2007年 · 第120卷第11期

Abstract：Objective To review the recent studies about microRNAs and advances in malignant lymphoproliferative disorders.Data sources Published articles (2001-2006) about microRNAs and malignant iymphoproliferative disorders were selected using MEDLINE.Study selection After independent review by two observers, 43 of 421 originally identified articles were selected that specifically addressed the stated purpose.Results Two observers independently assessed studies using explicit methodological criteria for evaluating microRNAs in malignant lymphoproliferative disorders. Recent work has revealed a class of small noncoding RNA species,microRNAs, which affect various biological processes. MicroRNAs inhibit the expression of protein encoding genes at the posttranscriptional level in a variety of eukaryotic organisms. In this review, we focused on the biogenetic pathways of microRNAs (miR-15a, miR-16-1, miR-155, miR-17-92 cluster, miR-142) and discussed the implications for human malignant lymphoproliferative disorders.Conclusions microRNAs are involved in tumorigenesis and mediate gene regulation as a fundamental genetic program at the posttranscriptional level. Further study of microRNAs may lead to novel concepts in the diagnosis and treatment of malignant lymphoproliferative disorders。

Abstract：Background Podocyte has inflammatory role in the development of diabetic nephropathy (DN). Mycophenolate mofetil (MMF), an anti-inflammatory agent, can suppress macrophage infiltration and reduce renal injury in streptozotocin-induced diabetic rats. Angiotensin Ⅱ receptor blocker (ARB), another renal protecting agent, can decrease podocyte loss in DN. In this study, we detected the expression levels of monocyte chemoattractant protein-1 (MCP-1) and nephrin to evaluate podocyte's role in inflammatory reaction in DN, observe and compare the effect of MMF alone and in combination with valsartan, on preventing podocyte loss in streptozotocin (STZ) induced diabetic rats.Methods Diabetic model was constructed in uninephrectomized male Wistar rats by single peritoneal injection of STZ (65 mg/kg). The successfully induced diabetic rats were randomly divided into four groups: diabetes without treatment group (DM), valsartan treated group (DMV), MMF treated group (DMM), and combined therapy group (DMVM). Normal rats of the same sibling were chosen as control (NC). At the end of the 8th week, serum biochemistry, 24-hour urinary protein (UP) and the ratio of kidney weight/body weight (RWK/B) were measured. The rats were sacrificed for the observation of renal histomorphology through light and electron microscope. Nephrin, desmin and MCP-1 levels were detected by semi-quantitative immunohistochemical assays. Real-time quantitative PCR was used to detect the mRNA levels of nephrin and MCP-1.Results Compared with group NC, serum glucose level, 24-hour UP and RWK/B in group DM were significantly higher (P＜0.01), and the nephrin mRNA level in DM group was significantly lower (P＜0.05). The nephrin mRNA expression levels in group DMV, DMM and DMVM were all higher than that of DM group (P＜0.05) and no significant differences were found among the three treatment groups (P＞0.05). Treatment with MMF, valsartan or their combination could significantly decrease the 24-hour UP and RWK/B, and suppress glomerulosclerosis and interstitial fibrotic lesions in diabetic rats. In diabetic rats, the high expressions of desmin and MCP-1 in kidney were suppressed by valsartan, MMF or their combination.Conclusions Podocytes are involved in the inflammatory reaction of diabetic rats. MMF could suppress MCP-1 and desmin expression, enhance nephrin expression, and attenuate proteinuria in diabetic rats. The combined therapy of valsartan and MMF did not show any superiority over monotherapies on renal protection. MMF may have renoprotective effect in early stages of diabetic nephropathy through preventing podocytes loss and anti-inflammatory activity。

