OBJECTIVETo explore the effect and action mechanism of moxibustion on healing of cutaneous wound in rats.

METHODSTwenty-four SD rats were selected and made into linear full-thickness skin injury model. With randomized digital table, rats were randomly divided into a treatment group and a model group, 12 cases in each one. Then according to treatment time, each group was again divided into a 1d group, a 3d group and a 7d group, 4 cases in each one. The moxibustion at injured skin was applied in the treatment group, 30 min per time, once a day. Hematoxylineosin (HE) staining method was adapted to measure growth status of capillary and number of vascular endothelial cell; immunohistochemical method was used to measure the expression of vascular endothelial growth factor (VEGF).

RESULTSThe wound healing indices in the treatment 7d group were higher than those in the model 7d group on both the 4th day and 8th day after treatment (both P < 0.05). The number of capillary in the treatment 1d group and 3d group was higher than that in the model 1 d and 3 d groups (both 1 < 0.05). The number of capillary in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The number of vascular endothelial cell in the treatment 3d group was higher than that in the model 3d group (P < 0.05). The number of vascular endothelial cell in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of number of vascular endothelial cell between the treatment 1d group and model 1d group was not significant (P > 0.05). Positive cells accumulated score of V EGF expression in the treatment 3d group was higher than that in the model 3d group (P < 0.05). Positive cells accumulated score of VEGF expression in the treatment 7d group was lower than that in the model 7d group (P < 0.05). The difference of positive cells accumulated score of VEGF expression between the treatment 1d group and model 1d group was not significant (P > 0.05).

CONCLUSIONMoxibustion could improve the healing of skin wound in rats, which could be related with regulating vascular endothelial cell and VEGF in wound tissue at different time.

Animals ; Endothelial Cells ; metabolism ; Female ; Humans ; Male ; Moxibustion ; Rats ; Rats, Sprague-Dawley ; Skin ; injuries ; metabolism ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Wounds and Injuries ; genetics ; metabolism ; therapy

0.05).There was excellent correlation between the lesion size on EC Ⅲ-60 enhanced T_1-weighted MR images and histomorphometry(r=0.999,P

OBJECTIVETo observe the effects of carboxymethyl chitosan calcium (CCC) on concentration of lead, calcium and zinc, and the liver antioxidative capacity in lead poisoned mice.

METHODSMice were randomly divided into 7 groups, including normal group, calcium carbonate group, lead-model group, and three experimental groups treated with CCC in three different doses, and the CaNa2EDTA positive control group. The lead poisoned mice model was established by giving water contained with lead acetate. CCC was administrated to mice i.g. once a day. Thirty days later, mice were killed and the concentrations of lead, calcium and zinc in blood, liver, brain and femur were determined by atomic absorption spectrophotometer. Maleic dialdehyde (MDA), total antioxidative capacity (T-AOC), superoxide dismutase (SOD), and glutathione peroxidase (GSH-Px) activities in liver were measured by using assay kit.

RESULTSCCC significantly reduced the concentration of lead in blood, brain, liver and femur from about 1.56 microg/g, 13.38 microg/g, 16.15 microg/g, 1011.62 microg/g to about 0.50 microg/g, 5.57microg/g, 5.64 microg/g, 457.86 microg/g, and markedly increased the concentration of calcium in femur in lead poisoned mice. CCC had no significant side-effects on concentration of zinc in lead poisoned mice. The antioxidative profile was favorably changed as manifested by decreasing the level of MDA, increasing the activities of SOD, GSH-Px and T-AOC in livers of the in lead poisoned mice.

CONCLUSIONCCC might significantly advance the excretion of lead, increase the concentration of calcium in femur and the antioxidative capacity in lead-loaded mice.

Animals ; Brain Chemistry ; Calcium ; metabolism ; Chitosan ; analogs & derivatives ; pharmacology ; Female ; Femur ; chemistry ; Lead ; metabolism ; Lead Poisoning ; metabolism ; Liver ; chemistry ; Mice ; Mice, Inbred Strains ; Zinc ; metabolism

Objective To investigate the prevalence and characteristics of visual hallucination among patients with Parkinson's disease (PD), and analyse the potential risk factors. Methods One hundred and twenty-eight patients with PD were administered self-prepared visual hallucination questionnaires, and prevalence of visual hallucination was surveyed. The differences in sex, age, disease duration, Mini Mental State Exam (MMSE) scores, Hoehn & Yahr stage, types of medicine used and levodopa equivalent doses (LDE) were compared between the patients with visual hallucination and those without visual hallucination. The prevalence of rapid eye movement sleep behavior disorder (RBD) was investigated in patients with visual hallucination. Results Eighteen patients (14.06%) experienced visual hallucination, among whom 10 (55.56%) experienced visual hallucination no less than one time per day, 11 (61.11%) "saw" the shadow of human figure and 15(83.33%)were complicated with RBD. There were significant differences in MMSE scores, Hoehn & Yahr stage, female proportion and usage of dopamine agonists between patients with visual hallucination and those without visual hallucination(P<0.05). Conclusion Visual hallucination is a common non-motor symptom among patients with PD, and cognitive function, disease severity and usage of dopamine agonista may be related to visual hallucination.

