Objective:Explore decoy oligonucleotide (ODN) technology targeted blocking STAT3 activation way effects on colorectal cancer HT29 cell growth,to provide experimemt basis for gene therapy of colorectal cancer.Methods:Using decoy ODN in vitro transfect colorectal cancer HT29 cell line,to targeted blocking STAT3 activation way.Fluorescence microscope and flow cytometry detect transfection conditions and efficiency,and MTT method detect the cell growth inhibition,Realtime RT-PCR and Western blotting test STAT3 in cells,the Bcl-xl and Caspase3 mRNA and protein expression.Results:STAT3 decoy ODN can efficient transfection into colorectal cancer cells,and is due to the nucleus.HT29 cell in the STAT3 decoy ODN,growth inhibition rate increased significantly,and mRNA and quantity of protein expression of STAT3 significantly reduce (P＜0.05),while caspase3 increase significantly (P＜0.05).Conclusion:Named Decoy ODN targeted after blocking STAT3 activation way,which can effectively cut colorectal cancer cells HT29 STAT3 gene expression,and inhibit the growth of the cells.The mechanism may be related to lower the expression of STAT3 and the Bcl-xl and Caspase3.

Corneal collagen cross-linking (CXL) is a photochemotherapy for keratoconus, keratectasias and infectious keratitis.Corneal collagen fiber infiltrated with riboflavin, a photosensitizer which generates reactive oxygen species when activated by UVA at 370nm, was induced to form the crosslinks in corneal stroma.Corneal collagen cross-linking alone or combined with medicine therapy could be used to treat fungal keratitis, improve the cure rates, and reduce the complications and the demands of corneal transplantation surgery.The purpose of this paper is to review the basic principle, procedure, laboratory researches, clinical applications and the safety about this treatment.

AIM: Currently available anti-scarring regimens for glaucoma filtration surgery have potentially blinding complications and safer alternatives would be beneficial. This experiment is to investigate the effect of TGF-β2 antisense oligodeoxynucleotide on differentiation, proliferation of subconjunctival fibroblast following glaucoma filtration surgery.METHODS: Glaucoma filtration surgery were performed on both eyes of 28 rabbits. TGF-β2 antisense oligodeoxynudeotide was subconjunctivally injected in the right eyes (A group), and TGF-β2 missense oligodeoxynucleotide (B group)or PBS(C group) was used at the same method in the left eyes as controls. Rabbits were killed at 4,7,14 and 28 days after surgery. Intraocular pressure (IOP), bleb characteristics were recorded at different time point. Subconjunctival fibroblasts were examined by immunohistochemistry and electron microscopy.RESULTS: The IOP of rabbits in group A was significantly lower at 14 days (6.74± 1.18 mmHg) and 21 days (8.15± 1.97mmHg) after operation than the IOP in group B (8.53± 1.04,9.72± 1.09 mmHg)(P ＜0.01) and group C(8.79± 1.21, 9.43±1.27 mmHg) (P ＜0.05). The mean bleb survival time was longer (17.2 days) in group A than that of group B (14.5 days) and group C (13.5 days)(P＜0.05). The population of the cells expressing α -smooth muscle actin(α -SMA) and proliferating cell nuclear antigen (PCNA) was significantly reduced in group A compared with the group B and C. The ultrastructure of fibroblast was not altered by TGF-β2 anti-sense oligodeoxynucleotide.CONCLUSION:TGF-β2 antisense oligodeoxynucleotide can prevent the scar formation after glaucoma surgery by inhibit the differentiation and proliferation of subconjunctival fibroblast. It could be a potentially useful anti-scarring alternative for the prevention of late surgical failure.

