Background Previous studies showed that microRNA-184 (miR-184) might inhibit the biological function of vascular endothelial cells.In ophthalmology,to determine the inhibitory effect of miR-184 on angiogenesis is of clinical significance for the prevention and treatment of corneal inflammation,retinal and choroidal neovascularization.Objective This study was to verify the influence of miR-184 transfection on the proliferation,migration and angiogenic ability of HUVECs and explore its mechanism.Methods HUVECs line was cultured in vitro and divided into 4 groups.No transfection regent and RNA molecule were added in the medium of the blank control group;only transfection regent was added in the blank transfection group; hsa-miR-184 mimic/negative was added in the negative control group,and Cy3 labeled hsa-miR-184 mimic-lipofectamine 2 000 mixture was added in the miR-184 transfection group.The proliferation of HUVECs (absorbance,A490) was detected by MTT.The transmembrane cell number was counted by Transwell insert to evaluate the migration ability of HUVECs.A three dimensional cultural system of cells was constructed on the matrigel,and tube formation number of HUVECs was assessed.Results Positive HUVECs for Cy3 labeled siR-RiboTM + hposome showed red fluorescence,with the optimal transfecting concentration ＞ 100 nmol/L.The relative expression levels of miR-184 were 1 524.10±385.89 and 1.00±0.05 in the miR-184 transfection group and the negative control group,showing a significant difference (t=-7.894,P＜0.01).The A490value of HUVECs was 0.50±0.04 and the number of migrating cells through the transwell membrane was (55.40±5.86)/field in the miR-184 transfection group,and they are significantly reduced in comparison with (0.62±0.04) and (83.40±5.59)/field in the negative control group (t =5.639,7.730,all at P＜ 0.01).A significant difference was found in the total branch points of capillary tubes between the miR-184 transfection group and negative control group ([13.33 ± 2.08]/field versus [22.00 ± 2.00]/field) (t =5.511,P＜ 0.05).Conclusions MiR-184 can suppress the proliferation,migration and tubing of HUVECs after transfection in vitro.MiR-184 participates in the regulation of angiogenesis.

[ ABSTRACT] AIM:To investigate the therapeutic effects of lentivirus-mediated shRNA targeting growth hormone secretagogue receptor 1a (GHSR1a) on colorectal cancer cell line SW480 both in vitro and in vivo.METHODS: Human GHSR1a sequence was used for the design of shRNA targeting GHSR1a, which was introduced to lentivirus, followed by transfection into SW480 cells.CCK-8 assay was performed to detect cell viability.The mRNA expression of GHSR1a and PTEN in colorectal cancer cells was detected by RT-PCR.The protein levels of GHSR1a, ghrelin, PTEN, p-AKT and p53 were determined by Western blot.Stable GHSR1a silencing in SW480 xenografts in nude mice was established.After the mice were sacrificed and weighted, immunohistochemistry was used to detect the positive expression of Ki-67 and PTEN in the tumors.RESULTS:GHSR1a was over-expressed in the malignant cells in comparison with the normal cells in vitro. The tumor specific lentivirus-mediated shRNA targeting GHSR1a gene and GHSR1a knockdown SW480 cells were success-fully constructed.After transfection with GHSR1a shRNA, the expression of GHSR1a at mRNA and protein levels was markedly inhibited in the SW480 cells.Furthermore, GHSR1a silencing by specific shRNA showed increased PTEN, inhi-bition of AKT phosphorylation and promotion of p53 release in the SW480 cells.In vivo results demonstrated that down-re-gulation of GHSR1a in the SW480 cells significantly decreased the expression of Ki-67 and increased the expression of PTEN in the tumor tissues, leading to a marked reduction in tumor weight in comparison to blank control or negative control
tumors.CONCLUSION:Down-regulation of GHSR1a by shRNA technique inhibits the growth of colorectal cancer cell line and xenograft tumor through activation of the PTEN/PI3K/AKT signaling pathways.

