Objective To evaluate the feasibility, safety and efficacy of selective portal vein embolization (SPVE) in treatment of liver tumor in rats and to provide the groundwork for its future clinical applications. Methods 24 healthy rats underwent the embolization. Pre and post SPVE portogram and liver chemical profile were obtained. Four rats were sacrificed at 10 min, 7,14, 21 and 28 days respectively following follow up portography. The liver, heart, lungs and kidneys were examined macroscopically and microscopically. Fifteen rats implanted with Walker 256 tumor sized from 3 to 10 mm in liver were scanned with MRI and portography pre SPVE taken. Post SPVE 3 rats were examined with MRI for each group at the same interval as above and the lives were examined microscopically. Results (1) The blood flow to the target portal branches were immediately halted after SPVE. These vessels remained occluded without collateral formation up to 28 days. (2) The liver indexes and BUN level increased after embolization, but returned to normal within 21 d. Macroscopic and microscopic changes were not found in the heart, lungs or kidneys. (3) In the healthy rats, the affected segment was atrophic and the remaining liver underwent compensatory hypertrophy. Histologic examination revealed that the targeted portal veins were coagulated, the endothelium were degenerated and the local hepatocytes were necrotic after embolization. (4) In the rats with implanted liver tumor, the affected segment including the tumor was necrotic and atrophic. The tumors were completely necrotic, and no viable tumor cell was seen under microscope in 12 among the 15 rats. Three tumors 10 mm in diameter were not completely necrotic. Part of tumor cells were still alive and infiltrated into the surrounding liver. Conclusion SPVE with ethanol is effective in the treatment of small liver tumor in rats. However,in case of bigger tumors involving several segments, SPVE should be combined with other treatment.

BACKGROUND:The technique of vacuum sealing drainage, initialy used for better wound healing, has been widely applied to al kinds of refractory wounds. OBJECTIVE:To summarize the research progress in vacuum sealing drainage for treatment of severe soft tissue injury. METHODS: An electronic retrieve was performed in Wanfang database, VIP database and PubMed database from January 1989 to August 2014 was performed for papers regarding the vacuum sealing drainage technology with the key words of “vacuum sealing drainage technology; injury; gentamicin; hyperbaric oxygen; nerve growth factor; chymotrypsin” in Chinese or English. Finaly 48 articles were involved in the final analysis according to the inclusion criteria. RESULTS AND CONCLUSION:Vacuum sealing drainage is a highly effective treatment technique to promote wound healing which can promote wound blood circulation, reduce sweling, inhibit bacterial growth, stimulate growth of granulation tissue by mechanical stress, inhibit cel apoptosis, thereby helping wound healing. Compared to conventional open-dressings, vacuum sealing drainage has a faster wound healing speed, lower infection rate, and fewer dressing change frequency. With recent advances in medicine, vacuum sealing drainagetechnology cannot be simply used for treatment of skin soft tissue damage, but in combination with gentamicin, hyperbaric oxygen, nerve growth factor and chymotrypsin to achieve better therapeutic effects.

In order to observe the replication and expression of HGV RNA genome in HepG2 cells and establish a cell model of HGV infection, HGV RNA genome was prepared in vitro and transfected HepG2 cells with lipofec-tamin. HGV RNA-positive supernatants were used to infect fresh HepG2 cells. RT-PCR, immunohistochemistry and Western blot assays were carried out to detect the replication and expression of HGV in HepG2 cells. Both positive and negative strands of HGV RNA could be detectable in cell culture supernatants and cells at 24h post-transfection. During the culture periods of 90 days, the cells were maintained by changing the medium every 3 or 5 days, and cultured for more than 20 passages. Both strands of HGV could be detectable in culture supernatants and cells. Immunohistochemistry and Western blot results also confirmed that HGV E2 protein could be expressed in the infected HepG2 cells. HGV RNA could also be detectable in the frozen-thawed HepG2 cells infected with HGV RNA genome. Therefore, HGV RNA genome can replicate and express in HepG2 cells, this HGV RNA genome transfected cells model could be used as a cell model in the studies of replication and infection of HGV.

OBJECTIVETo investigate the effect of miRNA-101 on the expression of the enhancer of zeste homolog 2 (EXH2) in human androgen-independent prostated cancer LNCaP cells.

