OBJECTIVETo investigate in situ visualization using near-infrared quantum dots (QDs) conjugated with arginine- glycine-aspartic acid (ROD) peptide fluorescent probes in oral squamous cell carcinoma (08CC).

METHODSQDs with emission wavelength of 800 nm (QD800) were conjugated with RGD peptides to produce QD800-RGD fluorescent probes. Human OSCC cell line BcaCD885 was inoculated in nude mice cheeks to establish OSCC mouse models. Frozen BcaCD885 tumor slices were immunofluorescence double stained by using QD800-RGD and CD105 monoclonal antibody and were observed using a laser scanning confocal microscope. QD800-RGD was injected into the OSCC models through the tail veins, and the in situ visualization was analyzed at different time points. The mice were sacrificed 12 h after injection to isolate tumors for the ex vivo analysis of probe localization in the tumors.

RESULTSQD800-RGD specifically targeted the integrin avβ3 expressed in the endothelial cells of tumor angiogenic vessels in vitro and in vivo, producing clear tumor fluorescence images after intravenous injection. The most complete tumor images with maximal signal-to-noise ratios were observed 0.5 h to 6 h after injection of the probe and significantly reduced 9 h after the injection. However, the tumor image was still clearly visible at 12 h.

CONCLUSIONUsing intravenously injected QD800-RGD generates high quality OSCC images when integrin avβ3, which is expressed in the endothelial cells of tumor angiogenic vessels, is used as the target. The technique offers great potential in the diagnosis and individual treatment of OSCC.

Animals ; Arginine ; Aspartic Acid ; Carcinoma, Squamous Cell ; Cell Line, Tumor ; Fluorescent Dyes ; Glycine ; Humans ; Mice ; Mice, Nude ; Mouth Neoplasms ; Oligopeptides ; Peptides ; Quantum Dots

Objective To study the effects of endotoxin on aldosteron secretion and nuclear factor-κB P65(NF-κB P65)mRNA expression in rat hepatic stellate cell(HSC).Methods Cultured rat HSC were treated with endotoxin of different concentrations. Aldosteron secretion of HSC was detected by radio-immunoassay. NF-κB P65 mRNA expression of HSC was determined by semiquantitative reverse transcription-polymerase chain reaction(RT-PCR).The data were analyzed by variance analysis,t-test and Pearson linear regression analysis. Results Aldosteron secretions of HSC groups treated with 0.01,0.1,1.0 and 10.0 mg/L endotoxin[(4.95±0.35),(5.52±0.32),(6.04±0.60)and(5.16±0.46)μg/L, respectively] were all significantly higher than that in control group(3.655±0.51)μtg/L(t=2.9745,5.8725,6.8465 and 3.2065,respectively;all P＜0.05).NF-κB P65 mRNA expressions of HSC groups treated with 0.01,0.1,1.0 and 10.01 mg/L endotoxin (0.82±0.06、1.07±0.07,1.23±0.06 and 0.96±0.05.respectively)were also significantly higher than that in control group 0.43±0.04(t=5.4776,6.8084,7.9382 and 7.5136,respectively;all P＜0.01).Both aldosteron secretion and NF-κB P65 mRNA expression in HSC groud treated with 10.0 mg/L endotoxin were significantly lower than those in HSC group treated with 1.0 mg/L endotoxin(t=4.3865,3.7246;both P＜0.05).In these treated HSC,aldosteron secretion was positively correlated with NF-κB P65 mRNA expression(r=0.886,P＜0.01).Conclusions Aldosteron secretion and NF-κB P65 mRNA expression in rat HSC could be up-regulated by stimulation with endotoxin, which shows a certain dose-response relationship. This may be one of the important factors of hepatic fibrosis development.

Objecfive To investigate the effects of endotoxin on nuclear factor-κB p65(NF-κB p65)mRNA expression and ahtosteron secretion in rat hepatic stellate cells(HSCs).Methods Cultured rat HSCs(HSC-T6)were divided into endotoxin-treated group and control group.Cells in endotoxin-treated group were exposure to 1 mg/ml.endotoxin.Aldosteron secretions of HSCs were determined by radioimmunoassay,and NF-κB p65 mRNA expressions of HSCs were detected by one-step RT-PCR.Results At 6,12,24 and 48 h,aldosteron secretions in endotoxin-treated group were significantly hisher than those in the control group(t=3.063,4.577,6.847 and 9.317,P<0.05),and the expressions of NF-κB p65 mRNA in endotoxin-treated group were also higher than those in control group(t=5.155,6.095,7.875 and 9.313,P<0.01).Aldosteron secretions and NF-κB p65 mRNA expressions in HSCs displayed a positive correlation(r=0.886,P<0.01).Conclusion Endotoxin can up-regulate the aldosteron secretion and NF-κB p65 mRNA expression in rat HSCs,which may be one of the mechanisms of liver fibrosis induced by endotoxin.

