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Objective To observe the differences of cerebral activation pattern with resting state functional magnetic resonance imaging (rs-fMRI) between patients with spasmodic torticollis (ST) and healthy controls,thus to investigate the pathogenesis of ST.Methods Nineteen ST patients and 21 age,sex and education-matched healthy controls,recruited from the Department of Neurology,First Affiliated Hospital of Guangxi Medical University between November 2012 and January 2016,were included in this study.rs-fMRI and factional amplitude of low frequency fluctuation (fALFF) were used to obtain differences between patients with ST and healthy controls,and correlative analysis was made on fALFF values of abnormal brain regions and ST patients' symptom severity (Tsui scores).Results Compared with healthy controls,patients with ST had significantly increased fALFF in the left cerebellum and significantly decreased fALFF in the left posterior cingulate cortex/precuneus,right posterior cingulate cortex/precuneus,left middle temporal gyrus,right angular gyrus,left post-central gyrus,right supplementary motor area (t =-5.714-5.920,P ＜0.01),and abnormal brain regions' fALFF values had no correlation with patients' age of onset,disease course,symptom severity (P ＞ 0.05).Conclusion Abnormal sensorimotor area,default mode network and cerebellum dysfunction may play a role in the pathophysiology of ST.
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This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4(+) helper T lymphocytes (Th1/Th2) and human leukocyte antigen (HLA)-DR expression on CD14(+) monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10) and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein (CRP) were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14(+) monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.
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This study examined the effect of intensive insulin therapy on immune function and inflammatory factors at the early phase after severe trauma. At day 1, 3, 5, 7 after admission, subsets of CD4(+) helper T lymphocytes (Th1/Th2) and human leukocyte antigen (HLA)-DR expression on CD14(+) monocytes were flow cytometrically measured. Levels of cytokines, including tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-10 (IL-10) and other immunity markers, such as IgA, IgG, IgM, C3, C4 and C reaction protein (CRP) were examined in two groups. The results showed that TNF-α, IL-6 and CRP levels in the intensive insulin therapy group were significantly lower than those in the conventional therapy group, whereas IL-10 levels were substantially increased after intensive insulin therapy. C3 level at day 3, 5, 7 and C4 levels at day 5, 7 were lower in the intensive therapy group than in the conventional therapy group. Th1/Th2 ratios decreased gradually over time in both groups, and were much lower at day 3, 5, 7 in intensive therapy group. There were significant differences among day 3 to day 7 after admission in HLA-DR expression in CD14(+) monocytes. It was concluded that the intensive insulin therapy could decrease pro-inflammatory cytokines and increase anti-inflammatory cytokines in the elderly suffering from severe trauma, at the same time, with complement recovery being delayed. Moreover, intensive insulin therapy promoted immune suppression and, therefore, measures need be taken to address the issue.
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Aged , Aged, 80 and over , Female , Humans , Male , Cytokines , Allergy and Immunology , Hyperglycemia , Drug Therapy , Allergy and Immunology , Hypoglycemic Agents , Therapeutic Uses , Immunity, Innate , Allergy and Immunology , Immunologic Factors , Allergy and Immunology , Insulin , Therapeutic Uses , Treatment Outcome , Wounds and Injuries , Drug Therapy , Allergy and ImmunologyABSTRACT