METHODSWe divided LNCaP cells into a blank control, a negative control, and a miRNA-l01 transfection group, constructed the vector by transfecting synthetic miRNA-101 mimics into the LNCaP cells, and evaluated the efficiency of transfection by fluorescence microscopy. Then we determined the expression level of EZH2 mRNA by qRT-PCR in the three groups of cells and that of the EZH2 protein in the negative control and transfection groups by Western blot.

RESULTSGreen fluorescence signals were observed in over 70% of the LNCaP cells in the transfection group after 24 hours of transfection. At 72 hours, the expression of miRNA-101 was significantly upregulated in the transfected cells (P < 0.01), that of EZH2 mRNA was remarkably lower in the transfection group (0.01 ± 0.10) than in the blank control (0.95 ± 0.40) and negative control (0.86 ± 0.30) groups (both P < 0.01), and that of the EZH2 protein was increased in the negative control but decreased in the transfection group with the extension of culture time.

CONCLUSIONmiRNA-101, with its inhibitory effect on the expression of EZH2 in LNCaP cells, is a potential biotherapeutic for prostate cancer.

Androgens ; Cell Line, Tumor ; Enhancer of Zeste Homolog 2 Protein ; Genetic Vectors ; Humans ; Male ; MicroRNAs ; physiology ; Polycomb Repressive Complex 2 ; genetics ; metabolism ; Prostatic Neoplasms ; metabolism ; RNA, Messenger ; metabolism ; Transfection

Objective: To establish a curve of dosage gradient-liver signal intensity of superparamagnetic iron oxide (SPIO) in normal rats and to find an appropriate dose for enhancement. Methods: Seventy-two SD rats, 4 rats a group at random, underwent MR enhancement with 0, 2, 5, 8, 10, 12, 15, 20, 30, 40, 50, 60, 70, 80, 100, 140, 210, 280 μmol/kg SPIO respectively after plane examination. The signal-to-noise ratio of liver was measured and a curve of dosage gradient-liver signal intensity was made. Results: (1)With the increase of SPIO dose, the signal intensity of both T1 and T2 weighted images of liver declined. (2)T2 weighted images was more sensitive than T1 weighted images in small dose, the ED50 of T1 weighted was 8 μmol/kg, and the ED50 of T2 weighted was 5 μmol/kg. (3)When the dose was greater than 15 μmol/kg, the signal intensity of T1 weighted images declined more rapidly than T2 weighted, the effect of enhanced T1 weighted images resembled T2 weighted ones, and its images was with more fine resolution. (4)At the dose of 40 μmol/kg, the signal intensity of T1 weighted images approached the background noise, and at the dose of 15 μmol/kg, the signal intensity of T2 weighted images approached the background noise. Conclusion: Good effect of T1 and T2 enhanced MR imaging can be acquired at the dose of 20-10 μmol/kg SPIO respectively, the best contrast-to-noise ratio is found on T2 weighted enhanced MR image.

Objective To study the significance and value of imaging typing by analyzing the correlation between contrast-enhanced ultrasound perfusion imaging typing and vascularization in patients with primary hepatocellular cancer.Methods The early enhanced arterial phase reflecting angioarchitecture,and the size and edge of enhanced tumor closely related to microvessel distribution, position and density, were observed and analyzed in 89 patients (113 lesions).All the cases were pathologically proved, and some received immunohistochemisty staining and the microvessel density were recorded.Results The perfusion imagings were classified into 4 types according to changes on contrast-enhanced ultrasound,dendritic,mixed,circular and net-like type.Atypia vessels were commonly seen in dendritic type,and Ⅲ and Ⅳ accounted for 79.1%.The microvessel density was higher than the other types.The circular type was relatively regular and Ⅲ and Ⅳ accounted for 12.5%.The microvessel density was lower than the other types.Combined with the contrast-enhanced ultrasound perfusion imaging and compared with pathological grading,growing methods and microvessel density, the dendritic type was characterized by infiltrative growth, and had strong invasion tendency.The circular type was characterized by expansive growth.And the other two types were characterized by transitional type.Conclusions Contrast-enhanced ultrasound can reflect the angioarchiteeture, the microvessel density and the position,and it is related to the pathological grading and growing methods.

