2006年 · 第119卷第05期

Abstract：Background Human cytomegalovirus (HCMV) infects a number of organs and tissues in vivo. The different symptoms and tissue tropisms of HCMV infection perhaps result from genetic polymorphism. A new region of DNA containing at least 19 open reading frames(ORFs) (denoted UL133 to 151) was found in the low-passage HCMV clinical strain, Toledo, and several other low-passage clinical isolates, but not present in the HCMV laboratory strain, AD169. One of these genes, UL143, was studied to explore the sequence variability of UL143 ORF in HCMV clinical isolates and examine the possible association between gene variability and the outcome of HCMV infection.Methods The UL143 gene of the strains obtained from suspected congenitally HCMV-infected infants was amplified by polymerase chain reaction (PCR) and sequenced.Results Nineteen sequences of the strains were divided into 2 major groups, G1(n=16) and G2(n=3). All of the sequences had frame-shift mutation compared to Toledo. Nucleotide polymorphisms conferred substantial amino acid substitutions when compared with Toledo. All 16 UL143 putative proteins of the strains in G1 had a new myristylation site and loss of two PKC sites owing to missense mutations. No convincing relationships were observed between the presence of HCMV disease and the UL 143 sequence group.Conclusions HCMV-UL143 existed in low passage isolates. Sequence variability caused by frame-shift mutation was found in all HCMV clinical strains. No obvious linkage was observed between UL143 polymorphisms and the outcome of suspected congenital HCMV infection。

Abstract：Background Non-adhesive liquid embolic agents are increasingly gaining importance in the embolization of cerebral arteriovenous malformations (AVMs). We investigated the use of poly (N-isopropylacrylamide)(PNIPAM) as a non-adhesive embolic agent in swine rete mirabile.Methods The PNIPAM hydrogel was mixed with iohexol and embolization was performed in swine rete mirabile in 30 animals. The microcatheter was examined after embolization. Follow-up angiography was performed for embolic efficacy after embolization. Embolized retia were examined histopatholgically, and the alterations of inside rete and surrounding tissue were observed.Results The copolymer hydrogel was used for rete embolization in 30 swine, 28 swine survived the procedure,2 swine died, 1 swine died of cerebrum infarction and the other died of embolic agent reflux into the occipital artery. The inside wall of the microcatheter was smooth, without copolymer adhering to it. Follow-up angiography was performed in 22 swine, there was no rete recanalization in 20 swine and partial rete recanalization in 2 swine because of the trunk embolization of ascending pharyngeal arteries. Histopatholgically,the copolymer was found diffused into vessels of 100-150 μm in diameter. In acute group, neutrophils scattered surrounding the copolymer and endothelial integrity was observed, without endothelial denuding and necrosis. In subacute and chronic groups, the copolymer was found inside retia, a few mononuclear cells and eosinocytes scattered inside and surrounding it. The muscular layer was loosened with most muscular nuclei degraded.Conclusion Experimental rete embolization with PNIPAM, made radiopaque with iohexol, is technically feasible in swine. Because of its properties, PNIPAM has great potential as a therapeutic non-adhesive embolic agent。

