血管内皮生长因子对佐剂性关节炎滑膜细胞基质金属蛋白酶3及基质金属蛋白酶9表达的影响

目的:观察血管内皮生长因子对佐剂性关节炎滑膜细胞质金属蛋白酶3及质金属蛋白酶9表达的影响,并探讨其意义。方法:实验于2006-02/12在桂林医学院实验中心完成。①实验材料:清洁级8周龄雄性Wistar大鼠6只,血管内皮生长因子为Peprotech EC LTD公司产品,基质金属蛋白酶3(扩增369bp)上游引物、下游引物,基质金属蛋白酶9(扩增405bp)上游引物、下游引物均购自晶美公司。②实验干预:先用弗氏完全佐剂造Wistar大鼠模型;造模20d后取Wistar大鼠右后足滑膜细胞进行原代培养。③实验分组:实验分为血管内皮生长因子组:取P2代细胞接种于6孔培养板,分别加入终浓度为5,25,50μg/L血管内皮生长因子;对照组:不加血管内皮生长因子。④实验评估:取病理切片观察滑膜细胞形态学改变;采用半定量反转录聚合酶链反应检测Wistar大鼠佐剂性关节炎滑膜细胞基质金属蛋白酶3及基质金属蛋白酶9的m-RNA表达。结果:①培养细胞的形态学观察:原代培养14d滑膜细胞从组织块边缘逸出,21d密集生长开始传代;传代细胞48h可明显分辨出树突样细胞、巨噬细胞样细胞和成纤维细胞样细胞;传至21代,细胞生长及特性稳定。②病理切片:滑膜组织有中性粒细胞、单核细胞、淋巴细胞浸润,滑膜细胞增生、排列紊乱,纤维素渗出,胶原纤维沉着,纤维素样坏死,呈滑膜炎表现。③基质金属蛋白酶3、基质金属蛋白酶9的mRNA表达:对照组基质金属蛋白酶3mRNA表达相对灰度值与血管内皮生长因子终浓度5,25,50μg/L组比较,差异有显著性意义(0.32±0.03,0.77±0.06,1.12±0.12,1.59±0.02,P<0.05);对照组基质金属蛋白酶9mRNA表达相对灰度值与血管内皮生长因子终浓度5,25,50μg/L组比较,差异有显著性意义(0.47±0.07,0.50±0.10,0.91±0.10,1.31±0.06,P<0.05);基质金属蛋白酶3和基质金属蛋白酶9mRNA表达随血管内皮生长因子浓度的加大表达增加。结论:血管内皮生长因子以剂量递增的方式对体外培养的佐剂性关节炎滑膜细胞基质金属蛋白酶3及基质金属蛋白酶9的表达有促进作用。     

The National Marrow Donor Program

As the number of successful marrow transplants has increased, the lack of HLA-identical sibling donors for 60 to 70 percent of transplant candidates has become a serious problem. Pilot studies established that marrow transplantation between phenotypically HLA-identical, unrelated individuals can be accomplished successfully. Therefore, the National Marrow Donor Program was established to develop a large file of volunteer marrow donors and to serve as a center for the coordination of the donor search and donor-recipient matching processes. By November 1991, 63 months after the program was established, 457,205 potential marrow donors typed for HLA-A and -B antigens had agreed to be listed in the marrow donor registry. A donor search had been initiated for 8481 patients. At least one potential donor matched for at least three of the four HLA-A and -B antigens was located for 99.8 percent of patients. Among the 3156 searches that were completed, 940 (29.8%) resulted in a transplant. The median time in which to locate a matched donor, complete all predonation evaluations, and obtain donor consent was 208 days. The most common diagnosis in patients who underwent transplantation was chronic myelogenous leukemia (42.0%). When this analysis was completed in November 1991, the National Marrow Donor Program was operating a national network of 99 donor centers and 53 transplant centers. The donor file was increasing rapidly, and a follow- up system was in place to determine the effects of donation on the donors and the outcome in the patients who underwent transplantation. This national network of donor and transplant centers exists and is now facilitating unrelated-donor marrow transplants. The National Marrow Donor Program made it possible to locate donors for many patients in need of a transplant and helped to determine the role of unrelated- donor marrow transplants in the treatment of many diseases.     

