Cardiotoxic effects of adriamycin were investigated in cultured heart cells. Three-day cultured myocardial cells were treated with the different doses of adriamycin (0.3mg/L, 1 mg/L, 3 mg/L) , and incubated in verieus times times ( 4 h, 8 h, 24h ) , and, then the content of lactate dehydrogenase ( LDH ) in media and malondialdehyde ( MDA ) in cells were measured. The results showed that the content of LDH in media increased with the time in culture. The content of LDH in media and MDA in cells increased markedly with the increase in adriamycin concentration. It suggested that the adriamycin has direct damage to myocardial cells, which is time-dependent and dose-dependent.

AIM: To investigate the role of catecholamine, angiotensin converting enzyme(ACE) and adenosine triphosphatase in ischemic preconditioning in isolated rat hearts. METHODS: Isolated perfused rat heart was subjected to global ischemia for 40 min followed by reperfusion for 10 min (I/R). Preconditioning (PC) was induced by 5 min of ischemia and 10 min of reperfusion. The tissue concentrations in NE, and ACE, ATPase activities were determined in left ventricle in the PC and I/R groups by fluorometry and spectrophotometry. RESULTS: There were no significant difference in NE and ACE between PC and I/R groups. PC hearts showed improved recovery of the contractile function after 40 min ischemia/10 min reperfusion, but activities of the myocardial total ATPase, Mg2＋-ATPase, Na＋K＋-ATPase were inhibited markedly compared with I/R group. CONCLUSION: The inhibited myocardial ATPase may be involved in the mechanism of ischemic preconditioning protection in the isolated rat heart. Endogenous myocardial norepinephrine and ACE activation are not essential for ischemic preconditioning in the isolated rat heart.

Objective To investigate the clinical significance of thyroglobulin antibody (ATG) and thyroid peroxidase antibody (ATPO) in patients with vitiligo. Methods Venous blood samples were obtained from 87 patients with vitiligo and 90 age- and sex-matched normal human controls. Chemiluminescence was applied to measure the serum levels of ATG, ATPO, free triiodothyronine, free tetraiodothyronine and thyroid stimulating hormone (TSH). Results There was a significant increase in the positivity rates of ATG (23.0% vs 6.7%, P < 0.01) and ATPO (24.1% vs 7.8%, P < 0.01) as well as the serum level of TSH (3.4 ± 2.4 vs 2.4 ± 1.2 pmol/L, P < 0.05) in the patients with vitiligo compared with the normal human controls. It is worth mentioning that all patients positive for ATG or ATPO were diagnosed with vitiligo vulgaris. The positivity rates of ATG and ATPO in patients with vitiligo aged from 11 to 20 years and 21 to 40 years were significantly higher than those in age-matched normal controls (all P < 0.05). Also, female patients had a higher positivity rate of ATG and ATPO than female controls did (34.1% vs 8.5%, χ2 = 8.90, P < 0.01; 34.1% vs 10.6%,χ2 = 7.29, P < 0.05). The highest positivity rates of both ATG and ATPO were 53.3%, which were observed in vitiligo patients aged from 11 to 20 years, followed by patients from 21 to 40 years (ATG 34.5%, ATPO 34.5%). In patients with vitiligo positive for both ATG and ATPO, the occurrence of autoimmune thyroid disease was 70% (14/20), significantly higher than that in ATG- and ATPO- positive healthy controls (16.7%, χ2 = 5.4, P < 0.05). Conclusions ATG and ATPO were observed in young female patients with vitiligo vulgaris, and they may be associated with the development of autoimmune thyroid diseases.

