Establishment of a new rat model of complete spinal cord transection and defect / 中国组织工程研究

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.

Effect of ningxinjieyu capsule on contents of monoamine neurotransmitters and their metabolites in cerebral cortex of chronic depression model rats / 中华行为医学与脑科学杂志

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.

Detection and significance of the main nutritional ingredients of tree shrew’ s milk / 中国比较医学杂志

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.

The expression and significance of MCM7 protein in hepatocellu-lar carcinoma tissues of human, rat and tree shrew / 中国肿瘤临床

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 (P0.05).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 (P0.05). 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.

Effect of Astragali Radix injection on myocardial cell mitochondrial structure and function in process of reversing myocardial cell hypertrophy / 中国中药杂志

<p><b>OBJECTIVE</b>To study pathological and therapeutical problems concerning myocardial cell mitochondria changes during myocardial cell hypertrophy by culturing rat primary myocardial cells.</p><p><b>METHOD</b>Primary 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.</p><p><b>RESULT</b>In 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.</p><p><b>CONCLUSION</b>During 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.</p>

Variational trend of vascular endothelial growth factor in rat models of lower limb ischemia / 中国组织工程研究

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.

Biological characteristics of human bone marrow-derived mesenchymal stem cells at different passages: Third to sixth passages are recommended for clinical application / 中国组织工程研究

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.

Effects of tetramethylpyrazine on angiotensin II -induced proliferation and type I collagen synthesis of rat cardiac fibroblasts / 中西医结合学报

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.

Effects of Qingkailing effective components on nuclear factor-kappa B in an ischemia-reperfusion injury model of rat brain microvascular endothelial cells in vitro / 中西医结合学报

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.

Effects of Radix notoginseng extracts drug-containing serum on expressions of bcl-2, Bax and p21WAF1 proteins in MNNG transformed GES-1 cells / 中西医结合学报

OBJECTIVE: To investigate the effects of Radix notoginseng extracts drug-containing serum on the expressions of apoptosis-regulating proteins including Bax, bcl-2 and p21WAF1 in precancerous gastric cells. METHODS: The N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) transformed eternalized human gastric mucosa epithelium GES-1 cell line (MC cell) was used in vitro as a model of gastric precancerous lesion. The medicated canine serum was prepared by feeding to the adult Beagle dog with Radix notoginseng extracts and obtaining the serum after 2-hour medication. MC cells were cultured with medicated canine serum (medicated serum group) or non-medicated canine serum (normal control group) for 72 hours. Expressions of Bax, bcl-2 and p21WAF1 proteins were detected by immunocytochemical assay and the average optical density of the cells was determined by an image analysis system. RESULTS: Compared with those of the normal control group, Bax and p21WAF1 expressions in medicated serum group were significantly enhanced (P<0.01), while the expression of bcl-2 was significantly reduced (P<0.01). CONCLUSION: Radix notoginseng extracts may inhibit the proliferation and promote the apoptosis of precancerous gastric cells through altering expressions of the bcl-2, Bax and p21WAF1 genes.

Changes of spatial learning and memory ability in rats with heart failure after myocardial infarction / 中国组织工程研究

