Objective To investigate the expression of radiation-induced IER5 protein and screen its potential interaction proteins that may participate in DNA repair process.Methods HeLa cells were irradiated with 4 Gy ionizing radiation.IER5 protein expression in whole cell lysate and in nuclear fraction were detected by Western blot at different timepoints after irradiation.3 × Flag-IER5 pCMV plasmid was constrcuted and the Flag tagged-IER5 expression was verified by Western blot.293T cells were transfected with 3 × Flag-IER5 pCMV plasmid.After irradiation the cells were collected and proteins were extracted.The IER5 interaction proteins were purified using immunoprecipitation and separated by 12％ SDS-polyacrylamide gel electrophoresis.Then the binding proteins were cut from the gel and analyzed by Mass spectrometry.Results The expression of IER5 protein began to increase 4 hour post-irradiation and its peak level was observed at 12 hour post-irradiation,and it lasted until 48 hour after irradiation.The expression level of IER5 protein in whole cell lysate and nuclear fraction were both increased.With the mass spectrometry analysis,a total of 347 proteins and 256 proteins were identified in irradiated and nonirradiated groups,respectively.Fourty one differential proteins were obtained,where 10 proteins were associated with DNA metabolic process and DNA rapair in the irradiated group and the poly(ADP-ribose) polymerase 1 (PARP1) protein was further confirmed by Western blot.Conclusions IER5 protein is an DNA damage related protein,and it may participate in DNA repair process.

Objective To discuss the changes of correlative ultrasonic parameter index of normal hips and abnormal hips with developmental dislocation of the hip (DDH) in infants of different months in order to provide objective information for the diagnosis. Methods Three-hundred and seventy-eight normal hips and 244 hips with DDH among 622 hips of 311 infants were detected by ultrasonography(US). The morphology and structure information of hips were observed, and the values of ultrasonic parameter index,including angle α,angle 3, acetabular index( AI), femoral head percentage of cover(FHC) of normal hips and abnormal hips were measured. The values of each parameter index were collected by being divided into different groups (3 months a group) ,then the correlation was analyzed. Results Morphology and structure,position relation between femoral head and acetabulum of the hips were demonstrated by US. Normal or abnormal hips,the degrees of abnormal hips and the types of hips could be judged according to the findingsof US. Analysis of values of parameter index of normal hips:①There was significantly statistical significance in the values of ultrasonic parameter index, such as angle α, angle β, AI, FHC of normal hip between the groups of different age (P＜0.01). ②There was correlation between the age and the values of each parameter index, among which angle α, FHC had positive correlations with age ( r = 0. 537, 0. 554,respectively ) while angle β and Al negative correlations ( r = -0. 465, -0.424, respectively ). ③There was correlation between the values of different parameter index. Both angle β and AI had negative correlation with angle α,among which the latter correlation was closely ( r = - 0. 794). No statistical significance was found between the ultrasonic values of each group under different ages of different type abnormal hips( P ＞0.05) ,but closely negative correlations still existed between angle α and AI. ConclusionsUS can be viewed as an early definite and a screening method of diagnosing DDH.For older infants (above 6 months) it will be more accurate to analyze the ultrasonic parameter index together with the age of infants.

Aconitum, as a kind of common traditional Chinese medicine, contains multiple biological active substances, with a very high medicinal value but high toxicity. Its major toxic ingredients are aconitine, mesaconitine and hypaconitine, which are also efficient ingredients. Therefore, the safety of its clinical application has aroused wide attention. With the constant deepening of drug development studies, people want to learn about its toxic mechanism and the regularity of its emergence and development of its toxicology, so as to make a scientific and rational assessment for its safety. Therefore, toxicokinetics and metabonomics have gradually become important content in the new drug assessment. During the development of drug performance, it is crucial to establish a scientific, objective and standardized Aconitum safety evaluation system and correctly assess and utilize its toxicity. Having summarized studies on metabonomics and toxicokinetics of Aconitum drugs in recent years, authors proposed to strengthen the studies on Aconitum drug safety assessment and establish a scientific and standardized safety evaluation system as soon as possible, in order to make the national treasure more useful.
Aconitum ; chemistry ; toxicity ; Animals ; Drugs, Chinese Herbal ; pharmacokinetics ; toxicity ; Humans ; Metabolomics