According to the designed specific primers of gene fragment based on the Salvia miltiorrhiza transcriptome data, with the method of reverse transcription polymerase chain reaction (RT-PCR), this study cloned full-length cDNA sequence of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase gene from Salvia miltiorrhiza bge.f.alba, this sequence is named as SmHDS and its GenBank registration number is KJ746807. SmHDS, 2 529 bp long, contains an ORF of 2 229 bp, encodes 742 amino acids, including 5' UTR 170 bp and 3' UTR 130 bp. Using bioinformatics software, having made a homology analysis of the obtained sequence, we can have a conclusion that SmHDS have a close genetic relationship with HDS of Salvia miltiorrhiza. Analysis result of prokaryotic expression revealed that in Escherichia coli, SmHDS expressed target proteins which in size are comparable with the protein predicted. Meanwhile, the 4 factors which can influence the protein expression were optimized, the 4 factors are inducing temperature, inducing time, IPTG concentrations and density of inducing host bacterium (A600). The optimal expression conditions of SmHDS were 30 degrees C until the A600 is 0.6, and add IPTG to a final concentration of 0.2 mmol x L(-1), and the induction time of 20 h. It provides theoretical basis for the further study of the function of 1-hydroxy-2-methyl-2-(E)-butenyl-4-diphosphate synthase in the biosynthesis of tanshinone compounds.
Cloning, Molecular ; DNA, Complementary ; genetics ; Diterpenes, Abietane ; biosynthesis ; Enzymes ; biosynthesis ; genetics ; Escherichia coli ; metabolism ; Plant Proteins ; biosynthesis ; genetics ; Salvia miltiorrhiza ; enzymology ; genetics

OBJECTIVETo construct four helper plasmids which are necessary in reverse genetic system of a cell adapted rabies virus strain.

METHODSNucleoprotein gene, phosphprotein gene, glycoprotein gene and the viral RNA polymerse gene are amplified by RT-PCR. The four gene fragments were cloned into an expression vector respectively after sequencing to construct four helper plasmids.

RESULTSThe four structral genus were amplified successfully, the sequence were correct. And all the structral protein gene fragments were cloned into expression vectors, the results were correct identified by cleavage reaction.

CONCLUSIONConstruction of the four helper plasmids have been done successfully, and it is the basis for rescuing the adapted cell rabies virus strain from cloned cDNA.

Animals ; Cloning, Molecular ; methods ; Genetic Vectors ; genetics ; Humans ; Mice ; Plasmids ; genetics ; Rabies ; virology ; Rabies virus ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism

OBJECTIVETo study the expression of lipoprotein related genes in subchondral bone of early experimental os-teoarthritis, which may play an important role in the pathogenesis of osteoarthritis.

METHODSAnimals are equally divided into two groups: experimental group and control group, both of which contain fifteen rats of similar weight. The right knee joints of experimental group underwent surgery,which involved in both medial collateral ligament(MCL) transaction and medial meniscectomy, while the control group was only carried out with a sham operation. Rats were killed at 1, 2 and 4 weeks postsurgery to obtain the right knee joints. Total RNA of the subchondral bone was extracted,and then hybridized to Agilent Whole Rat Genome Microarray. Differentially expressed genes analysis was used to study the chemokine signaling pathway.

RESULTSApoa5 expression was down-regulated at 1, 2 weeks post-surgery, Apoc2 expression was up-regulated at 1 week after surgery, Apol3 expression was up-regulated at 1 and down-regulated at 4 weeks post-surgery, Lrp1 expression was down-regulated at 1, 2 weeks after surgery. Lrp5 was down-regulated at 2 weeks after surgery. Gpihbp1, Lpl, Tfpi and Vldlr were up-regulated at 1 weeks after surgery. Lrpap1 and RGD1309808 were down-regulated at 4 weeks after surgery.

CONCLUSIONDysregulation of lipoprotein related genes plays an important role in pathogenesis of early experimental osteoarthritis.