BACKGROUND: Chronic congestive heart failure occurs at the late stage of many heart diseases. Cognitive disability exists in chronic congestive heart failure which has been reported in most clinical studies, and measures that can successfully improve heart function can improve cognitive function.OBJECTIVE: To probe into the effect of chronic congestive heart failure after myocardial infarction on spatial learning and memory ability,and to observe the correlation between heart failure and cognitive function.DESIGN: A randomized controlled observation based on the experimental animals.SETTING: Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine), Ministry of Education.MATERIALS: This experiment was performed at the Key Laboratory of Traditional Chinese Medicine (Beijing University of Chinese Medicine),Ministry of Education. in November 2001. Totally 100 healthy male rats aged 10 to 12 weeks, weighing 200 g to 220 g purchased from Beijing Weitong Lihua Experimental animal Technical Co. Ltd were enrolled and randomly divided into the operation group (n=60) and sham-operation group (n=40).METHODS: Model rats in the operation group were infarcted by ligation of the left anterior descending coronary arteries to produce acute myocardial infarction models. Those in the sham-operation group left a loose tie without ligation. The rats that showed myocardial infarction-like changes through electrocardiogram were taken as the rats for operation (n=38), and those survived rats without abnormal changes and without ligation were taken as the sham-operation group (n=25). The two groups were subdivided into 10-day gro up, 30-day group and 60-day group. Morris water maze test was performed 10, 30 and 60 days after the operation followed by hemodynamic monitoring. Among the 10-day group, 30-day group and 60-day group, rats that did not complete the whole Morris swimming test due to heart failure with poor power and died 2 days after the operation or after hemodynamic monitoring were set as heart failure end period group.Rats in the same group as those died with similar body mass were set as the control group.MAIN OUTCOME MEASURES: ① Escape logarithm latency and the good rate if cognitive ability; ② stroke volume, cardiac output per minute,heart rate and cardiac index of hemodynamics, in which cardiac index was the main index for responding heart functionRESULTS: Totally 33 rats met the criteria in the operation group and 25rats in the sham-operation group. All the animals entered the stage of result analysis. ①Results of hymodynamic index: Stroke volume, cardiac output per minute and cardiac index of the rats in the 10-day group, 30-day group and 60-day group were significantly lower than those of the homochronous sham-operation group (P ＜ 0.01), and those indexes were significantly lower in the heart failure end period group than in the end period control group and each period of the operation (P ＜ 0.01-0.05). As compared with homochronous control group, the cardiac index in 10-day group, 30-day group, 60-day group and end period group was 51.21%,50.58%,55.84% and 33.91% respectively, and all were 60% lower than the normal value. ② Results of Morris water maze: Logarithm latency of 10-day operation group after training of 8 times was obviously longer than that of the 10-day sham-operation group [F(1,28) =5.997,P=0.021]. As for cognitive function, it was obviously worse in the 10-day operation group than that of the 10-day sham-operation group (x2=8.142 ,P ＜ 0.005). There was no significant difference between the 30-day operation group and the 30-day sham-operation group, and there was no significant difference in the logarithm latency as well between the 60-day operation group and the 60-day sham-operation group. As for searching strategies, the good rate of general cognitive ability in the 60-day operation group was worse than that in the 60-day sham-operation group (x2=4.988 ,P ＜ 0.05). Logarithm latency prolonged significantly in the heart failure end period group than in the end period control group [F( 1,6)=19.567 ,P=0.004], and the good rate of general cognitive ability was worse in the heart failure end period than that in the end period control group(x=1 1.82,P ＜ 0.01).CONCLUSION: Spatial learning and memory ability after heart failure in rats with myocardial infarction was injured obviously 10 days after the operation, and it recovered to the normal level 30 days after the operation.Cognitive injury occurred to some extent 60 days after the operation, and the cognitive function was obviously worse at the end period of congestive heart failure.

Effect of promoting blood flow therapy on HSP70 and ptk2 in gastric mucosa of precancerous lesion of CAG in rats / 中华中医药杂志

Objective:To investigate the differentially expressed genes of gastric mucosa of precancerous lesion of chronic atrophic gastritis (CAG) in rats intervented by promoting blood flow therapy of TCM,and to analyze its mechanism at genetic level. Methods:The model of precancerous lesion of CAG rats was induced by spring insertion and intragastric administration of NaCl solution and hot paste. To detect the differentially expressed genes of precancerous lesion of CAG rats by gene chip technology,and the main differentially expressed genes were indentified by real-time PCR. Results:Compared with blank group,HSP70 and ptk2 significantly reduced in natural recovery group,the HSP70 expression increased signifi cantly after being treated with supplementing qi for promoting blood flow,and ptk2 expression increased obviously after being treated with regulating qi and promoting blood flow. The results of real-time PCR in expression level of HSP70mRNA and ptk2mRNA were the same as that of the gene chip technology. Conclusion:Supplementing qi for promoting blood flow can upregulate HSP70,thus inhibit precancerous lesion of CAG in rat gastric mucosal cell apoptosis and promote mucosal repair; Regulating qi and promoting blood flow can up-regulate ptk2,sequentially regular remodeling of actin cytoskeleton and promote repairing and healing of injury gastric mucosal. These may be one of mechanism that supplementing qi for promoting blood flow and regulating qi and promoting blood flow treat precancerous lesion of CAG.

Effect of high glucose on expression of human brain microvascular endothelial cellular adhesion molecule and formation of endogenetic toxins / 中华中医药杂志

Objective:To explore the effect of high glucose on formation of endogenetic toxins and expression of intercellular cell adhesion molecule-1(ICAM-1) and vascular cell adhesion molecule-1(VCAM-1) of human brain microvascular endothelial cells.Methods:Cultured HBMEC was used as target cells,anhydrous glucose was added to the cell medium to prepare the injury model of human endothelial cell.Methyl thiazolyl tetrazolium method was used to examine the cell activity in a high glucose environment.Immunocytochemistry method was used to detect the expression of ICAM-1 and VCAM-1 of HBMEC.Results:The expressions of ICAM-1 and VCAM-1of HBMEC were enhanced by high glucose after pretreating HBMEC for 24,48 and 72h(P

