Objective: To explore the clinical efficacy of Xiaoshuan enteric-coated capsule (XSECC) in treating cerebral infarction and its potential mechanism of action. Methods: Patients with acute ischemic stroke (AIS) of the qi deficiency and blood stasis type were randomly assigned to the control and observation groups. They were evaluated using the National Institutes of Health Stroke Scale (NIHSS), Activities of Daily Living (ADL), Hachinskilnchemic Scale (HIS), Barthel Index (BI), clinical efficacy scores, and TCM syndrome scores on days 0, 14, 30, and 90. Furthermore, VEGF and BDNF levels were measured on days 30 and 90. Finally, we analyzed the changes in each scale score and vascular neurological factor in both groups. Results: After 14 days of treatment, the difference values in NIHSS, ADL, and BI were higher, and TCM syndrome and clinical efficacy scores were increased in the observation group compared with those of the control group (all P < .05). After 30 days, the NIHSS, ADL, HIS, and TCM syndrome scores were decreased compared with those of the control group, while BI and clinical efficacy scores were increased (all P < .05). After 90 days, the difference value in ADL was higher, and TCM syndrome score was increased in the observation group compared with that of the control group (P = .047, P = .005, respectively). The levels of VEGF and BDNF were higher in the observation group than in the control group on days 14, 30, and 90 (all P < .05). VEGF and BDNF levels on day 0 were associated with prognosis of patients with AIS; therefore, they have a predictive value for the prognosis of acute cerebral infarction. Conclusions XSECC therapy can improve clinical outcomes in patients with acute and recurrent cerebral infarctions. Its mechanism of action may be associated with the secretion of VEGF and BDNF.

BACKGROUND:Rat models of complete spinal cord transection are common models for neural tissue engineering. After transecting the spinal cord by the previous methods, gap length of broken end cannot keep relatively uniform, so we cannot objectively evaluate effects of various treatments or tissue engineering materials. OBJECTIVE:The spinal cord transection models were established by using double edged micro scissors, andthe feasibility of this new model was explored by comparing with the conventional method. METHODS: A total of 42 adult female Sprague-Dawley rats were divided randomly into group A (n=6), group B (n=18) and group C (n=18). Group A only received laminectomy. In the group B, the spinal cord was transected with a sharp-pointed knife. Knife point should touch anterior wal of spinal canal and sidewal bone surface. Complete spinal cord transection models were prepared by repeated cutting. In group C, complete spinal cord transection models were established by using self-made double edged micro scissors. RESULTS AND CONCLUSION: (1) At 1 week after model establishment, in the groups B and C, complete paralysis of the hind limbs was found, and BBB scores were similar. However, significant differences in the spacing of broken end were detected. (2) At 4 weeks after model establishment, hind limb functions could restore to different degrees in both groups, but no significant difference in BBB scores was found. (3) At 8 weeks after model establishment, significant differences in hindlimb motor function scores were detectable between both groups. Biotin glucosamine tracer display: In group B, a few labeled axon fibers were observed at the caudal side of the injured spinal cord. In group C, spinal cord was completely transected, and labeled axon fibers cannot be found at the caudal side. (4) Results suggested that the modeling method of self-made double edged micro scissors could effectively eliminate individual differences, contribute to quantitative analysis and comparative study of therapeutic effects.

