Acupuncture Points ; Acupuncture Therapy ; methods ; Hemiplegia ; therapy ; Humans

Acute Disease ; therapy ; Adult ; Bronchiectasis ; therapy ; Female ; Humans ; Moxibustion

BACKGROUND:Exercises play an important role in the recovery of neurological function after stroke. Few studies concerned the amount of exercise in rats after stroke. Hippocampus is strongly associated with cognitive function, but no reports addressed the expression of repulsive guidance molecule A in the rat hippocampus after ischemia and reperfusion. OBJECTIVE:To explore the effects of exercise on repulsive guidance molecule A expression in the hippocampus on the ischemic side in rats after cerebral ischemia-reperfusion injury. METHODS:120 Sprague-Dawley rats were equal y and randomly divided into normal group, sham-operation group, and 7-, 14-, 28-day model groups. The model of right cerebral ischemia-reperfusion injury was induced by ligation with nylon monofilament in rats of 7-, 14-, 28-day model groups. Low exercise group received treadmil training of 5 m/min, 5 minutes;7 m/min, 5 minutes;9 m/min, 20 minutes. Moderate exercise group received treadmil training of 8 m/min, 5 minutes;10 m/min, 5 minutes;13 m/min, 20 minutes. High exercise group received treadmil training of 8 m/min, 5 minutes;11 m/min, 5 minutes;20 m/min, 20 minutes. RESULTS AND CONCLUSION:Repulsive guidance molecule A mRNA and protein expression was highest in the ischemic side of the hippocampus in the 7-day model group without excercise. Moreover, repulsive guidance molecule A relative expression gradual y reduced over time. Compared with non-exercise, repulsive guidance molecule A mRNA and protein expression significantly decreased in the 14-and 28-day model groups during moderate exercise (P<0.05), but repulsive guidance molecule A mRNA and protein expression increased during high exercise. Above data confirmed that moderate exercises could decrease repulsive guidance molecule A expression in the affected side of the hippocampus of rats with cerebral ischemia-reperfusion injury.

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).

Objective To explore the toxic effect of platelet-derived growth factor receptor (PDGFR)-αantisense oligonucleotide (αASODN) on the retina. Methods Twenty-four healthy adult colored rabbits were selected and randomly divided into four groups in six for each group. Intravitreous injections of 0.1ml different density diluents containing PDGFR-αASODN and liposome were performed in the right eyes in 3 groups. The other group was injected with 0.1 mL balanced salt solution (BSS) as the control group. The left eyes of all animals were not rejected. Slit lamp examination, indirect ophthalmoscopy and electroretinogram (ERG) examination were performed at 1, 7, 14 and 28 days after the injection. On the day 28, the right eyes were harvested, and HE、immunohistochemistry and transmission electron microscopy of retinal tissue were performed . Results The slit lamp, indirect ophthalmoscope examination of all groups were normal in each time. ERG examination yielded no difference in the amplitude of b wave between the treated groups and the normal control group. Pathological changes of the retinal tissue were not observed in the examinations of HE and immunohistochemical at the day 28 after injection. Electron microscope observation of retinal photoreceptor cells in the group D showed the parts of gaps between membranous discs were expanding, parts of were fusing, a few of clearances around cell nucleus were slightly enlarged,and the shapes of the cell nucleus were slightly irregular. Conclusions To inject 0.1 mL PDGFR-αASODN/lip2000 into the vitreous chamber, PDGFR-αASODN can be relatively safe when its concentration is less than 1.5μmol/L.

Objective To detect the effect of olfactory bulb(OB)lesion on proliferation,migration and differentiation of the neural stem cells(NSCs)in the subventricular zone(SVZ).Methods Forty-two healthy female SD rats were enrolled and randomly divided into normal control group,isotonic saline group and OB lesion at day 3,at weeks 1,2,3 and 4 groups,six rats per group.OB lesion was induced by N-methyl-D-aspartic acid(NMDA)injection.Bromodeoxyuridine(BrdU)was injected intraperitoneally to label NSCs.Immunohistochemical staining was used to detect the proliferation of SVZ NSCs.In addition,another 18 rats were randomly divided into normal control group,isotonic saline group and lesion group,six rats per group.BrdU was injected intraperitoneally one week after OB lesion and then the animals were sacrificed four weeks after BrdU injection to detect the migration and differentiation of NSCs with immunohistochemistry and immunofluorescence.Results Three days after OB lesion,BrdU-positive cells in SVZ began to increase(26.33 ± 2.58,P ＜0.01),reached the maximum at week 1 (35.33 +3.01,P＜0.01)and still sustained a high level at week 4(19.50+ 2.26,P＞0.05).Five weeks after the OB lesion,the rostral migratory-stream(RMS)and the BrdU-positive cells in OB were significantly increased(86.50 + 5.09,P ＜ 0.01)and(52.83 + 3.87,P ＜ 0.01),respectively.By using fluorescence double staining,most of the BrdU-positive cells were co-localized with the neuronal nuclear antigen(Neun),with a portion of BrdU-positive cells expressing the glial fibrillary acidic protein (GFAP).Conclusions OB lesion can improve the proliferation of NSCs in SVZ and migration of NSCs to OB.The newborn cells can differentiate into not only neurons,but also gliocytes and may be involved in lesion repair.