Abstract：Background Mutation or deletion in the phosphatase and tensin homologue deleted on chromosome ten (PTEN)gene has been identified as an important cause of endometrial carcinoma; stromal cell derived factor-1α (SDF-1α)exerts growth-promoting effects on endometrial cancer cells through activation of the PI-3 kinase/Akt pathway and downstream effectors such as extracellular-responsive kinase (ERK). In this study, a plasmid containing the PTEN gene was transfected into Ishikawa cells to investigate the difference in growth and signal transduction between Ishikawa-PTEN and Ishikawa cells after SDF-1α stimulation, and to study mechanisms of the involvement of PTEN protein in endometrial carcinoma development.Methods Ishikawa cells were transfected with a plasmid (pLXSN-PTEN) containing the PTEN gene and a plasmid (pLXSN-EGFP) with enhanced green fluorescent protein (EGFP). Cells were then screened to obtain Ishikawa-PTEN cells and Ishikawa-neo cells that can both stably express PTEN protein and EGFP. Expression of PTEN protein, phosphorylation levels of AKT and ERK (pAKT and pERK) and growth differences in Ishikawa-PTEN, Ishikawa-neo and Ishikawa cells before and after SDF-1α stimulation were then determined by Western blots and MTT assays.Results Western blot analysis showed that Ishikawa cells produced PTEN after transfection with the PTEN gene. At 15 minutes after SDF-1α stimulation, the pAKT level of Ishikawa-PTEN cells was lower than that of Ishikawa-neo cells and Ishikawa cells. There was no significant difference in pERK levels among the three cell lines. The positive effect of SDF-1α on Ishikawa-PTEN cells growth was markedly less than the effect on Ishikawa-neo and Ishikawa cells. However, in the absence of SDF-1α stimulation (baseline), the pAKT level in Ishikawa-PTEN cells was less than that in Ishikawa cells. There was a significant difference in growth between the Ishikawa-PTEN cells and the Ishikawa-neo cells.Conclusions PTEN gene transfection can regulate the level of pAKT but not pERK in Ishikawa-PTEN cells.PTEN protein may suppress the growth-promoting effect of SDF-1α on endometrial carcinoma by inhibiting the PI-3K/AKT signal transduction pathway。

Abstract：Background Leigh syndrome is an inherited neurodegenerative disease that emerges in infancy and childhood and presents with a clinically heterogeneous variety of neuromuscular and non-neuromuscular disorders. It can result from the inheritance of mutations in either nuclear or mitochondrial DNA. In the current study, we performed a retrospective study in 65 patients in order to investigate the clinical and genetic characteristics of Leigh syndrome in Chinese patients.Methods Sixty-five unrelated cases (35 men and 30 women) who were hospitalized in the past 12 years were reviewed. Diagnosis was based on both the clinical presentation and the characteristic neuropathologic findings of bilateral symmetric necrotizing lesions in the basal ganglia and brain stem as detected using cranial computed tomography (CT) scan or magnetic resonance imaging (MRI). The differential diagnosis of organic acidurias and fatty acid β-oxidation defects were performed. Specific point mutations and deletions in mitochondrial DNA (T8993G, T8993C, T9176C, A8344G, A3243G) were screened by PCR-restriction analysis and Southern blot.The SURF1 gene was sequenced. Skeletal muscle biopsies were performed in 17 (26.2%) of the patients. The diagnosis was confirmed by autopsy in 6 (9.2%) patients.Results The patients had various forms of metabolic encephalomyopathy. Fifty-nine (90.8%) of the patients had the typical neuroradiological features of Leigh syndrome, including symmetrical necrotizing lesions scattered within the basal ganglia, thalamus and brain stem. Twenty (30.8%) patients were confirmed by genetic,biochemical analysis and autopsy. Specific point mutations in mitochondrial DNA were found in 5 cases (7.7%).Of these, the A8344G mutation was detected in 2 patients. The T8993G, T8993C, and A3243G point mutations were identified in 3 other patients, respectively. SURF1 mutations associated with cytochrome c oxidase deficiency were identified in 8 (12.3%) families by DNA sequencing. A G604C mutation was identified in 6(9.2%) patients. The genotypes of 52 patients remained unknown.Conclusions Leigh syndrome presents as a diverse array of clinical features and can result from specific mutations in nuclear or mitochondrial DNA. In this study, SURF1 mutations associated with cytochrome c oxidase deficiency were identified in 8 (12.3%) out of 65 patients with Leigh syndrome. It indicates that SURF1 mutations might be a common cause of Leigh syndrome in China. The etiology of Leigh syndrome in Chinese patients represents a persistent challenge to clinicians。