Objective:To express and purify recombinant human collagen type Ⅲ and evaluate its properties.Methods:The recombinant genetic engineering strain pET30a(+)-1880/pACYCDuet-hy726/bL21(DE3) was constructed to stably co-express recombinant human type Ⅲ collagen (rhCol) and prolyl hydroxylase. rhCol was prepared and purified by E. coli high-density fermentation, salting out and column chromatography protein purification technology. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis was used to determine the purity of rhCol. The N-terminal amino acid sequences of rhCol were determined by automatic protein polypeptide sequencing instrument. The hydroxyproline content of rhCol was determined by ultraviolet spectrophotometry. The cellular compatibility of rhCol was evaluated by MTT assay. Results:The final wet weight of high-density fermentation was about 200 g/L. The expression level was about 3 g/L. The purity of rhCol by affinity chromatography was over 95%. The results showed that the hydroxyproline content of rhCol was 11.44%, and the rhCol products have good water solubility and cell compatibility.Conclusions:RhCol can be widely applied to the field of skin care and biomedicine as an excellent biological material.

Objective To investigate the effects of Thomas solution in combination with nitroglycerine of different concentrations on rat cardiac preservation for 8 h. Methods Thirty Wistar rats were selected,and Langendorff heart perfusion models were established.Rats were randomly divided into Thomas solution group,Thomas solution+4?10-6 mol/L nitroglycerine group and Thomas solution+4?10-5 mol/L nitroglycerine group(n=10).After hypothermic preservation(4 ℃)for 8 h,the cardiac function of each group was determined.Tissues of left ventricular wall were obtained,and changes of myocardial ultrastructure were observed by electron microscopy. Results Compared with Thomas solution group,the cardiac function was significantly improved and the myocardial ultrastructure injury was less severe in Thomas solution+4?10-5 mol/L nitroglycerine group after hypothermic preservation(4 ℃)for 8 h,while there was no significant difference in myocardial ultrastructure injury and cardiac function between Thomas solution group and Thomas solution+4?10-6 mol/L nitroglycerine group. Conclusion Thomas solution in combination with 4?10-5 mol/L nitroglycerine can significantly improve the preservation effects for isolated hearts in rats.

OBJECTIVETo investigate clinical characteristics of posterior cruciate ligament (PCL) injuries.

METHODSTo review all 326 patients with PCL injuries from January 2006 to June 2013 retrospectively, which were grouped according to injury patterns. The survey included the gender, age, sides, duration from injury to surgery, cause of injury and combined injuries. Compare the patient-specific factors between two groups.

RESULTSMale patients were the majority of PCL injuries (73%), and 30 to 50 years old patients accounted for 59%. 62.6% of all patients went for a doctor within one month after trauma. Traffic accidents were the main cause of PCL injuries (59%), especially motorcycle accidents (74.3%). Anterior cruciate ligament (ACL) injuries were the most common combined injuries (46.9%),followed by medial collateral ligament (MCL) (29.8%) and posterolateral corner (PLC) (26.1%). The meniscal tears accounted for 30.4% of total cases. The in-substance PCL injuries (71.5%) were far more than avulsion fractures (28.5%), and the former had more combined injuries than the later (P < 0.05). The causes of injuries were statistically different between two groups (P < 0.05). The prevalence of isolated avulsions was higher than isolated in-substance injuries (P < 0.05).

CONCLUSIONIn the general population, in-substance PCL tears were dominant on PCL injuries. The prevalence of combined injuries was quite high, among which ACL injuries were the most common. Insertion avulsion fractures emerged mainly at the tibial side, which had less combined injuries than those suffered in-substance PCL injuries. The incidence of isolated insertion avulsion fractures was higher than isolated in-substance injuries.

