1.Effect of mesenchymal stem cell used on prevention and treatment for acute pulmonary injury induced by paraquat poisoning in rats
Yanmin ZHANG ; Zewu QIU ; Guangxian LIU
Chinese Journal of Emergency Medicine 2011;20(1):39-43
Objective To explore the possible mechanism and protective effect of mesenchymal stem cell (MSC) on rats with paraquat-induced acute lung injury. Method The solution of 20% paraquat (PQ) in dose of 18 mg/kg was injected intra-peritoneally into rats to induce poisoning,and phosphate buffered solution (PBS) was given to rats instead of PQ in rats of control group. Eighty specific pathogen free (SPF) Wistar rats were randomly divided into four group: PQ plus PBS group (n = 20), PQ plus MSCs group (n = 20), MSCs plus PBS group (n=20), normal group (n = 20). The forth generation of MSCs were transfected with Ad5-EGFP virus vector, and then the MSCs-EGFP was delivered to rats through the tail vein of rats 4h after PQ. Five rats of each group were sacrificed 1 d, 3 d, 7 d and 28 days after MSCs administration, and lung tissues of rats were taken to make sections for histological observation of the migration of MSCs under fluorescence inverted microscope. The lung tissues of rats sacrificed on the 28 th day after PQ poisoning were taken for detecting pulmonary coefficient and the content of hydroxyproline (HYP) in the lung tissue homogenate, and at the same time, the levels of serum transforming growth factor-β1(TGF-β1) were assayed. Results The pathological changes of lung tissue showed that the pulmonary fibrosis and consolidation in the MSCs treatment group were milder than those in PQ poisoning model group. In the MSCs treatment group, the levels of serum TGF-β1 and lung tissue HYP, and pulmonary coefficient were lower than those of PQ poisoning model group (P<0.05). Conclusions The use of MSCs for treatment of paraquat intoxication can protect pulmonary structure by decrease in TGF-B1 and inhibiting the fibroblast migration, suppressing the production of collagenous protein.
2.Relationship between independent risk factor of ischemic cerebrovascular disease in youth and genic mutation of methylene tetrahydrofolate reductase
Guangxian NAN ; Mingji CUI ; Songyan LIU
Chinese Journal of Tissue Engineering Research 2006;10(4):184-186
BACKGROUND: The increase of concentration of plasma homocysteine is the independent risk factor of atherosclerotic and thrombotic cerebral infarction. Genic mutation of methylene tetrahydrofolate reductase (MTHFR), which is the metabolic enzyme of homocysteine in thansulfuration and remethylation,can induce the elevation of the concentration of plasma homocysteine.OBJECTIVE: To explore the relationship of hyperhomocysteinemia and genic mutation of MTHFR of homocysteine with ischemic cerebrovascular disease in youths.DESIGN: Case-control observation,SETTING: Department of Neurology, China-Japan Friendship Hospital,Jilin UniversityPARTICIPANTS: 100 young patients with cerebral infarction, who were hospitalized within 2 days after episode at the Department of Neurology,China-Japan Union Hospital, Jilin University between April 2003 and December 2004, were enrolled as case group, 73 males and 27 females, aged 27-45 years old with an average of (42±5) years. 100 cases in control group were healthy people receiving health examination in the same period, 70 males and 30 females, aged 18-45 years old with an average of (39±4) years.METHODS: The homocysteine in fasting plasma of testees was detected with high performance liquid chromatograpy (HPLC). C677T site and A1298C site of MTHFR gene were analyzed with polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and detected with armplification refractory mutation system (ARMS).MAIN OUTCOME MEASURES: Detection of MTHFR C677T and A1298C gene; Relationship between concentration of plasma homocysteine and MTHFR genotype.RESULTS: A total of 100 inclusive patients and 100 normal control people were involved in the result analysis. ①Detection of MTHFR C677Tand A1298C gene: Distribution of genotype, frequency of homozygote and frequency of allele of MTHFR C677T in the case group and control group had significant difference (P < 0.01 ) while the distribution of genotype,frequency of homozygote and frequency of allele of MTHFR A1298C gene in the case group and control group had insignificant difference (P > 0.05 ). ②Relationship between the concentration of plasma homocysteine and MTHFR genotype: The concentration of plasma homocysteine between MTHFR C677T and A1298C genotype had significant difference (P< 0.001 ). The mutant result LSD-t check of the 2 sites showed that mean difference of homozygote and heterozygote, homozygote and concentration of wild type homocysteine had statistical significance (P < 0.05). The mean difference of MTHFR C677T and A1298C heterozygote and concentration of wild type homocysteine in plasma had no statistical significance (P> 0.05 ).CONCLUSION: The mutation of MTHFR C677T and A1298C leads to the marked increase in the concentration of plasma homocysteine. The MTHFR C677T polymorphism site is the independent risk factor of is chemic cerebrovascular disease in youths. The genic mutation of MTHFR A 1298C has no correlation with the attack of ischemic cerebrovascular disease of youths.