Effect of PTS on praxiology, pathomorphology and serum IL-6 in rats with middle cerebral artery occlusion / 中华中医药杂志

Objective:To investigate the effect of PTS on the related pathophysiological changes and serum IL-6 level at different time points after the middle cerebral artery occlusion (MCAO) in rat’s brain.Methods:The model of focal cerebral ischemia of rat was established by the suture-occluded method.The effect of PTS on the behavioral disturbance,the pathological change of ischemia tissue and the level of serum IL-6 at different time points(3d,7d,28d) after MCAO and cerebral ischemia in rats were studied.ResultsAfter MCAO,different degrees of motion disturbances were observed in all rats.The infarct spot can be observed in all groups except the normal.The nerve cell necrosis can be found in all rats after 24 hour of MCAO.The result showed that PTS could improve the degree of motion disturbance,reduce the infarct spot obviously;after MCAO,the serum IL-6 were increased obviously but could be reduced by PTS.Conclusion:PTS can protect the neurons,reduce and eliminate the infiammatory reaction of cerebral ischemia cascade reaction effectively,which may relate to its inhibiting effect on cell factors such as IL-6 production after cerebral ischemia.

Effect of new QingKaiLing injection on adhesion molecules expression of an in vitro ischemia and reperfusion injury model of rat brain microvascular endothelial cell / 中华中医药杂志

Objective: To observe the expression of cell adhesion molecules ICAM-1 and VCAM-1of cultured rat brain microvascular endothelial cells(MVEC),expecting to explore the mechanisms of new QingKaiLing injection protecting brain from injury of inflammatory cascade in cerebral ischemia diseases.Methods: Rat cerebral MVEC were extracted by separating microvessel sections and collagenase enzymatic digesting,an in vitro ischemia reperfusion model was established(Kreb,95%N2+5%CO2),the protein and mRNA expression of ICAM-1 and VCAM-1 were detected by using immunocytochemical stain and RT-PCR method.Results:The expression of adhesion molecules of model group were significantly higher than those of noral group(P

Apoptosis-promoting effect of Panax notoginseng extracts on MNNG-transformed GES-1 cells / 中西医结合学报

OBJECTIVE: To study the apoptosis-promoting effect of the serum from Panax notoginseng extracts-fed dog on precancerous gastric cells by means of flow cytometry. METHODS: In the experiment, we adopted eternalized human gastric mucosa epithelium GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) (MC cells) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at two different points of time (2 and 6 hours) after feeding the dog with Panax notoginseng extracts for experiment. The MC cells were cultured in mediums with different concentrations of the medicated serum at 2- or 6-hour point of time for 72 hours. By means of flow cytometry, we examined the apoptosis-promoting effects of the serums on the MC cells. RESULTS: The medicated serums at these 2 points of time had significant effects in promoting MC cell apoptosis. The proportions of apoptotic cells in culture mediums with medicated serums had a significant increase as compared with those in culture mediums with non-medicated serums (serum obtained before administration of extracts to the dog) under the same conditions (P<0.05). The number of MC cells in G(0)/G(1)phase was decreased (P<0.05) and that in G(2)/M phase increased (P<0.05), while no consistent changes were observed in S phase. CONCLUSION: The medicated serums obtained at the two different points of time have significant apoptosis-promoting effects on MC cells. They decrease the number of MC cells in G(0)/G(1) phase and increase the number of MC cells in G(2)/M phase. This is probably responsible for the effects of Panax notoginseng extracts in inhibiting the proliferation of MC cells and promoting its apoptosis.

Treatment of lower limb ischemia with combination of traditional Chinese medicine and transplantation of autologous bone marrow mononuclear cells: a report of 23 cases / 中西医结合学报

OBJECTIVE: To evaluate the therapeutic effect of transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine for the treatment of limb ischemia. METHODS: Twenty-three patients with limb ischemia were treated. G-CSF was used to stimulate the bone marrow. The mononuclear cells were separated from the aspirated bone marrow fluid in the stem cell studio. The cell amount was above 1x10(9). The transplantation was performed by the way of intra-muscular multi-injection. Traditional Chinese medicine for replenishing qi to activate blood was prescribed from the first day after operation. The pain, poikilothermia, ulcer or necrosis and ankle/brachial index (ABI) of the ischemic limb were evaluated before and after the treatment. RESULTS: The pain score and poikilothermia score decreased one month after the transplantation, with distinct differences as compared with the scores before the treatment (P<0.05). The ABI increased gradually after the treatment, and one month after the treatment, it was 0.15 higher than that before the treatment. CONCLUSION: Transplantation of autologous bone marrow mononuclear cells combined with traditional Chinese medicine can decrease the symptoms and signs of severe lower limb ischemia effectively, and improve the circulation of the ischemic area.

