnostic sensitivity and specificity were 91.9% and 86.7%, respectively. ④The risk factors of IMLN metastasis inculded univariate analysis showed tumor length, tumor volume, axillary lymph node long diameter, axillary lymph node metastasis, and clavicle lymph node metastasis. There was a statistically significant difference in the pathology of the lower lymph nodes between the two groups ( P<0.05). Multivariate analysis showed that the long diameter of the tumor and the metastasis of the axillary lymph nodes were independent risk factors of IMLN metastasis. Conclusions:The metastatic IMLN mostly manifest as no lymphatic hilum structure or cortical thickening (≥1.9 mm), and multiple IMLN can help diagnose metastasis.Ultrasound can better assess breast cancer IMLN metastasis, and the diagnostic efficiency of IMLN long-diameter combines structure type is higher.Independent risk factors for IMLN metastasis include tumor size and axillary lymph node metastasis.
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Objective:To affirm the importance of clinical trial kick-off meeting and improve the meeting quality.Methods:According to the literature review of latest laws and regulations, combined with working experiences in clinical trial management, this article tried to identify possible problems that might occur before, during and after the kick-off meeting, analyze how to better manage the kick-off meeting, and propose suggestions for possible solutions.Results:Newly approved clinical trial institutions may face challenges such as lack of training, unclear responsibilities, lack of attention to quality control of professional department and post-meeting management. Thus, a series of suggestions were proposed, including more substantial communication and tailored training should be in place before the kick-off meeting, responsibilities for each stakeholder should be clarified during the meeting with specific concern of the professional quality control, in addition, meeting minutes and related materials should be well documented after the meeting.Conclusions:Clinical trial kick-off meeting is crucial that clinical trials institutions should pay more attention and work for better administration of such meetings.
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Clinical residual biological specimens are invaluable for medical research and can be reused for medical research. This paper expounded the possibility, necessity and applied range of the medical research reuse of clinical residual biological specimens and put forward some suggestions on how to standardly supervise clinical residual biological specimens for medical research reuse. The authors raised four aspects of concern: how to strengthen the management of ethical review; how to establish strict privacy protection and information confidentiality system; how to keep samples reasonably to ensure clinical examination; and how to ensure the compliance treatment of residual biological specimens after reuse, so as to promote clinical residual biological specimens more normatively and effectively used in medical research.
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OBJECTIVE: To explore the interference correction reduction method for the determination of cadmium(Cd), chromium(Cr), manganese(Mn), nickel(Ni) and lead(Pb) concentration in workplace air by inductively coupled plasma-atomic emission spectrometry(ICP-AES). METHODS: The interference sources in the determination of metals in workplace air with ICP-AES was analyzed. The results before and after the background correction or the internal standard calibration were compared. Linear regression is performed on the interference element mass concentration by interference error to obtain a table of interference factors. The measurement results of actual samples were corrected using background subtraction, internal standard correction, and interference factor method. RESULTS: The relative deviation of results and theoretical values without background subtraction were 219.5% and 131.5% for Cd and Pb, respectively. The relative deviation between the measured values and theoretical values was less than 10.0% after background subtraction. When the total metal ion concentration was >800.00 mg/L, the relative deviation between measured result and the theoretical values was-21.0%--11.0% without internal standard correction, and the absolute value of them were less than 10.0% after internal standard correction. The interference correction method was used to correct the concentration of Cd, Cr, Mn, Ni, Pb in welding smoke. The mass concentrations of Cd before and after interference correction were 3.90-32.50 and 1.20-7.10 μg/L, respectively. The mass concentrations of nickel before and after interference correction were 111.00-1 220.00 and 99.00-1 120.00 μg/L, respectively. The mass concentration of Cr,Mn and Pb remained unchanged before and after the interference correction. CONCLUSION: The background correction, internal standard correction and interference factor table can eliminate the interference for determination of metals in workplace air by ICP-AES and ensure the accuracy of results.