ObjectiveTo investigate the clinical efficacy of acupuncture in treating Tourette's syndrome.MethodEighty patients were allocated, using a random number table, to a treatment group of 40 cases and a control group of 40 cases. The treatment group received acupuncture and the control group, Western drug treatment.ResultThe YGTSS score decreased significantly in the two groups of patients after treatment(P<0.01). After treatment, the range of decrease in the YGTSS tic score was larger in the treatment group than in the control group; there was a statistically significant difference (P<0.05). The total efficacy rate was higher in the treatment group than in the control group (P<0.05). The incidences of adverse reactions were significantly lower in the treatment group than in the control group.ConclusionThe effect and safety of treatment for Tourette's syndrome are higher in the treatment group than in the control group.

OBJECTIVETo investigate the change of JNK and c-Jun in lung injury associated with paraquat poisoning of rats.

METHODS46 Rats were randomly divided into four groups: PQ group (n = 12), control group (n = 10), PQ + ZnPP group (n = 12) and PQ + Hm group (n = 12). The rats were injected with 2% PQ (25 mg/kg, ip) in PQ group. ZnPP and Hemin (10 mg/kg, 10 mg/ml) were injected through inguinal vein before intraperitoneal administration of 2% paraquat in PQ + ZnPP group and PQ + Hm group respectively. The rats were injected NS (1 ml/kg, ip) in control group. HE dyeing of lung tissue and MDA content of plasma were used for estimating the injury of lung tissue. The content of CO in the lung tissue was determined. The expression of HO-1 mRNA of the lung tissue was detected by the reverse transcription-polymerase chain reaction. The phosphorylation of JNK and c-Jun was evaluated by Western blot analysis.

RESULTSThe degree of lung injury in PQ group and PQ + ZnPP group was higher than that in control group and PQ + Hm group. But in PQ + Hm group the degree of lung injury was lower. The content of MDA in PQ group and PQ + ZnPP group was higher than that in control group and PQ + Hm group (P < 0.01). The content of MDA in PQ + Hm group was higher than that in control group (P < 0.05). The content of CO in lung tissue in PQ group, PQ + ZnPP group and PQ + Hm group was and (1.08 +/- 0.15 mg/L) respectively, and higher than that in control group (P < 0.01). The content of CO in lung tissue in PQ + Hm group was significantly higher than that in PQ + ZnPP group (P < 0.01). The expression of HO-1 and the phosphorylation of JNK (55.24 +/- 9.34, 38.15 +/- 10.71, 128.55 +/- 19.43) and c-Jun (23.16 +/- 4.85, 15.49 +/- 3.13, 44.89 +/- 10.37) were increased remarkably in PQ group, PQ + ZnPP group and PQ + Hm group. Those in PQ + Hm group were higher significantly than PQ group and PQ + ZnPP group (P < 0.01). Those in PQ + ZnPP group were lower than PQ group (P < 0.05).

CONCLUSIONThe increase of CO of lung tissue in rats at the lung injury associated with paraquat poisoning reduces the acute lung injury of rats. The level of JNK and c-Jun phosphorylation increases obviously, especially after Hemin is utilized.

