1.Experimental Study on the Effect of KangXin oral solution on brain mitochondrial DNA deletion mutation in aged Balb/c mice
Hai-Ling LIANG ; Xiao-Ru ZHANG ;
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(05):-
Objective: To explore the effect of KangXin oral solution on brain mitochondrial DNA(mtDNA) deletion mutation in aged Balb/c mice.Methods: The Balb/c mice were divided into the young group(6weeks),middle-aged group(6months) and aged group(14 months),each group has 10 mice.Brain mtDNA were obtained and polymerase chain reaction(PCR) technique was used to examine the fragment deletion of brain mtDNA,thus,to confi rm there was deletion of mtDNA in aged mice,s brain.The aged Balb/c mice were divided into two groups: the aged blank control group being given 0.9% normal sodium,KangXin oral solution group.After being treated for four months,the brain mtDNA were obtained and polymerase chain reaction(PCR) technique was used to amplify wild-type and deletion from of mtDNA.Gel imaging meter was used to detect optical density,then,to compare the optical density ratios of deletion from mtDNA/ wide type mtDNA in two groups.Results: There were 304bp mtDNA deletion in brain mitochondrial of aged Balb/c mice,but same mtDNA deletions were not detected in brain mitochondrial of young and middle-aged mice.Compared with aged blank control group,the mtDNA deletion of aged Balb/c mice in KangXin oral solution group decreased obviously(P〈0.001).Conclusion: mtDNA deletion mutation accumulates with the increase of age.KangXin oral solution can inhibit mtDNA deletion of aged mice.
2.Effect of Digital Auditory Activation and Touching Intervention on Infants with Cerebral Dysfunction
Li-ru LUO ; Ling ZHANG ; Li-qiong SHI
Chinese Journal of Rehabilitation Theory and Practice 2006;12(10):842-843
ObjectiveTo observe the effect of digital auditory activation and touching intervention on infants with cerebral dysfunction.Methods388 infants with perinatal high-risk factors and abnormal result of Denver Development Screening Test (DDST) were randomly divided into group A (n=135), group B (n=128) and group C (n=125), and treated with digital auditory activation combined with touching (group A), simple touching (group B) and simple drug (group C) with 10 days as a course. All infants were tested with DDST after one therapeutic course, and tested again with DDST after the infants of group B and group C treated continuously for six therapeutic courses; and all infants were assessed with the Gesell development quotient (DQ) after six months.ResultsAfter one therapeutic course, normal rate of DDST was 71.11% in group A, 26.69% in group B and 20.00% in group C. After six therapeutic courses, that was 90.37 % in group A, 62.50 in group B and 40.00% in group C. After six months, the children with the Gesell DQ over 86 scores was 125 (92.60%) in group A, 90 (70.31%) in group B and 62 (49.60%) in group C. There were significant differences among three groups ( P<0.01).ConclusionThe digital auditory activation combined with touching has short therapeutic course and high efficacy, so it is a good therapeutic method for infants with cerebral dysfunction.
3.Detection of Serum S-100? in Children with Acute Carbon Monoxide Poisoning and Its Clinical Significance
yu-hong, CAO ; guang-yun, ZHANG ; guo-cheng, ZHANG ; cui-ling, DING ; ru-ying, LI
Journal of Applied Clinical Pediatrics 2006;0(18):-
Objective To explore the changes serum S-100? in children with acute carbon monoxide poisoning and its clinical significance.Methods The levels of serum S-100? of 28 children with acute carbon monoxide poisoning and those of 20 healthy children were mea-sured by enzyme-linked immunosorbent assay.Results The serum S-100? levels of the study group and control group were(0.517?0.346)and(0.037?0.014)?g/L respectively,there was significant difference between two groups(t=6.197 P
4.Reconstruction of corneal surface layer using human amniotic epithelial cells modified by lentiviral vector-mediated EGFP gene
Ling, JIN ; Jian, CHEN ; Jing, WU ; Jin-tang, XU ; Qing, ZHOU ; Ru-shan, YE ; Hong, ZHANG
Chinese Journal of Experimental Ophthalmology 2011;29(8):685-689
