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In the past three decades, we have witnessed the great promise of gene therapy in treating a variety of inherited and acquired retinal diseases. The large number of preclinical studies in retinal gene therapy has led to over sixty clinical trials at different phases and the approval of Luxturna, a gene therapy product that treats type 2 Leber congenital amaurosis (LCA2) caused by mutations in the RPE65 gene. While current gene delivery systems have led to some success in clinical studies, safer and more efficient gene delivery calls for improved vectors and alternative intraocular administration routes. Progress has been made in large gene delivery and less invasive vector administration with the advancement of novel adeno-associated viral (AAV) vector technologies. Clustered regularly interspaced short palindromic repeats (CRISPR)-based technology has enlarged the toolbox of gene therapy and will inevitably find wide applications in the treatment of retinal diseases. Although gene therapy faces a number of challenges, in the coming years, it will become a viable treatment option for a number of currently incurable retinal disorders.
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Objective:To establish the emergency medical equipment rental center, not only can reduce the repeated purchase for medical equipment, but also can achieve resource sharing, reduce equipment maintenance cost, and improve utilization efficiency.Methods: To establish emergency medical equipment rental center, which adopted lease mode and centralized management mode to use equipment. Department of medical engineering managed this center and this department is responsible to ensure every clinical department can use equipment by lease method.Results: The center has optimized the resource allocation of hospital, obviously reduced equipment input cost, improved utilization rate of medical facility since it were established. This mode provided reliable service for clinical departments.Conclusion: The established rental center of emergency medical equipment has changed the traditional management mode to a new feasible mode which can improve centralized management for medical devices, reduce waste for medical resource and achieve resource sharing.
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Objective To investigate the diagnostic and prognostic values of 18 F?FDG PET/CT in pa?tients with cervical nodal metastases from carcinoma of unknown primary site (CCUP). Methods A total of 137 consecutive patients(95 males, 42 females, age range 24-84 (55.77±11.71) years) with histological proven CCUP who underwent 18 F?FDG PET/CT imaging to find primary tumors from March 2010 to June 2015 were retrospectively analyzed. The gold standard was the final pathological diagnosis or clinical long?term fol?low?up (≥6 months) results. The diagnostic sensitivity, specificity, accuracy, positive predictive value and negative predictive value of PET/CT imaging in detecting primary tumors were calculated. Kaplan?Meier anal?ysis was conducted for survival analysis and to identify the presence of any prognostic factors, such as age, gender, presence or absence of distant metastasis, region of involved cervical lymph nodes, left (right)?side/both sides of lymph node involvement, histopathologic tumor type and primary site found or not by PET/CT imaging. Results Primary tumors were confirmed in 96 patients, of which 87 patients were diagnosed correct?ly by 18 F?FDG PET/CT imaging. The diagnostic sensitivity, specificity, accuracy, positive predictive value and negative predictive value were 90.6% (87/96), 80.5% (33/41), 87.6% (120/137), 91.6% (87/95) and 78.6% (33/42) respectively. Primary tumors were found most commonly in the head, neck and lung. However, the false positive and negative lesions were also mainly seen in those sites. Distant metastasis and the inferior region of cervical lymph node metastasis were associated with the worst survival (χ2=20. 990 and 12?277, both P6.5 were of higher mortality risk than those with SUVmax≤6.5 (χ2=7.120, P<0.01) . Conclusions Whole?body 18 F?FDG PET/CT scan is a significant and valuable tool in the identification of the primary tumor site in patients with CCUP . Further? more, according to the distant metastasis and region of cervical lymph nodes metastasis, 18 F?FDG PET/CT scans are of value in prognostic evaluation even if without the need to identify the primary tumors.
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Objective: To investigate the effects of extract of Bulbus Allii Caespitosi on cardiocyte viability of swines with myocardial reperfusion injury by analyzing the 18F-fluorodeoxyglucose ((18)F-FDG) position emission tomography (PET) imaging. Methods: Twenty-four swines were randomly divided into sham-operated group, untreated group, trimethazine group and extract of Bulbus Allii Caespitosi group. Myocardial reperfusion injury was induced by plugging the anterior descending coronary artery of swine with sacculus. Bulbus Allii Caespitosi or trimetazidine was given twice daily for 28 days. Then myocardial perfusion was detected with (18)F-FDG PET/CT and the radioactivity distribution was evaluated. Results: Compared with the untreated group, Bulbus Allii Caespitosi and trimetazidine could improve the activity of myocardial cells after myocardial infarction (P0.05). Conclusion: Bulbus Allii Caespitosi can improve myocardial metabolism after ischemia and reperfusion in swines.