Objective To evaluate the clinical value of CT perfusion (CTP) imaging for providing quantitative information about angiogenesis in patients with lung carcinoma and investigate the correlation of CTP enhancement parameters and histological microvessel density (MVD) with lymphatic involvement in peripheral lung carcinoma. MethodsFifty-three patients with pathology-proved peripheral lung carcinoma underwent CT perfusion scan before operation. The enhancement parameters of CTP were calculated based on the time-density curves (TDC) of fist pass phase. All cases were classified into two groups according to pathologic results: tumor with and without lymph node involvement. Two-sample t test was used for the statistics. The ROC curve was used to assess the efficiency of the enhancement parameters of CT perfusion and MVD for predicting lymphatic involvement.Results Tumors with lymph node involvement had significantly higher value of MVD than those without lymph node involvement (64.69±16.34 and 42.67± 16.78, respectively,t=4.84,P<0.01). Tumors with lymph node involvement had significantly higher value of CTP enhancement parameters (PH, M/A, PV) than those without lymph node involvement [PH= (41.79±15.50) and (29.99±10.91) HU,M/A =0.24±0.09 and 0.15±0.06, PV=(2.14±1.09) and (1.27±0.53) ml·min~(-1)·ml~(-1), t=3.21,3.95, 3.66, P<0.01, respectively]. The CTP enhancement parameters (PH, M/A, PV) of lung cancer correlated positively with the MVD, the highest correlation coefficient was between the PV and MVD (r=0.716, P<0.01). MVD and PV had higher values for predicting lymph node involvement in ROC curve analysis.The sensitivity, specificity and accuracy for predicting lymph node involvement were 80.8%, 81.5% and 81.1% or 84.6% ,85.2% and 84.9% respectively if MVD>52/0.74 mm~2 or PV>1.52 ml·min~(-1)·ml~(-1). ConclusionThe CT perfusion PV and histological MVD have good correlation with lymph node involvement in peripheral lung carcinoma and are important predicting parameters before operation.
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@#Objective To explore the value of high frequency echocardiography to evaluate cardiac structure and function of rat with acute myocardial infarction (AMI).Methods The rat AMI model was established by ligating the proximal left anterior descending coronary artery. The cardiac structure and function of model rats were examined with high frequency echocardiography in the 2nd and 8th week after ligation successfully. Another 20 healthy rat were selected as the sham operative group.Results The survival rat is 82.0% beyond 24 hours. At 2nd week, the model animals had an increased end-diastolic diameter (EDD), end-systolic diameter (ESD), end-diastolic volume (EDV), and further end-systolic volume (ESD), compared with the sham operative group. Ejection fraction (EF) and fraction shortening (FS) of model rats decreased. The ratio of E/A was higher. At 8th week, these parameters deteriorated left atrial diameter (LA) increased ( P<0.05). EF and FS were relevant to dp/dtmax.Conclusion High frequency echocardiography can evaluate left ventricle remodeling of rat dynamically.
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@#Objective To investigate the influence of rehmannia glutinosa oligosaccharide(RGOs)on apoptosis of human adipose tissue-derived mesenchymal stromal cells(ADMSCs)induced by hydrogen peroxide(H2O2).Methods Cultured human ADMSCs were randomly divided into three groups as the normal group(group N,without any treatment),H2O2 group(group H,treated with 0.1 mmol/L H2O2),and RGOs group(group R,treated with 0.2g/L RGOs plus 0.1 mmol/L H2O2).After 1 h,6 h and 24 h,morphological changes of ADMSCs apoptotic were observed,the apoptotic rate was determined by flow cytometry.Results After 1 h,6 h and 24 h,the apoptotic rate of group H was significantly higher than that of the group N and group R(P<0.05),and the apoptotic rate of group H was significantly higher than that of the group R(P<0.05).Conclusion RGOs can attenuate apoptosis of human ADMSCs induced by H2O2.
