Background and purpose:The reason for hepatitis B virus (HBV) with hepatocyte specificity is PreS1 enchased on the hepatitis B virus envelope (HBVE). So HBVE may have a potential application in liver targeting gene transfer. In this study, we investigated whether HBVE has the ability to target liver cancer cells. Methods:HBVE was obtained from the supernatant of Hep G 2.2.15 cells through PEG8000 system and ?-propiolactone method. The pIRS2-EGFP was packed with HBVE and resulted in the product HBVE-GFP. HBVE-GFP was transfected into HepG2, A549, HeLa and FB cells. The green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results:The GFP could be observed in the four groups, but the HepG2 group had a higher fluorescent intensity than the other 3 groups. The transfected rate of HepG2 group was (71.35?0.03)% , much higher than other groups(P

Objective To evaluate the effect of limited arthroscopic debridement under local anesthesia for knee osteoarthritis. Methods A retrospective analysis was made including a total of 63 patients (77 knees ) with knee osteoarthritis.There were 11 male and 52 female,with an average age of 56.5 years old(range from 38~71 years old).The limited arthroscopic debridement included removal of loose bodies hyperplastic synovum,degenerated cartilage and the injured meniscus.The average follow-up period was 2.1 years (from 6 month to 4 year).Outcomes were assessed by JOA knee function score and rate of satisfactory. Results During the follow-up,the patients showed significant improvement of knee function score from 41 point to 78 point.Clinical outcomes of 77 knees were excellent in 31 knee,good in 28 knee,fair in 12 knee,poor in 6 knee.The rate of good and excellent results was 76.6%. Conclusions Limited arthroscopic debridement under local anesthesia for knee osteoarthritis is a relative simple and effective method.It could alleviate pain and improve function of the paticents.

OBJECTIVE:To observe clinical efficacy and safety of linezolid,teicoplanin and vancomycin in the treatment of hospital-acquired pneumonia caused by methicillin-resistant Staphylococcus aureus (MRSA). METHODS:120 patients diagnosed as hospital-acquired MRSA pneumonia were divided into linezolid group,teicoplanin group and vancomycin group according to therapeutic regimen,with 40 cases in each group. Linezolid group received Linezolid injection 600 mg,ivgtt,bid;teicoplanin group received Teicoplanin injection 0.4 g,ivgtt,bid;vancomycin group received Vancomycin injection 1 000 mg,bid,ivgtt. 3 groups received 2 weeks of treatment. Clinical efficacy and bacterial clearance effective rate of 3 groups were observed as well as serum levels of inflammatory factors before and after treatment. ADR of 3 groups were compared. RESULTS:The clinical effective rates of linezolid group,teicoplanin group and vancomycin group were 90.0%,72.5% and 67.5%;the effective bacterial clearance rates were 85.0%,60.0% and 57.5%,respectively. The clinical effective rate and the effective bacterial clearance rate in linezolid group were significantly higher than those in teicoplanin group and vancomycin group,with statistical significance (P<0.05). There was no statistical significance in above indexes between teicoplanin group and vancomycin group (P>0.05). There was no statistical significance serum inflammatory factors among 3 groups before treatment (P>0.05). CRP and PCT of 3 groups de-creased significantly after treatment,and those of linezolid group were lower than teicoplanin group and vancomycin group,with statistical significance(P<0.05). There was no statistical significance in serum inflammatory factors between teicoplanin group and vancomycin group before and after treatment (P>0.05). There was no statistical significance in the incidence of ADR among 3 groups (P>0.05). CONCLUSIONS:For hospital-acquired MRSA pneumonia,linezolid is better than teicoplanin and vancomycin in pneumonia control with good safety.

To study the effects of MRF4 transfection on differentiation and expression of myogenic regulatory factors of human rhabdomyosarcoma RD cells, the plasmid-MRF4 cDNA was transfected into cultured rhabdomyosarcoma RD cells with lipofectin method. The myogenic regulatory factors MRF4 and MyoD mRNA were measured with in situ hybridization and the expressions of myosin heavy chain(MHC) and a-actin in the cells were assayed with immunocytochemical method. The cell growth and morphology were observed at the same time. It was found that the morphology of differentiation increased and the growth was suppressed in RD cells after transfection. The expression of MHC and a-actin were significantly increased in RD cells after transfection, while the expressions of MRF4 and MyoD mRNA were up-regulated. It is suggested that transfection of MRF4 can induce differentiation of RD cells and up-regulate the expression of MyoD.