Objective To explore the influence of hyperbaric oxygen(HBO)on the life-spans of tumor-bearing mice. Methods Twenty-eight male Balb/c mice were randomly and evenly divided into:a celiac-tumorgroup(inoculated with tumor cells in the abdominal cavity);a celiac-tumor-HBO group(inoculated and then ex-posed to hyperbaric oxygen);a back-tumor group(inoculated under the skin of the back);and a back-tumor-HBOgroup(inoeulatedunder the skin of the back and then exposed to hyperbaric oxygen).S-180 carcinoma cells were in-oeulated,and then HBO was administered once a day.The life-spans and any skin ulceration were observed. Re-sults The average life-spans were(27.6±4.5)days in the celiac.tumor group,(24.0±2.9)days in the celiac-tumor-HBO group,(63.0±21.8)days in the back.tumor group and(35.0±8.9)days in the back-tumor-HBOgroup.The difference in average life-span between the celiac-tumor group and the celiac-tumor-HBO gmup was not significant.The difference in life-span between the back-tumor group and the back.tumor.HBO group was significant.Ulcers occurred in 4 mice in the back-tumor group and 4 in the back-tumor-HBO group.The times of ulcer formation were significantly different between the back-tumor group and the back-tumor-HBO group. Conclusions The life-spans of mice in the back-tumor-HBO group were shortened by HBO exposure,but life-spans in celiac-tumor-HBOgroup were not.Ulcer formation was postponed by HBO in the back-tumor-HBO group.

The finite element method(FEM)is a numerical computation method based on computer tech-nology, and has been gradually applied in the fields of medicine and biomechanics. The finite element analysis can be used to explore the loading process and injury mechanism of human body in traffic in-jury. FEM is also helpful for the forensic investigation in traffic injury. This paper reviews the develop-ment of the finite element models and analysis of brain, cervical spine, chest and abdomen, pelvis, limbs at home and aboard in traffic injury in recent years.

Objective To investigate the clinical value of a newly designed surgical therapy for familial polyposis coli by severing the superior mesenteric artery&vein in order to make a complete lysis of the mesentery and an ileum pouch and the anal anastomosis within the entire muscular sheath of the rectum.Methods Six patients with familial polyposis coli(5 males and 1 female,aged 24-36 years)were admitted and underwent the procedure which was consisted of:(1)An incision was made in the left middle and lower parts of the rectus abdominis;(2)The greater omentum was retained and the large intestine was removed;(3)At the juncture of the sigmoid colon and the rectum,the muscular sheath was dissociated 0.5cm,the mucous membrane of the rectum was stripped in a revolving manner,the nourishing artery and vein in the membrane were exposed,and clamped and cut in sequence up to the anocutaneoue line;(4)The rectal mucous membrane was completely removed;(5)Under the right colonic artery,the superior mesenteric artery and vein were severed;(6)An N-,J-or W-shaped pouch was made in the ileum accordingly:(7)An anastomosis of the ileum pouch and the anal canal was made within the entire muscular sheath of the rectum,and a drainage was placed;(8)The mesostenium was fixed on the fight posterior abdomen,the small intestines were spread out to the right side,and the mesostenium was covered on the coarse surface of the colon bed:(9)A tube was placed in the left lower abdomen for a vacuum aspiration for 2 days after operation,combined with the suction drainage,to eliminate the pelvic effusions;and(10)The abdomen was closed.Results Patients were able to discriminate stools and flatus 3-7 days after operation.and the formed stools occurred 7-10 days after operation.Five patients were followed-up for 3-17 years,with averagely one defecation a day,with no night defecation and seepage.Urination was normal;In another one patient who underwent the procedure 4 months ago the defecation was twice a day,with no night defecation.All the 6 patients had normal autonomic nerve function and sexual function as well as normal defecation and urination,with no recurrence of polyposis coli or infection.The small bowel functions well with no ischemia related symptoms.Conclusion Cutting the superior mesenteric artery and vein and then making anastomosis of the ileum pouch and the anal canal within the muscular sheath of the rectum is a new surgical approach to familial polyposis coli.It is safe and significantly improves the patients' life quality.