Abstract：Background Lipoxins (LXs), endogenous anti-inflammatory and pro-resolving eicosanoids generated during various inflammatory conditions, have novel immunomodulatory properties. Because dendritic cells (DCs) play crucial roles in the initiation and maintenance of immune response, we determined whether LXs could modulate the maturation process of DCs and investigated the effects of lipoxin A4 (LXA4) on lipopolysaccharide (LPS)-induced differentiation of RAW264.7cells into dendritic-like cells.Methods RAW264.7 cells were cultured in vitro with 1 μg/ml LPS in the absence or presence of LXA4 for 24 hours, and cellular surface markers (MHC-Ⅱ, CD80 (B7-1), CD86(B7-2)) were measured by flow cytometry (FCM). Mixed lymphocyte reaction was performed to evaluate the allostimulatory activity. Cytoplastic IκB degradation and nuclear factor kappa B (NF-κB) translocation were detected by Western blotting. Luciferase reporter plasmid was transiently transfected into RAW264.7 cells, and luciferase activity was determined to measure the transcriptional activity of NF-κB.Results LXA4 reduced the ratio of LPS-treated RAW264.7 cells to DCs with morphological characteristics and inhibited the expression of MHC Ⅱ. LPS-induced up-regulation of CD86 was moderately suppressed by LXA4 but no obvious change of CD80 was observed. Moreover, LXA4 weakened the allostimulatory activity of LPS-treated RAW264.7 cells.These alterations of LPS+LXA4-treated cells were associated with a marked inhibition of IκB degradation, NF-κB translocation and then the transcriptional activity of NF-κB.Conclusions LXA4 negatively regulates LPS-induced differentiation of RAW264.7 cells into dendritic-like cells.This activity reveals an undescribed mechanism of LXA4 to prevent excessive and sustained immune reaction by regulating maturation of DCs。

Abstract：Background Candida albicans (C. albicans) infection, often occurring in genital candidiasis, has increased dramatically recently. Developing an efficient C. albicans typing method may contribute to understanding its epidemiological characteristics and guiding efficient treatment. We used rapid microsatellite genotyping assay for interstrain differentiation of C. albicans isolates and explored some characteristics of its spread.Methods DNA was extracted from C. albicans isolates from gentalia, recta and mouths of 39 female cases and 27 male cases of genital candidiasis. Three fluorescent primers for the microsatellite markers in conserved genes (CDC3, EF3and HIS3) of C. albicans were used to amplify the isolates DNA by PCR. Fluorescent signals were read with an automatic sequencer and analyzed with GeneScan software.Results Analysis of the three microsatellites markers showed 18 gene allelic associations in genital C. albicans infected patients: 10 allelic associations in female and 11 allelic associations in male, of which 3 allelic associations shared by both genders covered 71% of infections. The most dominant allele association of pathogenic strains for both genders was 116:124, 122:131,160:200 that covered about 50% of infection. Gentalia and recta shared the same strains in 80%of female patients, but in only 3.8% of male patients. There were 2.7% female patients, but no males, with same strain in both gentalia and mouths. Five of seven genital C. albicans infected couples had the same allelic associations of which 4were the dominant pathogenic C. albicans susceptible for both genders.Conclusions The predominant allelic association of the pathogenic strain in genital C. albicans infection is 116:124,122:131, 160:200. Vaginal pathogenic strains are probably maintained from the rectal reservoir. Pathogenic strains of male patients are probably from frequent sexual intercourse. The aggressiveness of some strains varies with gender。

Abstract：Background Dopamine exerts its effects mainly in nervous system through D1, D2 or D3 receptors. There are few reports dealing with the effects of dopamine on leukaemia cells. However, some dopamine agonists or antagonists do show biological effects on some types of leukaemia cells. Here, we report the effects of dopamine on the proliferation,differentiation and apoptosis of K562 leukaemia cells.Methods Proliferation was determined by MTT assay and cell counting both in liquid and semisolid cultures.Differentiation was verified by morphology, benzidine staining and flow cytometry. Apoptosis was checked by Hoechst 33258 staining and flow cytometry. The two groups were untreated group and treated group (dopamine 10-9 mol/L-10-4mol/L).Results In liquid culture, MTT assay and colony assay, dopamine inhibited proliferation of K562 cells. Inhibition rate was 29.28% at 10-6 mol/L and 36.10% at 10-5 mol/L after culture for 5 days in MTT assay. In benzidine staining and CD71 expression, dopamine induced K562 cells toward erythroid differentiation by increased 155% at 10-6 mol/L and by 171% at 10-5 mol/L after culture for 5 days in benzidine staining. In Hoechst 33258 staining and flow cytometry,dopamine induced K562 cells toward apoptosis. The sub G1 peak stained by PI was 14.23% at 10-4 mol/L dopamine after culture for 3 days compared with the control (0.81%) in flow cytometry.Conclusion Dopamine inhibites proliferation and induces both differentiation and apoptosis of K562 leukaemia cells。