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BACKGROUND: Studies confirm that ischemia/reperfusion (I/R) injury can induce myocardial apoptosis. The loss of mitochondrial membrane potential (MMP) after reperfusion is the inevitable pathway of apoptosis. Protection of MMP may reduce apoptosis.OBJECTIVE: To observe the effect of naloxone on MMP of hypoxic myocardial cells and apoptosis in neonatal rats, and investigate the protective effect of naloxone on hypoxic myocardial cells.DESIGN: Observation and controlled trial.SETTING: Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: Detection of apoptosis of myocardial cells was carried out in the Laboratory of Pathophysiology, General Hospital of Chinese PLA in December 2004. Ten neonatal rats were used, and detection of MMP of myocardial cells was carried out in the same laboratory in March 2006 and 20 rats were used. All the involved rats were provided by the Animal Center of the General Hospital of Chinese PLA on the day of birth, were involved in this trial. Reagents: Naloxone hydrochloride injection (0.4 g/L, Beijing Sihuan Pharmaceutical Factory, Batch No. 0206272); the lowest and essential medium (Dulbecco, DEME,GIBCO company); phosphate buffer solution and fetal bovine serum (PBS and FBS, SIGMA Company).METHODS: The neonate rats were cut open along the median line of chest bone after local sterilization with iodine tincture on the day of birth. 1/3 ventricular myocardium before cardiac apex was harvested. On the 4th day of the culture,culture flasks of cells in good growth status ( ＞ 106 cells/bottle) were selected and divided into 3 groups: control group (normal culture, n =3 bottles), hypoxia group (hypoxia/reoxygenation, n =15 bottles) and naloxone group (hypoxia/reoxygenation, and treated by naloxone, n =15 bottles). Three time points were set in hypoxia group and naloxone group according to different time of hypoxia and reoxygenation: hypoxia 2 hours/reoxygenation 0 hour; hypoxia 2 hours/reoxygenation 2 hours; hypoxia 2 hours/reoxygenation 4 hours, 5 bottles at each time point. In the hypoxia group, DEME medium, which was pre-filled with 0.95 volume fraction of N2 and 0.05 volume fraction of CO2, containing 0.01 volume fraction of FBS, was used and 0.95 volume fraction of N2 and 0.05 volume fraction of CO2 was also filled to replace the air in the culture flasks. The culture flasks were enveloped for incubation at 37 ℃. The cells in the hypoxia group were incubated at normal condition (0.95 volume fraction of air and 0.05 volume fraction of CO2) at set time. In the naxolone group, hypoxia/reoxygenation treatment was the same as above, and naloxone hydrochloride was added at the sametime and the final concentration of naloxone in culture flasks was 5 μmol/L (The volume for naloxone ≤ 0.5% of the total medium). In the control group, hypoxia/reoxygenation and naloxone treatment were not given, but the same volume of normal saline was added, and this time served as time point of hypoxia 0 hour/reoxygenation 0 hour. After intervention,myocardial MMP changes and apoptosis were detected with fluorescent staining-flow cytometer at different time after hypoxia/reoxygenation.MAIN OUTCOME MEASURES: ① MMP changes of hypoxia group and naloxone group at different time points; ② Comparison of survival, apoptosis and necrosis of cells at hypoxia 2 hours/reoxygenation 4 hours in each group.RESULTS: ① After hypoxia, MMPs of the cells in hypoxia group and naloxone group were decreased. MMP of the cells in the naloxone group was higher than that in the hypoxia group at each time point (P ＜ 0.01). The great amplitude of decrease of MMP occurred in hypoxia period, but not in reoxygenation period. ② At hypoxia 2 hours/reoxygenation 4 hours, the apoptotic and necrotic rates in the hypoxia group were significantly higher than those in the naloxone group [(9.88±0.98)% vs. (2.41±0.52)%; (5.10±0.29)% vs. (3.56±0.56)%, both P ＜ 0.01]. The apoptotic rate was significantly higher than the necrotic rate in the hypoxia group (P ＜ 0.05).CONCLUSION: Early application of naloxone can significantly alleviate and postpone the decrease of myocardial MMP after I/R, and reduce apoptosis and necrosis.