Acute Lung Injury ; chemically induced ; metabolism ; Animals ; Disease Models, Animal ; JNK Mitogen-Activated Protein Kinases ; metabolism ; Lung ; metabolism ; Male ; Paraquat ; poisoning ; Proto-Oncogene Proteins c-jun ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective To explore the influence of home synthesize magnetic iron oxide (called Fe2O3-PLL) labeling on peripheral blood endothelial progenitor cells (EPCs) bionomics to provide experimental foundation for MR imaging ex and in vivo. Methods Fe2O3 was incubated with PLL for 2 hours to obtain a complex of Fe2O3-PLL. Rabbit peripheral blood mononuclear cells were isolated and EPCs were selected by adherence method. Fe2O3-PLL was used to label EPCs. Prussian blue stain and electron microscope was used for showing intracellular iron. MTT assay was assessed to evaluate the difference of growth curve between unlabeled and labeled with 25 mg/L Fe2O3-PLL. Flow cytometry was performed to analyze cell cycle, cell apoptosis and the expression of surface markers of labeled and unlabeled cells. Expressions of Enos, KDR and Vwf at Mrna levels among unlabeled and labeled EPCs were detected by real-time polymerase chain reaction. Calcium ion channel and membrane fluidity were observed and analyzed by laser confocal microscopy. Statistical analyses were used with ANOVA and t test. Results Almost 100% cells were labeled by Fe2O3-PLL, iron-containing vesicles were intracytoplasma. There was no statistical difference in cells growth curve, cell life cycle [(93.74±3.52)% ,(94.57±3.66)% ] and cell apoptosis rate(12. 89±1.81) %, (11.67±1.18) %) between labeling with Fe2O3-PLL at a concentration of 25 mg/L and unlabeled cells (t = 0. 283, P > O. 05 ; t = 0. 977, P > 0. 05). There was also no statistical difference in relative amount of Enos, KDR and Vwf at Mrna levels and the expression of sudace phenotypic markers (CD34, CD106, CD146 and KDR) between two groups (P > 0. 05). In addition,Labeling had little influence on calcium ion channel and didn't significantly alter cell membrane fluidity.Conclusions The rabbit peripberal blood EPCs can be effective labeled with Fe2O3-PLL and without significant influence on cells bionomics at a low concentration of 25 mg/L. Almost every cell can be labeled and the labeled cells can be used further.

BACKGROUND:Steroid-induced osteonecrosis of the femoral head (SONFH) is a common bone disease characterized as high morbidity and poor prognosis,but the pathogenesis is unclear.Oxidative stress treatment is closely related to the occurrence and development of SONFH,and has tremendous potential in the treatment of SONFH,which can be realized by Nano-Se.OBJECTIVE:To observe the protective effect of porous Se@SiO2 nanocomposite on chondrocytes by antioxidant stress,and to further explore its mechanism of protection and treatment of SONFH.METHODS:(1) In vitro experiment:The rat chondrocytes were isolated,cultured and identified.Then,the chondrocytes were cultured with porous Se@SiO2 nanocomposite to suppress the production of reactive oxygen species (ROS).(2) In vivo experiment:A total of 36 rats were randomly divided into three groups.Steroid-induced group and experimental group were treated with intraperitoneal injection of lipopolysaccharide and intramuscular injection of methylprednisolone to induce SON FH models.Seven days after modeling,the experimental group was intraperitoneally injected with porous Se@SiO2 nanocomposite.No intervention was done in control group (blank control).At 8 weeks after modeling,rat bilateral femoral heads were taken for hematoxylin-eosin staining and Micro-CT scanning.RESULTS AND CONCLUSION:Results from the ROS detection and TUNEL apoptosis tests showed that the level of ROS in the chondrocytes was significantly reduced after intervention with Se@SiO2 (P ＜ 0.05).Micro-CT scanning findings showed that the bone mineral density,bone volume,bone area/bone volume,trabecular number,trabecular thickness,and trabecular separation in the steroid-induced and experimental groups were significantly different from those in the control group (P ＜ 0.05).Hematoxylin-eosin staining results showed smooth femoral head,normal bone cells,chondrocytes and trabecular bone,as well as few empty bone lacunae and fat cells in the control group,while in the steroid-induced group,there was bone trabecular fracture,fat cell hypertrophy fusion,a large number of empty bone lacunae and obvious osteonecrosis.These manifestations were significantly improved in the experimental group.To conclude,the porous Se@SiO2 nanocomposite has good antioxidative stress ability,suppresses the ROS production and exerts therapeutic effects on SONFH.