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OBJECTIVE: To establish a method for detection of 2-methylpropanal,2-methyl butanal and 3-methyl butanal in workplace air by portable gas chromatography-mass spectrometry( GC-MS). METHODS: The workplace air was collected by portable GC-MS sampling probe. After enrichment by Tri-bed tube,the thermal desorption sample was analyzed by retention time and characteristic ion characterization. The characteristic ion peak area was quantified. RESULTS: The linear ranges of 2-methylpropanal,2-methyl butanal and 3-methyl butanal were 0. 079-15. 880,0. 080-16. 080 and 0. 161-32. 120 mg/m~3,respectively. All the correlation coefficients were greater than 0. 999. The detection limits were 0. 012,0. 007 and0. 013 mg/m~3,and the quantitation limits were 0. 041,0. 023 and 0. 044 mg/m~3,respectively. The recovery rates of2-methylpropanal,2-methyl butanal and 3-methyl butanal were 92. 2%-104. 4%. The within-run relative standard deviation( RSD) was 5. 8%-9. 1%. The between-run RSD of different batches samples was 9. 3%-19. 6%. CONCLUSION: This method has high accuracy and is precise,which is suitable for rapid qualitative and quantitative detection of2-methylpropanal,2-methyl butanal and 3-methyl butanal in workplace air.
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OBJECTIVE: To establish a method for simultaneous detection of 16 kinds of gas and particle-phase polycyclic aromatic hydrocarbons(PAHs) in workplace air by high performance liquid chromatography with ultraviolet and fluorescence detector(HPLC-UV-FLD).METHODS: PAHs in workplace air were collected by polytetrafluoroethlene filter with XAD-2 tube. The samples were added with 5 mL of acetonitrile. The samples were extracted by ultrasound and eluted with acetonitrile-water gradient. Ultraviolet with tandem fluorescence detector was used for determination.RESULTS: Eleven kinds of PAHs showed a good linear range of 1.0-400.0 μg/L. Five kinds of PAHs showed a good linear range of 10.0-2 000.0 μg/L and the linear correlation coefficient was greater than 0.999 0. The minimum detection concentration was 0.000 01-0.000 67 mg/m~3 (75 L air sample). The average elution efficiency of filter on granular PAHs except for naphthalene and acenaphthene ranged from 71.0% to 99.0%. The average desorption efficiencies of adsorbent tube was 43.0%-90.0%. The within-run relative standard deviations(RSD) was 0.3%-8.3%, and the between-run RSD was 0.0%-9.2%.CONCLUSION: This method is simple, sensitive, has good linear range and precise. It is suitable for simultaneous detection of PAHs in workplace air.
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Objective To investigate the expression of SATB2 and P53 protein in colorectal cancer and to analyze their relationship with the clinical pathologic characteristics. Methods The expression of SATB2 and P53 protein in 30 normal colorectal tissues and 120 colorectal cancer tissues were detected by immunohistochemical method,then analyzed their correlation. Results The expression of SATB2 in colorectal cancer was lower than that in normal colorectal tissues(P<0.05);and its expression correlated with the tumor size,differentiation,inva-sion,depth,lymph node metastasis and TNM stages(P<0.05),while not with age and gender.The expression of P53 in colorectal cancer was significantly higher than that in normal colorectal tissues(P<0.01),and its expres-sion correlated with differentiation,invasion depth and TNM stages(P<0.05),while not with age,gender,tumor size and lymph node metastasis. The expression of SATB2 and P53 in colorectal cancer have negative correlation (P < 0.01). Conclusion Compared with in normal colorectal tissues,the expression of SATB2 declined and the expression of P53 increased in colorectal cancer,while they have negative correlation. The expression levels of SATB2 and P53 protein have important significance to evaluate the prognosis of colorectal cancer.
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Objective To invesigate the reproducibility of automated breast volume scanner ( ABVS) in detection of breast lesions . Methods Thirty‐five patients with breast lesions diagnosed by conventional hand‐hold ultrasound were involved in the study . Every patient underwent ABVS twice by two different doctors . The number ,location and size of findings were documented and analyzed . Intra‐class correlation coefficients were used to evaluate the reproducibility . Results The number of findings was 67 and 68 respectively ,and the cronbach′s ɑwas 0 .997 . The ɑ of distance from nipple ,distance from skin ,and clock face location were 0 .343 ,0 .883 ,and 0 .949 . The ɑ of length superior‐inferior ,medial‐lateral ,and anterior‐posterior were 0 .938 ,0 .936 ,and 0 .948 . Conclusions The reproducibility of ABVS in detection of breast lesions is perfectly high . ABVS may be a good modality in monitoring benign lesions and evaluating neoadjuvant chemotherapy in the future .