Abstract：Background Verapamil-sensitive, idiopathic left ventricular tachycardia (ILVT) with right bundle branch block configuration and left-axis deviation is known to be due to re-entry mechanism but the exact nature of reentrant circuit in ILVT is not fully elucidated. Radiofrequency (RF) ablation was applied during ventricular tachycardia (VT) and termination of the VT or abolishing the inducibility of the tachycardia was used as an endpoint for successful RF. In this study, the left posterior fascicular block in surface electrocardiogram (ECG)was used as a new endpoint of ablation to cure ILVT.Methods Electrophysiological studies and radiofrequency ablation were performed in 39 consecutive patients [30 men, 9 women; age ranging from 10 to 64 years, mean (29± 16) years] with verapamil-sensitive ILVT and structurally normal hearts. VT could be terminated by the intravenous administration of verapamil in all patients.The target site was the midseptum of LV where the earliest Purkinje potentials were recorded during VT. RF current was applied to the target site with or without late diastolic potential (LDP) during sinus rhythm in 37patients and during VT in 2 patients to meet the ablation endpoint: the left posterior fascicular block in the surface ECG.Results Thirty-seven patients with ILVT had been treated by RF ablation during sinus rhythm and two had been treated during VT. All of them met the endpoint of the left posterior fascicular block. Thirty-eight cases were symptom-free without medications during the follow-up period (range from 3 to 95 months, median 17months). One patient developed a clinical recurrence and the left posterior fascicular block in surface ECG disappeared. The patient received another treatment. The endpoint was met and the procedure was successful.Conclusions The left posterior fascicular block in surface ECG used as an endpoint of RF ablation to treat ILVT is effective. It is important especially in those patients whose VT can not be induced or the inducible condition is unstable. The effective endpoint implied that the left posterior fascicle might be a critical part of the re-entrant circuit。

Abstract：Background Cilostazol is a newly developed antiplatelet drug that has been widely applied for clinical use. Its antiplatelet action appears to be mainly related to inhibition of intracellular phosphodiesterase activity. Recently,cilostazol has been used for antiplatelet therapy after coronary bare metal stent implantation for thrombosis and restenosis prevention. This prospective randomized and double blind trial was designed to investigate the safety and efficacy of cilostazol for the prevention of late restenosis and acute or subacute stent thrombosis.Methods One hundred and twenty patients who underwent elective stent were randomly assigned to treatment group with cilostazol 200 mg/d (n = 60), clopidogrel 75 mg/d and aspirin 100 mg/d or to control group with clopidogrel treatment 75 mg/d (n = 60) and aspirin 100 mg/d. Follow-up coronary angiography was performed 6-9 months later.Results Nine months major adverse cardio-cerebral event (MACCE) were lower in treatment groups (P＜0.05).The quantitative coronary angiography (QCA) at 6 months follow-up showed that minimum lumen diameter(MLD) was higher in treatment group than that of control group [(2.14±0.52)mm vs (1.82 ±0.36)mm, P＜0.05].Late lumen loss (LL) [(0.82 ± 0.42)mm vs (1.31 ± 0.58)mm; P＜0.01 ], restenosis rate (RR) (14% vs 32%; P＜0.05)and target lesion revascularizaion (TLR) rate (5% vs 17%; P＜0.05) were lower in treatment group than in control group.Conclusion Cilostazol therapy is an effective regimen for prevention not only stent thrombosis but also RR and TLR through reducing MLD without the risk of increasing bleeding。

