2006年 · 第119卷第09期

Abstract：Background T-cell receptor (TCR) plays an important role in the development of autoimmune diseases.Recently, it was reported that immunization of animals with TCR peptide derived from the pathogenic cells could prevent autoimmune diseases. The aim of this study was to investigate whether vaccination with a synthetic peptide from the hypervariable region of TCR Vβ 8.3, an experimental autoimmune uveoretinitis (EAU)-associated gene, was able to prevent the disease.Methods EAU was induced in Lewis rats by immunization with IRBP R16 peptide emulsified in complete Freund's adjuvant (CFA). The clinical and histological appearances were scored. Delayed type hypersensitivity (DTH) and lymphocyte proliferation were detected. Cytokine levels of aqueous humour, supernatants of cells from spleen and draining lymph nodes were measured by enzyme linked immunosorbent assay (ELISA). Gene expression of TCR Vβ 8.3 on CD4+ T cells was examined by real time quantitative polymerase chain reaction (PCR).Results After vaccination, the intraocular inflammation was significantly mitigated, antigen specific DTH and lymphocyte proliferation responses were suppressed, interleukin (IL)-2 in aqueous humour, interferon (IFN)-γand IL-2 produced by the spleen and draining lymph node cells were significantly decreased, whereas the production of IL-4 and IL-10 were increased. The response of draining lymph node cells to TCR Vβ 8.3 peptide was enhanced after vaccination. Inoculation with CFA alone did not affect the severity of EAU and the above parameters. The suppression of EAU was much stronger in the group of four fold inoculations than the group of two fold inoculations. The expression of TCR Vβ 8.3 gene was significantly reduced in the group of fourfold inoculations.Conclusion Vaccination with the synthetic TCR Vβ 8.3 peptide could remarkably inhibit the development of EAU。

Abstract：Background Cockayne syndrome (CS) is a rare human genetic disorder characterized by increased UV sensitivity, developmental abnormalities and premature aging. Cells isolated from individuals with CS have a defect in transcription-coupled DNA repair. Despite the repair defect, there is no any increased risk of spontaneous or UV-induced cancer for CS individuals. The strategy of RNA interfering was used here to explore the potential radiosensitizing and anticancer activity of targeting CS group B (CSB) gene.Methods The vectors encoding CSB-specific siRNAs were constructed by inserting duplex siRNA encoding oligonucleotides into the plasmid psilencer TM 3.1. The cell lines expressing the CSB-siRNA were generated from HeLa cells transfected with the above vectors. Colony-forming ability was used to assay cell survival. Cell cycle was analyzed by FACScan flow cytometry. The apoptosis was measured by detecting the accumulation of sub-G1 population as well as by fluorescence staining assay. Reverse transcriptase polymerase chain reaction (RT-PCR)was used to semi-quantify mRNA expression. Protein level was detected by Western blotting analysis.Results Two constructs encoding CSB-specific siRNA were generated, both of them resulted in remarkable suppression on CSB expression in HeLa cells, and led to an increased sensitivity to γ-ray and UV light.siRNA-mediated silencing of CSB decreased cell proliferation rate, increased spontaneous apoptosis as well as the occurrence of UV- or cisplatin-induced apoptosis by 2 to 3.5 fold. A significant S phase blockage and a remarkable reduction of G1 population were induced in control HeLa cells at 18 hours after being exposed to 10J/m2 of UV light. The S phase blockage was also observed in UV-irradiated CSB-siRNA transfected HeLa cells,but the extent of increased S phase population was lower than that in the UV-irradiated control cells. No or a relative weak reduction on G1 phase population was observed in UV-irradiated CSB-siRNA transfected HeLa cells. In addition, siRNA-mediated silencing of CSB promoted the elimination of G2/M phase cells after UV light radiation.Conclusions siRNA-mediated silencing of CSB causes cells to proliferate more slowly, sensitize cells to genotoxicants, and modify UV radiation-induced cell cycle changes. siRNA-mediated inactivation of CSB could be an attractive strategy for ameliorating cancer therapy, which can be fulfilled via the combination of gene therapy and sensitization of radiotherapy or chemotherapy。