3.Significance of nucleolar organizer regions associated proteins of T lymphocytes in diagnosis and mornitoring of cancer
Guangxian LIU ; Xiantao KONG ; Jing ZHANG
Chinese Journal of Laboratory Medicine 2001;0(03):-
Objective To analyze the Ag NORs in T cells of different patients to find clues for the diagnosis and monitoring of cancer.Methods A KL 2 imaging system was used to analyze the areas of Ag NORs in T cells. T cells from peripheral blood were cultivated and stimulated, and silver staining was performed. The areas of Ag NORs were analyzed.Results A total of 1 046 nomal adults, 393 patients with non malignant diseases and 706 patients with malignant diseases were analyzed. Their average IS% values were (7.81?0.69)%, (7.85?0 72)% and 5.17?0.87 respectively. The IS% values above 6% were 99.8%, 94.7% and 10% to 35% respectively. No differences were observed in different kinds of cancers except breast cancer and rectum cancer whose IS% were (5.61?0.86)% and (6.10?0.92)% respectively, 75% and 26% of their IS% were less than 6% respectively. 83% to 90% of the IS% in the patients with other cancers were below 6%. In comparison with serum CEA, AFP and the like, Ag NORs in T cells were more powerful in diagnosis and monitoring of cancers.Conclusion The value of Ag NORs of T cells was significantly lower in patients with cancers and was statistically different from that in nomal adults and patients without malignant diseases. The results demonstrated that the analysis of Ag NORs might play an important role in the diagnosis, differenciation and monitoring of cancer.
4.Influence of Ultra-micro-LiuweiDihuang Decoction on GH and SS of Intrauterine Growth Retardation Mice
Xiaoping PAN ; Baiyan LIU ; Guangxian CAI
Journal of Medical Research 2006;0(05):-
Objective To observe the effect of Ultra-micro-LiuweiDihuang Decoction on the embryo brain development and the influence on brain growth hormone(GH),Somatostatin(SS)in prengnant mice of passive smoking,then to demonstrate the relationship between brain and kidney in TCM.Methods Animals were divided into 5 groups at random:normal group(A),model group(B),traditional Liuwei Dihuang Decoction group(C),1/3 dosage of Ultra-micro-LiuweiDihuang Decoction group(D)and all dosage of Ultra-micro-LiuweiDihuang Decoction group(E).The IUGR model was established by passive smoking.On the 19th day of gestation,all mice were anatomized to scale the body and brain weight of lively embryos,then a part of brain tissue was homogenated.The levels of GH and SS in brain tissues were measured by ELISA.Results Passive smoking could decrease the body and brain weight,influence the level of GH and SS in brain tissues.Compared with A and B groups,the C,D and E groups could increase body and brain weight,regulate the level of GH and SS,improve the brain development of IUGR mice obviously(P
5.Comparative Study of Clinical Efficacy of Ultrafine Extracted Granule Preparation and Traditional Herbal Decoction of Li Zhong Tang
Qingping LIU ; Junfeng YAN ; Dongbo LIU ; Man LI ; Guangxian CAI
Chinese Journal of Information on Traditional Chinese Medicine 2014;(4):34-36
Objective To compare the clinical efficacy of different doses of ultrafine extracted granule preparation (EGP) and traditional herbal decoction (THD) of Li Zhong Tang in treatment of epigastric pain. Methods Sixty cases of epigastric pain patients in accordance with the diagnostic criteria of deficiency and cold pattern of spleen and stomach in TCM were randomly divided into THD group, 1/3 dose group and 1/5 dose group, and were given THD, 1/3 dose of ultrafine EGP and 1/5 dose of ultrafine EGP, respectively. The clinical efficacy of the three groups after one course of medication was comparatively analyzed. Results There were no significant differences in age, course of disease, symptom score before treatment, epigastric pain efficacy and syndrome curative effect among the three groups, the differences had no statistical significance (P>0.05). The severity, frequency and duration of epigastric pain were all reduced in the three groups, with significant differences between before and after treatment (P<0.05). Conclusion There are no significant differences in the clinical efficacy on epigastric pain among THD, 1/3 dose of ultrafine EGP and 1/5 dose of ultrafine EGP. In addition, the effect of 1/3 dose group is very close to the THD group.