Objective To explore the basic ingredients of the tree shrew’ s( Tupaia belangeri) milk and compare with the dairy ingredients of other milks.Methods We select ten seed tree shrews after delivery ( 1 ~21 ) d with lactation mother tree shrews, and use artificial passive breastfeeding method let the young tree shrews suck breast milk,we took the milk from the young tree shrews in the stomach, directly using aseptic operation with a syringe immediately, once every two days, for consecutive three to five times, and a total of 18 mL milk was taken from each seed tree shrew.Then the milk was detected according to the national standard method for component testing.Results The total solid content of the tree shrew’ s milk was 43.63%, including 26.01%of fat, 10.41%of protein, 0.45% of lactose and 0.99%of ash content.Compared with cow's milk, the tree shrew’ s milk contained 3.36 times of total solid contents, 1.24 times of ash, 2.74 times of protein, 6.67 times of fat, and 0.09 times of lactose.Compare with baby formula milk, the tree shrew’ s milk contained 1.44 times of total solid contents, 0.20 times of ash, 0.58 times of protein, 1.53 times of fat, and 0.06 times of lactose.The trace mineral composition of the tree shrew’ s milk showed that the calcium, phosphorus, potassium, sodium, magnesium, and iron contents were 1.83 times, 2.73 times, 1.25 times, 1.93 times, 1.28 times, and 1.48 times higher than those in the cow's milk, and were 0.66 times, 0.85 times, 0.34 times, 0.26 times, 0.85 times, 0.24 times lower than those in baby formula milk.Conclusions The main nutrients of tree shrew’ s milk is of high fat, high protein and low sugar, and it can provide a basis for tree shrews artificial brood and breeding work.

Objective To observe the impact of ningxinjieyu capsule on contents of monoamine neurotransmitters and their metabolites in cerebral cortex of chronic depression model rats.Methods The chronic stressed depression model rats were established by chronic unpredictable stress and separation.the model rats randomly divided into model group,ningxinjieyu high-dose group(10.8 mg/(kg · d)),mid-dose group(3.6 mg/(kg · d)),low-dose group(1.2 mg/(kg · d)) and Fluoxetine group(3.6 mg/(kg · d)) and the normal rats as the control group.Finally the changes of Noradrenaline (NE),Dopamine (DA),5-hydroxy tryptamine (5-HT),3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA) would be observed using HPLC-ECD technique.Results Compared with the control group,the contents of NE,DA and 5-HT were significantly lower in the model group (P＜0.01).Compared with the model group,the contents of DA were increased in the different concentrations of ningxinjieyue capsule groups,and the contents of 5-HT were higher in the low-dose and high-dose groups ((272.15± 129.89) ng/g,(445.08± 127.56) ng/g,(375.33±52.38) ng/g) ; the contents of NE were increased in the low-dose and mid-dose groups ((408.08 ± 89.55) ng/g,(530.12± 149.87) ng/g,(542.53 ± 96.10) ng/g) ; the contents of HVA were increased in the mid-dose and high-dose groups; the contents of DOPAC were increased in the low-dose group; the contents of 5-HT,DA and NE were increased in the fluoxetine group(P＜0.01 or P＜0.05).Conclusion Ningxinjieyu capsule has antidepressant effect,the mechanisms might be regulated to the contents of monoamine neurotransmitters and their metabolites in cerebral cortex.

Objective:To test the expression of Minichromosome maintenance complex component 7(MCM7) protein in hepato-cellular carcinoma(HCC) of different species including human, rat and tree shrew (tupaia) by cross-species oncogenomics approach, and to investigate the relationship between the expression of MCM7 and the development of hepatocellular carcinoma and its clinical significance. Methods:Western blot and Immunohistochemistry were applied to detect the expression levels of MCM7 protein in HCC tissues,corresponding HCC-adjacent liver tissues and normal liver tissues collected from different species including human, rat and tree shrew, respectively. The clinicopathologic factors were also analyzed with the results of Immunohistochemistry. Results:Western blot analysis showed that the expression of MCM7 protein in HCC tissues of human and rat were higher than that in corresponding HCC-ad-jacent liver tissues and normal liver tissues, respectively and significantly (P<0.05). However, the expression of MCM7 protein in HCC tissues of tree shrew were also higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, but no significant difference was found among three types of tissues (P>0.05).There was also no significant difference between HCC-adjacent liver tis-sues and normal liver tissues in three species (P>0.05). Immunohistochemical analysis showed that MCM7 protein was mainly ex-pressed in nucleus of HCC cells, and the positive rate of MCM7 protein in HCC tissues of human, rat and tree shrew were significantly higher than that in corresponding HCC-adjacent liver tissues and normal liver tissues, respectively (P<0.05). However, no significant difference was found between HCC-adjacent liver tissues and normal liver tissues (P>0.05). Moreover, the protein level of MCM7 was intimately related to patient's HCC stage, extrahepatic metastases and postoperative recurrence (P<0.05). Conclusion:MCM7 protein might play a pivotal role in hepatocarcinogenesis. In addition, it was probably related to patient's HCC stage, extrahepatic metastases and postoperative recurrence. It seems very likely that MCM7 may be applied as a new molecular target in HCC prevention and treat-ment.