Objective To explore the clinical effect of detection of serum procalcitonin (PCT), C-reactive protein (CRP)and white blood cell (wBC)on diagnosis and treatment of severe pneumonia in children.Methods A total of 189 cases of pediatric patients with pneumonia including 51 severe pneumonia cases (severe pneumonia group)and 138 common pneumonia cases (common pneumonia group)treated from Mar 2014 to Dec 2014 and 30 healthy cases (control group)were enrolled,and the level of PCT,CRP and wBC of all cases were detected.Results The levels of PCT before treatment in severe pneumonia group,common pneumonia group,and control group were 1.12(0.44 ~3.07)ng /ml,0.14 (0.09 ~0.26) ng /ml and 0.03(0.01 ~0.06)ng /ml respectively,and there were significant differences among three groups (P ﹤0.05,respectively).The levels of CRP before treatment in severe pneumonia group,common pneumonia group,and control group were 82.2(25.9 ~120.3 )mg /L,10.8 (5.3 ~23.9)mg /L and 3.2 (2.1 ~6.9) mg /L respectively,and there were significant differences among three groups (P ﹤0.05,respectively).The counts of wBC before treatment in severe pneumonia group,common pneumonia group,and control group were 10.1 (9.1 ~14.1 )×109 /L,8.8(6.8 ~1 1.7)×109 /L and 6.2(4.8 ~7.9)×109 /L respectively,and there was significant difference only between severe pneumonia group and control group (P ﹤0.05 ).The level of PCT and CRP significantly decreased in severe pneumonia group after one week of treatment[PCT:0.15(0.09 ~0.24)ng /ml,CRP:9.9(3.6 ~19.0)mg /L](P ﹤0.05),but there was no significant differ-ence of wBC counts in severe pneumonia group between after and before treatment[8.5(6.3 ~9.8)×109 /L vs.10.1 (9.1 ~14.1 )×109 /L](t =1.312,P =0.205 ).After two days of anti-inflammatory treatment in severe pneumonia group,serum PCT dropped to 44% of the level before treatment,and smoothly dropped to nearly 10% of the basic value every two days.Serum PCT was correlated with serum CRP in children with pneumonia(R2 =0.550 4,P ﹤0.05).Conclusion The combined detection of PCT and CRP could provide important guidance for the differential diagnosis,treatment and prognosis for severe pneumonia in children.

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.

Objective To investigate the clinical efficacy and safety of six-hole moxibustion box therapy for stable chronic obstructive pulmonary diseases(COPD).Methods Sixty patients with thoracic facet joint disorder were randomly allocated to treatment and control groups, 30 cases each. The control group inhaled tiotropium bromide inhalation powder spray and the treatment group received six-hole moxibustion box therapy in addition. The COPD Assessment Test (CAT) score and the dyspnea score were recorded and peripheral blood oxygen saturation was measured in the two groups before and after treatment. The adverse reactions were monitored in the two groups. Results There were statistically significant pre-/post-treatment differences in the CAT score and the dyspnea score in the two groups (P<0.05). There were statistically significant post-treatment differences in the CAT score and the dyspnea score between the treatment and control groups (P<0.05). There were statistically significant differences in pre-/post- treatment CAT score difference value and dyspnea score difference value between the two groups (P<0.05). There was a statistically significant pre-/post-treatment difference in peripheral blood oxygen saturation in the treatment group (P<0.05). There was a statistically significant post-treatment difference in peripheral blood oxygen saturation between the two groups (P<0.05).Conclusion Six-hole moxibustion box therapy plus tiotropium bromide inhalation powder spray is safe and effective in treating stable COPD.

Objective:To study the role of adhesion molecules in the pathogenesis of polymyositis. Methods:The abnormal expression of adhesion molecules on T cells in peripheral blood and muscle fibers from patients with myositis was analyzed by two colour immunofluoresence and RT PCR methods respectively. Results:The expression of adhesion molecules including lymphocyte function associated antigen 1(LFA 1 ),very late antigen 4(VLA 4) on T cells in peripheral blood and intercellular adhesion molecule l(ICAM 1) on muscle fibers from patients with myositis was markedly higher than that in the healthy control group. Conclusion: These findings suggested that adhesion molecules may be responsible for the migration of T cells and destraction of muscle fibers.