BACKGROUND: The clinical effect of drug-eluting stent implantation in treatment of coronary artery disease is good, but there may be some differences in the therapeutic effect between different stents. OBJECTIVE:To compare the clinical effect of different stent implantation in treatment of coronary artery disease. METHODS:A total of 278 patients with coronary artery disease were enroled and al underwent coronary artery stent implantation treatment, of which 91 patients underwent implanted rapamycin eluting stent, 92 patients underwent implanted paclitaxel eluting stent, and 95 patients underwent implanted bare metal stents. Al the patients were folowed up for 12 months after stents implantation. The occurrence of adverse cardiac events such as death, myocardial infarction, and the occurrence of coronary restenosis and host reaction were recorded. RESULTS AND CONCLUSION:The rates of coronary artery stenosis, acute myocardial infarction, coronary artery bypass grafting or second-time percutaneous coronary interventional therapy in the paclitaxel eluting stent and rapamycin eluting stent groups were al lower than those in the bare metal stent group (P< 0.05). There were no significant differences in the above indicators between paclitaxel eluting stent and rapamycin eluting stent groups (P> 0.05). There was no significant difference in the mortality between these three groups (P> 0.05). There was no stent shift, shedding, breakage, bad implantation position, leukocytosis, thrombocytopenia and hemolysis in these three groups. These results demonstrate that the therapeutic effects of these two kinds of drug-eluting stents are al good enough, and better than those of bare metal stents.

ObjectiveTo evaluate the effects of simvastatin on patients with transient ischemic attack (TIA) following carotid atherosclerotic plaque.Methods96 cases of TIA patients with carotid atherosclerosis plaque were randomly divided into simvastatin (statins) group and control group, 48 cases in each group. The statins group took simvastatin except routine therapy for 6 months, while the control group took Xuezhikang. The ultrasonic examination of carotid artery intima-media thickness (IMT) and atherosclerosis area were carried out before and after treatment for both groups. The incident rates of cerebral vascular diseases within 6 months were compared between two groups.ResultsThe ultrasonic examination showed significantly thinning of carotid IMT and reducing of plaque area in statins group (P<0.05), while there wasn't significant difference in control group(P>0.05). The cerebral vascular incident rates in statins group were lower than in control group(P<0.05).ConclusionSimvastatin may be more effective for antiatherosclerotic function with bigger dosage and decrease ischemic cerebral vascular incidence.

BACKGROUND: The study indicated that directed neural differentiation of bone marrow stromal cells (BMSCs) was associated with changes in local micro-environment in injured spinal cord following cerebral ischemia, in particular neurotrophic factor induction. Group pre-test has confirmed that acupuncture can increase the expression of vadous cytokines and neurotrophic factors to promote nerve regeneration and repair.OBJECTIVE: To observe the effects of electro-acupuncture combined with BMSC kansplantaUon on recovery of neurological function in rats with spinal cord injury.DESIGN, TIME AND SETTING: Randomized controlled animal experiments were performed at the Laboratory of Cell Biology,Harbin Medical University from March 2005 to July 2006.MATERIALS: A total of 80 pure healthy Sprague Dawley rats were selected. Eight were used for the isolation and culture of BMSCs, and the remaining 72 rats were equally and randomly divided into 4 groups: blank control group, cell transplantation group, electro-acupuncture group, and combination group. KWD-80811 pulse-type apparatus was produced by the third radio factory in Jiangsu Wujin production.METHODS: In vitro isolation and culture of the third passage of BMSCs were obtained and labeled by BrdU at hour 72 before transplantation, and BMSCs at 1 ×10 11/L were harvested for usa. Rat models of spinal cord injury were established in each group.Following model establishment, BMSC suspension was injected into the injured spinal cord near the junction of the gray matter and white matter in the cell transplantation group, totally 6 × 10<'5> cells. The same volume of phosphate buffer saline was infused into rats of the blank control group by the same method. In the electro-acupuncture group, at hour 24 after the successful model establishment, Jiaji Electroacupuncture Electroacupuncture instrument treatment was performed by pulse electro-acupuncture apparatus at 3.0-4.0 mm from median line in the upper and lower end of the spinous process vertebral body adjacent to open space for 20 minutes, once a day. In the combination group, rats underwent BMSC transplantation + Jiaji electro-acupuncture treatment. MAIN OUTCOME MEASURES: After transplantation for 3, 7, 14 days, neurological conditions were assessed by combined behavioral scores in rats with spinal cord injury. Glial fibrillary acidic protein and neuron-specific enolase expression was measured in BrdU-labeled BMSCs by immuno-double-labeling method.RESULTS: 3, 7, 14 days after injury, compared with the blank control group, significant differences in combined behavioral scores were determined in the call transplantation group, electro-acupuncture group, and combination group (P <0.05, 0.05, 0.01).The recovery of neurological function was significantly greater in the combination group than in the cell transplantation and electro-acupuncture groups (P < 0.05). No significant difference was detected between the cell transplantation group and the electro-acupunctura group (P > 0.05). Compared with blank control group, spinal cord structure was relatively complete in the cell transplantation group and electro-acupuncture group. The structure was more integrity in the combination group. Bone marrow stromal cell transplantation presented the organization BrdU-labeled cells in the lesion and the surrounding area, with obvious aggregation and survival. At day 14 following cell transplantation, positive rates of neuron-specific enolase and glial fibrillary acidic protein were respectively 7.2% and 1.5% in the call transplantation group, 7.9% and 2.1% in the combination group.CONCLUSION: The BMSCs after transplantation can survive in the host body. Electro-acupuncture could promote the differentiation of BMSCs into neural cells. Combination of electro-acupuncture and BMSC transplantation can significantly improve the motor and sensory function in rats with spinal cord injury.