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Objective This research is aimed to investigate the efficacy and toxicity of icotinib for lung adenocarcinoma pa-tients with poor performance status and unknown EGFR gene status .Methods A total of 27 lung adenocarcinom patients with poor Eastern Cooperative Oncology Group-Performance status(ECOG-PS) and unknown EGFR gene status referred to Chongqing Canc-er Institute from August 2012 to August 2014 were analyzed .Icotinib (125 mg) was orally administered three times per day .Asess the efficacy and adverse reaction ,calculate survival rates .Results Among the 27 patients ,the objective response rate(ORR) and disease control rates(DCR) were 29 .6% and 81 .5% ,respectively .The median progression free survival time was 6 .0 months .A to-tal of 70 .4% of patients had an significant improvment in ECOG-PS scores ,following icotinib treatment (Z= - 2 .157 ,P= 0 .031) . Fatigue ,anorexia and diarrhea were the most frequent adverse reaction ,which defined as grade 1 to 2 rashes .Conclusion Lung ade-nocarcinoma patients with poor performance status and unknown EGFR gene status may benefit from icotinib therapy ,and patients were tolerated well .
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Purpose To explore the effect of miR-100 in rat liver development and the occurrence of liver cancer. Methods The liv-er tissues of neonate SD rats, adult SD rats and 2-AAF/PH rats were collected, then real-time fluorescence quantitative polymerase chain reaction ( RT-PCR) and FISH methods were used to detect the expression of miR-100 in the liver tissues. Results The expres-sion level of miR-100 in adult SD rats liver tissues was significantly higher than that in neonate SD rats liver tissues (P<0. 05). Com-pared with the adult rats, the expression level of miR-100 in 2-AAF/PH rats liver tissues was obviously decline (P<0. 05). Conclu-sion miR-100 can as a maker of liver mature, and its deletion maybe related to the tumor formation.
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Objective To investigate the relationship between glutathione S-transferases P1(GSTP1)Ile105Val and glutathione S-transferases M1(GSTM1)single nucleotide polymorphisms(SNP) and the sensitivity to chemotherapy among patients with ad-vanced non-small cell lung cancer(NSCLC) .Methods We used gene sequencing analysis to determine the SNP of GSTP1 Ile105Val and PCR analysis to GSTM1 in DNA from peripheral lymphocytes of NSCLC patients .Totally 89 patients with NSCLC were trea-ted with platinum-based chemotherapy ,and clinical response was evaluated after 2 cycles .The association between GSTP1 Ile105Val and GSTM1 SNP and chemosensitivity were analyzed .Results The overall response rate was 29 .2% .Chemotherapy re-sponse did not show statistically significant differences between the wild genotypes and the variant genotypes for the GSTP1 Ile105Val and GSTM1 gene(P>0 .05) .Conclusion The polymorphisms of GSTP1 Ile105Val and GSTM1 may be not associated with sensitivity to chemotherapy in NSCLC patients .
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Objective To evaluate the application value of virtual touch tissue quantification(VTQ) in the differential diagnosis of small breast lesions.Methods Fifty-six patients with sixty-nine small breast lesions whose largest diameter were less than or equal to 1 cm were evaluated with VTQ.The shear wave velocity (SWV) of the lesions and normal breast tissue were measured to analyze the diversity of different pathological types lesions of SWV and to find the optimal cut-off points using ROC curve to predict small breast cancer.Resuits The mean SWV(Vm) and the ratio (mean of lesions and normal tissue) of SWV (Vm1/Vm2) in benign group was (2.93 ± 1.32)m/s,1.79 ± 0.63,respectively,and that in malignant group was (5.65 ± 2.63) m/s,3.61 ± 1.35,respectively,there were statistically significant differences between benign group and malignant group (t test,P <0.05).The cut-off point of Vm was determined as 4.43 m/ s,the sensitivity,specificity and accuracy of diagnosing breast cancer were 80.5%,89.3%,92.8%,respectively.The cut-off point of Vm1/Vm2 was determined as 2.37,the sensitivity,specificity and accuracy of diagnosing breast cancer were 76.9%,81.2%,89.9%,respectively.Conclusions VTQ technique can provide helpful information in the differential diagnosis of small breast lesions.