OBJECTIVETo study pathological and therapeutical problems concerning myocardial cell mitochondria changes during myocardial cell hypertrophy by culturing rat primary myocardial cells.

METHODPrimary myocardial cells were seperated and cultured together with angiotensin II (Ang II) for 72 or 96 hours. The total protein content with the BCA method and the photography and measurement of cell diameter with inverted microscope reflected myocardial cell proliferation. The mitochondrial membrane potential (Delta Psi m) with fluorescence microscope, the mitochondrial single amine oxidase (MAO) activity with spectrophotometer, the mitochondrial cytochrome oxidase (COX) activity and the injury percentage of mitochondrial outer membrane with microplate reader and the contents of ATP, ADP, AMP with high performance liquid chromatography reflected the injury and energy metabolism of myocardial cell mitochondrial structure and function when being cultured together with Ang II. On that basis, cells were treated with Astragali Radix injection and valsartan for observing pharmacological effects on mitochondrial structure and function in restructured myocardial cells.

RESULTIn 72 h and 96 h, compare with the control group, the model group showed significantly increased total protein content and enlarged myocardial cell diameter. During the course of proliferation, the myocardial cell MAO activity and the injury percentage of mitochondrial outer membrane were significantly increased, with significant decrease in mitochondrial COX activity, mitochondrial Delta Psi m and the content of ATP, ADP and rise in the content of AMP. Astragali Radix injection and valsartan reduced myocardial cell total protein content and cell diameter caused by Ang II, decreased myocardial cell MAO activity, significantly increased mitochondrial COX activity and the content of ATP and ADP, and decreased the content of AMP.

CONCLUSIONDuring the process of myocardial hypertrophy, the injury of mitochondrial structure and function and the changes in myocardial cell energy metabolism injury occurred after the injury of mitochondria. Astragali Radix injection and valsartan can reverse myocardial cell mitochondrial structure and function during myocardial cell hypertrophy caused by Ang II. Reversion of myocardial cell hypertrophy and restructuring of myocardial cells helps improve energy metabolism of the myocardial cells.

Animals ; Astragalus Plant ; chemistry ; Cells, Cultured ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Hypertrophy ; drug therapy ; Injections ; Male ; Membrane Potential, Mitochondrial ; drug effects ; Mitochondria, Heart ; drug effects ; Myocytes, Cardiac ; drug effects ; pathology ; Rats ; Rats, Sprague-Dawley

BACKGROUND: Mesenchymal stem cells are few in human bone marrow, and their number will decrease with aging or body weakening, so a large amount of amplification is necessary. However, the biological characteristics of human mesenchymal stem cells of each passage remain poorly understood.OBJECTIVE: To analyze and compare the biological characteristics of each passage of bone marrow-derived mesenchymal stem cells (MSCs) so as to provide a basis for clinical demands of tissue engineering.DESIGN,TIME AND SETTING: Cytological observation in vitro. The experiment was performed at the Department of Orthopedics and Central Laboratory, Dongzhimen Hospital, Beijing University of Chinese Medicine from March to October 2008.MATERIALS: From bone marrow of patients with non-hematopoietic disease, MSCs were provided by Department of Orthopedics, Dongzhimen Hospital, Beijing University of Chinese Medicine.METHODS: Bone marrow was collected form posterior superior iliac spine of patient, MSCs were isolated and cultured by Percoll method. When the cells were confluent at 90%, they were trypsinized and observed by inverted miscroscopy. The second passage of cells were collected for index detection.MAIN OUTCOME MEASURES: Cell morphological characteristics and immunophenotype; cell activity was detected by MTT; cell division and apoptosis in the proportion of necrosis were analyzed by flow cytometry analysis.RESULTS: The passaged MSCs exhibited uniform appearance in fusiform shape, and their growth was slowed down after 9 passages, exhibiting cytoplasm vacuolization and body enlarging. The second passage of MSCs was positive for CD44, CD106,and CD105, but negative for CD34 and CD45. MTT values peaked at passage 9, and gradually decreased since passage 10. At passage 11, the number of MSCs at division stage was increased, but from the sixth passage, the number of apoptotic cells increased significantly, reaching more than 60% at passage 8.CONCLUSION: According to biological characteristics analysis of MSCs at each passage, the third to the sixth passage cells are recommend for clinical therapy.

