In order to assess whether gene transfection could be mediated by ultrasound in association with P85 and find the appropriate parameters of ultrasound irradiation, the effects of ultrasound with or without P85 on gene transfection of HepG2 cells were examined. The HepG2 cells were irradiated by ultrasound at 1 MHz, 0.4-2.0 W/cm(2) and 50% duty cycle with plasmid encoding enhanced green fluorescent protein (EGFP) as a report gene. Forty-eight h later, the expression of EGFP was detected under the fluorescence microscopy. Transfection efficacy was quantitatively assessed by flow cytometry, and cell viability was evaluated by trypan blue exclusion. The results showed that the transfection efficacy was increased with the increases in ultrasound output power and the ideal transfection efficacy was achieved in HepG2 cells irradiated by ultrasound at 0.8 W/cm(2) for 30 s. The transfection efficacy in ulstrasound+P85 group was three times higher than in single ultrasound group [(17.63+/-1.07)% vs (5.57+/-0.56)%, P<0.05]. The cell viability was about 81% and 62% in ultrasound group and ultrasound+P85 group respectively. It was concluded that ultrasound in combination with P85 could mediate the gene transfection of HepG2 cells, ideal transfection efficacy was achieved by ultrasound irradiation at 0.8 W/cm(2) for 30 s, and P85 could somewhat increase the damage to cells caused by ultrasound.
Cell Survival/genetics ; Green Fluorescent Proteins/genetics ; Hep G2 Cells ; Poloxalene/*pharmacology ; Transfection ; Ultrasonics

The value of color Doppler flow imaging (CDFI) and intravenous contrast-enhanced ultrasound (CEUS) for assessing the transplanted liver and early diagnosing complications by examining hemodynamic changes was discussed. Seventy-five patients with orthotopic liver transplantation (OLT) underwent CDFI. The following parameters were measured: peak systolic velocity (PS), resistance index (RI) and Doppler perfusion index (DPI) of the hepatic artery (HA), time average velocity (TAV) of portal vein (PV) and velocity of hepatic vein (HV) in different stages postoperation. And 11 patients of them received CEUS. Thirty healthy subjects were enrolled as controls. The results showed that: (1) In 23 patients without obvious complications, TAV of PV within 15 days post-operation was significantly higher than in controls (P<0.05), PS and DPI of HA within 7 days postoperation were lower, but RI was higher than in controls (P<0.05); (2) When the hepatic artery thrombosis (HAT) occurred, PS and DPI of HA were obviously decreased, but TAV of PV significantly increased like a high saw-tooth wave; (3) While rejection occurred, both TAV of PV and PS of HA were decreased with the increase in RI of HA, and the triphasic wave of HV disappeared and displayed as saw-tooth wave; (4) The incidence of biliary complications in liver transplantation was increased when DPI was reduced; (5) Seven cases of hepatic carcinoma relapse after OLT demonstrated hyperecho in the arterial phase and hypoecho in the portal and later phase on CEUS; (6) In 2 cases of HA thrombus, there was no visualized enhancement in arterial phase of CEUS, but enhancement during the portal vein and parenchymal phase. It was concluded that the hemodynamic changes of PV, HA and HV in the transplanted liver are valuable for assessing the transplanted liver and early diagnosing complications on CDFI and CEUS.
Bile Ducts/pathology ; Contrast Media/*administration & dosage ; Hemodynamics ; Hepatic Artery/pathology ; Infusions, Intravenous/*methods ; Liver Transplantation/*adverse effects ; Liver Transplantation/*methods ; Liver Transplantation/ultrasonography ; Perfusion ; Postoperative Complications ; Ultrasonography/*methods ; Ultrasonography, Doppler/*methods

This study examined the effect of P85 (a pluronic block copolymer) and microbubble (MB) ultrasound contrast agents under ultrasound irradiation on gene transfection and expression. The pEGFP plasmids that can encode enhanced green fluorescent protein (pEGFP) served as a report gene and were mixed with different concentrations of MB/0.05% (w/v) P85. Then the plasmids were transfected into human hepatoma G2 (HepG2) cells. The HepG2 cells treated with MB/P85 or without treatment were exposed to ultrasound (US parameters: 1 MHz, 1.0 W/cm(2), 20 s, 20% duty cycle). Twenty-four hours later, the transfection efficiency was assessed by fluorescence microscopy and fluorescence activated cell sorting (FACS) analysis. The cell viability was evaluated by Trypan blue exclusion test. The results showed that the gene transfection efficiency in HepG2 cells under ultrasound irradiation was significantly higher than that without ultrasound irradiation. HepG2 cells in the MB or P85 group in the absence of ultrasound expressed less amount of green fluorescent protein. The expression efficiency reached (22.14±3.06)% and the survival rate was as high as (55.73±3.32)% in the 30% MB plus P85 group. It was concluded that MB and P85 in the presence of ultrasound can enhance gene transfection and expression.

Objective To investigate the value of postprocessing technique of multi-slice helical CT for the occult fracture located in complicated structure. Methods One hundred-thirteen patients with acute trauma but negative on plain X-ray film were reviewed retrospectively. All of the patients underwent MSCT ,and original data were reformed with reconstruction. Results Of one hundred-thirteen injuried patients , fractures were demonstrated with MSCT imaging in forty-eight cases. Conclusion The combination use of axial image of MSCT , reconstruction and post-processing is of high value in definite detection of occult fracture , and play an important role in selecting therapy project and evaluating prognosis.