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Objective To explore the clinical application value of virtual touch tissue quantification (VTQ) technique in Hashimoto's thyroiditis(HT).Methods Fifty-three patients with HT and 46 health subjects were examed by conventional ultrasonography and VTQ, their shear were velocity (Vs) was measured.A receiver-operating characteristie(ROC) curve was drew according to Vs which can obtain the value of optimal operating point for diagnosing HT.Results The 2D ultrasonic appearance of 53 cases HT were classified into five species, but there was no significant difference in the Vs (F = 0.424, P = 0.790).The area under the ROC curve was 0.976,which showed a high statistical significances(P = 0.000).ROC curve displayed that Vs of 2.53 m/s could be used to diagnose HT,the sensitivity and specificity were 87%and 100% ,respectively.Fifty-three patients with HT were classified into three groups according to thyroid function, 10 patients with hyperthyroidism, 25 patients with hypothyroidism and 18 patients with euthyroidism.The Vs of HT among the three groups were no significant difference(P = 0.884), but their Vs were all obviously higher than that of normal thyroid Vs(P = 0.000).Conclusions VTQ technique can evaluate the hardness of thyroid and has some help for differential diagnosis of HT.
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Objective: To study the apoptosis of T lymphocyte subsets of peripheral blood in systemic lupus erythematosus(SLE)and the correlated pathogenesis.Methods: The percentages of T lymphocyte subsets,the positive expression rates of Fas/FasL on T lymphocyte subsets,and the apoptosis of T lymphocyte subsets were determined by three-colorflow cytometry.The level of serum IL-10 was analyzed by ABCELISA.IL-10 antibody and FasL antibody were added respectively into the 48 hours' culture of 10 SLE patients' PBMCs in vitro,whose serum IL-10 significantly increased, and then, the changes of T lymphocyte subsets of SLE PBMCs were determined by three-colodlow cytometry.Results: 1)In SLE, mainly in hyperactive SLE, the apoptesis of CD4~+ T lymphocytes increased significantly(P<0.05),and the ratio of CD4~+/CD8~+ decreased compared with normal controls,and the abnormal apoptesis of CD4~+ T lymphocytes had a positive correlation with the increased expression rates of Fas/FasL on CD4~+ and CD8~+ T lymphocytes(P<0.05).2)The serum IL-10 level increased significantly in SLE (P<0.01), expecially in hyperactive SLE, and serum IL-10 had a positive correlation with the level of serum anti-dsDNA antibodies, CD4~+/CD8 + ratio, and the positive expression rate of FasL on CD4 * T lymphocytes(P<0.05).3)According to a tracking experiment on two different groups of SLE patients,we found that:with the stabilization of the illness,the level of serum IL-10 decreased significantly, the positive expression rate of Fas/FasL on T lymphocytes,particularly on CD4~+ T lymphocytes, reduced evidently,and the apoptosis of CD4~+ T lymphocytes decreased(P<0.05).Conclusion: The abnormal activities of T lymphocytes, particularly of CD4~+ T lymphocytes from patients with SLE,which lead to AICD,cause the percentage of CD4~+ T lymphocytes and the ratio of CD4~+/CD8~+ to decrease,disorder immune response, and accelerate the process of SLE.As a key factor in modulating the positive expression rate of Fas/FasL on CD4~+ T lymphocytes,IL-10 plays an important role in causing SLE.