Preclinical characterization of WAY-211612: a dual 5-HT uptake inhibitor and 5-HT1A receptor antagonist and potential novel antidepressant

Background and purpose

As a combination of 5-HT selective reuptake inhibitor (SSRI) with 5-HT_1A receptor antagonism may yield a rapidly acting antidepressant, WAY-211612, a compound with both SSRI and 5-HT_1A receptor antagonist activities, was evaluated in preclinical models.

Experimental approach

Occupancy studies confirmed the mechanism of action of WAY-211612, while its in vivo profile was characterized in microdialysis and behavioural models.

Key results

WAY-211612 inhibited 5-HT reuptake (K_i = 1.5 nmol·L⁻¹; K_B = 17.7 nmol·L⁻¹) and exhibited full 5-HT_1A receptor antagonist activity (K_i = 1.2 nmol·L⁻¹; K_B = 6.3 nmol·L⁻¹; I_max 100% in adenyl cyclase assays; K_B = 19.8 nmol·L⁻¹; I_max 100% in GTPγS). WAY-211612 (3 and 30 mg·kg⁻¹, po) occupied 5-HT reuptake sites in rat prefrontal cortex (56.6% and 73.6% respectively) and hippocampus (52.2% and 78.5%), and 5-HT_1A receptors in the prefrontal cortex (6.7% and 44.7%), hippocampus (8.3% and 48.6%) and dorsal raphe (15% and 83%). Acute or chronic treatment with WAY-211612 (3–30 mg·kg⁻¹, po) raised levels of cortical 5-HT approximately twofold, as also observed with a combination of an SSRI (fluoxetine; 30 mg·kg⁻¹, s.c.) and a 5-HT_1A antagonist (WAY-100635; 0.3 mg·kg⁻¹, s.c). WAY-211612 (3.3–30 mg·kg⁻¹, s.c.) decreased aggressive behaviour in the resident-intruder model, while increasing the number of punished crossings (3–30 mg·kg⁻¹, i.p. and 10–56 mg·kg⁻¹, po) in the mouse four-plate model and decreased adjunctive drinking behaviour (56 mg·kg⁻¹, i.p.) in the rat scheduled-induced polydipsia model.

Conclusions and implications

These findings suggest that WAY-211612 may represent a novel antidepressant.     

Considerations in establishing an oral disease clinical research center

Oral Diseases (2010) 16 , 586–591 High quality clinical research is necessary to improve oral health and translate research findings to the practice of dentistry. This has led many academic institutions to consider establishing a formal clinical research center. This is not a trivial undertaking and requires that the center have an appropriate physical infrastructure, trained investigators with recognized expertise in the planning and conduct of high quality clinical research, and very importantly, a financial plan to assure its long‐term sustainability. The purpose of this paper is to provide some guidance and practical advice with respect to factors that should be considered in developing and maintaining a successful oral disease clinical research center.     

Polyacrylamide Gel Affinity Electrophoresis for Separation of Enzyme Isoforms

Affinity electrophoresis of differently glycosylated isoforms of enzymes using lectin as affinity ligand has been reported on support media such as cellulose acetate membrane (CAM) or agarose gel. We report a method for affinity electrophoresis in polyacrylamide gel (PAG) using wheat germ agglutinin (WGA). WGA is added to acrylamide-Bis mixture and incubated for 10 minutes at room temperature. This causes WGA to react covalently with acrylamide and Bis. Polymerization is initiated with addition of N,N,N,N-tetramethylethylene diamine (TEMED) and ammonium persulphate to give polyacrylamide gel with immobilized lectin. This gel has been found to effectively separate differently glycosylated isoforms of alkaline phosphatase (ALP). Concanavalin – A, similarly immobilized, did not give effective separation of ALP isoforms. The immobilization of lectin on polyacrylamide as support media requires less amount of lectin in comparison to CAM and agarose. Additional advantage of affinity electrophoresis on PAG is separation of biomolecules according to size.Key Words: Affinity electrophoresis, Alkaline phosphatase, Isoforms separation, Lectin, Polyacrylamide gel     

