Objective:To explore the establishment of an animal model of abdominal aortic aneurysm(AAA),and elaborate the role of iNOS inhibitor in the smooth muscle apoptosis of abdominal aortio aneurysm in rats,to find a new theoretical basis for the clinical treatment of small drugs AAA.Methods:SD rats underwent intra-aortic elastase (25U/mL) perfusion to induce AAAs,the positive control group (30) and experiment group (30) use elastase perfusion while the negative control group(30) gives the saline perfusion.After operation the positive and negative control groups were treated with intraperitoneal injections of saline,experimental group injects the iNOS inhibitor Aminoguanidi hydrochloride;Postoperative second,7,and 14 days,The NO content in the serum and specimen of abdominal aortic aneurysm was detected by iNOS Immuno histochemistry and Terminal Transferase-mediated dUTP Nick End-Labeling (TUNEL) to evaluate distribution of smooth muscle apoptosis in abdominal aortic aneurysm.Results:Underwent intra-aortic elastase perfusion to induce AAAs have a high-success-rate.Rate of AAA formation in positive control group 10％,60％,80％,respectively.The treatment group was 0％,10％,20％,and the negative control group was not formed.The treatment group and the negative control group were lower than the positive control group,there were significant differences.In the positive control group,NO content increased gradually from second days,7 days to reach the peak and maintained at a higher level,the treatment group serum NO content was lower than the other two groups,there was significant difference (P＜0.05),iNOS was strong expression in the positive control group,in the other two groups of mild expression.TUNEL results showed that a lot of apoptotic cells in the positive control group,after 7 days showed a significant increase trend,to observe the end (2 weeks) gradually increased.,The positive control group was higher than the negative control group and the negative control group,there was significant difference(P＜0.05).Conclusion:iNOS inhibitors significantly decreased the content of NO in serum,reduced the apoptosis of smooth muscle cells,and inhibited the formation of abdominal aortic aneurysm.To provide theoretical basis for clinical application of iNOS inhibitors in the treatment and control of AAA.

Adolescent ; Adult ; Child ; Female ; Humans ; Male ; Middle Aged ; Myringoplasty ; Risk Factors ; Tympanic Membrane Perforation ; etiology ; surgery ; Young Adult

OBJECTIVE: To investigate the effect of serglycin knockdown on the sensitivity to cisplatin in nasopharyngeal carcinoma highly metastatic cells. METHODS: Stable transfected nasopharyngeal carcinoma highly metastatic cell lines was established. The alteration of serglycin expression was examined by qRT-PCR and Western blot assay established. The IC50 of cisplatin in stable cells were measured by MTT assay. The effect of serglycin knockdown on the proliferation of stable cells treated with cisplatin were measured by MTT assay. Colony formation assay were used to show cloning capacity. Annexin V/PI staining were used to detect apoptosis. qRT-PCR analysis was performed for stem cell-associated genes in stable cells. RESULTS: Stable transfected cell lines were successfully established and cellular morphology of stable transfected cells were changed. When treated with cisplatin, the IC50 and the colonies of serglycin knockdown cells were significant lower than controlled cells, however, the inhibition of proliferation and the percentage of apoptosis were significant higher than controlled cells. The stem cell-associated genes were reduced in serglycin knockdown cells. CONCLUSION: Stable interference of serglycin makes nasopharyngeal carcinoma highly metastatic cells more sensitive to cisplatin.

Objective To explore clinical effect of repairing soft tissue defect in forearm, hand and foot with free super-thin anterolateral thigh perforator flaps. Methods At first the site of perforator vessels were determined by Doppler, then the flaps were designed and harvested with the site as center; the fascia lata and subcutaneous fat were removed by sandhill-likely only the 4.0 cm × 3.0 cm - 3.0 cm×2.5 cm disc-like fascia lata and dermis layer were reserved. 15 traumatic soft tissue defects including forearm, hand and foot were repaired with the ree super-thin antemlateral thigh perforator flaps. Results No vascular crisis happened and all skin grafts survived in donor sites. 2.0 cm×1.2 cm of the distal of flap was necrosis in 1 case and it was healed by dress changing. 15 cases were followed up 3 months-2 years and the average is 6 months. The contour and texture of all flaps were good and two point discrimination (2-PD) was about 8-10 mm of. Conclusions The contour and texture of free super-thin anterolateral thigh perforator flap are good, the feeling of recipient site recovered well, it's less injury for donor site and there is no reshaping for flap. It is a fineness donor site for repairing soft tissue defects in hand and foot.

Objective To explore the alterations of protein phosphatase-2A (PP-2A) in lymphocytes in mild cognition impairment (MCI) and Alzheimer's disease (AD).Methods The activity PP-2A of was measured by ~(32)p liquid seintillography for incorporated radioactivity in control group(n=11) , the MCI group(n=11),and the AD group(n=11).The expression of PP-2A was determined by Western blot.Results In the control group,the activity of PP-2A (1.01?0.09) and the expression of PP-2A (0.96?0.07) were high while in the MCI group,the activity of PP-2A (0.71?0.12) and the expression of PP-2A (0.80?0.05) were decreased (both P