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BACKGROUND:Systemic erythematosus lupus (SLE) is closely associated with activity of dendritic cells (DCs) function.DCs derive from hematopoietic precursor cells (HPCs) in vivo and many factors might affect the differentiation and maturation of DCs.The cytokine network was in disturbance and the levels of various cytokines were anomalous in SLE serum.Previous studies mainly addressed effects of single factors.OBJECTIVE:To investigate the effects of joint action of serum interferon (IFN)-α and interieukin (IL)-6 in the serum of SLE patients on the differentiation and maturation of DCs derived from CD34~+ HPCs.DESIGN,TIME AND SETTING:The grouping,controlled and completely random designed study was performed at the Department of Microbiology and Immunology,Nanjing Medical University from May 2006 to October 2008.MATERIALS:Peripheral blood samples were collected from 50 SLE patients from the Department of Rheumatology,First Affiliated Hospital of Nanjing Medical University and 30 healthy volunteers from Nanjing Medical University.Cord blood samples were collected from 15 full-term normal delivery neonates from Nanjing Maternity and Children Health Care Hospital Affiliated to Nanjing Medical University.Informed consent was obtained from all patients and their family members.METHODS:Peripheral blood samples were collected from SLE patients and healthy volunteers.According to serum IFN-α and IL-6 concentrations of normal person,95% reference value range of serum IFN-α and IL-6 concentrations were calculated by the method of percentiles.Above this range represented abnormal increase.Cord blood mononuclear cells were harvested by Ficoll-Hypaque density gradient centrifugation.CD34~+HPCs were purified from cord blood by magnetic cell sorting system (MACS),and cultured to differentiate to DCs.There were 6 groups in this study.Normal serum,SLE serum with elevated levels of IFN-α,SLE serum with elevated levels of IL-6,SLE serum with elevated levels of IFN-α and IL-6,SLE serum with anti-IFN-α and anti-IL-6 neutralizing antibodies,or normal serum with exogenous IFN-α and IL-6 were respectively added to the culture medium in each group.10% volume fraction serum was used in each group,for totally 14 days incubation.MAIN OUTCOME MEASURES:The phenotype of DCs was analyzed by flow cytometry (FCM).Cytokine production was assessed by ELISA.The capacity of DCs to stimulate allogenic T lymphocyte proliferation and to regulate T cell differentiation was evaluated in a mixed lymphocyte reaction (MLR).RESULTS:Compared with normal serum,HLA-DR,CD80 and CD86 expression was significantly increased in the medium containing SLE serum with elevated levels of IFN-α,SLE serum with elevated levels of IL-6,SLE serum with elevated levels of IFN-α and IL-6 and normal serum with exogenous IFN-α and IL-6 (P<0.05),whereas IL-12 levels were significantly decreased (P<0.05),and capacity of DCs to stimulate allogenic T lymphocyte proliferation was significantly enhanced (P<0.05).Above-mentioned indexes recovered to a normal level in the medium containing SLE serum with anti-IFN-α and anti-IL-6 neutralizing antibodies.Compared with normal serum,proportion of both CD3~+CD8~-IFN-γ~+ and CD3~+CD8~+IFN-γ~+ T cell subsets was significantly reduced in the medium containing SLE serum with elevated levels of IL-6 (P<0.05).However,proportion of both CD3~+CD8~-IFN-γ~+ and CD3~+CD8~+IFN-γ~+ T cell subsets was significantly increased in the medium containing SLE serum with elevated levels Of IFN-α,SLE serum with elevated levels of IFN-α and IL-6,normal serum with exogenous IFN-α and IL-6 (P<0.05).CONCLUSION:The joint action of IFN-α and IL-6 in SLE serum promotes the differentiation and maturation of DCs derived from CD34+HPCs and affects the DC-mediated T cell differentiation,which might contribute to the pathogenesis of SLE.
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Objective To explore the clinical value of tissue synchronization imaging(TSI) in assessment of the characteristics of myocardium asynchrony in patients with ischemic cardiomyopathy(ICM). Methods Thirty-eight patients with ICM were divided into two groups: 26 patients without left bundle branch block(LBBB) in group A, and 12 patients with LBBB in group B. The control group consisted of 26 normal healthy subjects. According to 2D color,the characteristics of myocardium motion in each group were compared by TSI. The time for reaching systolic velocity peak(Ts) and early diastole velicity peak(Te) were measured separately in 12 segments. Standard deviations of Ts(Ts-SD) and Te(Te-SD) as well as difference of Ts(Ts max-min) and Te(Te max-min) of all 12 segments were calculated. The myocardium synchronization were assessed in each group. Results ①91.8% segments in the control group showed normal, with only 8.2% segments were slightly prolonged. TSI of all the ICM patients showed an obvious uneven chroma distribution from green to yellow and then to red. There were far more yellow or red segments than in the control group. ②Ts-SD.Ts max-min of LV 12 segments were significantly prolonged in all ICM groups compared with that in the control group ( P <0. 05). Systolic asynchrony was more obvious in group B than in group A. ③Te-SD,Te max-min of LV 12 segments were significantly prolonged in ICM groups than in the control group ( P < 0. 05). Diastolic asynchrony was more obvious in group B than in group A( P < 0. 05). ④Compared with that in the control group, postsystolic shortening(PSS) was common in patients with ICM. 32. 4% segments of group A were detected PSS.and 56. 0% segments of group B were detected PSS. Conclusions Left ventricular systolic and diastolic mechanical asynchrony is obvious in patients with ICM and PSS is common among them. Systolic and diastolic mechanical asynchrony is more severe in ICM patients with LBBB. TSI is the optimal means to evaluate the characteristics of asynchronous myocardial contraction.