Objective To assess the clinical value of three dimensional dynamic contrast enhanced MR angiography (3D DCE MRA) in the detection for intracranial aneurysm. Methods 3D DCE MRA was performed in 54 patients highly suspected with intracranial aneurysms. Then conventional digital subtraction angiography (DSA) and feasible endovascular treatment were performed simultaneously. A three dimensional fast imaging with steady state precession (3D FISP) was used for 3D DCE MRA(Gd DTPA dose, 0.2 mmol per kilogram for body weight; acquisition time, 10 seconds). The source images were subtracted from mask images and transferred to computer workstation. All images were subsequently post processed using three dimensional reconstruction. 3D DCE MRA images and DSA images were compared for demonstration of the aneurysm, its neck, and relationship with parent artery, and the usefulness for endovascular treatment was evaluated. Results There were 39 cases with 45 intracranial aneurysms. The sensitivity, specificity, and accuracy of 3D DCE MRA were 96%, 73%, and 90%, respectively. Aneurysm and its neck depiction at 3D DCE MRA was significantly better than that at DSA, especially for aneurysms adjacent to the cavernous sinus and near the PICA of vertebral artery. 3D DCE MRA could guide neurosurgeons to the desired DSA projection, and helped them make plan for interventional or surgical treatment in advance. But the diagnosis should be very carefully made for small aneurysms located in the periphery and the arterial bifurcation. Conclusion 3D DCE MRA is a fast, noninvasive and efficient technique for diagnosing intracranial aneurysms. Its three dimensional information is helpful for DSA demonstration and treatment planning. Any uncertain diagnosis requires DSA confirmation.

Aged ; Female ; Humans ; Male ; Melanosomes ; Metaplasia ; Middle Aged ; Nasopharynx ; cytology ; pathology ; Oxyphil Cells ; cytology

[ ABSTRACT] AIM: To investigate the protective effect of autophagy on oxidized low density lipoprotein ( ox-LDL)-induced macrophage apoptosis and the underlying molecular mechanisms .METHODS:The RAW264.7 macropha-ges were pretreated with 3 mmol/L 3-methyladenine (3-MA), 1 μmol/L rapamycin (Rap) or 4 mmol/L 4-phenylbutyric acid ( PBA) respectively for 1 h and then treated with ox-LDL (100 mg/L) for 12 h.The cell viability and apoptosis were determined by MTT assay and flow cytometry with Annexin V-FITC/PI staining, respectively.The activities of lactate de-hydrogenase ( LDH) in the medium and caspase-3 in the cells were determined by detection kits .The protein levels of bec-lin-1 (a molecular marker of autophagy ), glucose-regulated protein 78 (GRP78, an endoplasmic reticulum stress marker) and C/EBP homologous protein ( CHOP, a key-signaling component of endoplasmic reticulum stress-induced apoptosis ) were examined by Western blot .Microtubule-associated protein 1 light chain 3 (LC3, another molecular marker of autoph-agy) was observed under laser scanning confocal microscope .RESULTS: Treatment of the RAW264.7 macrophages with ox-LDL at 100 mg/L for 12 h resulted in significant decrease in cell viability , and dramatic elevation in LDH leakage , cell apoptosis and caspase-3 activity, which were promoted by 3-MA (an autophagy inhibitor) and inhibited by Rap (an autoph-agy inducer ) .ox-LDL induced autophagy in the macrophages as assessed by beclin-1 upregulation and frequent granulation of LC3, which were inhibited by 3-MA and promoted by Rap.Interestingly, 3-MA enhanced, while Rap blocked, the CHOP upregulation induced by ox-LDL.Moreover , PBA ( endoplasmic reticulum stress inhibitor ) significantly inhibited ox-LDL-induced GRP78 upregulation and autophagy as determined by the attenuation of beclin-1 upregulation and frequent granula-tion of LC3.CONCLUSION: Endoplasmic reticulum stress mediates ox-LDL-induced autophagy in macrophages , and moderates activation of autophagy may protect macrophages from ox-LDL-induced apoptosis by inhibiting CHOP expression .