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BACKGROUND: Adipose tissue-derived mesenchymal stem cells (ADMSCs) have the multilineage differentiation potential, and are relatively easier to be obtained, thus they have attracted more and more attention as a new seed cell for cell engineering.OBJECTIVE: To observe the in vitro culture conditions of ADMSCs isolated from rat's subcutaneous adipose tissue, and identify them using immunohistochemical staining.DESIGN: An animal experiment.SETTING; Department of Cardiology, the General Hospital of Chinese PLA.MATERIALS: One healthy male Wistar rat of clean degree, 4 months old, weighing 200 g, was used. DMEM, fetal bovine serum were from GIBCO; Monoclone antibodies of rabbit-anti-rat CD13, CD34, CD44, CD45, CD105, D-related human leucocyte antigen (HLA-DR), factor-Ⅷ, vov Willebrand factor (VWF), Myosin, SABC kits and DAB staining kit from Wuhan Booster Biological Engineering, Co.,Ltd; Adeno-associated virus encoding green fluorescent protein from Vector Gene Technology Company Limited (Beijing).METHODS: The experiments were carried out in the Department of Internal Medicine, the General Hospital of Chinese PLA in October 2006. ① Cell isolation and culture: 0.3 g adipose tissue was cut from subcutaneous adipose tissue of Wistar rat's groin under aseptic condition, then minced and digested before culture, DMEM was changed at 2-3 days after plenty of fusiform-shap ed attached cells were observed under microscope, and the cell growth was observed. The cell concentration was adjusted to 2×107 L-1, then seeded into 96-well plate, and 100 μL for each well. From the second day, 3 wells were randomly selected every day, the cells were released with tripsin, and counted with blood cell counting chamber under inverted microscope. ② Cell viability assay: ADMSCs of passages 3 to 8 were added to DMSO freeze medium, and thawed after 2-4 weeks, and the cell viability was assessed by trypan blue dye exclusion. ③Immunohistochemical staining and identification: 2 ×107 L -1 cells were seeded to culture plate, then the immunohistochemical (SABC method) identification and Oil red O staining were performed to determine the cell surface antibodies of CD13, CD34, CD44, CD45, CD105, HLA-DR, factor-Ⅷ, HLA-DR and VWF. ④Lineage-specific differentiation and identification: The ADMSCs were plated on multi-well chamber and induced with lineage-specific media supplementation at least two weeks and identified by histologic/immunohistochemical assay of Oil red O for adipogenisis, alkaline phosphatase (ALP) stain for osteogenisis and Myosin monoclonal antibody for myogenisis. ⑤Transfected adenovirus carried green fluorescence protein (AD-GFP) medium: The fourth generation of ADMSCs were seeded on 96-well plate, 3 000 cells for each well, serum-free DMEM was changed after 24 hours, and added by AD-GFP at the same time, then transfected with different multiplicity of infection (MOI) of 1∶50, 1∶100, 1∶150 and 1∶200respectively, and then the transfection was observed.MAIN OUTCOME MEASURES: ① Results of cell isolation and culture; ② Cell viability after freezing and thawing; ③Results of immunohistochemical staining and identification; ④ Results of lineage-specific differentiation and identification;⑤ Results of transfected adenovirus carried AD-GFP.RESULTS: ① About 3.6×105 attached cells were obtained from 0.3 g subcutaneous adipose tissue, and these cells could be subcultured for passages in vitro with stable population doubling time. ② The cells were thawed after freezing for 2-3 weeks, and the trypan blue staining showed that the cell viability was above 90%. ③ The immunocytochemical staining showed that CD13, CD44, CD105 were positive and CD45, factor-Ⅷ, HLA-DR and VWF negative in different generations. ④ From the second generation, a few Oil red O positively stained cells were observed, which were obviously increased after prolonging the refreshing. After lineage-specific differentiation, the cells were all positive by Oil red O staining, ALP staining and Myosin immunohistochemical staining. ⑤ 72 hours after transfection, it was observed under fluorescence microscope that most cells were green fluorescence when the MOI value was 1∶200, the transfection was successful, and it was generally determined that the transfection rate was above 90%.CONCLUSION: A large number of ADMSCs with multilineage differentiation potential can be easily obtained from rat adipose tissue, osteoblast, myoblasts, they can be expanded in large quantity and stored in vitro for long time, AD-GFP were also successfully transfected.
