Objective To investigate the effect of estrogen on expression of matrix GLA protein (MGP)in ovariectomized Sprague-Dawley(SD)rats and the role of estrogen in improving postmenopausal osteoporosis.Methods Thirty-six SD female rats were allocated into 3 groups randomly,every 12 rats in ovariectomized group(OVX group),estrogen group(E group)and control group(sham group).Rats in OVX and E group all underwent bilateral ovariectomy,those rats in E group were given by estradiol benzoate intramuscularly after 3 weeks of ovariectomy.Rats in sham group underwent bilateral lipectomy near the ovary.All rats were kept the urine and the serum every three weeks and were sacrificed after 15 weeks.The pathology changes of uterus,lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density(BMD)of lumbar vertebra of rats was determined by dual energy X ray absorptiometry(DEXA).The content of MGP in serum and urine was determined by ELISA.Expression of underearboxylated matrix GLA Protein(MGP)was detected by immunohistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerase chain reaction.Results(1)After 15 weeks of ovariectomized,the endometrium of uterus and lumbar vertebra exhibit remarkable pathologic changes in OVX group.The serum estrogen of(454±66)pmol/L in OVX group were lower than in(527 ±77)pmol/L in sham group and(556 ±80)pmol/L in E group significantly (P ＜ 0.05).The BMD of lumbar vertebra of(0.263 ± 0.030)g/cm2 in OVX group were lower than (0.295 ±0.024)g/cm2 in sham group and(0.279 ±0.024)g/cm2 in E group significantly(P ＜0.01).(2)The serum MPG protein in OVX group and E group showed decreased trends after ovariectomized,which were(104 ±64)ng/L in OVX group and(134 ±6)ng/L in E group at 9 weeks,which reached statistical difference(P ＜ 0.05).However,MGP in urine in sham group did not exhibit significant difference after 15 weeks of surgery(P ＞0.05).The MGP in urine of E group showed increased trends after 12 weeks of surgery,which reached(110.0 ±3.4)ng/L at 15 weeks,in the mean time,it was found that(86.5 ±2.5)ng/L of MGP in urine in OVX group,which showed significant difference(P ＜ 0.05).(3)MGP could be observed in lumbar vertebra in OVX group by immunochemistry staining.In the other two groups,the expression of MGP was not dominant.(4)Relative quantification of MGP mRNA expression in lumbar vertebra was defined as 1 in OVX group,when compared with 0.289 ±0.260 in E group and 0.103 ±0.098 in sham group,the difference showed statistically significant(P ＜ 0.01).Conclusion Estrogen could increase the expression of MGP mRNA and protein in ovariectomized rats and might play an important role in improving postmenopausal osteoporosis.

ObjectiveTo observe the effect of parathyroid hormone on expression of matrix GLA protein (MGP) in ovariectomized SD rats and primary osteoblast,and to study the role of MGP on the possible mechanism of postmenopausal osteoporosis.MethodsThirty-six Sprague-Dawley female rats were allocated into 3 groups,12 in each:sham operation group,ovariectomized group( OVX group),ovariectomized and parathyroid hormone treatment group.Animals in the parathyroid hormone group were injected parathyroid hormone (20 μg/kg,three times a week for 12 weeks) three weeks after ovariectomy.All rats were sacrificed after 18 weeks.Urine and serum were collected every three weeks.Lumbar vertebral bones were observed by immunohistochemistry.Bone mineral density (BMD) of lumbar vertebra of rats was determined.The content of MGP in serum and urine was determined by enzyme-linked immunosorbent assay (ELISA).Expression of undercarboxylated Matrix GLA Protein (ucMGP) was detected by immunochistochemistry.Relative quantification of MGP mRNA expression in lumbar vertebra bone was detected by Fluorescent real-time quantitative polymerause chain reaction.Results ( 1 ) 18 weeks after ovariectomy,BMD of lumbar vertebra in OVX group was lower than those in sham group and parathyroid hormone group significantly ( P＜0.05 ).(2) The content of MGP in serum and urine was dynamic variation after treatment hy parathyroid hormone,and it was significant compared with OVX group ( P＜0.05 ).( 3 ) Immunohistochemical localization of ucMGP was seen in lumbar vertebra in OVX group.(4) Relative quantification of MGP mRNA expression in lumbar vertebra in OVX group was increased significantly compared with other groups ( P＜0.01 ).( 5 ) parathyroid hormone ( 1-34 ) in 10-7mol/L,10-8mol/L,10-9 mol/L up-regulated MGP mRNA expression in primary osteoblasts about 6.78,5.31,and 2.23 times than control respectively.It was in a dose-dependent manner.ConclusionThe effect of parathyroid hormone on the expression of matrix gla protein may play an important role in mechanism of postmenopausal osteoporosis