BACKGROUND:In the initial stage of ischemia,organisms complete angiogenesis and maintain the blood supply of organization through compensatory regulation and repair,in which process a variety of cytokines,especially vascular endothelial growth factor (VEGF),have played a key role.OBJECTIVE:To observe the characteristics and rules of VEGF level changes in both ischemic tissues and blood serums at different time points after establishing rat models of lower limb ischemia.DESIGN,TIME AND SETTING:A randomized controlled animal experiment was performed at the Loboratory for Key Subjects in Dongzhimen Hospital and the Laboratory of Molecular Biology in Beijing University of Chinese Medicine from April to November in 2007.MATERIALS:A total of 42 male SD rats of 4 weeks were divided by random digits table into 6 groups,namely a control group and a model group which was subdivided into 5 groups at the time points of hour 4,day 3,weeks 1,2 and 4 post modeling respectivly.METHODS:Rat models of lower limb ischemia were established in the model group by performing ligation and mutilation operation to left femoral arteries of rats that were anesthetized with 100 g/L chloral hydrate. At the time points of hour 4,day 3,weeks 1,2 and 4 following modeling respectively,rats were selected to extract their abdominal aorta blood samples whose supernatant was then obtained through 10 minutes of 3 000r/min centrifugalization. Gastrocnemius tissues in left legs of rats were isolated at the same time. Samples in control group were obtained directly from anesthetized rats.MAIN OUTCOME MEASURES:Western blotting method was used for detecting protein expression of VEGF in ischemic tissues and ELASA method for VEGF level in serum.RESULTS:The protein expression of VEGF in ischemic tissues began to increase immediately at hour 4 following ischemia and reached a peak at day 3,after which it reduced gradually till week 2 when it reached its minimum. Then it began to increase again and reached the level close to that of normal tissues by the end of week 4 following ischemia. As for the VEGF level in serum,it decreased immediately after ischemia,with the maximum decrease amplitude between immediate and hour 4 following ischemia,a smaller one between hour 4 and week 1;From week 1 following ischemia on,it began to increase but was still lower than the normal level by the end of week 4 (P＜0.01 ).The VEGF level in serum changed with the tendency of immediate decrease from hour 4,minimum at week 1 and graduate increase after week 1 following ischemia.CONCLUSION:After ischemia in lower limb,VEGF level in ischemic tissues changes in the direction of increase-decrease-increase;VEGF level in serum changed in the direction of decrease-increase. In another words,VEGF levels after ischemia shows the characteristics of being low in serum and high in local ischemic tissues.

To establish an ischemia-reperfusion injury model of rat cerebral microvascular endothelial cells (MVECs) in vitro, and to explore the relationship between nuclear factor-kappa B (NF-kappaB) and the protective effects of Qingkailing effective components (hyocholic acid, taurocholic acid, baicalin, jasminoidin, Pinctada martensii) on MVECs.

To observe the effects of tetramethylpyrazine (TMP) on the proliferation and type I collagen synthesis of rat cardiac fibroblasts (CFBs) induced by angiotensin II (Ang II), and to explore the mechanism of TMP in treating myocardial fibrosis.