Abstract：Background Angiopoietin-1 (Ang-1) is an endothelial-specific growth factor that can promote angiogenesis.Studies demonstrated that Ang-1 can inhibit apoptosis of umbilical endothelial cells, but so far little is known about its effects on apoptosis of microvascular endothelial cells. With the apoptotic model of murinecerebral-derived microvascular endothelial cells (bEnd.3) induced by serum-free culture,we attempted to clarify the molecular mechanism of bEnd.3 apoptosis, particularly its relation to cytochrome C (Cyt C).Methods The cultured microvascular endothelial cell strain, bEnd.3 cell, was employed. An apoptotic model of bEnd.3 was established by serum-free culture. Flow cytometry after Annexin labeling and PI staining were used to assess the apoptotic effects of Ang-1 on bEnd.3, and the expression of Bax/Bcl-2, caspase 8, caspase 3, and Cyt C were detected with Western blotting and ELISA.Results The apoptotic rate of bEnd.3 cells after stimulation with Ang-1 (100 ng/L) in serum-free medium was significantly higher than that in control group. Ang-1 inhibited early-stage apoptosis more than late-stage apoptosis provided by propidium iodide (PI) and AnnexinV double staining. The inhibition of Ang-1 on bEnd.3cell apoptosis was strengthened with the increase in concentration (0-400 ng/ml). Ang-1 could decrease the expression of Bax, caspase3 and 8, and increase that of Bcl-2. The results of ELISA indicated that Ang-1significantly decreased CytC content in cytoplasm and increase that in mitochondria.Conclusions Ang-1 could inhibit bEnd.3 apoptosis induced by serum-free medium culture. The apoptosis was associated with decreased Bax expression, increased Bcl-2 expression, which result in Cyt C transferring from mitochondria to cytoplasm, and then caspases activation are reduced and cell apoptosis is suppressed。

Abstract：Background The mechanisms responsible for the pathogeneses of gestational hypertension and preeclampsia are unclear. Tumor necrosis factor-α (TNF-α) is a pro-inflammatory Th1-type cytokine. TNFA gene is located in the human leukocyte antigen (HLA) class III region of the major histocompatibility complex (MHC) on chromosome 6. The high TNF-α mRNA expression may be associated with the TNF2 (A) allele, which is the polymorphism of TNF-α at position -308 in promoter region. This study assessed whether the TNF2 (A) allele at position -308 plays a role in the alteration of blood pressure (BP) and urinary protein excretion during pregnancy.Methods The original prospective cohort study comprised 1623 pregnant women from January 2000 to October 2001. The G/A polymorphism was done by restriction fragment length polymorphism (RFLP) analysis with Nco I enzyme.Results The distributions of the G/A polymorphism of TNF-α in the promoter region at position -308 were wild-type 72.4% and variant 27.6%, respectively. The frequency of TNF2 (A) allele was approximately 0.15 for Caucasian pregnant women in the study. It was not significantly different in the distributions of genotypes and G/A allele frequencies among the three groups of pregnant women with gestational hypertension, preexisting hypertension and normal blood pressure (P＞0.05). The maternal blood pressure in the third trimester was significantly higher in the group of women possessing the TNF2 (A) allele compared to homozygous for the TNF1 (G) allele (systolic BP, P＜0.01 and diastolic BP, P＜0.05). The elevated blood pressure in the TNF2 (A)group was accompanied by higher urinary protein excretion in the third trimester (P＜0.05). The blood pressure and urinary protein excretion did not change apparently between the two groups in the first and second trimesters (P＞0.05).Conclusions Maternal TNF2 (A) allele of TNF-α promoter region at position -308 could play a role in the alteration of blood pressures and/or enhancement of urinary protein excretion during pregnancy, and might play an important role in the development of both gestational hypertension and preeclampsia。