Objective: To construct a liver targeting gene transfer vector using hepatitis B virus envelope particles. Methods: Hepatitis B viruses were obtained from the supernatant of HepG 2.2.15 cells by a PEG8000 system and were inactivated by ?-propiolactone to prepare hepatitis B virus envelope. The hepatitis B virus envelope was used to pack 5.3 kb pIRES_2-EGFP to assess their packing ability. Subsequently, the products were studied with ELISA, PCR, SDS-PAGE, and electron microscopy. Finally, the product was used to transfect HepG2 cells and the green fluorescent protein (GFP) expression was observed under a fluorescent microscope. The rate of GFP positive cells was determined by flow cytometer.Results: The acquired hepatitis B virus envelope retained the surface protein HBsAg+pre S_1+pre S_2, but with no virus DNA. The prepared envelope had high packing ability for GFP and the packed GFP had a high transfection rate in HepG2 cell. Conclusion: Hepatitis B virus envelope has been successfully obtained from the supernatant of HepG 2.2.15 cells with a PEG8000 system and ?-propiolactone.

Objective:To observe the transfection efficacy and targeting efficiency of hepatitis B virus envelope(HBVE) as a gene transfer vector for liver cancer cells.Methods:HBVE was obtained from the supernatant of HepG2.2.15 cells with a PEG8000 system and ?-propiolactone.The pIRS2-EGFP was packed with HBVE to obtain HBVE-GFP and was packed with liposome to obtain Liposome-GFP.HBVE-GFP and Liposome-GFP were used to transfect human hepatoblastoma cell line HepG 2 to study the transfection efficiency.HepG 2,A 549,HeLa and FB cells were transfected with HBVE-GFP to appraise the targeting ability of HBVE-GFP.GFP protein expression was observed under a fluorescent microscope and the ratio of GFP positive cells was determined by flow cytometry.Results:(1) Transfection efficiency:The GFP protein was observed in both the liposome group and the HBVE group under the fluorescent microscope;the fluorescent intensity in the HBVE group was 3-4 times that of liposome group as determined by flow cytometry(P

Objective:To observe the effect of the proteininhibitor of activated STAT 3 (PIAS3) on the proliferation and apopto-sis of U251 glioma cells after PIAS3 expression was inhibited by RNAi. Methods: Three RNAi expression vectorstargeting PIAS3 were constructed and transfected into CHG-5 cells by liposomein vitro. The most efficient RNAi vector was subsequently selected by examiningthe mRNA expressions of PIAS3 in the transfected cells by semi-quantitativeRT-PCR. The selected RNAi vector was then transfected into U251 cells. After 48h of transfection, the mRNA and protein expressions of PIAS3 in glioma cellswere examined by semi-quantitative RT-PCR and western blot. Apoptosis wasobserved by flow cytometry using a double-staining method with FITC-con-jugatedannexin V and PI. Flow cytometry was also applied in cell cycle assay. Results:RNAidownregulated the mRNA (P<0.01) and protein (P<0.01) expressionsof PIAS3 in transfected cells.RNAi promoted the resistance of U251 cells to apoptosisand subsequently al-tered the cell cycle. A high percentage of G2 phaseand a low percentage of Sphase were observed in U251 cells. Conclusion:The down-regulation of PIAS3arrested U251 cells in the G2 phase andinduced the resistance of U251 cells to apoptosis.