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BACKGROUND: At present, many problems deserve to be solved before clinical utilization of adipose tissue-derived mesenchymal stem cells (ADMSCs) such as complete differentiation from ADMSCs into cardiomyocytes and ADMSCs with or without specific function of cardiomyocytes. It is of significance to solve the problems including how to elevate the differentiation rate of ADMSCs into cardiomyocytes and how to elevate the homing and survival rate after transplantation for clinical utilization of stem cells.OBJECTIVE: To observe the differentiation of ADMSCs into cardiomyocytes after in vitro culture and induction.DESIGN: Randomized controlled observation.SETTING: Laboratory of Cardiology, General Hospital of Chinese PLA.MATERIALS: Adipose tissue was collected from abdominal operative patients at Department of General Surgery of General Hospital of Chinese PLA with the agreement of patients. Iscove's Modified Dulbecco's Medium (IMDM), type Ⅰ collagenase, 5-azacytidine (5-aza) and polyclonal antibody of specific anti-myocardial Troponin T (TnT) were purchased from Hyclon company, Gibco company, Sigma and Fujian Maixin Biotechnology Company, respectively.Monoclone antibodies of CD44, CD45, CD34, HLA2DR and factor-Ⅷ, Desmin, anti-α-striated muscle actin and anti-myosin heavy chain (MHC) were bought from Beijing Zhongshan Golden Bridge Biotechnology Limited Corporation. DAB stain, reverse transcription-polymerase chain reaction (RT-PCR) kit and atrial natriuretic peptide (ANP) radioimmunity kit were purchased from Beijing Zhongshan Golden Bridge Biotechnology Limited Corporation,Invitrogen and Radioimmunity Research Institute of Science and Technology Development Center of General Hospital of Chinese PLA, respectively.METHODS: The experiment was performed at the Laboratory of Cardiology, General Hospital of Chinese PLA from March 2005 to April 2006. ADMSCs were isolated and cultured by digestion and attachment culture method. The third generation of cells were determined by immunocytochemical method for surface molecule CD44, CD13, CD105, CD45,CD34, HLA-DR, factor Ⅷ and induced by addition of 5-aza into culture medium. On the 7th, 14th, 21st and 28th days, gene of GATA4 and Nkx2.5 were tested by reverse transcription-polymerase chain reaction (RT-PCR), and the concentration of atrial natriuretic polypeptide (ANP) were also examined by radioimmunoassay.MAIN OUTCOME MEASURES: Determination of cell surface marker, gene of GATA4 and Nkx2.5, and ANP concentration.and factor Ⅷ negative for the cultured cells. After induction for 7 days with 5-aza, no cells expressed Desmin,α-Sarcomeric Actin, Myocin heavy chain (MHC) and TnT, most cells were positive stained for CD44. After induction for 14 days, small amounts of cells displayed positive for Desmin,α-Sarcomeric Actin and MHC but still negative for TnT,while partial cells expressed positive CD44. After induction for 21 days, most cells were positive for Desmin,α-Sarcomeric after induction and no significant difference was found compared with that at the 7th day [(0.022±0.01) ng/L (P＞0.05].ANP concentration was respectively (7.92±0.21) and (8.12±0.50) ng/L at the 21st day and the 28th day (P＞0.05), but there was significant difference compared with that at the 7th day (P＜0.05).
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Objective To explore the diagnostic value of MR diffusion weighted imaging (DWI) in the differential diagnosis between encephalitis and acute cerebral infarction.Methods The MR DWI appearances of 23 cases of encephalitis including 14 cases with viral encephalitis, 9 cases with demyelinative encephalitis and 30 cases with acute cerebral infarction were retrospectively analyzed.Results The 30 cases of acute cerebral infarction appeared bright signal in DWI with a ADC values of (0.46?0.13)?10 -3mm2/s. 7 cases with viral encephalitis and 9 cases with demyelinative encephalitis appeared slightly high or high signal intensity in DWI with a ADC values of (0. 98?0.18)?10 -3mm2/s and(0. 89?0.07)?10 -3mm2/s respectively. The ADC values of viral encephalitis and demyelinative encephalitis were higher comparing to acute cerebral infarction(?0.05). However, the ADC values showed higher or lower in different areas in 4 cases, and lower ADC values appearing in 3 cases with viral encephalitis comparing normal parenchyma.Conclusion MR DWI is useful in the differential diagnosis between encephalitis and acute cerebral infarction.