6.Effect of Buyang Huanwu Decoction on vascular endothelial growth factor and its receptor Flk1 in rats after focal cerebral ischemia
Baiyan LIU ; Guangxian CAI ; Wei LIU ; Xuemei CHEN
Chinese Traditional and Herbal Drugs 1994;0(03):-
Objective To explore the effect of Buyang Huanwu Decoction (BYHWD) on vascular endothelial growth factor (VEGF) and its receptor, Fetal liver kinase 1 (Flk1) in rats after focal cerebral ischemia. MethodsRats were randomly divided into following groups including Sham group, model group, Nimodipine group, and BYHWD group. The model of focal cerebral ischemia in rats was repro-duced by middle cerebral artery occlusion. Rats were ig administered and killed after operated, then the expression of VEGF and Flk1 and protein level of VEGF in brain tissue of every groups were measured by immunohistochemisty and enzyme linked immunosorbent assay, and the nervous function deficit scores were evaluated. ResultsThere were a few VEGF and Flk1 positive cells in normal brain tissue of rats. After focal cerebral ischemia the VEGF and Flk1 positive cells were increased. Compared to model group, Nimodipine and BYHWD significantly improved the neurological behavior performance, increased the numbers of VEGF and Flk1 positive cells, and enhanced the VEGF protein level (P
7.Effect of Bu Yang Huan Wu decoction(补阳还五汤) on basic fibroblast growth factor in rats with cerebral ischemia
Fang LIU ; Xuesong BAI ; Baiyan LIU ; Guangxian CAI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2006;0(01):-
Objective To investigate the effect of Bu Yang Huan Wu decoction(BYHWD,补阳还五汤) on the expression of basic fibroblast growth factor(bFGF) and its mechanism of anti-cerebral ischemia.Methods Eighty-five Sprague-Dawley rats(body weight 280-300 g) were randomly divided into five groups: normal control group(n=5),sham-operated group,model group(n=20),traditional BYHWD group and ultra-fine powder BYHWD group.Then the latter four groups were further divided into four subgroups of 1,3,5 and 7 days(each n=5).The cerebral ischemic rat model was made by occluding the middle cerebral artery(MCA) with a thread.The traditional decoction and ultra-fine powder decoction groups were ingested with the liquid medicine of BYHWD after 2 hours of the operation(2 ml per rat per day,according to the surface area,it was equivalent to three times the dosage of 70 kg adult(human)).All groups were executed at the 1,3,5 and 7 days after the operation.At the same time there were 5 rats as the normal control group.The brain tissue was taken for the preparation of specimens.The distribution and expression difference of bFGF in the ischemic brain tissues of rats in different groups had been investigated with immuohistochemistry technique.The contents of bFGF in the brain tissue were detected with enzyme linked immunosorbent assay(ELISA).Results The positive expression of bFGF at peri-infarct area after ischemia increased obviously,but at central-infarct area there was less such positive expression.There was no expression at contra-lateral side to the ischemic area.The expression of bFGF immune positive cell increased at 1 day,peaked at 3 days,and returned to baseline at 7 days.The positive cells of bFGF in tradition decoction and ultra-fine powder decoction groups were obviously more than those in the model group at 3,5 and 7 days(all P
8.Association of killer immunoglobulin-like receptors with leukemia
Jia LIU ; Quanli WANG ; Guangxian LIU ; Xuedong SUN ; Yan TAN
Journal of Third Military Medical University 2003;0(09):-
Objective To study the correlation between the genes of killer immunoglobulin (Ig)-like receptors (KIR) in natural killer cells and leukemia. MethodsPrimers of 16 KIR genes, including KIR2DL1-5, KIR3DL1-3, KIR2DS1-5, KIR3DS1 and 2 pseudogenes 2DP1 and 3DP1, were designed and synthesized. A polymerase chain reaction with sequence-specific primers (PCR-SSP) was developed to detect the mRNA expressions of the 16 genes. Totally 46 healthy individuals and 46 leukemic patients (15 cases of acute myelocytic leukemia, 13 cases of chronic myelocytic leukemia and 18 cases of acute lymphocytic leukemia) were detected and the frequencies of the KIR genes were analyzed. ResultsThe frequency of 2DL4, 3DL2, 3DL3 and 2DP1 in healthy and leukemic patients were 100%, and were detected in all patients and healthy individuals. The other KIR genes had similar frequencies except those of 2DL2 and 2DS2, which were 47.8% of the healthy individuals and 89.1% in leukemic patients. There was a significant difference between the 2 groups.ConclusionThere may be an association between pathogenesis of leukemia and 2DL2 and 2DS2 genes.
