OBJECTIVETo evaluate the effect of antisense monocyte chemotactic protein-1 (A-MCP-1) nanoparticles (NPs) as gene carrier on gene transfer in two kinds of animal models.

METHODSPoly (lactic acid-co-glycolic acid) (PLGA) was used to make the NPs loaded with A-MCP-1 through a double-emulsion/solvent evaporation technique. NPs size was assessed by dynamic laser defractometer. The particle morphology was observed by scanning electron microscopy. DNA content in the NPs was measured by dissolving known amounts of NPs in chloroform and extracting DNA with water. In vitro release was performed in tris-EDTA buffer at 37 degrees C using double-chamber diffusion cells. The receiver buffer was replaced daily. The A-MCP-1 NPs was transfected into the cultured smooth muscle cells. PCR was used to evaluate the transfection of A-MCP-1. Cationic lipid (Lipofectamine) was used to transfect A-MCP-1 as control. After 48 hours incubation, cells were digested and examined by polymerase chain reaction. Twenty New Zealand white rabbits under jugular vein to artery bypass grafting procedure were divided into four groups: the first group received grafts treated with A-MCP-1 NPs, the second group received grafts treated with cationic liposome (dioleoyl trimethyl ammonium propane)-A-MCP-1, the third group received grafts treated with plasmid DNA, and the fourth group received grafts without transfection as control. Fourteen days after surgery the grafts were harvested. The expression of A-MCP-1 and its effect on MCP-1 in vein grafts were detected by dot blotting. The morphology of the grafts was investigated. To establish abdominal aortic aneurysms rats model, rats were randomly divided into three groups: A-MCP-1 NPs injection group, shame NPs injection group and control groups (without injection). Two weeks after surgery, diameter of abdominal aorta was measured and aortic tissue was obtained for PCR analysis to evaluate the A-MCP-1 expression. Western blot were applied to detect the inhibitory effect to the expression of MCP-1 mRNA and CD68 protein by A-MCP-1 NPs.

RESULTSNPs size ranged 198nm to 205nm with average around 201.4 nm. DNA content in the NPs was 4.14%. NPs showed steady release rate in vitro in Tris-EDTA solution. It released faster in the first week then maintained a slowly sustained release up to 16 days. In cell culture A-MCP-1 gene successfully transfected into smooth muscle cells by NPs vector. In vein grafting animal model, A-MCP-1 expression was detected in the vascular walls of NPs and cationic lipid treated groups. The degree of vascular hyperplasia in the gene NPs treated group was significantly lower than that in control group. There was no significant difference in the inhibition of intimal hyperplasia between NPs and cationic lipid treated groups. Two weeks after transfection in abdominal aortic aneurysm rats models, the abdominal aortic diameter of A-MCP-1 NPs injection group was (1.79 +/- 0.12) mm, significantly smaller than that of control groups [shame NPs group was (2.58 +/- 0.21) mm, and saline group was (2.63 +/- 0.29) mm] (P < 0.01). The expressions of MCP-1 mRNA and CD68 protein in A-MCP-1 NPs injection group were 12.5 +/- 1.5 and 17.6 +/- 2.1, which were much lower than those in control group [in shame NPs group, which were 35.7 +/- 4.5, 42.3 +/- 5.7 (P < 0.01), and saline group which is 32.4 +/- 3.9, 39.8 +/- 4.8 (P < 0.01)]. Specific band of A-MCP-1 was detected only in the A-MCP-1 NPs injection group by PCR.

CONCLUSIONA-MCP-1 gene NPs can be successfully used in rabbit vein grafting model and abdominal aortic aneurysm rats models, and may be potentially applied in clinical practice.