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OBJECTIVE:To provide references for the application and management of anti-infectives.METHODS:Data on sale volume,DDDs etc.of anti-infection drugs in6hospitals of Guang'an city from2001to2004were statistically analyzed.RESULTS:The DDDs of oral lomefloxacin dominated the first or2nd places for the4years;The DDDs rank orders of isoniazid and rifampin were No.4and No.5respectively in the4years;The DDDs of penicillin for injection dominated the first place on the injectable preparation lists(2001～2003);The latecomer levofloxacin assumed the tendency of surpassing the early starters not only in sale volume but also the DDDs.CONCLUSION:The proportion for anti-infectives in sales volume to the sum total drug consumption of the above mentioned area were close to those of the other medical institutions in southwest area,there are less new drug varieties and the grade of drugs used in the area was low.
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BACKGROUND: Recombinant adeno-associated virus 2(rAAV-2) has attracted considerable attention due to its nonpathogenic nature in contrast to other viral vectors such as adenoviral and retroviral vectors in gene therapy attempts.OBJECTIVE: To explore rAAV-2 transduction to bone marrow mesenchymalstem cell(BMSC) in vitro and evaluate the possibility of using rAAV-2 as a vector for gene therapy of acute myelogenous leukemia(AML).DESIGN: An open experiment with cells as the observational subjects.SETTING: Department of Hematology, Nanfang Hospital, Southern Medical University.MATERIALS: The experiment was conducted in the Department of Hematology, Nanfang Hospital, Southern Medical University from February to July 2004. We used passages 3 to 5 BMSCs derived from six de novo AML patients and four healthy volunteers in this study.METHODS: BMSC was isolated from 6 to 10 mL of bone marrow aspirates obtained from the iliac crests of the patients who had been diagnosed as having de novo AML and from those of healthy volunteers. The acquired BMSC was infected by rAAV-2 which contained enhanced green fluorescent protein (rAAV-2-eGFP) at different multiplicity of infection(MOI) (MOI = 1 × 102,1 × 103, 1 × 104, 1 × 105, 1 × 106, 1 × 107) . Then we observed through phase contrast fluorescent microscope and flow cytometer to evaluate green fluorescent protein(GFP) expression 10 to 14 days after transduction. GFP expression was observed as the rAAV-2-eGFP transduced BMSC cultured in vitro. We also observed the in vitro gene expression profile of GFP in rAAV-2-eGFP transduced BMSC which was selected by neomycin ( G418). First, we confirmed GFP expression in BMSC through phase contrast fluorescent microscope, then on flow cytometer to detect the percentage of GFP expression.MAIN OUTCOME MEASURES: The efficiency of rAAV-2-eGFP transduction to BMSC. GFP expression was observed through phase contrast fluorescent microscope and flow cytometer at different time points after transduction.rAAV-2-eGFP to BMSC derived from normal volunteers and AML patients had no significant differences. GFP began to express 10 to 14 days after transduction, and the transduction efficiency ranged from 0. 3% to 1.4%. By changing infection condition, we could not make a higher transduction efficiency( P ＞ 0.05) . One round infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was ( 1. 030 ± 0. 034) %, 3 rounds of infection of BMSC by rAAV-2-eGFP at a MOI of 1 × 105 was (1. 140 ±0. 036)%, and coinfected by LipofectAMINE was (1. 380 ± 0. 054)%. However, 293 cell line which was the package cell of rAAV-2 could be efficiently transduced by AML patients transduced by rAAV-2-eGFP at MOI = 1 × 105: The percentage of GFP expression cell gradually decreased from 1.14% at day 12 after transduction to 0. 6% as cell passaged from 2 to 3, and maintained at a level of 0. 5% to 0. 6% later on till 61 days after transduction. After selected by neomycin(G418) 1 month later, rAAV-2-eGFP transduced BMSCs could maintain a long-term GFP expression at a level of 6.0% in vitro without significant decay within 100 days of observation period after transduction.CONCLUSION: The advantages of rAAV-2 mediated gene transduction lie in safety, no immune response to the host, and long-term expression maintained by the target gene. rAAV-2 and BMSC can be used for in vitro gene therapy, and as a systemic gene delivery system, it might be an alternative for systemic gene therapy in the future.
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In order to supply a gap of PACS which is unidirectional,frame transmission and lag,a demonstration and teaching system which can be used as the tech support of the academic exchanging,teaching,studying,and emulating on spot is developed.By this,the medical audio and video signals and images including the video of the operation process,the images from endoscopes,electron microscopes,and the pathological images etc.in the clinical buildings and in the terminal units including research and teaching(training and exchanging) centers,lecture theaters and the director's offices etc.can be transmitted synchronously and bi-directionally in real time.