A STUDY OF HYPERINSULINAEMIA AND DYSLIPIDAEMIA IN HYPERTENSIVE PATIENTS OF ARMED FORCES

All patients attending the outpatient department were screened for hypertension. An attempt was made to correlate presence of hyperinsulinaemia (HI), dyslipidaemia and anthropometric characteristics in these hypertensives. Effect of angiotensin converting enzyme inhibitor (ACEI) and beta blockers on serum insulin was also studied. 85 patients with blood pressure (BP) ≥ 160/90 mm Hg and 94 controls with a BP of < 130/85 mm Hg were studied. All underwent clinical examination, anthropometric measurements (body mass index (BMI), waist hip ratio (WHR), skin fold thickness (SFT) and laboratory investigation (serum insulin, glucose, lipid profile) and post oral glucose load (POGL) for insulin and glucose. Serum insulin was estimated by I¹²⁵ radio immuno assay. Patients were randomly divided into group A (Tab enalapril) and group B (Tab atenolol). In 51 patients who completed the study, fasting and POGL insulin and fasting lipid profile were estimated two months after treatment. Mean age of cases was 38.91 years. 50% of patients had stage II hypertension. BMI was increased in 36 cases (42.35%) as compared to 9 in (9.57%) controls. Increased WHR was found in 40 cases as compared to 26 in controls. The SFT was more in cases compared to controls. 47 (55.29%) of 85 cases had abnormal lipid profile as compared to 25 (26.60%) in 94 controls. The fasting and POGL insulin levels (13.85 and 60.05 micro u/ml respectively) in cases were significantly higher than in controls (6.87 and 16.16 micro u/ml respectively). The mean POGL insulin values were much higher in obese compared to non-obese hypertensives. The decrease in mean fasting and POGL insulin values in patients taking ACEI and beta blockers were similar. Abnormal lipid profile was significantly more in cases than controls. Increased total cholesterol (TC), Low density lipoprotein cholesterol (LDLC) and total cholesterol (TC)/high density lipoprotein (HDL) ratio were the most frequent abnormality. The mean insulin (both fasting and POGL) levels were higher in obese hypertensives and those with abnormal lipid profile. Both drugs had equal efficacy in reducing the insulin values.KEY WORDS: Angiotensin converting enzyme inhibitor, Beta-blockers, Body mass index, Hyperinsulinaemia     

ERCC6 founder mutation identified in Finnish patients with COFS syndrome

Jaakkola E, Mustonen A, Olsen P, Miettinen S, Savuoja T, Raams A, Jaspers NGJ, Shao H, Wu BL, Ignatius J. ERCC6 founder mutation identified in Finnish patients with COFS syndrome. Cerebro‐oculo‐facio‐skeletal (COFS) syndrome is an autosomal recessive disorder characterized by microcephaly, congenital cataracts, facial dysmorphism, neurogenic arthrogryposis, growth failure and severe psychomotor retardation. We report a large consanguineous pedigree from northern Finland with six individuals belonging into four different sibships and affected with typical COFS syndrome phenotype. Two deceased patients have been published previously in 1982 as the first cases exhibiting cerebral calcifications typical for this disorder. Two living and one of the deceased patients were all shown to possess a novel homozygous mutation in the ERCC6 [Cockayne syndrome B (CSB)] gene, thereby confirming the diagnosis on molecular genetic level even for the earlier published cases. Genealogical investigation showed a common ancestor living in a northeastern village in Finland in the 18th century for all six patients implying a founder effect.     

利用T_1加权DCE-MRI技术对脑胶质瘤分级

应用T1加权动态对比度增强磁共振成像(DCE-MRI)技术对脑胶质瘤进行术前分级,验证其用于脑胶质瘤分级的准确性和可重复性。对26例脑胶质瘤患者团注Gd-DTPA对比剂后,以1.5TSiemens Syngo MRI扫描仪采集DCE T1加权图像。首先基于改进的Tofts两室药物动力学模型,从采集到的动态组图像建立病灶区感兴趣区内的平均信号强度-时间曲线,然后应用非线性最小二乘拟合方法对该曲线进行拟合,获取定量参数。这些参数包括:初始增强率ER、回流速率常数Kep、排泄速率常数Kel、曲线下面积AUC、峰值高度PH、到达峰值的时间TTP,最后与临床组织病理学结果进行比较,建立这些定量参数与脑胶质瘤分级之间的定量关系。采用独立样本t检验比较任意两个分级之间各定量参数的统计学差异,P0.05作为统计学差异标准。ER、AUC和PH随着脑胶质瘤的病理分级由低到高地增加;低级别脑胶质瘤中的Ⅰ与Ⅱ级之间以及高级别中的Ⅲ与Ⅳ级之间的比较结果显示,ER、AUC和PH的值均无明显统计学差异(P0.05);低级别与高级别级之间的比较结果显示,仅参数ER的差异具有统计学意义(P0.05)。ER是术前准确界定脑胶质瘤低级别与高级别的最具敏感性和特异性的参数,其界定阈值约为0.68。