Animals ; Aortic Aneurysm ; genetics ; Chemokine CCL2 ; genetics ; metabolism ; Gene Transfer Techniques ; Genetic Vectors ; Lactic Acid ; chemistry ; Models, Animal ; Nanoparticles ; Oligonucleotides, Antisense ; genetics ; Polyglycolic Acid ; chemistry ; Polymers ; chemistry ; Rabbits ; Rats ; Transfection

Objective To investigate the therapeutic effects of simvastatin on experimental autoimmune encephalomyelitis(EAE)and explore its mechanisms. Methods Fifty-five Wistar rats were randomly divided into EAE group (n=15),.STATINS group(n=15),triptolide(TP)group(n=15)and normal control group(n=10).In STATINS group,the rats were given simvastatin and the changes in the expressions of P53,interleukin-6(IL-6),tumor necrosis factor-α(TNF-α)and transforming growth factor-β(TGF-β)were observed,with triptofide as the positive control.Results Compared to the EAE group,the rats in STATINS group had significantly lowered incidence of EAE,mild symptoms,reduced body weight loss and lesion foci number,and prolonged latency of EAE onset(P＜0.05).The rats in the TP group also exhibited significantly milder symptoms and fewer lesion foci than the EAE group(P＜0.05),but the body weight changes.,latency or incidence of EAE had no significant difference between the two groups(P＞0.05).Simvastatin significantly suppressed the expression of IL-6 and TNF-α and increased the expressions of P53 and TGF-β in rats with EAE,whereas TP only resulted in significant suppression of TNF-αexpression(P＜0.05).The expressions of P53 and TGF-β were significantly higher in STATINS group than in TP group(P＜0.05),but the expressions of TNF-α and IL-6 were comparable between the two groups(P＞0.05). Conclusions Simvastatin Can suppress EAE more effectively than TP by suppressing the expressions of the inflammatory factors such as IL-6 and TNF-α and promoting the expressions of P53 and TGF-β.

The different biological functions were studied in mouse bone marrow-derived endothelial progenitor cells isolated by differential time attachment to obtain the optimal adherent time in this study. Density gradient centrifugation-isolated bone marrow mononuclear cells were seeded on the fibronectin-coated dish. The 1-day cultured unattached cells were seeded on the second dish for 2 more days. Then unattached cells in the second dish were seeded on the third dish. The cells on 3 dishes were defined as 1-day adherent cells, 3-day adherent cells and 3-day unattached cells, respectively. After 20-day culture, the biological functions, such as the percentage of biomarkers, the ability of adhesion, and the ability of forming tubes in vitro were analyzed. The results showed that the percentages of positive CD34, FLK-1, and CD34/FLK-1 expressions in 1-day attached cells were significantly increased compared to those in the 3-day adherent or unattached cells (P < 0.01), which showed the strongest adhesion ability. The expression of eNOS in 1- or 3-day adherent cells was significantly higher than that in 3-day unattached cells (P < 0.01). The expression of VEGF in 3-day adherent cells was significantly higher than that in 1-day adherent cells or 3-day unattached cells (P < 0.01). These results suggest the biological functions of 1-day adherent cells are significantly stronger than that of 3-day adherent or unattached cells. VEGF expression in 3-day adherent cells is higher than that in 1-day adherent cells or 3-day unattached cells. The expression of eNOS in 1-day adherent cells or 3-day adherent cells is higher than that in 3-day unattached cells. The optimal adherent time to obtain mouse bone marrow-derived endothelial progenitor cells is 1-3 d.
Animals ; Bone Marrow Cells ; cytology ; Cell Culture Techniques ; methods ; Cell Differentiation ; Cell Separation ; methods ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; Leukocytes, Mononuclear ; cytology ; Male ; Mice ; Mice, Inbred C57BL ; Nitric Oxide Synthase Type III ; metabolism ; Stem Cells ; cytology ; metabolism ; Time Factors ; Vascular Endothelial Growth Factor A ; metabolism

OBJECTIVETo investigate the safety and efficiency of the disposable circumcision suture device (DCSD) in the surgical treatment of phimosis and redundant prepuce.

METHODSWe randomly assigned 249 outpatients with phimosis or redundant prepuce to be treated with DCSD (n = 129) and by conventional circumcision (CC, n = 120), respectively. Then we compared the safety and efficiency of the two strategies.