MicroRNAisatypeofsingle-strandednon-codingRNA,directlybondingtothecomplemen-tary target gene mRNA.That leads to the inhibition of mRNA translation of the target molecule in order to reduce expression of the target gene.MicroRNAs are not only involved in the regulation of inflammatory reac-tion,but also play important roles in the formation of neoplasm.The researches showing that the inflammatory reaction interacting with miRNAs results in the differential expressions of microRNAs in lung cancer have been widespreadly concerned.Along with the deepening of the research on the mechanisms of inflammatory reaction and differential expression of microRNAs,it will provide new strategies for the prevention,diagnosis and treat-ment of lung cancer.

Implant-associated infection is a serious problem in the clinical application of biomaterials.which greatly restricts the generalizatjon of cardiovascular biomaterials,while bacterial adhesion is the initial cause for implant-associated infections of cardiovascular biomaterials.Therefore.the prevention of bacteriaI adhesion to the biomaterials plays a crucial role in the cure of biomaterial center infection.It has reported that human plasma protein influences the bacterial adhesion to biomateriais.Some plasma proteins inhibit bacterial adhesion.whereas some promote jt.Study on the correlation between plasma protein and hacterial adhesion to the biomaterials is of great importance in the prevention and cure of biomaterials center infection.

Objective To evaluate the safety and efficacy of multi-band mucosectomy (MBM) for early esophageal cancer and precancerous lesions.Methods Data of 28 patients with early esophageal cancer or precancerous lesions undergoing MBM were reviewed in regarding of procedure complications and follow-up results.Results A total of 32 lesions were resected successfully by MBM in one session,with mean procedure time of 28.3 minutes.The mean diameter of specimens was 12mm.No residual neoplasm was found at the base of any resected specimens.The post-MBM pathological findings consisted of 2 cases of intramucosal cancer,1 case of submucosal cancer,and 25 cases of moderate-severe dysplasia.No perforation,delayed hemorrhage or subcutaneous emphysema occurred.Intraoperative bleeding occurred in 23 cases,including 3 cases of pulsatile bleeding,which were controlled with metal clip,and 20 cases of minor bleeding which were managed with APC or halted automatically at the end of procedure.Chest pain after the procedure occurred in 5 cases and were relieved soon.The patient with submucosal cancer underwent subsequent surgical resection,with no residual cancer in surgical specimen or lymph node metastasis.Twenty seven other cases were followed up endoscopically for 2-12 months.Esophageal stricture occurred in 2 cases,and were successfully relieved by dilatation with stent or bougienage.No recurrent lesion or metastasis were revealed.Conclusion MBM is a relatively safe and effective endoscopic technique for treatment of early esophageal intramucosal cancer and precancerous lesions,but further studies are needed to evaluate the long-term results.

Objective To determine the effect of calcitonin gene-related peptide (CGRP) on triggering receptor expressed on myeloid cells-1 (TREM-1) in the lipopolysaccharide (LPS)-induced macrophages and its signal transduction pathway. Methods The levels of TREM-1 mRNA in the macrophages were observed by reverse transcription-polymerase chain reaction (RT-PCR), and flow cytometry was performed to detect TREM-1 protein expression levels in the macrophages. Results CGRP had no regulating effect on the expression of TREM-1 in the macrophages; LPS could up-regulate macrophages to express TREM-1; CGRP increased TREM-1 mRNA expression in LPS-induced macrophages in dose and time-dependent manner; CGRP increased TREM-1 protein expression in LPS-induced macrophages, which could be partially reversed by H-7 or H-89 (P<0.05). Conclusion CGRP can regulate the LPS-induced macrophages synthesis and secretion of TREM-1, and the intracellular signal transduction pathway is related to PKA and PKC.

Cerebral venous thrombosis, as a rare extraintestinal manifestation of inflammatory bowel disease, is often neglected in the course of clinical diagnosis and treatment. This article reviews the advances in the treatment of inflammatory bowel disease related cerebral venous thrombosis.

Objective To investigate the effect of caspases-3 on doxepin-induced apoptosis in rat neurons.Methods The PC12 cells seeded in culture plates were randomly divided into 4 groups (n =10 each) using a random number table:normal control group (group C);doxepin group (group D);caspase-3 inhibitor Z-DEVD-FMK group (group Z);doxepin + Z-DEVD-FMK group (group DZ).In group C,the cells were continuously incubated for 24 h.In group D,doxepin was added with the final concentration of 120 μmol/L,and the cells were continuously incubated for 24 h.In group Z,Z-DEVD-FMK was added with the final concentration of 10 μmol/L,and the cells were continuously incubated for 24 h.In group DZ,doxepin and Z-DEVD-FMK with the final concentrations of 120 and 10 μmol/L,respectively,were added,and the cells were continuously incubated for 24 h.After 24 h of incubation,the cell viability was detected by methyl thiazolyl tetrazolium assay,the cell morphology was observed under inverted microscope,and the neuronal apoptosis was measured by flow cytometry.Apoptosis rate was calculated.Results Compared with group C,the cell viability was significantly decreased,and apoptosis rate was increased in D and DZ groups (P＜0.01),and no significant change was found in the parameters mentioned above in group Z (P ＞ 0.05).Compared with group D,the cell viability was significantly increased,and apoptosis rate was decreased in group DZ (P＜ 0.01).The morphological changes were significantly mitigated in group DZ as compared with group D.Conclusion Caspases-3 may mediate doxepin-induced apoptosis in rat neurons.