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BACKGROUND: Blood stem cells (BSCs) are the most primitive cells in the immune system and can be differentiated into many kinds of cells. As the regulatory cells of immune response, dendritic cells (DCs) have been attracted more and more attention in the field of autoimmune diseases. Due to different resources of precursor cells, DCs have different cytokines, ideal cytokine matching,applying orders, and experimental cultured conditions. Furthermore, development, phenotypic expression, and mature degree are still different.OBJECTIVE: To investigate the influence of tumor necrosis factor- α (TNF- a ) and interleukin-4 (IL-4) on the culture system and provide improved method for inducing functional DCs in vitro, derived from cord blood CD34+ hematopoietic precursor cells (HPC).DESIGN, TIME AND SETTING: Observational study, which was performed in the Department of Microbiology and Immunology, Nanjing Medical University from March 2005 to November 2005.MATERIALS: The cord blood was collected from neonatal umbilical cord in the 81 Hospital of Nanjing City. CD34 monoclonal antibody coated magnetic bead system (MACS) was provided by Miltenyi Biotec Company, Germany; human recombinant granulocyte-macrophage colony-stimulating factor (rhGM-CSF), human recombinant interleukin-4 (rhlL-4), and human recombinant tumor necrosis factor-α (rhTNF-α) by Pepro Tech Company, USA.METHODS: CD34+HPC were isolated and purified from umbilical cord blood by using a high-gradient magnetic cell sorting system (MACS). Then the cells were cultured with different culture medium which contained different combinations of cytokines:GM-CSF and TNF-α (GT) or GM-CSF, TNF-α and IL-α (GTI) in order to be induced to differentiate into dendritic cells (DCs).The DCs derived from CD34+HPC were identified for their morphology and phcnotype by FACS and laser scanning con_focal fluorescence microscopy, and also for their abilities of inducing proliferation of allogenic T cells by 3H-TdR incorporation assay.RESULTS: The purity of selected CD34+ cells with MACS was more than 90%. DCs could be obtained from CD34+HPC by the culture in presence of GM-CSF and TNF-α or GM-CSF, TNF- o and IL-4. With the time of culture lasting, the cells expressed lower level surface antigen of CD34 and HLA-DR (P < 0.05), and possessed the phenotypes of DCs characterized by higher expression of CD80, CD86, CD83 and CDIa. At 13-15 days, the cells possessed higher level of phenotypes of DCs compared to 7-9 days and 10-12 days. DCs induced with GTI culture system expressed higher levels of surface antigen CD80, CD86, CD83,CD 1 a and a lower level of CD 14 than those induced with GT culture system. DCs displayed typical morphology and property and expressed higher level surface antigen of CD86 and CD80, especially expressed CD86 when they were induced with proper cytokines of GM-CSF and TNF- α added at 0 hour, and of IL-4 added at 48 hours.CONCLUSION: DCs can be generated from CD34+HPC by proper culture system, the design of GM-CSF + TNF- α + IL-4(GM-CSF and TNF- α were added at 0 hour, IL-4 was added at 48 hours) is preferred.
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As an important force in the process of China’s public medical education, private higher medical education is faced with severe challenges at the same time the development is made. On the basis of analyzing the development situation of private higher medical colleges and the deep space of development, the article promotes suggestions of the healthy development of private medical colleges, by macro-coordination from the government, private schools running orderly, improving the quality of education, and social support.