9.Effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide in chronical stressed mice
Baiyan LIU ; Jian YI ; Xuemei CHEN ; Xiaoyuan LIN ; Guangxian CAI
Chinese Journal of Behavioral Medicine and Brain Science 2011;20(4):292-294
Objective To explore effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide(VIP) in the hypothalamus, spinal cord and duodenum of chronical stressed mice. Methods Mice were randomly divided into normal, stress and Simotang group( n= 10 in each group), and given a variety of unpredictable chronic mild stress. After 21 days gastric emptying and intestinal propulsion function were measured,the expression of VIP was detected by immunohistochemistry and RT-PCR. Results Compared with mice in normal group( (49.81 ± 8.56)%; (51.02 ± 5.11 )% ), chronic stress increased gastric residual rate( (61.53 ±8.71 ) %; P < 0.05 ) and reduced small intestine propulsion rate ( ( 31.79 ± 2.38 ) %; P < 0. 05 ). There were differences in expressions of VIP positive cells and mRNA in duodenum( (8.8 ± 1.1 )/mm2 and(0. 58 ±0.03) ),hypothalamus ( ( 12.9 ± 1.5 )/mm2 and (0.81 ± 0. 07 ) ) and spinal cord ( ( 12.1 ± 1. 2)/mm2 and (0.76 ± 0.02) )in chronic stress group compared with normal group( (6.5 ± 0. 9)/mm2 and (0.43 ± 0. 04);( 10.8 ± 1.3 )/mm2and (0.57 ± 0.03 ); (9.3 ± 1.5 )/mm2 and (0.53 ± 0. 02 ) respectively). There was not difference in gastric residual rate (52.93 ± 9.15 )%, small intestine rate(48.98 ± 4.38 )% and expressions of VIP positive cells and mRNA in duodenum ( (6.7 ± 0.9)/mm2 and (0.48 ± 0. 05 ) ), hypothalamus ( ( 10. 6 ± 1.4 )/mm2 and ( 0. 61 ± 0. 05 ) )and spinal cord ( (9. 1 ± 1.3)/mm2 and(0.55 ± 0.05 ) ) in Simotang group compared with those in normal group (P > 0.05 ), but there were decreased compared with those in chronic stress group (P < 0.05 ). Conclusion Simotang can regulate expressions of VIP in duodenum, hypothalamusand spinal cord in chronically stressed mice.
10.Effects of Buyang Huanwu decoction on protein kinase B1 and c-Jun amino terminal kinase 1/2 in rats after ;focal cerebral ischemia
Fang LIU ; Tianlei YIN ; Feiyue DAI ; Liangying LIAO ; Guangxian CAI
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2013;(5):275-278
Objective To explore the effect of Buyang Huanwu decoction(BYHWD)on protein kinase B1 (AKT1)and c-Jun amino terminal kinase 1/2(JNK1/2)in rats after focal cerebral ischemia. Methods According to the random number table method,48 Sprague-Dawley(SD)rats were randomly allocated to four groups:normal control group,sham-operated group,model group,traditional BYHWD group(each n=12). The rat model of right focal cerebral ischemia was established by the method of middle cerebral artery occlusion(MCAO). The rats in BYHWD group were ingested with the decoction of BYHWD 14.2 g/kg after 2 hours of the operation(the main ingredients of BYHWD including astragalus mongholicus 120 g,Chinese angelica 6 g,radix paeoniae rubra 4.5 g, rhizoma ligustici wallichii 3 g,safflower 3 g,peach kernel 3 g,earthworm 3 g),once a day for 7 days. Other groups of animals were given the same amount of normal saline orally. After operation,on the 7th day,the animals were killed,and their brains were taken out. The reverse transcription-polymerase chain reaction(RT-PCR)assay was used to detect AKT1 mRNA expression,and immunohistochemical method was applied to measure JNK1/2 protein expression. Results Compared with normal control and sham-operated groups,the level of AKT1 mRNA expression〔absorbance(A)〕was decreased obviously(0.48±0.08 vs. 0.63±0.11,0.61±0.09,both P<0.05),and the number of JNK1/2 positive cells(cell/mm2)was increased significantly(34.13±4.57 vs. 16.15±1.09,16.23±2.05,both P<0.05)in model group;compared with model group,the AKT1 mRNA expression in brain tissue(0.93±0.11)and the number of JNK1/2 positive cells(45.04±5.68)was increased significantly in BYHWD group,the differences being statistically significant(P<0.05 or P<0.01). Conclusion BYHWD can up-regulate expressions of AKT1 mRNA and JNK1/2 positive cells in ischemic brain tissue that is one of the mechanisms in the protection of brain.