Animals ; Disease Models, Animal ; Knee Joint ; metabolism ; Lipoproteins ; genetics ; Male ; Oligonucleotide Array Sequence Analysis ; Osteoarthritis ; genetics ; Rats ; Rats, Sprague-Dawley ; Transcriptome

Objective To establish a determination method for leuprolide acetate microspheres. Methods HPLC was performed on Inertsil ODS-SP (150×4.6 mm×5μm) with mobile phase consist of 0.1 mol/L of Ammonium Dihydrogen Phosphate(adjust its pH to 7.00±0.05 with Ammonium Hydroxide) and Acetonitrile in ratio of 3:1(V/V),and the flow rate was 1.0 mL/min.The wavelength was 220 nm and column temperature was 30℃. The injection volume was 20 μL. Results The linear range of leuprolide acetate was 20.0-160.0μg/mL (r=0.9999) with an average recovery of 99.80%,RSD=0.63%(n=9). Conclusion The method of HPLC was accurate,reliable and specific, which could be used to determinate the assay of leuprolide acetate microspheres and for quality control of microspheres.

Objective To evaluate the accuracy of nostril width method in intubation model choose in children with cerebral pal‐sy FSPR surgery .Methods Three hundred and fifty six cases of cerebral palsy FSPR surgery were selected ,the endotracheal intu‐bation were selected by nose width method and modified weight method ,and then we made a comparison between the two methods . Kappa consistency test and rank correlation analysis were both used to analyze the consistency .Results With Kappa=0 .81 ,rank correlation coefficient 0 .905 ,we could know that there was high degree of consistency between the two methods (P<0 .01) ,of which 298 cases (83 .7% ) children selected the same model intubation strengthen model in different method ,58 cases (16 .3% ) pe‐diatric intubation choice of two methods differ by one model .Conclusion Nostril width method was simple and reliable to select in‐tubation in pediatric surgery to strengthen FSPR model .

Four strains of aerobic denitrifying bacteria were isolated from biomembrane of the laboratory biofilter by selective medium. The denitrifying rates of these four strains were found more than 80% under aerobic condition for 40 hours. The denitrifying rate of A1 was highest, which was 99.05%. When measuring the course of nitrogen element changing, it is found that they accumulated nitrite expect the strain A1. The strain A1 was gram positive and spherical. It is identified as Pseudomonas putida based on its biochemical and morphological characters and phylogenetic analysis of 16S rDNA sequences(genbank accession NO.DQ836052.1). For the strain A1, the optimum beginning pH was 7.0 around, and the optimum tempera- ture was 30 ℃ around, The change of DO did not influence the effect of denitrification when it was more than 2.0 mg/L.

Objective To evaluate the effect of point-of-care hemoglobin/hematocrit (POC HGB/HCT) devices and intraoperative blood salvage on the amount of perioperative allogeneic blood transfusion and blood conservation in clinical practice.Methods A total of 46 378 medical records of 22 selected hospitals were reviewed. The volume of allogeneic red blood cell and plasma, number of patients transfused, number of intraoperative autologous blood salvage, total volume of autologous blood transfusion, and amount of surgery in the year of 2011 and 2013 were tracked. Paired t-test was used in intra-group comparison, while t-test of two isolated samples carried out in inter-group comparison. P<0.05 was defined as statistically significant difference.Results In the hospitals where POC HGB/HCT device was used (n=9), the average allogeneic blood transfusion volume per 100 surgical cases in 2013 was significantly lower than that in 2011 (39.86±20.20 vs. 30.49±17.50 Units, t=3.522, P=0.008). In the hospitals without POC HGB/HCT meter, the index was not significantly different between 2013 and 2011. The average allogeneic blood transfusion volume was significantly reduced in 2013 than in 2011 in the hospitals where intraoperative autologous blood salvage ratio [autologous transfusion volume/(autologous transfusion volume+allogeneic transfusion volume)] was increased (n=12, t=2.290, P=0.042). No significant difference of the above index was found in the hospitals whose autologous transfusion ratio did not grow.Conclusion Intraoperative usage of POC HGB/HCT devices and increasing autologous transfusion ratio could reduce perioperative allogeneic blood transfusion.