Objective To study the microRNA ( miRNA) regulation on the expression of housekeeping and non-house-keeping human genes from the systemic level , and to reveal the influence of conservation at 3′UTRs region for the density difference of miRNA regulatory between the two classes of genes .Methods We obtained miRNA regulation informations for the two classes of genes through the integration analysis of three sequence-based target gene prediction softwares and the conjoint analysis based on gene expression profile data .We also used phastCons-score based evolution analysis to study the conservation of 3′UTRs regions between the two classes of genes .Results Our results showed that housekeeping genes have a significantly higher density of miRNA binding sites and their 3′UTR are relatively conservative .Conclusion These findings highlight the importance of miRNA regulation in housekeeping genes , suggesting that investigating the roles of miR-NAs on the robustness of housekeeping genes expression is highly necessary .

Objective To provide reference on the difference in blood glucose metabolism between different mouse strains by comparing blood glucose changes under food and /or water deprivation.Methods C57BL/6J mice, BALB/c mice,and ICR mice used in this study were divided into four groups:control group, food deprivation group, water depriva-tion group and food &water deprivation group .Their blood glucose was measured within 12 hours under various stresses . Results and Conclusion There are different blood glucose regulation patterns in different mice strains .The blood glucose level of C57BL/6J mice is usually higher than that of the BALB/c and ICR mice, while the of blood glucose level BALB/c mice stays relatively stable , and the ICR mice have the widest range of blood glucose levels upon these stresses .Accord-ing to our data, the difference in blood glucose metabolism between different mouse strains should be taken into considera -tion in blood glucose related studies for proper interpretation and design of experiments .

Objective To provide reference for clinical application of combined antoxidants , by evaluating the com-bined and individual antioxidant activities of three commonly used antioxidants :edaravone,glutathion(GSH) and vitamin C ( VC) .Methods The multiple and individual antioxidant activities of the three antioxidants were compared by determining total antioxidant activity assay , reductive activity assay , superoxide anion scavenging assay and hydroxyl radical scavenging assay.Results and Conclusion Results showed that the combined antioxidants acted synergistically in scavenging free radicals and increased remarkably antioxidant activity at the same molar concentration .The rationality of different antioxi-dant combination should be taken into consideration in subsequent clinical application .

OBJECTIVE: To investigate the effect of trichostatin A (TSA),a specific inhibitor of histone acetyltransferase,on apoptosis of human hepotoma cell line HepG2,the expression of fragile histidine triad (FHIT) and apoptosis-related proteins in order to study apoptosis mechanisms.METHODS: Human hepatoma cell lines HepG2 were cultured and treated with different concentrations of TSA(125,250,500,1 000,2 000 nmol?L-1) for 24 and 48 hours.Human hepatoma cell lines HepG2 survival and apoptosis were determined by MTT assay with absorbance vale and inhibitory rate as index.Apoptotic percentage of HepG2 treated with 250 and 1 000 nmol?L-1 TSA were determined using TUNEL assay.The expressions of FHIT and caspase-3,bax,bcl-2 were analyzed by immunocytochemistry with solvent as control.RESULTS: As compared with control group,absorbance vale of human hepatoma cell lines HepG2 were decreased after treated with different concentration of TSA in dose-dependent and time-dependent manners.Positive cell rate in TUNEL was increased (P0.05).CONCLUSION: TSA may inhibit proliferation and promote apoptosis of hepatoma HepG2 cell by up-regulating the protein expression of FHIT,caspase-3 and bax.

The changes of Fas.FasL and Bcl-2 expression in thyrocytes of patients with Graves'disease were investigated before and 2 weeks after 131Ⅰ administration. The results showed that 131Ⅰ couhl induce thyrocytes to express the apoptotic protein Fas, FasL and the anti-apoptotic protein Bet-2 in patients with Graves'disease. A statistically significant correlation was found between the dose of 131Ⅰ and the expression levels of Fas and FasL but not Bet-2 ,suggesting that early onset of hypothyroid after 131Ⅰ administration may be due to the increased expression of Fas and FasL in thyrocytes.