Abstract：Background Low molecular weight heparin (LMWH) was more effective than unfractionated heparin (UFH)in treating acute coronary syndrome (ACS). However, it remains uncertain whether LMWH can be used in patients undergoing percutaneous coronary intervention (PCI) instead of UFH. This study aimed to evaluate the efficacy and safety of using enoxaparin instead of UFH in the intervention treatment of patients with coronary heart disease (CHD)Methods From October 2003 to Febuary 2005, 966 patients with CHD were enrolled into this study. Among 966 patients, 455 patients received the PCI, including 283 patients with Non-ST segment elevation ACS (NSTEACS), 511 patients did not received PCI due to mild, moderate lesions or were suitable for coronary artery bypass graft (CABG). The 966 patients were randomized to enoxaparin group (484 patients) and UFH group (482patients). Patients in the enoxaparin group were given enoxaparin at least twice subcutaneously (1 mg/kg, q12 h)before catheterization. Plasma anti-Xa activity was determined 1-8 hours after the last dose of enoxaparin was determined. The catheterization was performed within 8 hours after the last dose of enoxaparin. The sheath was removed immediately after the procedure. Patients in the UFH group were given UFH 25 mg intravenously before coronary angiography. Additional 65 mg was given intravenously if PCI was to be performed. The sheath was removed 4 hours after the procedure.Results A total of 227 patients in the enoxaparin group and 228 patients in the UFH group received PCI. In the enoxaparin group, one patient developed acute thrombosis during PCI and resulted in acute myocardial infarction (AMI), no acute or subacute thrombosis was found during hospitalization. In the UFH group, no acute or subacute thrombosis occurred during PCI procedure and hospitalization. Therefore, the incidence of major adverse cardiovascular events (MACEs) during the hospitalization was 0.44% in the enoxaparin group and 0 in the UFH group. In the enoxaparin group, the sheath was removed immediately after the procedure and 8 patients had hematoma on the puncture site. In the UFH group, the sheath was removed 4 hours after the procedure and 20 cases had hematoma on the puncture site. The incidence of hematoma on the puncture site was significantly higher in the UFH group than that in the enoxaparin group (P＜0.05). Anti-Xa activity was determined in 174 patients in LMWH group. The mean anti-Xa activity was (0.87±0.23) U/ml, and 94.8% of them had anti-Xa activity ＞0.5 U/ml and 6.9% of the patient ＞1.2 U/ml. There was no death and AMI occurred in enoxaparin group,but one patient had AMI caused by subacute thrombosis in UFH group during 30-day follow-up. MACE rate at 30-day follow-up was 0 in enoxaparin group and 0.43% in UFH group.Conclusions The results of the study suggest that it is safe and efficient to give enoxaparin at least twice before the PCI procedure, and the sheath can be removed immediately after PCI. For NSTEACS patient who has received enoxaparin more than twice during the hospitalization can undergo PCI directly and no UFH is necessary before or during PCI。

Abstract：Background It has been identified that in patients with pre-eclampsia, several factors were released into the serum, which can regulate the proliferation of vascular smooth muscle cells and stimulate the synthesis of extracellular matrix (ECM). However, the signal transduction pathway of the growth factors and cytokines synthesized by endothelial cells leading to the proliferation and ECM synthesis of human umbilical arterial smooth muscle cell (HUASMC) is unknown. The aim of this study was to research the effects of protein kinase C (PKC) inhibitor, polymyxin B (PMB), on the proliferation, nuclear factor-kappa B (NF-κB) activation, and expression of procollagen Ⅰ and Ⅲ mRNA in HUASMC cultured with pre-eclamptic umbilical sera.Methods Normal HUASMCs were treated with pre-eclamptic umbilical serum (pre-eclamptic group), preeclamptic umbilical serum plus PMB (PMB group), or normal umbilical serum (normal group). The expression of I-κB, NF-κB was detected by Western blotting after the HUASMC was incubated for 2 hours. The proliferation of HUASMC was evaluated by MTT, the cell cycle was analyzed by flow cytomerty, and the expression of procollagen Ⅰ, Ⅲ mRNA was measured by RT-PCR assay after HUASMC was incubated for 48hours. ANOVA was used for statistical analysis.Results Umbilical sera in pre-eclampsia could stimulate the proliferation, the DNA synthesis, the transition of G0+G1 phase or G2/M phase to S and phase, the activation of NF-κB, and the expression of procollagen Ⅰ mRNA of HUASMC as compared with normal umbilical sera (P＜0.01). PMB could inhibit the proliferation, the DNA synthesis, the transition of G0+G1 or G2/M phase phase to S phase, the activation of NF-κB, and the expressions of procollagen Ⅰ mRNA of HUASMC stimulated by umbilical sera in pre-eclampsia (P＜0.01).Conclusion PKC-NF-κB signal transduction pathway may play a key role in SMC phenotype modulation,which is more important in the pathogenesis of placental blood vessel in pre-eclampsia。