OBJECTIVETo study the effect of nitric oxide synthase inhibitor, S-methyl thiocarbamate (SMT), on proteoglycan metabolism in repaired articular cartilage in rabbits.

METHODSTwenty-four male New Zealand white rabbits, aged 8 months and weighing 2.5 kg+/-0.2 kg, were used in this study. Cartilage defects in full thickness were created on the intercondylar articular surface of bilateral femurs of all the rabbits. Then the rabbits were randomly divided into 3 groups (n=8 in each group). The defects in one group were filled with fibrin glue impregnated with recombinant human bone morphogenetic protein-2 (rhBMP-2, BMP group), in one group with fibrin glue impregnated with rhBMP-2 and hypodermic injection with SMT (SMT group) and in the other group with nothing (control group). All the animals were killed at one year postoperatively. The tissue sections were stained with safranine O-fast green and analyzed by Quantiment 500 system to determine the content of glycosaminoglycan through measuring the percentage of safranine O-stained area, the thickness of cartilages and the mean gray scale (average stain intensity). Radiolabelled sodium sulphate (Na(2)(35)SO(4)) was used to assess the proteoglycan synthesis.

RESULTSAt one year postoperatively, the percentage of safranine O-stained area, the mean gray scale and the cartilage thickness of the repaired tissues in SMT group were significantly higher than those of BMP group (P<0.01) and the control group (P<0.05). Result of incorporation of Na(2)(35)SO(4) showed that the proteoglycan synthesis in SMT group was higher than those of BMP group and the control group (P<0.01).

CONCLUSIONSSMT, a nitric oxide synthase inhibitor, can significantly increase the content of glycosaminoglycan and proteoglycan synthesis, and computer-based image analysis is a reliable method for evaluating proteoglycan metabolism.

Analysis of Variance ; Animals ; Biopsy, Needle ; Cartilage Diseases ; drug therapy ; pathology ; Cartilage, Articular ; drug effects ; pathology ; Disease Models, Animal ; Immunohistochemistry ; Isothiuronium ; analogs & derivatives ; pharmacology ; Male ; Nitric Oxide Synthase ; antagonists & inhibitors ; pharmacology ; Probability ; Proteoglycans ; drug effects ; metabolism ; Rabbits ; Random Allocation ; Reference Values ; Sensitivity and Specificity

This study was aimed to investigate whether difference exists between real time RT-PCR results of nucleated cells isolated by lysis method and mononuclear cells isolated by gradient concentration method. 14 bone marrow samples from leukemia patients (7 samples of AML-M(2), 1 of AML-M(4), 1 of AML-M(4)EO, 1 of AML-M(6), 1 of APL and 3 of CML) were collected. Each sample was divided into 2 parts, and was used to isolate mononuclear cells by Ficoll-Hypaque gradient centrifugation, and other was used to isolate nuclear cells by lysis method. The RNA extraction and detection of internal reference gene ABL for all of samples were performed by RT-PCR, and mRNA expression levels in 3 samples of BCR/ABL, 6 of AML/ETO, 1 of CBFβ-MHY11, 1 of WT1 and 6 PRAME were detected by RT-PCR. The results showed that ABL copies of all samples were over 3 × 10(4), and there was significant difference in ABL copies between each pair of samples by lysis and gradient centrifugation method (p > 0.05). There was also no significant difference in every detected mRNA levels between 14 pair samples (p > 0.05). It is concluded that the lysis method may be useful one for extracting RNA from nuclear cells, and can adopt as a routing detection method for simultaneously extracting RNA from many samples to detect leukemia specific mRNA by using RT-PCR.
Adolescent ; Adult ; Child ; Female ; Humans ; Leukemia ; genetics ; Male ; Middle Aged ; RNA, Messenger ; analysis ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; methods ; Young Adult

Up to now,studies among the world have put forward various hypotheses about the pathophysiology and future research direction of Takotsubo cardiomyopathy (TTC).Large amounts of diagnosis and differential diagnosis have been done on TTC,and consensuses have been reached on it's definition,epidemiology,clinical manifestations and prognosis.In this review,we will summarize the recent progress in the study of TTC,and make a comprehensive analysis and perspective on it's clinical research and forensic value.

The human UDP-glucuronosyltransferase 1A1 (UGT1A1), one of the most essential conjugative enzymes, is responsible for the metabolism and detoxification of bilirubin and other endogenous substances, as well as many different xenobiotic compounds. Deciphering UGT1A1 relevance to human diseases and characterizing the effects of small molecules on the activities of UGT1A1 requires reliable tools for probing the function of this key enzyme in complex biological matrices. Herein, an easy-to-use assay for highly-selective and sensitive monitoring of UGT1A1 activities in various biological matrices, using liquid chromatography with fluorescence detection (LC-FD), has been developed and validated. The newly developed LC-FD based assay has been confirmed in terms of sensitivity, specificity, precision, quanti-tative linear range and stability. One of its main advantages is lowering the limits of detection and quantification by about 100-fold in comparison to the previous assay that used the same probe substrate, enabling reliable quantification of lower amounts of active enzyme than any other method. The precision test demonstrated that both intra- and inter-day variations for this assay were less than 5.5％. Further-more, the newly developed assay has also been successfully used to screen and characterize the regu-latory effects of small molecules on the expression level of UGT1A1 in living cells. Overall, an easy-to-use LC-FD based assay has been developed for ultra-sensitive UGT1A1 activities measurements in various biological systems, providing an inexpensive and practical approach for exploring the role of UGT1A1 in human diseases, interactions with xenobiotics, and characterization modulatory effects of small mole-cules on this conjugative enzyme.