Background Studies demonstrated that human amniotic epithelial cells (AECs) have some characteristics of embryonic stem cells and they were used to re-establish the surface of eyes. Human AECs may serve as new seed cells in tissue engineering for corneal epithelium reconstitution in the future. Objective The present study was to investigate the application value of human amniotic epithelium cells transfected by lentiviral vectormediated enhanced green fluorescent protein (EGFP) gene as new seed cell source for engineering the corneal surfacelayer. Methods Lentiviral vector carrying the objective gene EGFP was transfected into human amniotic epithelial cells (pLenti6/V5-DEST),and the transient expression of the transgene in the human amniotic epithelial cells was observed under the fluorescence microscope. Flow cytometry was used to detect the positive expression rates of EGFP in transfected cells. The transfected human amniotic epithelial cells were seeded onto the fresh corneal stromal surface of New Zealand white rabbit and cultured in vitro. The stem cell deficiency ( SCD ) models were established by cutting off the limbus of cornea in 20 eyes of New Zealand white rabbits, and the model rabbits were then divided into 2 groups randomly. The transplanted grafts carrying the pLenti6/V5-DEST-EGFP gene-transferred human amniotic epithelium cells were regarded as the pLenti6/V5-DEST-EGFP group, and the corneal stroma graft without any epithelial cell served as the control group. The opacity of stroma and corneal conjunctivalization and vascularization were observed daily. The rabbits' eyes were extracted one month after operation. The expression of EGFP in the cornea was detected under the fluorescence microscope, and the expression of CK8, CK18 and CK12 in cornea was detected by immunohistochemical staining. Results The shape of the transferred human amniotic epithelial cells resembled normal human amniotic epithelial cells. 48 hours after the transient transfection of EGFP presented with the highest expression level throughout the observation duration, with a positive expression rate of EGFP of 61.5% ,showing significant differences in comparison with that of 12 ( 5.24% ) , 24 ( 38.27% ) or 96 ( 39. 10% ) hours ( P <0. 05) post-transfection; but no obvious difference was found in the positive rate of transiently transfected EGFP between 48 hours and 72 hours ( 58.36% ) ( P>0. 05 ). Six cornea grafts were clear in 1 month and two corneas were rejected during the observation period in the pLenti6/V5-DEST-EGFP group. A few new blood vessels were seen around the graft. Ten corneas of the control group became opaque and cloudy with new blood vessels growth around the grafts. Imunohistochemistry revealed the positive expressions of CK8, CK1 8 and CK12 in the corneal epithelial layer in the pLenti6/V5-DEST-EGFP group. However,the expression of CK12 was absent in the control group. Conclusion Human amniotic epithelium cells transfected with the pLenti6/V5-DEST-EGFP gene is a new and ideal feed cell type to reconstruct the corneal surface layer. Lentivirus is a relatively safe gene transfection vector.
5.Evaluation of Myocardial Ischemia of Kawasaki Disease at Convalescence Period by Radioactive Nuclide Myocardial Perfusion Imaging
huan-bin, LI ; qi, ZHANG ; ling, WANG ; mao-ping, CHU ; ru-lian, XIANG
Journal of Applied Clinical Pediatrics 1986;0(01):-
Objective To discuss the clinical value of 99Tcm-sestamibi(99Tcm-MIBI) myocardial perfusion imaging on detecting myocardial ischemia in children with Kawasaki disease(KD) at convalescence period.Methods Twenty-one children wih KD at convalescence period were divided into 2 groups according to results of echocardiography.Four cases with coronary artery dilation,17 cases without coronary artery dilation.All cases accepted dipyridamole stress myocardial perfusion imaging.These patients who had positive results were given rest myocardial perfusion imaging again next day.Results Among 21 cases,9 cases(42.8%) were positive in perfusion imaging.Four cases with coronary artery dilation showed myocardial ischemia in different degree detected by myocardial perfusion imaging.Among 17 cases without coronary artery dilation,5 cases(29.4%) were positive.Conclusions Compared to echocardiography,99Tcm-MIBI myocardial perfusion imaging can objectively evaluate the location,extent and degree of myocardial ischemia of children with KD.It will be a routine test in observing its phase development.