Abstract：Background Hypoxia-inducible factor-1α (HIF-1α) is a transcriptional factor that could improve the stimulation of angiogenesis and the metabolic adaptation of tumor cells to hypoxia. A recent study showed that HIF-1α could induce colon cancer cells epithelial-mesenchymal transition (EMT). However, no evidence indicates a similar correlation in human prostate cancer cells. This study was designed to evaluate the effect of HIF-1 α over-expression on the EMT in human prostate cancer cells.Methods We selected the appropriate cell line for HIF-1α induction from those EMT negative prostate cell lines through vimentin gene detection by RT-PCR. As the result, LNCaP cell line is the best one for further experiment. LNCaP cells were transfected with recombinant plasmid pcDNA3.1 (-)/HIF- 1 α and pcDNA3.1 (-)control vector by Lipofectamine 2000 system. The positive cell colonies were confirmed by indirect immunofluorescence labeling. Then Transwell polycarbonate filter was used to analyze the invasive potency. The expression of EMT associated proteins, E-cadherin and vimentin, was detected by Western blotting.Results Among four of the EMT negative cell lines, LNCaP was the only one expressed the vimentin gene but not the associated protein. The expression level of HIF-1α in LNCaP/HIF-1α was distinctly higher than that in LNCaP/pcDNA3.1 and LNCaP. The cell numbers of LNCaP/HIF- 1 α that penetrated through the Transwell filter were higher than that of LNCaP/pcDNA3.1 and LNCaP. Compared with the LNCaP/pcDNA3.1 and LNCaP cells,the expression of vimentin was up-regulated in LNCaP/HIF-1α, whereas the expression of E-cadherin was down-regulated.Conclusions Over-expression of HIF-1α stimulates the invasion potency of human prostate carcinoma cells through EMT pathway. The expression of E-cadherin and vimentin, playing established roles in EMT, could be regulated by HIF-1α in human prostate cancer cell line。

Abstract：Background In recent years, transfer of the spinal accessory nerve to suprascapular nerve has become a routine procedure for restoration of shoulder abduction. However, the operation via the traditional supraclavicular anterior approach often leads to partial denervation of the trapezius muscle. The purpose of the study was to introduce transfer of the spinal accessory nerve through dorsal approach, using distal branch of the spinal accessory nerve, to repair the suprascapular nerve for restoration of shoulder abduction, and to observe its therapeutic effect.Methods From January to October 2003, a total of 11 patients with a brachial plexus injury and an intact or nearly intact spinal accessory nerve were treated by transferring the spinal accessory nerve to the suprascapular nerve through dorsal approach. The patients were followed up for 18 to 26 months [mean (23.5 ±5.2) months] to evaluate their shoulder abduction and function of the trapezius muscle. The outcomes were compared with those of 26 patients treated with traditional anterior approach. And the data were analyzed by Student's t test using SPSS 10.5.Results In the 11 patients, the spinal accessory nerves were transferred to the suprascapular nerve through the dorsal approach successfully. Intact function of the upper trapezius was achieved in all of them. In the patients,the location of the two nerves was relatively stable at the level of superior margin of the scapula, the mean distance between them was (4.2±1.4) cm, both the nerves could be easily dissected and end-to-end anastomosed without any tension. During the follow-up, the first electrophysiological sign of recovery of the infraspinatus appeared at (6.8±2.7) months and the first sign of restoration of the shoulder abduction at (7.6±2.9) months after the operation, which were earlier than that after the traditional operation [(8.7±2.4) months and (9.9±2.8)months, respectively; P＜0.05]. The postoperative shoulder abduction was 62.8°± 12.6° after transfer of the spinal accessory nerve, better than that after the traditional (51.6°±15.7°). All the 11 patients could extend and externally rotate the shoulder almost normally.Conclusions The accessory nerve transfer through dorsal approach is a safe and reliable procedure for the treatment of brachial plexus injury. Its postoperative effect is confirmed, which is better than that of the traditional operation。

The proliferation, differentiation and fibrosis of lens epithelia cells (LECs) is mainly responsible for posterior capsular opacification (PCO). From the primary culture of LECs to the culture of lens capsular bag, the models of posterior capsular opacification have been developed. At present, the most commonly used model is cell culture in medium with serum. But the culture in pure ocular tissue has not been reported. Therefore, we established a new model of posterior capsular opacification-culturing bovine lens capsular bag in pure ocular tissue to exclude the role of serum. Our study established a new culture method to investigate the proliferation,differentiation and apoptosis of lens epithelia cells in the aqueous humor with or without lens cortex and vitreous humor. The purpose of the study is to model posterior capsular opacification in vivo as closely as possible and to discuss the influence of ocular tissue on posterior capsular opacification。

Persistent hepatitis B virus (HBV) infection is the most important reason for chronic hepatitis B,hepatic cirrhosis, and hepatocellular carcinoma.1 T lymphocytes, including CD4+ and CD8+ T cells, are major composition of host cellular immunity.Furthermore, CD8+ cells play a primary role in host immune reaction of anti-tumor and anti-infection。

Renal artery stenosis (RAS) is a frequently overlooked clinical entity that can cause uncontrolled hypertension and lead to a progressive deterioration of renal function.1 We observed 20 patients with RAS complicated with cerebral artery stenosis (CAS), who underwent cerebral and renal artery angiography and combined intervening treatment. Clinical follow-up was performed for more than 3 years。