Objective To observe the expression of matrix gla protein(MGP) mRNA in primary osteoblasts of Sprague-Dawley (SD) rat in vitro after treatment with anti-osteoporosis agents [vitamin K2,PTH,1,25 (OH)2D3,and alendronate],and to investigate the potential role of MGP in the pathogenesis of osteoporosis.Methods Primary osteoblasts(OBs) were derived from sequential trypsin/collagenase-digested calvaria isolated from newborn SD rat (postnastal day 1-3).OBs of the second generation were identified by Van Gieson collagen staining,alkaline phosphatase(ALP) staining and calcified nodules staining.OBs of the fourth generation were selected to interfere with vitamin K2,PTH,1,25 (OH)2D3,and alendronate,then cultured for 24 h in mediums which contained various concentrations of vitamin K2 (10-7,10-6,and 10-5 mol/L),PTH (10-9,10-8,and 10-7 mol/L),1,25 (OH) 2D3(10-10,10-9,and 10-8mol/L),alendronate(10-6,10-5,and 10-4mol/L).After being cultured for 24 h,total RNA was extracted and examined by real-time quantitative RT-PCR.Results The primary cultured cells had typical morphological characters of osteoblast.van Gieson collagen staining,ALP staining,and calcified nodules staining were all positive.Vitamin K2,PTH,1,25 (OH)2D3,and alendronate could modulate the expression of MGP mRNA in osteoblasts in a dose-dependent fashion.MGP mRNA expressions were 2.56-fold,2.12-fold,and 1.57-fold with 10-5,10-6,and 10-7 mol/L of vitamin K2 treatment,respectively.The expressions were 6.78-fold,5.31-fold,and 2.23-fold with 10-7,10-8,and 10-9mol/L of PTH(1-34) treatment,8.93-fold,6.95-fold,and 3.47-fold with 10-8 10-9,and 10-10mol/L of 1,25 (OH)2D3 treatment,and 3.47-fold,2.49-fold,and 1.98-fold with 10-4,10-5,and 10-6mol/L of alendronate treatment.Conclusion Vitamin K2,PTH,1,25 (OH)2D3,and alendronate all canregulate MGP mRNA expression in calvarial osteoblasts in a dose-dependent manner.MGP seems to be a potent target of anti-osteoporosis agents,and involved in the pathogenesis of osteoporosis.

AIM:To evaluate the percentage of cutting fluctuations of central corneal thickness (CCT) intraoperative used low concentration (0.02%) mitomycin C (MMC) after laser-assisted subepithelial keratomileusis (LASEK).METHODS:In this prospective study, low and medium myopia group(spherical equivalent≤6.0DS) has 138 patients (276 eyes).Low concentration MMC used topically in 69 patients(138 eyes)randomized after excimer laser ablation;the another traditional LASEK as control.High myopia group(6.0DS

Coronary Sinus ; abnormalities ; Humans ; Male ; Middle Aged ; Subclavian Vein ; Vena Cava, Superior

Mice and 3-day-old chickens were orally inoculated with the recombinant attenuated Salmonella typhimurium strain ZJ111 carrying pcDNA3-F expression plasmid encoding the fusion protein of Newcastle disease virus (NDV). The results showed that ZJ111/pcDNA3-F was relatively safe. The recombinant plasmid pcDNA3-F was stable within the host stain ZJ111 in vitro and in vivo as shown by restriction enzyme analysis and PCR identification of the F gene. In an experimental vaccination study, 3-day-old chickens were orally immunized with ZJ111/pcDNA3-F with a dose of 108 cfu per chicken and boosted two weeks later. At week 4 post boosting, all chickens were challenged with a lethal dose of a virulent NDV strain F48 E9. The results showed that oral vaccination with ZJ111/pcDNA3-F induced stronger humoral and cellular immune responses than intramuscular immunization with naked pcDNA3-F plasmid. It also exhibited higher protection rate than the latter (66.7% vs 50%). This study indicates that the DNA vaccine using attenuated Salmonella typhimurium as delivery carrier had good safety, stability and immunogenicity and exhibited good potential of low cost and convenience for poultry disease control.
Animals ; Chickens ; Immunity, Cellular ; immunology ; Immunity, Humoral ; immunology ; Mice ; Newcastle Disease ; immunology ; virology ; Plasmids ; Polymerase Chain Reaction ; Salmonella typhimurium ; genetics ; metabolism ; Vaccines, DNA ; adverse effects ; genetics ; immunology