6.The clinical application of the modified radiography of the tempro-mandibular joint by digital radiography
Ru-Yao ZHUANG ; Xian-Ling ZHANG ; Xian-En CHEN ; Mao-Xiong WU ;
Chinese Journal of Primary Medicine and Pharmacy 2006;0(09):-
Objective To investigate the clinical application value of the modified radiography by digital ra- diography of the tempro-mandibular joint in the tempro-mandibular joint radiography.Methods A digital radiogra- phy machine(Siemens Aristos MX)was used to the tempro-mandibular joint disorders of 68 patients with the meth- ods of the modified radiography of the tempro-mandibular joint,and the results were compared with those of 45 cases acquired with conventional radiography.Results The modified radiography by digital radiography provided high res- olution,precise location and excellent images,and the total structures of tempro-mandibular joint was clearly dis- played,with a success rate of 99%(67/68),while the results acquired by conventional radiography were not clear, only with a success rate of 60%(18/45).There is significant statistical differences between the modified radiography by digital radiography and conventional radiography(x~2 = 35.08,P
7.Effects of aspirin on the expression of nuclear factor-κB in a rat model of acute pulmonary embolism
Ling-Cong WANG ; Rong-Lin JIANG ; Wei ZHANG ; Li-Ling WEI ; Ru-Hui YANG
World Journal of Emergency Medicine 2014;5(3):229-233
BACKGROUND: Acute pulmonary embolism (APE) is a disorder involving the pulmonary circulation resulting from a blockage of the pulmonary artery. The present study aimed to investigate the effects of aspirin on the nuclear factor-κB (NF-κB) activity in a rat model of APE. METHODS: A total of 108 healthy male Sprague-Dawley rats were randomly assigned into six groups (n=18 rats per group): control group, sham operation group, APE model group, and low-, medium- and high-dose aspirin groups. Six, 24, and 72 hours after the induction of APE, rats in the low-, medium- and high-dose aspirin groups were given aspirin at a respective daily dose of 150, 300, and 600 mg/kg by gavage for three consecutive days. Rats in the other groups were treated with equal volumes of normal saline. Six rats in each group were anesthetized with 10% chloral hydrate solution at each time point, and then the lung tissues were colected and analyzed using immunohistochemical staining. RESULTS: Positive immunohistochemical staining was present in the bronchial epithelial cells, alveolar cells, macrophages, and surrounding bronchial smooth muscle cells. When compared with the APE model group, the number of positive cells was significantly lower in the other groups at each time point (P<0.001). Statistically significant differences were also observed among the aspirin-treated groups at 6 hours (P<0.05,P<0.001). Compared with the APE model group, NF-κB protein expression was reduced in the other groups at each time point (P<0.05,P<0.001). Rats from the APE model group had thrombosis, damaged alveolar walls, and pulmonary hemorrhage, along with different degrees of infl ammatory cellular infiltration at each time point. However, pathological changes such as pulmonary hemorrhage and infiltration of inflammatory cells were attenuated after the aspirin treatment. CONCLUSION: Aspirin can significantly inhibit NF-κB activity in the lung of rats with APE in a dose-dependent manner, and can alleviate lung injury after APE.
8.Effects of carbaryl on serum steroid hormone and the function of antioxidant system in female rats.
Yang QIU ; Jian-feng CHEN ; Ling SONG ; Jun HE ; Ru LIU ; Chun-win ZHANG ; Xin-ru WANG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2005;23(4):290-293
OBJECTIVETo investigate the effects of carbaryl on serum steroid hormone and function of antioxidant system in female Sprague Dawley rats.
METHODSCarbaryl was administrated to the adult female rats at doses of 0, 1.028, 5.140 and 25.704 mg x kg(-1) x d(-1) for 30 d. Vaginal smears of rats were taken to determine estrous cycle. Serum 17beta-estradiol (E(2)) and progesterone (P(4)) concentrations were measured by radioimmunoassay. The activities of superoxide dismutase (SOD) and glutathione-s-transferase (GST), and the levels of malondialdehyde (MDA) and glutathione (GSH) were measured by spectrophotometry.
RESULTSThe number of estrous cycle in exposed groups were obviously lower than in control group (P < 0.05, P < 0.01). Body weight gain in high dose group (25.704 mg x kg(-1) x d(-1)) was significantly lower than that in control. Meanwhile, the organ coefficient of ovary and uterus declined in a dose-dependent manner. Serum E(2) level [(19.93 +/- 2.21) nmol/L] in 25.704 mg group was lower than in control group [(28.76 +/- 6.12) nmol/L, P < 0.05], and P(4) level (1.21 +/- 0.40) nmol/L in 1.028 mg group was higher than that in control group [(0.63 +/- 0.39) nmol/L, P < 0.05]. The activity of SOD first reduced then rose in ovary, and first rose then reduced in serum. The contents of MDA increased in ovary, while decreased in the serum. GSH contents and GST activity increased in ovary, while in serum GSH contents decreased and GST activity first increased then decreased.
CONCLUSIONCarbaryl could disrupt estrous cycle and affect serum steroid hormone, and the function of antioxidant system in female SD rats.