AIM: To observe the effect of Shenmai injection,a chinese medicine,on apoptosis of cardiac myocytes after hypoxia.METHODS: Cardiac myocytes were separated from neonate rat heart and cultivated in vitro.Hypoxia condition was induced by mixture of 95%N2 and 5%CO2.Cells were exposed to hypoxia for 6 h or 12 h and treated with Shenmai injection(5 mL/L) from 24 h before hypoxia until the end of hypoxia.First,apoptosis was detected with Annexin V-FITC and PI staining by flowcytometry.Then,the activity of cardiac myocyte mitochondria was observed by MTT method.Mitochondria membrane potential and the activity of caspase 3,7 were also measured by laser scan microscopy and multi-detection microplate reader,respectively.RESULTS: The apoptotic cells became more and more with prolonged hypoxia.Shenmai injection enhanced mitochondria activity,kept membrane potential,inhibited the activation of caspase3,7 and then decreased apoptotic cells(P

Objective:To study the effects of nitrous oxide(N2 O)sedation in the management of dental fear(DF)for dental treat-ments of children.Methods:66 cases of pediatric patients(aged 6 to 14 years)were given N2 O sedation for dental treatments.The heart rate(HR)and oxygen saturation(SpO2 )were measured before and after N2 O inhalation.N2 O effective concentration was eval-uated by Ramsay sedation score and Houpt behavior score.Results:The effective concentration of N2 O sedation was 25% -70%(49.6% ±12.1%),the maximum endurance concentration 35%-70% (56.4% ±10.1%).The Ramsy scores of N2 O sedation was (2.3 ±0.6).After N2 O inhalation,all patients could receive verbal demand during the treatments.The HR decreased(P <0. 05)and the Houpt behavior score increased(P <0.01).Before and after N2 O inhalation SpO2 had no significant difference(P >0.05).Conclusion:N2 O inhalation at 25% -70% is safe and effective in the management of DF for dental treatments of children.

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.

Objective To investigate the reliability and validity of a novel hand-held dynamometer, OE-210, for muscles strength of low-er extremities. Methods From March 1st to August 30th, 2016, 38 young adults were tested the muscle strength of quadriceps and ham-strings with OE-210 dynamometer by 2 rators, and were retested by one of the raters three days later. The isokinetic test was also conducted on all the subjects one day afterwards. The intraclass correlation coefficient (ICC) of OE-210 test results and the Pearson's correlation coeffi-cient between results of OE-210 and isokinetic test were calculated. Results The ICC of test-retest were 0.718 to 0.924, and the ICC of in-ter-rater were 0.784 to 0.870. The correlation between muscle performance measured with 2 tools were significant (P<0.001), that was light to medium on quadriceps (r=0.270-0.413), and strong on hamstrings (r=0.582-0.668). Conclusion OE-210 dynamometer was reliable for muscle strength measurement on quadriceps and hamstrings, and the conditions for valid application need further research.

AIM: To study the effect of berberine(Ber) on invasion and migration of PG cells from a high metastatic human giant lung carcinoma cell line and to explore its mechanism. METHODS: Agarose drop method was used to detect PG migration; transwell cabin with FN in lower chamber was adopted to detect PG chemotaxis. PG adhesion to FN and martrigel was detected by MTT; PG invasive ability was determined by transwell cabin covered with martrigel. Expression of MMP2/TIMP2 protein and mRNA were detected by quantitative immunocytochemical method and RT - PCR respectively. RESULTS: After PG was treated by Ber( 10 mg/L) for 24 h: 1 ) migration distance of Ber- treated PG cells was markedly shorter than that of control cells (P ＜0. 01 ) and the number of passed membrane cells towards FN was much fewer than that of control cells ( P ＜ 0. 01 ); 2) PG adhesion to FN and martrigel was inhibited remarkably by Ber compared with control PG; 3) the migration of PG cells through the martrigel -coated transwell was significantly inhibited by the addition of Ber; 4) MMP2 expression was reduced significantly(P ＜0. 01 ), while the TIMP2 expression showed up - regulating tendency, but had no differences compared with control group (P ＞ 0. 05). The MMP2/TIMP2 ratio was decreased; 5 )the MMP2 mRNA/TIMP2 mRNA ratio was decreased by Ber. CONCLUSION: Inhibition of cell migration, adhesion to ECM and invasion into ECM of tumor cells and regulation of homeostasis between MMPs and TIMPs to maintain ECM integrity may be the basic mechanism of inhibitive effect of Ber on invasion and metastasis of tumors.