RESULTSComparisons between DCSD and CC showed that the operation time was (4.02 +/- 0.69) vs (30.8 +/- 4.05) min, blood loss was (1.07 +/- 1.29) vs (8.72 +/- 2.15) ml, intraoperative pain score was 0.81 +/- 0.81 vs 2.42 +/- 1.15, 24-hour postoperative pain score was 1.84 +/- 1.02 vs 4.99 +/- 1.36, postoperative complication rate was 13. 95% (18/129) vs 9.17% (11/120), wound healing time was (13.99 +/- 9.06) vs (17.48 +/- 3.49) d, satisfaction with the penile appearance was 98.4% (127/129) vs 95% (109/120), and treatment cost was (2215.62 +/- 17.67) vs (576.47 + 15.58) Y RMB. DCSD exhibited obvious superiority over CC for shorter operation time, less blood loss, milder intraoperative pain, sooner wound healing, and better penile appearance, but it also had a higher rate of postoperative complications (P > 0.05) and involved more treatment cost than the latter (P < 0.05).

CONCLUSIONThe disposable circumcision suture device affords ideal clinical effects and therefore deserves clinical popularization.

Circumcision, Male ; instrumentation ; Disposable Equipment ; Follow-Up Studies ; Humans ; Male ; Phimosis ; surgery ; Surgical Staplers ; Treatment Outcome

Objective To study the anatomic and biomechanical characteristics of the dorsal radial ligament and anterior oblique ligament in carpometacarpal (CPC) joints of the thumb,so as to provide references for ligament repair.Methods Forty fresh hand specimens of adult male cadavers were dissected to make specimen of trapezium bone-ligament-the first metacarpal bone,of which 20 cases retained the dorsal radial ligament and 20 cases retained the anterior oblique ligament,respectively.The ligaments were tested on biomechanical testing machine,and their length,width,thickness,the maximum load,elastic modulus and elongation rate were measured and calculated.Results For the dorsal radial ligament and anterior oblique ligament,their maximum load was (213.5 ±72.4) and (168.7 ±35.2) N,their elastic modulus was (17.2 ±6.7) and (9.3 ±2.5) MPa,their elongation rate was (116.2 ± 21.3) ％ and (92.7 ± 22.4) ％,respectively.The maximum load,elastic modulus and elongation of the dorsal radial ligament were larger than those of the anterior oblique ligament.Conclusions For the capsular ligament in CPC joints of the thumb,the dorsal radial ligament has a higher stiffness and stronger toughness,which plays an important role in maintaining stability of the joint.The anterior obligue ligament is easy to be damaged due to its smaller stiffness and poor toughness.The anterior oblique ligament is suggested to be reconstructed firstly to treat arthritis of CMC joints,and materials whose elastic modulus and elongation rate are similar with the dorsal radial ligament should be selected.

OBJECTIVETo investigate the expressions of X-linked inhibitor of apoptosis protein (XIAP)-associated factor-1 (XAFI) and heat-shock transcription factor 1 (HSF1) and their relationship in human gastrointestinal cancers.

METHODSImmunoblotting was used to analyze the expressions of HSF1 and XAF1 in gastric and colon cancer tissues and in gastrointestinal cancer cells. The gastrointestinal cancer cells were tranfected with a eukaryotic expression vector containing HSF1 gene fragment or subjected to RNA interference to induce up- or down-regulation of HSF1 expression, and the consequence changes in XAF1 expression in the cells was measured. XAF1 expression was also assayed in the cells after stress stimulation for HSF1 expression.

RESULTSThe expression of HSF1 was higher in gastrointestinal cancer tissues than in normal tissues. The expression of XAF1 and HSF1 was inversely correlated in the cancer cell lines, and stress stimuli of the cells up-regulated the expression of HSF1 but down-regulated XAF1 expression.

CONCLUSIONHSF1 expression is increased in gastrointestinal cancer tissues to result in suppressed expression of XAF1, which may be one of the reasons for the low expression of XAF1 in association with the defect of the apoptosis mechanisms in the cancer cells

Cell Line, Tumor ; Colonic Neoplasms ; genetics ; metabolism ; pathology ; DNA-Binding Proteins ; biosynthesis ; genetics ; Heat Shock Transcription Factors ; Humans ; Immunoblotting ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; biosynthesis ; genetics ; RNA Interference ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Transcription Factors ; biosynthesis ; genetics ; Transfection

OBJECTIVEX-linked inhibitor of apoptosis protein (XIAP) To gastrointestinal (GI) investigate the expression of XAF1 and heat-shock transcription factors 1 èHSF1éand their relationship in human gastrointestinal cancers.