Abstract：Background It is difficult and challenging to reconstruct hepatic venous outflow in adult right lobe living donor liver transplantation (LDLT) without the middle hepatic vein (MHV). Excessive perfusion of the portal vein and venous outflow obstruction will lead to acute congestion of the graft, ultimately resulting in primary nonfunction. Although various reconstruction patterns have been explored in many countries, there is currently no clear consensus. In this study we describe a technique to prevent "chocking" of the graft at the outflow anastomosis with the inferior vena cava (IVC) in LDLT using right lobe graft without the MHV.Methods A retrospective analysis was conducted on clinical data from 55 recipients undergoing LDLT using right lobe grafts without the MHV or reconstruction of hepatic venous outflow. The donor's right hepatic vein (RHV) was anastomosed with a triangular opening of the recipient IVC; the inferior right hepatic vein (IRHV), if large enough, was anastomosed directly to the IVC. The great saphenous vein (GSV) was used for reconstruction of significant MHV tributaries.Results No deaths occurred in any of the donors. Of the 55 recipients, complications occurred in 6, including hepaticvein stricture (1 case), small-for-size syndrome (1), hepatic artery thrombosis (1), intestinal bleeding (1), bile leakage (1),left subphrenic abscess and pulmonary infection (1). A total of three patients died, one from small-for-size syndrome and two from multiple system organ failure.Conclusions The multiple-opening vertical anastomosis was reconstructed with hepatic vein outflow. This technique alleviates surgical risk of living donors, ensures excellent venous drainage, and prevents vascular thromboses and primary nonfunction。

Fat-forming variant of solitary fibrous tumor (fat-forming variant of SFT), previously called lipomatous hemangiopericytoma (L-HPC), is a recently recognized rare variant of solitary fibrous tumor (SFT)with unpredictable biologic behavior. Fat-forming variant of SFT occurs predominately in the deep soft tissues of the lower extremities. Histologically, it is composed of an admixture of benign hemangiopericytomatous and lipomatous components. Up to now, only 37 cases have been reported in the English literature worldwide1-7 and the molecular analyses are sparse. Reported herein is the first Chinese case of fat-forming variant of SFT. Although one case of fat-forming variant of SFT in the mediastinum was briefly reported, this is the first case in the region with detailed histologic, immunohistochemical features and flow cytometric DNA analysis。

Polydactyly, one of the fairly common congenital deformities of the hands and/or feet, occurs as an isolated disorder or as part of a syndrome. It is not only observed sporadically, but also inherited with a mainly autosomal dominant inheritance. However, postaxial polydactyly in both hands and feet with 24 digits in total was rarely reported in China。

Both diabetes mellitus (DM) and hypertension are common in the aged population. The incidence of angiopathies and calcification in pulp vessels is more frequent in DM patients than nondiabetics.1 Some evidences have suggested an interaction between control of blood pressure and pain regulation. Zamir and Shuber 2found that hypertensive patients had a higher threshold for sensation of pain than normotensive controls in the tooth-pulp electricity test。

The islet transcription factor pancreatic duodenal homeobox-1 (PDX-1, also known as insulin promoter factor-1 or IPF-1) is an orphan homeodomain protein that plays an important role in the development, proliferation,differentiation and maturation of pancreatic cells。

Abstract：Background Promoter analysis is currently applied to detect the expression of the targeted gene in studies of signal transduction and transcriptional regulation. As a reporter gene, luciferase plays an important role and has been used widely in the promoter assay.Methods Human embryonic lung fibroblast cells (2BS), HeLa cells and MCF-7 cells were transfected with various genes embedded by lipofectamine. This study determined various factors that affect promoter activity determination,such as the selection of the reporter genes and internal references, the dose and the type of the vectors carrying the transcription factors, the host cells and the instruments.Results The sensitivity of the luciferase assay was much higher than that of enhanced green fluorescence protein (EGFP). Moreover, promoter activity is increased in a dose-related manner only in certain ranges outside of which the results may be reversed and the promoter activity is related to the expression vector which is carrying the cDNA.Otherwise, the length of the promoter, internal references and the host cell can also influence the promoter activity.Conclusions To detect the promoter activity accurately, a few factors including dose, vector, length and host cell which influence reporter gene assay aforementioned should be considered。