Animals ; Antioxidants ; metabolism ; Carbaryl ; toxicity ; Estradiol ; blood ; Female ; Glutathione ; blood ; Glutathione Transferase ; blood ; Malondialdehyde ; blood ; Progesterone ; blood ; Rats ; Rats, Sprague-Dawley ; Superoxide Dismutase ; blood
9.Construction of plant expression vectors with PMI gene as selection marker and their utilization in transformation of Salvia miltiorrhiza f. alba.
Ru TAO ; You-Can ZHANG ; Qian FANG ; Ren-Jiu SHI ; Yan-Ling LI ; Lu-Qi HUANG ; Gang-Ping HAO
China Journal of Chinese Materia Medica 2014;39(7):1209-1213
OBJECTIVETo construct plant expression pCAMBIA1301-PMI by substituting PMI for hygromycin resistance gene in pCAMBIA1301 and obtain transgenic Salvia miltiorrhiza f. alba using PMI-mannose selection system.
METHODThe 6-phosphomannose isomerase gene (PMI) of Escherichia coli was amplified by PCR. Sequence analysis showed that it shared 100% amino acids identities with the sequences of PMI genes isolates reported in the NCBI. Based on pCAMBIA1305, the plant expression pCAMBIA1305-PMI was constructed successfully by substituting PMI for hygromycin resistance gene in pCAMBIA1305. pCAMBIA1305-PMI was transformed into Agrobacterium tumefaciens LBA4404, and then the leaves of S. miltiorrhiza f. alba were inoculated in LBA4404 with pCAMBIA1305-PMI.
RESULTPlant expression pCAMBIA1301-PMI was successfully constructed and the leaves of S. miltiorrhiza f. alba inoculated in LBA4404 with pCAMBIA1305-PMI were selected on medium supplemented with a combination of 20 g x L(-1) mannose and 10 g x L(-1) sucrose as a carbon source. The transformation efficiency rate was 23.7%.
CONCLUSIONGenetic transformation was confirmed by PCR, indicating that a new method for obtaining transgenic S. miltiorrhiza f. alba plants was developed using PMI-mannose selection system.
Anti-Bacterial Agents ; pharmacology ; Biomarkers ; Cinnamates ; pharmacology ; Escherichia coli ; enzymology ; genetics ; Escherichia coli Proteins ; genetics ; metabolism ; Gene Expression ; Genetic Vectors ; genetics ; metabolism ; Hygromycin B ; analogs & derivatives ; pharmacology ; Mannose-6-Phosphate Isomerase ; genetics ; metabolism ; Plants, Genetically Modified ; drug effects ; genetics ; metabolism ; Salvia miltiorrhiza ; drug effects ; genetics ; metabolism ; Transformation, Genetic
10.Study on irritation of calcium oxalate crystal in raw Pinellia ternata.
Ling-Yun ZHONG ; Hao WU ; Ke-Wei ZHANG ; Qian-Ru WANG
China Journal of Chinese Materia Medica 2006;31(20):1706-1710
OBJECTIVEConfirm the irritation of needle-like calcium oxalate crystals in raw Pinellia ternata.
METHODComparing the irritations of raw P. ternate containing calcium oxalate crystals, the raw P. ternate no containing calcium oxalate crystals, the pure needle-like calcium oxalate crystals isolated from raw P. ternata, the extracts of water and various solvents from raw P. ternate. by using the model of rabbits' eyes. Studying the quantity effect relationship of different concentration suspensions of needle-like calcium oxalate crystal isolated from raw P. ternate on rabbits' eyes. Observing the shape and appearance of calcium oxalate crystals in raw P. ternate and raw India Madder Root by the electro microscope and comparing their irritation degrees with the same contents of calcium oxalate crystals.
RESULTCalcium oxalate crystals in raw P. ternata showed very strong irritation property. Under the same content of calcium oxalate crystals, the irritation of raw P. ternata and pure needle-like calcium oxalate crystals isolated from raw P. ternate had no significant difference. The concentrations of needle-like calcium oxalate crystals were do relative to the degree of irritation on rabbits' eyes and they showed undoubted quantity-effect relationship.
CONCLUSIONCalcium oxalate crystal is the irritation component in raw P. ternata.
Animals ; Calcium Oxalate ; chemistry ; isolation & purification ; toxicity ; Conjunctiva ; drug effects ; Cornea ; drug effects ; Crystallization ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; isolation & purification ; toxicity ; Edema ; chemically induced ; Eye Diseases ; chemically induced ; Female ; Iris ; drug effects ; Male ; Pinellia ; chemistry ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Rabbits ; Random Allocation ; Rubia ; chemistry