In China, the first report on transanal posterior myotomy of internal sphincter for ultrashort aganglionosis was in 1997,1 while reports of the transanal, single stage, Soave for Hirschsprung's disease (HD) began to appear only after Torre's paper,2 published in 1998. The transanal endorectal pull through for HD was soon widely used by Chinese paediatric surgeons even in many provincial hospitals and local modifications frequently reported in the literature. From 2001 to 2004, 85 papers on transanal pull through for HD were published, and from 1999 to 2004, 252 patients underwent transanal endorectal operation for HD in our department. After reviewing the literature and analysing the clinical data, we discuss the present status and possible further developments of transanal, single stage,endorectal pull through for HD in China。

Many health benefits have been ascribed to allicin, including vasorelaxant activities.1However, the molecular mechanisms underlying these effects remain unknown. To apply allicin in clinical anti-HCC treatment, different doses of allicin and Adriamycin (ADM) were led into transplanted tumor model established by injecting HCC BEL7402cells into subcutaneous tissue of nude mice.Treatment with allicin and ADM resulted in tumor regression with inducing of Bax and Fas ligand (FasL) mRNA。

The important aim in cancer treatment is the selective killing of cancer cells without damaging healthy cells and the most commonly used chemotherapy for eliminating cancer is achieved by activating the mitochondrial apoptotic pathway.1 Therefore, identification of new agents that can selectively kill cancer cells becomes crucial in cancer clinical treatment。

Imiquimod is an imidazoquinoline derivative. Some studies have shown that imiquimod modulates the Th1/Th2 response by inducing the production of Th1 cytokines IFN-γ and interleukin (IL)-12,1 and by inhibiting Th2 cytokines IL-4 and IL-5. These data suggest that imiquimod has therapeutic applications in atopic diseases such as allergic asthma that are associated with overexpression of Th2 cytokines。

Human papillomavirus (HPV), mainly types 16 and 18, are the most important initiating agents of cervical cancer.1 Prevention of high-risk HPV infections is a potentially effective approach to control HPV associated cervical cancer。

The prevalence of pleural effusion caused by hepatocirrhosis ranges from 0.4% to 30%. Some patients with hepatic hydrothorax (HH) fail to respond to aggressive medical management such as albumin application, diuretics, and thoracentesis,and remain refractory massive pleural fluid.1 The repeated suctions to relieve the symptoms such as respiratory distress are associated with significant hyponatremia and hypoalbuminemia caused by the large volumes of fluid loss. Review of the literature has revealed that, to date, no therapy is ideal. This report describes our experiments in treating HH of patients with cirrhosis and ascites by thoracoscopy and talc poudrage, to offer our efforts to approach the pathogenesis of HH and to explore the new therapeutic strategy。

Recently, with the rapid development of highway and high-rise buildings, trauma shows a tendency to increase continuously. The proportion of trauma patients with multiple injuries has increased and trauma arising from multiple injuries has become "the first public hazard"。

Gastrointestinal liposarcoma is rare, and the esophagus is the. least common location at which this kind of liposarcoma may occur. We reported a patient with a well-differentiated liposarcoma of esophagus, and discussed the diagnostic utility of these imaging techniques in patient management。

Behcet's disease (BD), a multisystem chronic autoimmune process of unknown etiology,usually leads to arterial impairment. Isolated case reports have described BD-related arterial dissections, pseudoaneurysms or aneurysms.1-4 Recently, we successfully treated a huge vertebral artery pseudoaneurysm (VAPA) in a patient with BD by stent-grafting with preservation of the affected vertebral artery。