BACKGROUND: In repair of nerve defect with allogenic nerve graft, to reduce immune rejection is one of the key problems. At present, the main approach is to reduce antigenicity of grafted nerve segment and apply generally immune inhibitor.OBJECTIVE: To observe the effects of freeze/thaw treatment and local application of transforming growth factor beta 1 (TGF-β1) plasmid on frozen nerve allograft.DESIGN: Randomized controlled animal experiment was designed.SETTING: Department of Hand Surgery, Union Hospital, Tongji Medical College Affiliated to Huazhong University of Science and Technology.MATERIALS: The experiment was performed in Tongji Medical College of Huazhong University of Science and Technology from January 2003 to December 2004, in which 40 Wistar healthy and adult rats were employed,from different delivery and were randomized into experimental group and control, 20 rats in each one.METHODS: Transforming growth factor-β1 (TGF-β1) plasmid and frozen allogenic sciatic nerve were prepared. In experimental group and control,sciatic nerve was cut off 2.0 cm in length, in the foramen 0.5 cm beneath piriformis. The nerve defect was repaired with pre-frozen allogenic nerve 2.0 cm in length. In experimental group, TGF-β1 plasmid was injected in local muscle and two broken ends of nerve. In the control group, physiological saline of equal volume was injected. In the 6th and 12th weeks, the samples were collected from 10 rats in each group for sectioning, staining,axonal counting and statistical analysis.RESULTS: No any animal was died in experiment and all of animals entered result analysis. In the 6th weeks, in the control group, mild edema appeared among axons on the grafted segment of nerve and in the experimental group, there was no edema among axons and the regenerated nerve numbers were close to the normal. In the 12th week, in the experimental group, the entire grafted nerve segment was basically filled up by the regenerated axons;myelinated nerve fiber was arranged in order and both axons and myelins were developed well. The regenerated axonal count in experimental group was more significantly than the control, indicating extremely significant difference [(98.6±4.8), (75.8±5.1) counts/μm2, t=2.962, P ＜ 0.01].CONCLUSION: Freeze/thaw treatment can decrease antigenicity of allogenic nerve, which provides the possibility of repair of nerve defect. Local application of TGF-β1 plasmid can provide immune inhibition locally and reduce immune rejection in the host.

BACKGROUND: Auto-neural transplantation is used widely on peripheral neurological defect, but it also has some difficulties. So some scholars try to use xenoma-neural transplantation; however, it is hard todeal with immunological rejection.OBJECTIVE: To study the effect of transforming growth factor-β1 (TGFβ1) used in local area on neural regeneration after transplantation of fresh nerve allograft.DESIGN: Randomized controlled study.SETTING: Hand Surgery Department of Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and TechnologY.MATERIALS: The experiment was conducted in the Union Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology between August 2001 and October 2002. Totally 60healthy adult Wistar rats from different confinements were randomly divided into three groups including experimental group, blank group and control group with 20 in each group.METHODS: TGF-β1 plasmid was prepared for using. Establishment of animal model: Sciatic nerve at the 0.5 cm deep of piriformis muscle of rats in the two groups was cut with disinfectant razor into chip regularly about 2.0 cm. The excisional nerve segment was exchanged to transplant plerosis neurological defect. TGF-31 was injected into the local muscles and bisection of nerve in the experimental group, and equal volume of saline was injected into rats in the blank group and the control group. In addition, rats in the experimental group and the blank group were not treated with any drugs, but cyclosporine A (15 mg/kg) was used to feed rats in the control group. Ten rats from each group were taken for section and staining at the 6th and the 12th week: ① Glees-luxot fast blue staining method; ② myelin sheath fast blue staining method. Axonal amount: Fields were randomly taken from the middle staining samples 12 weeks later and 1.0 mm2 interaxis-cylinder was counted under light microscope of 400 times. Comparisons among groups were analyzed with i test.MAIN OUTCOME MEASURES: Morphological observation and axonal amount of transplanted area in each group.RESULTS: Quantitative analysis of the experimental animals: Totally 60rats entered the final analysis without any loss. ① Infiltration of monocytes was observed widely in various areas of graft in the blank group;meanwhile, desiccation of myelin sheath and plenty of vacuolations were also observed, especially at the sixth week. The whole graft was infiltrated by monocyte with severe rejection. Few axis-cylinders were regenerated in the transplanted segment. At the 12th week, graft was slender, plenty of scar tissues were proliferated, edema was observed obviously, few Schwann cells and regenerated axis-cylinders were observed, and lots of regenerated axis-cylinders did not pass the whole graft. A few infiltrative monocytes were observed, and edema was observed obviously, but new vessel was formed in transplanted nerve, and regenerated axis-cylinders passed the whole graft in the experimental group and the control group.Lots of Schwann cells were observed at the 6th week; meanwhile, regenerated axis-cylinders passed the whole graft at the 12th week, a quantitative myelinization was formed, Schwann cells proliferated obviously, and edema between axis-cylinder was relieved. Numbers of peripherally regener ated axis-cylinder of nerve and remyelination in each ransplanted area were more than those in the central area, and edema between peripheral axis-cylinder was milder than that in the central area in the experimental group. ② Twelve weeks after operation, 5 rats in each group were selected to observe their fields, which were taken randomly from neural graft,under the microscope of 400 times to count 1.0 mm2 inter-axis-cylinders.Number of axis-cylinder was higher in the experimental group and the control group than that in the blank group, and the differences were significant [(78.3±4.6), (76.1±4.2) , (15.0±3.5) ,t=3.056, t=2.948, P ＜ 0.01];however, number in the experimental group was similar to that in the control group, and differences were not significant [(78.3±4.6), (76.1±4.2),t=1.982 P ＞ 0.05].CONCLUSION: TGF-β1 used in local area plays an immunosuppressive action locally, decreases host immunological rejection, increases the number of axis-cylinder, and accelerates growth of nerve.