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<p><b>BACKGROUND</b>To construct a series of recombinant herpes simplex viruses that can provide replicating and packaging functions for recombinant adeno-associated virus (rAAV), and to select the strains possessing stronger functions for large-scale production of rAAV.</p><p><b>METHODS</b>A set of cosmids that represents the whole genome of HSV-1 was used to generate recombinant HSV-1 expressing rep and cap proteins of AAV-2. An ATG-to-ACG mutation in the start codon of AAV-2 rep protein was generated by site-directed mutagenesis. rep and cap genes, under control of their native promoters, with or without the ATG-to-ACG mutation in the start codon of rep, were inserted into the Xba site of UL2 or UL44 genes, respectively, resulting in the recombinant cosmids cos6-rmc/ UL2, cos56-rc/ UL44 and cos56-rmc/ UL44. Homologous recombination among the HSV-1 fragment on the recombinant cosmids and the rest fragments of HSV-1 genome resulted in three strains of recombinant HSV-1. Together with the one was constructed previously, there were four strains of recombinant HSV-1named HSV1-rc/ UL2, HSV1-rmc/ UL2, HSV1-rc/ UL44 and HSV1-rmc/ UL44 respectively.</p><p><b>RESULTS</b>PCR detection confirmed the existence of rep- gene in the genomes of all four strains of the recombinant HSV-1. Recombinant AAV was produced after infecting the AAV vector cell line that carrying the GFP expression cassette with the four strains of recombinant HSV-1 respectively. However, HSV1-rc/ UL2 and HSV1-rmc/ UL2 produced much more rAAV than HSV1-rc/ UL44 and HSV1c/ UL44 did.</p><p><b>CONCLUSIONS</b>All the four strains of recombinant HSV-1 support rAAV replication and packaging. HSV1 UL2 and HSV1-rmc/ UL2 that provide much stronger functions may be useful for large-scale production of rAAV.</p>
Subject(s)
Animals , Cricetinae , Cells, Cultured , Cosmids , Genetics , Dependovirus , Genetics , Physiology , Genetic Vectors , Herpesvirus 1, Human , Genetics , Physiology , Recombination, Genetic , Transfection , Virus ReplicationABSTRACT
Objective In this experiment,we explored the effects of rAAV\|hGDNF on protecting spinal neurons From death. Methods rAAV\|hGDNF particals were produced by recombinant virus technolgy,and infected the culture spinal neurons which were exposed to glutamate.We counted the mortality rate and detected the expression of NOS mRNA by RT\|PCR. Results In the group transfering rAAV\|hGDNF the death rate was inhibited(50%?0^02,control 59^25%?0^023, P
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Objective To summerize the clinical experience of 23 cases of carotid cavernous fistula(CCF) and discuss the treatment strategy. Methods Retrospectively analyzed 23 patients(11 males and 12 females) of CCF,including 14 patients with Barrow type A,1 with type B,1 with type C and 7 with type D. Results Vascular approach for embolization was undertaken in 20 cases by transarterial access and 2 via venous route while 1 case was given up. Among them,9 cases obtained complete occlusion by simple balloon embolization and 5 were treated by direct occlusion of internal carotid artery. Another 5 cases used NBCA glue for the occlusion,1 case was embolized by gelatin-sponge particulates combined with compression of carotid artery,2 cases were treated by coil embolization of cavernous sinus and 1 case was only managed by carotid compression maneuver. We achieved satisfactory clinical results with 21 patients cured and 2 patients improved. Conclusion Complexity and refractory are the characteristic of CCF,and endovascular treatment should be the major choice of treatment. (J Intervent Radiol,2006,15: 323-326)
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Objective To evaluate the effect of endothelin A receptor antisense oligodeoxynucleotides (ETAR-ASODN)on the growth of human prostatic stromal cells. Methods Primary cultured prostatic stromal cells were derived from patients with benign prostatic hyperplasia (BPH).The cells of passages 4～6 were routinely used for this study after identification.ETAR-ASODN at the concentrations of 5,10 and 15 ?mol/L were added into the culture cells with lipofectin, and cell proliferation was measured by MTT assay. The expression of ETA receptor was tested by 125 I-ET-1 radioligand banding assay. Results MTT assays showed a significant decrease in cell proliferation in stromal cells after 5,10 and 15?mol/L ETAR-ASODN were added with the A 540 values being 0.304?0.082,0.296?0.008 and 0.194?0.061,respectively.The proliferative activity was significantly decreased compared with control group ( P
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The most common cause of glaucoma filtration surgery failure is cicatrization secondary to fibroblast proliferation and of collagen. The high-risk factors for filtration failure include young age, aphakia, inflammation, neovascularization, and previous failure of filtration operation. Studies have shown beneficial effects of a single dose intraoperative mitomycin C,an antimetabolite, in inhibiting fibroblast proliferation and slceral wound cicatrization, thereby promoting success in eyes of poor surgical prognosis. Corneal complications have been minor.