Transgenic Atropa belladonna with high levels of scopolamine was developed by metabolic engineering. A functional gene involved in the rate limiting enzyme of h6h involved in the biosynthetic pathway of scopolamine was over expressed in A. belladonna via Agrobacterium-mediation. The transgenic plants were culturing till fruiting through micropropogating and acclimating. The integration of the h6h genes into the genomic DNA of transgenic plants were confirmed by genomic polymerase chain reaction (PCR) analysis. Analysis of the difference of plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight was carried out using SPSS software. The content of hyoscyamine and scopolamine in roots, stems, leaves and fruits was determined by HPLC. The investigation of the expression levels of Hnh6h by qPCR. Both Kan(r) and Hnh6h genes were detected in five transgenic lines of A. belladonna plants (A8, A11, A12, C8 and C19), but were not detected in the controls. The plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight of transgenic plants did not decrease by comparison with the non-transgenic ones, and furthermore some agronomic characters of transgenic plants were better than those of the controls. The highest level of scopolamine was found in leaves of transgenic A. belladonna, and the content of scopolamine was also higher than that of hyoscyamine in leaves. The contents of scopolamine of leaves in different transgenic lines were listed in order: C8 > A12 > C19 > A11 > A8, especially, the content of scopolamine in transgenic line C8 was 2.17 mg x g(-1) DW that was 4.2 folds of the non-transgenic ones (0.42 mg x g(-1) DW). The expression of transgenic Hnh6h was detected in all the transgenic plants but not in the control. The highest level of Hnh6h expression was found in transgenic leaves. Overexpression of Hnh6h is able to break the rate limiting steps involved in the downstream pathway of scopolamine biosynthesis, and thus promotes the metabolic flux flowing toward biosynthesis of scopolamine to improve the capacity of scopolamine biosynthesis in transgenic plants. As a result, transgenic plants of A. belladonna with higher level of scopolamine were developed.
Atropa belladonna ; genetics ; metabolism ; Atropine ; metabolism ; Gene Expression ; Mixed Function Oxygenases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Solanaceae ; enzymology ; genetics

All three nitric oxide synthase (NOS) isoforms are found in the lungs. It has been demonstrated that eNOS-derived NO plays an important role in modulating pulmonary vascular tone and inhibiting pulmonary vascular remodeling. Histamine induces pulmonary vasoconstriction by activating H(1)-receptor on the smooth muscle cells and vasodilation by stimulating H(2)-receptor. It remains unclear whether histamine also modulates the pulmonary vascular tone by regulating eNOS gene expression and NO production in pulmonary artery endothelial cells. Therefore, the present study was performed on cultured primary porcine pulmonary artery endothelial cells (PAECs) to investigate the effects of histamine on eNOS gene expression, and to explore the role of CaMK II in eNOS gene expression. After treatment with different concentrations histamine for different times, the levels of eNOS mRNA and protein were measured by RT-PCR and Western blot, respectively. The results showed that histamine upregulated eNOS mRNA and protein levels in a concentration- and time-dependent manner. Incubation with 10 micromol/L histamine for 24 h could increase eNOS mRNA and protein level to 160.8+/-12.2% (P<0.05) and 136.2+/-11.2% (P<0.05), respectively, of the control values. These up-regulation effects were prevented by selective CaMK II inhibitor, KN-93 (10 micromol/L). To investigate whether or not histamine increases eNOS expression by upregulating eNOS gene transcription, PAECs were transiently transfected with 1.6-kb fragment of the human eNOS promoter driving a luciferase reporter gene. The results suggested that eNOS gene promoter activity was enhanced to 148.2+/-33.7% (P<0.05) of the control after PAECs were incubated with 10 micromol/L histamine for 24 h. The nitrite and nitrate content in culture media measured by colorimetric method after incubation with 10 micromol/L histamine for 24 h indicated that the NO production in PAECs was increased. These results suggest that histamine up-regulates eNOS gene transcription and enhances NO production in PAECs by a signaling pathway involving CaMK II, which might be one of the mechanisms of histamine modulating pulmonary vascular tone.
Animals ; Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; Calcium-Calmodulin-Dependent Protein Kinases ; pharmacology ; Cells, Cultured ; Endothelium, Vascular ; enzymology ; Histamine ; pharmacology ; Nitric Oxide Synthase Type III ; biosynthesis ; genetics ; Promoter Regions, Genetic ; Pulmonary Artery ; cytology ; enzymology ; RNA, Messenger ; biosynthesis ; genetics ; Swine ; Up-Regulation