1.Whether the hepatitis B virus envelope as a gene transfer vector has targeting ability for liver cancer
Dejian PAN ; Donglin WANG ; Zhengtang CHEN
China Oncology 2000;0(06):-
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
2.EFFECTS OF MRF4 TRANSFECTION ON DIFFERENTIATION AND EXPRESSION OF MYOGENIC REGULATORY FACTORS IN CULTERED HUMAN RHABDOMYOSARCOMA RD CELLS
Guanghui LI ; Donglin WANG ; Zhengtang CHEN
Medical Journal of Chinese People's Liberation Army 2001;0(10):-
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.
3.Clinical Observation of Linezolid, Teicoplanin and Vancomycin in the Treatment of Hospital-acquired MRSA Pneumonia
Yuefang WANG ; Wei ZHONG ; Donglin CHEN
China Pharmacy 2016;27(26):3708-3710
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.
4.Clinical result of limited arthroscopic debridement for knee osteoarthritis under local anesthesia
Haiyang YU ; Donglin FU ; Jian CHEN
Orthopedic Journal of China 2006;0(01):-
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.
5.Efficacy of rapamycin and paclitaxel eluting stent implantation in treatment of coronary artery disease
Jun YANG ; Donglin CHEN ; Yinchao CHEN ; Xinjing CHEN
Chinese Journal of Tissue Engineering Research 2015;(52):8517-8521
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.
6.Simvastation for Transient Ischemic Attack following Carotid Atherosclerosis
Donglin HAO ; Xiaonan CHEN ; Lihui WANG ; Baozhen CHEN
Chinese Journal of Rehabilitation Theory and Practice 2011;17(2):163-164
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.
7.Effects of PIAS3 silencing by RNAi on the proliferation and apoptosis of U251 glioma cells in vitro
Hua JI ; Wei YU ; Hong CHEN ; Guanghui LI ; Donglin WANG
Chinese Journal of Clinical Oncology 2014;(2):94-97
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.
8.Significance of cerebrospinal fluid matrix metalloproteinase-9, interleukin-6, and tumor necrosis factor-alpha protein in children with viral encephalitis
Donglin SHEN ; Yuanyuan DAI ; Jiao CHEN ; Ming LU ; Mingxia SUN
Chinese Journal of Applied Clinical Pediatrics 2014;29(22):1721-1723
Objective To explore the significance of changes of matrix metalloproteinase-9 (MMP-9),interleukin-6 (IL-6),and tumor necrosis factor-alpha (TNF-oα) protein level in the cerebrospinal fluid (CSF) of children with viral encephalitis (VE).Methods The concentration of neuron-specific enolase (NSE),structural proteins 100B (S100B),and MMP-9,IL-6,TNF-α in the CSF of VE children were detected by an enzyme linked immunosorbent assay,and the correlations of them were analyzed.Results NSE,S100B,MMP-9,IL-6,TNF-α protein expression could be found significantly higher than those in the control group,and there were significant differences according to statistics expression trends(all P <0.05).The NSE protein expression was significantly positive related with S100B in the VE group (r =0.467,P =0.009),and the concentration was markedly negative related with the duration of viral encephalitis (r =-0.472,P =0.008).MMP-9,IL-6 protein expression were significantly positive related with NSE,S100B respectively (r =0.698,P =0.00 ; r =0.559,P =0.00 ; r =0.812,P =0.00 ; r =0.664,P =0.00).TNF-α protein expression was positive related with CSF S100B(r =0.363,P =0.049),but there was no correlation between TNF-α and NSE (r =0.245,P =0.193).Conclusions The neurons and the neuroglial cells are damaged in the viral encephalitis children.MMP-9,IL-6,TNF-α protein may participate in the pathological damage process of nerve cells in VE children in different degrees.
9.Value of neutrophil CD64 in diagnosis of sepsis in adult patients:a meta-analysis
Huiye ZHANG ; Donglin PAN ; Jun YANG ; Tingyi ZHONG ; Yongchuan CHEN
Chinese Journal of Clinical Infectious Diseases 2015;(5):444-450
Objective To evaluate the application of neutrophil CD 64 in diagnosis of sepsis in adult patients.Methods Literature retrieval from PubMed, EMBASE, ISI Web of Knowledge, Cochrane Library, CBM, CNKI, CQVIP, Wanfang Data from the establishment of database to the year 2015 was conducted to identify all studies on CD 64 in diagnosis of sepsis .The quality of the literature was evaluated with the quality assessment of diagnostic accuracy studies ( QUADAS).Meta-Disc 1.4 and STATA 12.0 were used for meta analysis . Fixed-effects or random-effects model was performed based on the heterogeneity.The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and diagnostic odds ratio were calculated .Summary receiver operating characteristic curves ( SROC ) and area under the curve (AUC) were used to evaluate the diagnostic performance of CD 64 for sepsis.Results A total of 24 studies involving 3 198 patients were included for systemic review .The sensitivity, specificity, positive likelihood ratio, negative likelihood ratio and diagnostic odds ratio of CD 64 for diagnosis of sepsis were 0.79 (95 %CI:0.77-0.81), 0.86 (95 %CI:0.84-0.88), 7.40 (95 %CI:5.02-10.91), 0.15 (95 %CI:0.10-0.22) and 60.07 (95%CI: 29.19-123.60), respectively.The area under SROC of CD64 in diagnosis of sepsis was 0.95, and the Q* value was 0.88.Conclusion CD64 can be used to diagnose sepsis in adult patients , but it needs to be further confirmed by large multicenter studies .
10.Effects of electro-acupuncture combined with bone marrow stromal cell transplantation on recovery of neurological function in rats with spinal cord injury
Li ZHANG ; Donglin XIONG ; Kuihao CHEN ; Rongrong WANG ; Yingxiang MIAO
Chinese Journal of Tissue Engineering Research 2009;13(27):5217-5220
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.