Objective To investigate the expression and significance of Caspase-3 and Survivin at different stages of human hemangioma in children.Methods Fifty-five specimens of hemangioma tissue excised in operation and 8 normal skins removed in operation were harvested from People's Hospital of Gansu Province between Jan.2000 and Dec.2005.The pathologic diagnosis was divided into 3 groups according to Mulliken's standard under microscope:proliferated phase group(n=31),degenerated phase group(n=24),and control group(n=8).All these specimens were examined by immunohisto for Caspase-3 and Survivin expression.Results Caspase-3 positive rate in the proliferative phase and involuting phase were 35.5% and 79.2%,respectively.The positive rate in involuting phase was higher than that that in proliferative phase,the difference between the 2 groups was significant(P=0.045 9).A significant difference was not found between the proliferative phase and normal skin tissue(P=0.057 3).Survivin positive rate in the proliferating and involuting phase were 77.4% and 45.8%,respectively.The positive rate in proliferative phase was higher than that in involuting phase,the difference between the 2 groups was significant(P=0.008 5),the difference in involuting phase and normal skin tissue was not significant(P=0.059 3).Conclusions High level of Caspase-3 expression in vascular endothelial cells in involuting phase in contrast to that in proliferative phase,which indicates that Caspase-3 may play a positive role in the apoptosis of vascular endothelial cells.Survivin may inhibit the apoptosis of vascular endothelial cells in proliferative phase due to it's high expression.Caspase-3 and Surivivin involved in the hemangioma in the regulation of apoptosis.J Appl Clin Pediatr,2009,24(1):51-52

Objective To evaluate acute toxicity and genotoxicity of sapindus saponin and to provide toxicological basis for sapindus saponin ’s daily applications. Methods Acute oral toxicity test,mammalian erythocyte micronucleus test, bacterial reverse mutation test and in vitro mammalian chromosome aberration test were used to investigate the effect of the sapindus saponin on gene mutation and chromosome aberration in both prokaryotic and eukaryotic cells. Results The acute toxicity test showed that the LD50 of sapindus saponin was 4640 mg/kg for both male and female mice. Toxic symptoms were observed including salivation,mucus and other toxic manifestations. There was no significant difference between the each dose group and the negative control group in the results of mammalian erythocyte micronucleus test( P>0. 05). The results of bacterial reverse mutation test were also negative. In each dose group and strain with or without S9,the number of revertant colonies did not exceed 2 times than that of spontaneous revertant colonies( negative control). No dose-response relationship was observed. The vitro mammalian chromosome aberration test showed that the IC50 of sapindus saponin on CHL was 75 μg/ml,and the differences between each dose group and the negative control group were not statistically significant( P >0. 05 ). However,the positive control group differed from the negative control group in all tests( P <0. 01). Conclusion Under this experimental condition,sapindus saponin has no genotoxicity.

Adult ; Aged ; Aged, 80 and over ; Female ; Hepatic Encephalopathy ; diagnosis ; Humans ; Male ; Middle Aged ; Neuropsychological Tests ; Psychometrics ; methods ; Young Adult

OBJECTIVETo identify SNP in flos Lonicerae, and authenticate Lonicera japonica from its adulterants and the mixture by using bidirectional PCR amplification of specific alleles (Bi-PASA).

METHODSNP of L. japonica and its adulterants was identified by using ClustulW to align trnL-trnF sequences of the Lonicera genus from GenBank database. Bi-PASA primer was designed and the PCR reaction systems including annealing temperature optimized. Optimized result was performed in 84 samples to authenticate L. japonica, its adulterants and the mixture.

RESULTWhen the annealing temperature was 61 degrees C, DNA from L. japonica would be amplified 468 bp whereas PCR products from all of the 9 adulterants were 324 bp. The established method also can detect 5% of intentional adulteration DNA into L. japonica.

CONCLUSIONThe Bi-SPASA could authenticate L. japonica from its adulterants and the mixture.

Alleles ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; analysis ; genetics ; Flowers ; genetics ; Lonicera ; classification ; genetics ; Plants, Medicinal ; classification ; genetics ; Polymerase Chain Reaction ; methods ; Polymorphism, Single Nucleotide ; RNA, Transfer, Leu ; genetics ; RNA, Transfer, Phe ; genetics ; Reproducibility of Results ; Species Specificity