Polyethylene glycol-polybenzyl-L-glutamate copolymer (PEG-PBLG) was synthesized and paclitaxel-loaded core-shell type nano-micelles with amphiphilic copolymer PEG-PBLG was prepared by the dialysis method. The drug loading content and entrapment efficiency were determined by HPLC. The average size and its distribution were determined by dynamic light scattering method. The paclitaxel release rate in vitro from micelles was measured by HPLC. The cell cytotoxicity in vitro was observed with MTT assay. The anti-tumor activity of paclitaxel-loaded micelles were evaluated in tumor-inhibiting test of nude mice using human liver cancer HepG-2. The results indicated that paclitaxel could be entrapped in PEG-PBLG copolymer micelles and its size was in the range of 80-265 nm which increased with an increase in molecular weight of PBLG in copolymer; in vitro the paclitaxel could be released sustainably from the micelles. In high concentration of paclitaxel (>20 microg x mL(-1)) the paclitaxel-loaded PEG-PBLG micelles displayed much less cell cytotoxicity than paclitaxel injections with Cremophor EL (P<0.05); the tumor inhibiting activity of paclitaxel-loaded PEG-PBLG micelles was similar to that of paclitaxel injections with Cremophor EL in the same paclitaxel concentration. It was concluded that the paclitaxel-loaded PEG-PBLG micelles had more uniform size and size distribution, excellent drug sustainable-release behavior, less cytotoxicity, good anti-tumor activity similar to paclitaxel injections with Cremophor EL. So paclitaxel-loaded PEG-PBLG micelles would be a novel paclitaxel preparation in clinic for the treatment of tumor.
Animals ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Delayed-Action Preparations ; Drug Delivery Systems ; Humans ; Liver Neoplasms, Experimental ; pathology ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Micelles ; Nanoparticles ; Neoplasm Transplantation ; Paclitaxel ; pharmacology ; Particle Size ; Polyethylene Glycols ; chemistry ; Polyglutamic Acid ; analogs & derivatives ; chemistry ; Polymers ; Random Allocation

Objective To evaluate the application value of MALDI-TOF MS system in clinical routine separation microorganism by comparing two kinds of microorganism identification system MALDI-TOF MS and Vitek 2 Compact.Methods The bacterial species database of MALDI-TOF MS system and the Vitek 2 Compact system were compared,the new strains from MALDI-TOF MS library were screened out,and the number and frequency of new strains detected from May 2015 when MALDI-TOF MS was put in use to December 2016,and the frequency of detection of common pathogenic bacteria with high confidence were counted.Results There were 205 more new strains in the MALDI-TOF MS library,compared with Vitek 2 Compact.From May 2015 to December 2016,206 times were detected in clinical microbiological examination,and 286 cases if 4 kinds of clinical common pathogenic bacteria were identified by MALDI-TOF MS system with high confidence.Conclusion MALDI-TOF MS strain database is more accurate and large,which can meet the needs of clinical microbiological examination,and is suitable for wider application in clinical microbial identification.

Alleles ; Amelogenin/genetics* ; Chromosomes, Human, Y ; Humans ; Male ; Sex Determination Analysis

OBJECTIVETo investigate the effect of VEGF expression in osteosarcoma cell line and the target killing effect of HSV1-TK/GCV system on transfected osteosarcoma cells under hypoxia conditions.

METHODSEukaryotic expression plasmid with HRE promoter was constructed to express the antisense VEGF165 cDNA and Hygromycin phospho-transferase-thymidine kinase (HyTK) fusion gene. The recombinant vectors were then transfected into osteosarcoma cell line MG63 with lipofectin mediated gene transfer methods. PCR and RT-PCR were used to confirm the presence and expression of TK gene. The sensitivity of transfected cells to GCV and "bystander effect (BSE)" of HSV1-TK/GCV system under normoxia or hypoxia conditions were measured by MTT assay and mixed co-culture experiment. The expression of VEGF protein was detected by ELISA under hypoxia condition. Cell cycle phase distribution was determined by flow cytometry. In addition, electromicroscopy was used to document ultrastructural alterations.

RESULTSThe eukaryotic expression vector pBI-HRE-AsVEGF165 -HyTK was constructed successfully. The transfected cell line MG63TV was established and confirmed by PCR and RT-PCR of the presence of transgene and its mRNA expression. GCV was toxic to transfected cells in a concentration-dependent manner. The sensitivity to GCV toxicity was 100 times higher under hypoxia condition than that under normoxic condition. The mixed culture experiments showed that the "bystander effect" was enhanced significantly under hypoxia condition. VEGF expression of transgene cells under hypoxia condition decreased 50% compared to that of normal condition. Under hypoxia and GCV, DNA synthesis of MG63TV cells was inhibited along with an increase of cells at G0 approximately G1 phase, apoptosis and necrosis.

CONCLUSIONSAntisense VEGF expression driven by HRE promoter in combination with hypoxia can provide a target inhibition of VEGF expression in human osteosarcoma cells, with an enhanced selective killing effect and BSE of the HSV-TK/GCV system. The double-gene co-expression system in study provides experimental basis for therapy against osteosarcoma by a synchronous antiangiogenic and suicide gene approach.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Bystander Effect ; Cell Hypoxia ; Cell Line, Tumor ; Cell Proliferation ; DNA, Neoplasm ; biosynthesis ; Ganciclovir ; pharmacology ; Genetic Vectors ; Humans ; Hypoxia-Inducible Factor 1 ; genetics ; Oligodeoxyribonucleotides, Antisense ; Osteosarcoma ; metabolism ; pathology ; Phosphotransferases (Alcohol Group Acceptor) ; genetics ; Plasmids ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Recombinant Proteins ; genetics ; metabolism ; Thymidine Kinase ; biosynthesis ; genetics ; Transfection ; Vascular Endothelial Growth Factor A ; genetics ; metabolism