Adolescent ; Adult ; Aged ; China ; epidemiology ; Female ; Humans ; Incidence ; Knee Injuries ; epidemiology ; Male ; Middle Aged ; Posterior Cruciate Ligament ; injuries ; Retrospective Studies ; Young Adult

AIM: To investigate the protective effects and mechanisms of modified acidic fibroblast growth factor (maFGF) in anoxic reperfusion of rat hearts. METHODS: Using Langendorff apparatus, we established the model of anoxia/reperfusion of isolated hearts to compare the protective effects of maFGF and acidic fibroblast growth factor (aFGF). The changes of left ventricle development pressure (LVDP) and maximal rates of rise of ventricular pressure(dp/dt_~max ), maximal rates of decline of ventricular pressure (dp/dt_~min ) were determined, changes of LDH and MDA levels,SOD activity in efflux from coronary artery were also detected at different time point. RESULTS: Pretreatment with maFGF and aFGF produced a similar protective effect on myocardium during anoxia /reperfusion, including promoting obviously heart functional recovery after myocardial anoxia/reperfusion and reversing changes of LDH, MDA contents and SOD activity induced by anoxic/reperfusion. CONCLUSION: maFGF has a protective effect on anoxia/reperfusion heart, and the mechanism of this effect may be related to suppression of lipid peroxidation.

AIM: To investigate the mechanisms by which siduqing, a Chinese medicine, protects against lipopolysaccharide (LPS)-induced acute renal dysfunction. METHODS: Mice were divided randomly into control, LPS, siduqing treatment and siduqing groups, and treated intragastrically with siduqing at a dose of (1 000) g/L (0.2 (mL/10 g) body weight) or distilled water (0.2 (mL/10) g body weight) twice a day for 3 days, LPS (30 mg/kg) or normal saline was injected intraperitoneally on day 3, followed by intragastrical administration with siduqing at a dose of (1 000) g/L (0.2 (mL/10 g) body weight) or distilled water (0.2 (mL/10 g) body weight). Blood was collected for determining urea nitrogen (BUN) and creatinine (Cr) contents, renal tissue for examining superoxide dismutase (SOD) activity and malondialdehyde (MDA) content. In addition, electron microscopy was used to examine the ultrastructure changes in kidney, and RT-PCR was performed to detect renal intercellular adhesion molecule-1 (ICAM-1) mRNA expression. RESULTS: LPS significantly increased serum urea nitrogen (BUN) and creatinine (Cr) contents, and produced an obvious pathological change in renal ultrastructure, which were significantly attenuated by siduqing treatment. Moreover, siduqing treatment increased renal SOD activity, also markedly suppressed an increase in renal MDA production and ICAM-1 mRNA expression induced by LPS. CONCLUSION: These results suggest that siduqing protects against LPS-induced acute renal injury through inhibiting ICAM-1 mRNA expression, enhancing renal SOD activity and attenuating oxidant stress.

Objective To explore an algorithm to define the threshold value for tumor contouring on 18F-fluorodexyglucose (FDG) PET imaging. Methods A National Electrical Manufacturing Association (NEMA)NU 2 1994 PET phantom with 5 spheres of different diameters were filled with 18F-FDG. Seven different sphere-to-background ratios were obtained and the phantom was scanned by Discovery LS 4. For each sphere-to-background ratio, the maximum standardized uptake value ( SUVmax ) of each sphere, the SUV of the border of each sphere ( SUVborder ), the mean SUV of a 1 cm region of background (SUVbg) and the diameter (D) of each sphere were measured. SPSS 13.0 software was used for curve fitting and regression analysis to obtain the threshold algorithm. The calculated thresholds were applied to delineate 29 pathologically confirmed lung cancer lesions on PET images and the obtained volumes were compared with the volumes contoured on CT images in lung window. Results The algorithm for defining contour threshold is TH% = 33.1% + 46.8% SUVbg/SUVmax + 13.9%/D ( r = 0.994) by phantom studies. For 29 lung cancer lesions, the average gross tumor volumes ( GTV ) delineated on PET and CT are ( 7.36 ± 1.62 ) ml and (8.31 ±2.05) ml, respectively (t = -1.26, P＞0.05). Conclusion The proposed threshold algorithm for tumor contouring on PET image could provide comparable GTV with CT.