METHODSImmunoblotting was used to analyze the expression of HSF1 and XAF1 in either gastric or colon cancer tissue and GI cancer cell line. Transient expression of the HSF1-containing vector in GI cell lines and RNA interference were used to up/down-regulae the expression of the HSF1, and the subsequent expression of XAF1 was measured.

RESULTSThe expression of HSF1 was higher in GI cancers than in normal tissues. The expression of XAF1 and HSF1 was negatively correlated in GI cancer cell lines. Stress stimuli up-regulated the expression of HSF1 while the alteration of XAF1 expression was negatively correlated with HSF1 expression.

CONCLUSIONThe high expression of HSF1 in GI cancers is associated with suppressed expression of XAF1, which can be one of the mechanisms for low-expression of XAF1 and insufficient apoptosis in GI cancers.

Animals ; Base Sequence ; Cell Line, Tumor ; DNA-Binding Proteins ; genetics ; Gastrointestinal Neoplasms ; genetics ; metabolism ; pathology ; Gene Expression Regulation, Neoplastic ; Heat Shock Transcription Factors ; Humans ; Intracellular Signaling Peptides and Proteins ; Neoplasm Proteins ; genetics ; Oxidative Stress ; genetics ; Temperature ; Transcription Factors ; genetics

Objective To study the anatomic and biomechanical characteristics of the dorsal radial ligament and anterior oblique ligament in carpometacarpal (CPC) joints of the thumb, so as to provide references for ligament repair. Methods Forty fresh hand specimens of adult male cadavers were dissected to make specimen of trapezium bone- ligament-the first metacarpal bone, of which 20 cases retained the dorsal radial ligament and 20 cases retained the anterior oblique ligament, respectively. The ligaments were tested on the biomechanical testing machine, and their length, width, thickness, the maximum load, elastic modulus and elongation rate were measured and calculated. Results For the dorsal radial ligament and anterior oblique ligament, their maximum load was (213.5±72.4) and (168.7±35.2) N, their elastic modulus was (17.2±6.7) and (9.3±2.5) N/mm2, their elongation rate was (116.2±21.3)% and (92.7±22.4)%, respectively. The maximum load, elastic modulus and elongation of the dorsal radial ligament were larger than that of the anterior oblique ligament. Conclusions In the capsular ligament in CPC joints of the thumb, the dorsal radial ligament has a higher stiffness and stronger toughness, which plays an important role in maintaining the stability of the joint. The stiffness of the anterior oblique ligament is smaller, the toughness is poor, which is easy to be damaged. The anterior oblique ligament is suggested to be reconstructed firstly to treat arthritis of CMC joints, and material whose elastic modulus and elongation rate is similar with the dorsal radial ligament should be selected.

Objective·To compare the caries prevalence, unmet restoration needs and sealant prevalence between Shanghai 12-year-old native and migrant students from 2013 to 2015. Methods·From 2013 to 2015, a random cluster sample of 12-year-old native and migrant students was selected from each district in Shanghai. World Health Organization (WHO) indices were used to assess decayed-missing-filled teeth of permanent dentition (DMFT). The prevalence of caries and sealants, caries filling rate, unmet dental restoration needs and the value of DMFT were analyzed using the Mann-Whitney U test and chi-squared test. Results·A total of 2500 students were examined from 2013 to 2015, among which 36.8％ were migrants. There was no significant difference in caries prevalence and DMFT value between native and migrant students from 2013 to 2015. Sealant prevalence of migrants was significantly lower than that of natives in 2013 (0.8％ vs 6.6％) and 2015 (3.0％ vs 9.9％). Unmet restoration needs of migrants were significantly higher than those of natives in 2015 (60.5％ vs 46.4％). Conclusion·There was no significant difference in caries prevalence between 12-year-old native and migrant students in Shanghai from 2013 to 2015. But there were lower sealants and higher unmet restoration needs in 12-year-old migrant students than their native counterparts, even though free public dental health services were available to both groups.