Inhibiting effects of Panax notoginseng extracts on proliferation of GES-1 cells and MNNG-transformed GES-1 cells / 中西医结合学报

OBJECTIVE: Through cell cultivation, we studied the inhibiting effects of the serum of the dog fed with Panax notoginseng extracts on precancerous gastric cells, trying to find the best time points or periods when the extracts' function was the strongest after administration of the extracts to the dog. METHODS: The experiments adopted eternalized human gastric mucosa epithelium GES-1 cells and MC cells gained from GES-1 cells transformed by N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) as the model of precancerous lesions for study in vitro. We took the serum of a dog before and at different points of time after feeding the dog with Panax notoginseng extracts for experiment. By means of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we examined the inhibiting effects of the serum after culturing the GES-1 and MC cells for 72 hours with different concentration (8%, 4%, 2%) of medicated serum obtained from the dog at different points of time, so as to find that, at which points of time the medicated serum obtained, it would be the most effective. RESULTS: The results showed that the GES-1 and MC cells inhibition rates of medicated serum from the points of 2-hour and 6-hour were the highest, and the culture medium containing 8% of medicated serum from these two points had prominent inhibiting effects on both kinds of cells. The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 2-hour was 70.8% (P<0.01) and that of the MC cells was 45.3% (P<0.01). The GES-1 cells inhibition rate in culture medium containing 8% of medicated serum from the point of 6-hour was 88.5%(P<0.01) and that of the MC cells was 42.4% (P<0.01). CONCLUSION: The points of time with the strongest inhibiting effects are 2 hours and 6 hours after being fed with Panax notoginseng extracts. At these two points, the serum is most effective in inhibiting the proliferation of GES-1 and MC cells.

The detection and statistical analysis of blood lymphocyte immunophenotype / 中国免疫学杂志

Objective:To establish the reference values on blood lymphocyte immunophenotype by means of standardized quality control and proper statistical analysis.Methods:The whole bloods of 93 healthy Chinese adults were stained by fluorescent conjugated monoclonal antibodies of Simultest TM IMK lymphocyte Kit and then were analysed by FACS Calibur flow cytometer.The data were acquired and were analysed by SimulSET.Statistical analysis would be conducted after conforming the date of standardized quality control.Results:CD + 3,CD + 3CD + 4,CD + 3CD + 8 cells showed normal distribution but CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 showed skewness distribution .the 95% reference of confidence interval of CD + 3?CD + 3CD + 4?CD + 3CD + 8 cells were established by the method of normal distribution and they were respectively 49 28%～80 52%?25 29%～48 11%?11 93～44 77%;the 95% reference of confidence interval of CD - 3CD + 19 ,CD - 3CD + 16 and/or CD + 56 cells,ratio of CD + 3CD + 4 and CD + 3CD + 8 were established by the method of percentiles and they were respectively 4 74%～19 84%?10 63%～42 86%?0 65～3 05.Conclusion:To establish the reference values,it is considered that the methods of normal distribution and percentiles should be use comprehensively.It is also suggested that the reference values should be confirmed by means of standardized quality control and proper statistical analysis,so the reference values obtained from various laboratories of the same race but in different regions with the same reagents and instruments performed compatibility.

Effect of Ginsenoside Rb _1 on apoptosis induced by hypoxia/hypoglycemia and reoxygenation in cultured rat hippocampal neurons / 中国病理生理杂志

AIM: The aim of this work is to investigate the protective effects of Ginsenoside Rb 1(Rb 1) on apoptosis induced by hypoxia /hypoglycemia and reoxygenation in cultured rat hippocampal neurons. METHODS: Apoptosis were measured by flow cytometry; Morphological changes and neuronal necrosis were examined under microscope; The leakage of lactic dehydrogenase(LDH) and the product of nitric oxide(NO) were measured. RESULTS: hypoxia /hypoglycemia cultures for 5 h and reoxygenation induced neuronal apoptosis and necrosis,and significantly increased the leakage of LDH and the product of NO. The effects were enhanced with the extending time of reoxygenation. Rb 1 could significantly decrease the percentage of neuronal apoptosis and necrosis, and reduce the leakage of LDH and the product of NO. CONCLUSION: Rb 1 had an effect of anti-neuronal apoptosis. This effect might be related to the inhibition of the activity of NO synthase and NO production.