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

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

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.

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.

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 determine the dose-response relationship of sufentanil blunting responses to double-lumen endotracheal intubation when combined with propofol given by target-controlled infusion (TCI) in patients with pulmonary tuberculosis.Methods One hundred patients of both sexes with pulmonary tuberculosis,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,aged 24-58 yr,with body mass index ＜30 kg/m2,with Mallampati grade Ⅰ or Ⅱ,undergoing thoracic surgery under general anesthesia,were divided into Ⅰ-Ⅴ groups (n =20 each) using a random number table.Anesthesia was induced with iv sufentanil 0.35 μg/kg (group Ⅰ),0.40 μg/kg (group Ⅱ),0.45 μg/kg (group Ⅲ),0.50 μg/kg (group Ⅳ) and 0.55 μg/kg (group Ⅴ),and propofol TCI (target plasma concentration 3.5 μg/ml) and iv vecuronium 0.15 mg/kg.The patients were endotracheally intubated and mechanically ventilated.The response to double-lumen endotracheal intubation was defined as positive when mean arterial pressure increased by＞ 20％ of the baseline value and/or heart rate ＞ 90 bpm within 5 min after intubation.The median effective dose (ED50),ED95 and 95％ confidence interval (95％ CI) of sufentanil blunting the responses to double-lumen endotracheal intubation were calculated by Probit analysis.Results The ED50 (95％ CI) and ED95 (95％ CI) of sufentanil blunting the responses to intubation were 0.411 (0.370-0.441) μg/kg and 0.635 (0.556-0.888) μg/kg,respectively,when combined with propofol given by TCI.Conclusion When combined with propofol given by TCI (target plasma concentration 3.5 μg/ml),the ED50 and ED95 of sufentanil blunting the responses to double-lumen endotracheal intubation are 0.411 and 0.635 μg/kg,respectively,in patients with pulmonary tuberculosis.

BACKGROUND: At present, the repair by means of suture is still commonly used to repair the peripheral nerve injury and rupture, while the adhesion of the fibrin glue repairing peripheral nerve injury has been considered as a new topic of study.OBJECTIVE: To study the countertraction intensity of peripheral nerve and its dynamic changes after repaired with the adhesion of fibrin glue.DESIGN: A randomized controlled experimental study.SETTING and MATERIALS: The study was completed in the Laboratory of Biodynamics, Department of Orthopaedics, Union Hospital of Tongji Medical College, Huazhong University of Science and Technology. The healthy adult male Wistar rats weighing 250- 300 g were selected for the experiment.INTERVENTIONS: Totally 96 Wistar rats were completely randomized into the suture group and the adhesion group. Their sciatic nerves were cut, and the incisions were well lined. The fibrin glue was adopted in the adhesion group, while 11 -0 suture was adopted in the suture group. On the very day and 3, 7, 14, 21, 28 days after the operation, 8 rats were respectively taken each from the suture group and the adhesion group. The free sciatic nerves of them were detected immediately by the biodynamic test.MAIN OUTCOME MEASURES: The peak load and the power consumption were measured when the nerves ruptured and the nerve stress-strain curve was described.RESULTS: In normal countertraction intensity curve of the nerve, the elastic peculiarity can be manifested. Between the suture group and the adhesion group, there were no notable significances of the maximal countertraction intensity and power consumption on the very day and 14, 21, 28 days after the operation( P ＞ 0.05). While 3 days after the operation, the maximal countertraction intensity of the two groups was(1.35± 0. 27),( 1.97 ± 023) N/mm2 respectively, the power consumption was (0. 028 ± 0.007), (0.040 ± 0.003) J/mm2 respectively. Seven days after the operation, the maximal countertraction intensity was( 1.93 ± 0.26), (2.74± 0.30) N/mm2 respectively, the power consumption was(0.047±0.009), (0.063±0.007) J/mm2 respectively. The differences both had the notable significance ( P ＜ 0.05).CONCLUSION: The fibrin glue has enough countertraction intensity and can gratify the need of such nerve repairs.

Objective To investigate the effects of permissive hypercapnia ventilation on cerebral oxygen metabolism and postoperative cognitive function in elderly patients.Methods One hundred and twenty ASA Ⅰ-Ⅲ patients,aged 65-80 yr,undergoing elective lower abdominal surgery under general anesthesia,were randomly divided into 2 groups (n=60 each): routine ventilation group (group R) and permissive hypercapnia ventilation group (group H).In group H,VT=6-8 ml/kg,RR=12-14 bpm,I: E=1: 2,and PaCO2 was maintained at 45-65 mm Hg and pH value ＞ 7.2,while in group R,VT=10-12 ml/kg,RR=14-16 bpm,I:E=1:2,and PaCO2 was maintained at 35-45 mm Hg.Blood samples were taken from the radial artery and jugular bulb for blood gas analyses at 0,5,15 and 30 min after tracheal intubation (T0.3).Cerebral A-V O2 content differences (Da-jvO2)and cerebral O2 extraction rate (CERO2) were calculated at the same time.Cognitive function was assessed by Mini-Mental State Examination (MMSE) at 1 day before operation,and 24 h,48 h,1 week and 2 weeks after operation.Results Compared with group R,PEr CO2 andPaCO2 were significantly increased,and pH value,Da-jvO2 and CERO2 were significantly decreased at T1.3,MMSE score was significantly increased after operation,and the incidence of post-operative cognitive dysfunction was significantly decreased after operation (P ＜ 0.05 or0.01).Compared with the baseline value at T0,Da-jvO2 and CERO2 were significantly decreased at T1-3 in both groups,PETCO2 and PaCO2 were significantly increased,and pH value,Da-jvO2 and CERO2 were significantly decreased at T1-3 in group H,and MMSE score was significantly decreased at 24 h,48 h,1 week and 2 weeks after operation in both groups (P ＜ 0.01).Conclusion Permissive hypercapnia ventilation can improve the cerebral oxygen metabolism during operation,and reduce post-operative cognitive dysfunction in the elderly patients.

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.