Objective To evaluate the 1.5 T magnetic resonance imaging system to depict and track in vivo of magnetically labeled endothelial progenitor cells(EPCs),and to study the possibility for preventing the atherosclerotic plaque formation in New Zealand rabbit model of carotid arterial injury after transplantation.Methods New Zealand rabbit EPCs were isolated,confirmed,expanded and then incubated with home synthesized Fe_2O_3-PLL,Prussian blue stain was performed for showing intracellular irons.The model of carotid arterial injury was performed by 2.5F balloons,the group A of 8 rabbits received magnetically labeled EPCs,group B of 3 rabbits received fluorescent-labeled EPCs and the group C of 5 rabbits were given same volume saline injection after endothelial injury of the carotid artery.MR imaging and histology were performed and compared 4 days later for randomly chosen three rabbit,each from one of the three group;all the other rabbits were fed with high lipid diet and examed using MR imaging and histology after 15 weeks.Results Epcs labeling efficiency was more than 95% by Prussian blue stain, 4 days after transplantation of EPCs,only in group A,the injured endothelium of carotid artery had signal intensity loss in T_2 * WI,which were correlated well with the area where the most Prussian blue staining positive cells were found in histopathology analyses.The rabbits of group A and B which received EPCs transplantation exhibited fewer plaques formation than those of the group C(P

Objective To explore the relationship between gene polymorphism of neuropeptide Y (NPY) promoter and cerebral stroke subtypes according to TOAST (Trail of ORG 10172 in Acute Stroke Treatment) criteria in Chinese Han population. Methods The gene polymorphisms at position of-399T/C, -883Tgins/del and -602G/T in NPY promoter were detected by PCR method and gene sequencing in 190 cases of large-artery atherosclerosis stroke (LAA), 260 cases of small-artery occlusion (SAO), 60 cases of cardioembolism stroke (CE), 29 cases of stroke of other demonstrated etiology (ODE),10 cases of stroke of other undemonstrated etiology (UE) and 423 healthy control subjects. The PCR products were directly sequenced. Multivariate logistic regression was performed to analyze the relationship between gene polymorphism of NPY promoter and cerebral stroke subtypes according to TOAST by removing the confounding variables. Results Significant differences in the frequency of genotype CC and allele C at position of-399T/C were noted between the patients with SAO and controls (P=0.046, P=0.010). Compared with the control group, patients with LAA and SAO were more likely having high level of uric acid, hypertension, diabetes mellitus and heard disease (P＜0.05). No statistic differences in the frequency of genotype DD and allele D at position of-883Tgins/del were noted between patients with SAO and controls (P=0.0605, P=0.155). Gene polymorphisms of-399T/C,-883Tgins/del and -602G/T did not associate with an increased risk of having LAA, CE, ODE and UE.Conclusions The gene polymorphisms of promoter in position of-399T/C gene maybe associate with the happening of SAO; allele C at the position of-399T/C may raise the risk of the disease. There is no relationship between the gene polymorphisms of promoter at position of-399T/C, -883Tgins/del, -602G/T and the patients with LAA, CE, ODE and UE. High level of uric acid, hypertension, diabetes mellitus and heard disease history are the risk factors of LAA and SAO.

Nine triterpenes compounds were isolated from the male flowers of Eucommia ulmoides by recrystallization and chromatographic techniques over silica gel, Sephadex LH-20, and RP-18 gel. Their chemical structures were identified on the basis of spectral analysis and as 3-oxo-12-en-ursane-28-O-α-L-arabinofuranosyl (1 --> 6) -β-D-glucopyranoside (1), 2α, 3β-dihydroxyurs-12-en-28-oic acid(28 --> 1) -β-D-glucopyranosyl ester (2), ursolic acid (3), α-amyrin (4), uvaol (5), ursolic acid acetate (6), 3-O-acetate oleanoic acid (7), betulinic acid (8), and betulinol (9). Compound 1 was a new compound, and compounds 2, 4-7 were isolated from the Eucommiu genus for the first time. Cytotoxic activity was tested for all the compounds against K562 and HepG2 cells. The results showed that only compound 3, exhibited cytotoxic activity.
Antineoplastic Agents, Phytogenic ; pharmacology ; Eucommiaceae ; chemistry ; Hep G2 Cells ; Humans ; K562 Cells ; Triterpenes ; analysis ; pharmacology