Abstract：Background 11C-4-N-(3-bromoanilino)-6,7-dimethoxyquinazoline (11C-PD153035) has been reported as a tracer for imaging human tumors that overexpress epidermal growth factor receptor (EGFR). However it is still unclear whether 11C-PD153035 uptake correlates with EGFR expression levels. The objective of this study was to investigate the relationship between 11C-PD153035 accumulation and EGFR expression levels.Methods Synthesis of 11C-PD153035 was performed in the Tracerlab FXc system. Accumulation of 11C-PD153035 by MDA-MB-468, A549 and MDA-MB-231 cells was measured in vitro. There were six tumor-bearing mice in each group.11C-PD153035 uptake in tumors was determined by positron emission tomography/computed tomography (PET/CT).Tumor/normal muscle tissue (T/NT) analysis in PET images was applied to quantify the PET data. Sixty minutes after PET/CT scanning, the nude mice were sacrificed and the tumors were excised. The 11C-PD153035 accumulation in different tumors was determined by a gamma counter.Results Close correlation existed between the uptake and the level of EGFR expression both in vitro and ex vivo (r2=0.72, P＜0.001; r2=0.63, P=0.003). When the static T/NT analysis method was applied to analyze the PET data, the observed correlation was again excellent (r2=0.70, P=0.001).Conclusions The uptake of PET tracer 11C-PD153035 closely correlates with the EGFR expression levels in tumor cells. 11C-PD153035 has the potential to yield useful information for both cancer diagnosis and therapy。

Abstract：Background Pulmonary-vein isolation (PVI) is currently used for the treatment of chronic and paroxysmal atrial fibrillation and a major risk of PVI is thromboembolism. The purpose of this study was to observe embolic event rate in patients with persistent or paroxysmal atrial fibrillation (AF) undergone PVI.Methods Circumferential PVI (CPVI) was performed in 64 consecutive patients with persistent AF (42 men, aged (60.0±9.1) years) and in 84 consecutive patients with paroxysmal AF (53 men, aged (61.4±9.3) years). Warfarin was administrated in all patients before ablation for at least 3 weeks ((5.2±2.6) weeks) and continued for at least 3 months post ablation with international normalized ratio (INR) of 2.0-3.0. During CPVI, intravenous heparin was given at a dose of 5000-8000 U or 75-100 U/kg, followed by 1000 U or 12 U/kg per hour.Results In patients with persistent AF, 1 patient developed embolic event during ablation and 3 patients developed embolic events after ablation. In contrast, no thromboembolic event was observed in patients with paroxysmal AF (4/64vs 0/84, P=0. 033).Conclusion Thromboembolic event rate related to CPVI is significantly higher in patients with persistent AF than that in patients with paroxysmal AF。

Abstract：Background The methods for the treatment of postcatheterization femoral arteriovenous fistulas (AVF-s) - simple observation, ultrasound guided compression, covered stents implantation and coil embolization have poor outcome.Surgery is the standard method for treatment of femoral AVFs, but it is a traumatic operation. In this study, we report the results of the treatment of postcatheterization femoral AVFs by simple prolonged compressing bandage.Methods To treat iatrogenic femoral AVFs caused by transfemoral catheterization, prolonged binding with elastic or common bandage was applied in 16 cases. Catheterization was performed in 7 cases for radiofrequency current catheter ablation, in 4 for occlusion of congenital heart disease, in 3 for percutaneous coronary intervention, in 1 for coronary angiography and in 1 for right heart catheterization.Results All iatrogenic femoral AVFs were healed after simple binding with elastic or common bandage for 4-46 days (mean (15±10) days). During the period of binding, local skins ulceration occurred at puncture site in two cases and femoral vein thrombus was found in one patient. During 6-24 months (mean (11.8±3.6) months) followup with colour Doppler ultrasonography, no recurrent arteriovenous shunting or other complications were observed.Conclusion The results suggest that simple prolonged bandaging for postcatheterization femoral AVFs is an effective and economical procedure。

Achondroplasia (ACH) is the most common form of skeletal dysplasia characterized by disproportionately short stature, lumbar lordosis, relative macrocephaly and other skeletal anomalies resulting from a defect in the maturation of the chondrocytes in the growth plate of the cartilage. The combined frequency of the disease has been estimated to be 1 in 15 000 live births.1 ACH is inherited in autosomal dominant fashion with a complete penetrance, more than 80% of affected individuals have de novo mutations associated with increased paternal age。