Medical insurance service, the important part of national healthcare supporting system with a history dating back more than 100 years ago,remains a global challenge because of its high rates of compensation and difficulty in risk control. When developing the diabetes related, hospitalization insurance, we found that the risk loss of the diabetic inpatients does not follow a symmetrical unimodal distribution: in fact, it is hard to describe its risk loses distribution with a single probability distribution model. Therefore, we put forward a risk measurement method based on a mixed normal distributions model for medical insurance of inpatients with diabetes。

In 1957, Brantigan1 applied lung volume reduction surgery (LVRS) to the treatment of emphysema. In 1995, Cooper2 reused this operation and obtained remarkable benefit in patients with chronic obstructive pulmonary disease (COPD). Although there are still some doubts about the effect of LVRS,in the past 10 years, a lot of clinical information approved that LVRS can give benefit to more than 70% of COPD patients. At present, a series of researches in long-term investigation and function evaluation such as NETT3 have got elementary affirmative conclusion about some disputative problems。

Cystic neoplasms of the pancreas account for 10% to 15% of all cystic pancreatic lesions.The majority (85% to 90%) of cystic lesions of the pancreas are pseudocysts. Although cystic neoplasms of the pancreas are rare, they range from benign to malignant neoplasms. The clinical challenge is the differential diagnosis and management of the cystic neoplasms, which represent 10% to 25% of primary pancreatic neoplasms. Pancreatic neoplasms and tumour like lesions with cystic features have been recently reviewed. The incidence of pancreatic cystic neoplasms reported is variable. Because there is no large, systematic study on tne cases from China comparing the incidence and biology of cystic neoplasms of pancreas to that of Western series, we reviewed all the cases of cystic neoplasms from Zhongshan Hospital over 6 years. Most of the neoplasms in our series were classified according to the recent World Health Organization (WHO)classification.1,2

Abdominal aortic aneurysm (AAA) has a high propensity to rupture. Repair of AAA by conventional surgical replacement with a graft has been the standard of treatment since the mid 1960s.Because of advances in surgical, anaesthetic and intensive care techniques, the outcome of elective open AAA repair has improved constantly. However,with the improvement of endovascular techniques,the role of open repair is being challenged.1 More and more stents are being deployed in patients because of their less invasive characteristics.2 We reviewed our ten years of experience with elective open AAA repair and recent papers about endoluminal and open repair for AAA to discuss status and future of open repair for AAA in China。

Dystonia refers to a clinical syndrome in which sustained involuntary muscle contractions result in twisting and repetitive movements, or abnormal postures. Secondary dystonia is associated with acquired or exogenous causes, hereditary neurologic syndromes or neurodegenerative disorders. Tardive dystonia is a special type of secondary dystonia due to exposure to certain medicines such as neuroleptics, with a chronic and persistent extrapyramidal symptoms。

Takayasu arteritis (TA) is a nonspecific granulomatous inflammatory arteriopathy of unknown cause that results in occlusive obliteration or less commonly aneurysm degeneration of large and medium-sized elastic arteries. Most descriptions of this disease have emphasized the "pulseless"syndrome, however, less attention has been paid to involvement of other segments of the aorta, renal arteries, and in particular the visceral arteries. We reported a case of type Ⅱ TA, in which both the visceral and the bilateral renal arteries were involved。

Primary pigmented nodular adrenocortical disease (PPNAD), found in the mid 1980s, is a rare type of ACTH-independent hypercortisolism,can be part of Carney complex (CNC), an autosomal dominant multiple neoplasia syndrome characterized by spotty skin pigmentation, cardiac myxomas and endocrine tumors. One of the putative CNC genes,located on 17q22-24, has been identified as the regulatory subunit R1A of protein kinase A (PRKAR1A). Patients with isolated PPNAD and no familial history of CNC can also present a germline de novo mutation of PRKAR1A. In China, five patients with PPNAD underwent surgical treatment in our department from 2001 to August 2004.1

Coronary artery fistula (CAF), an uncommon form of congenital heart disease, is characterized by abnormal communication between a coronary artery and a cardiac chamber or vessel.1 This disease is often found incidentally in asymptomatic individuals,and leads to cardiac failure, myocardial ischemia and angina, infective endocarditis, and heart rupture in later life. Both surgical repair and transcatheter closure were effective and safe in the treatment of CAF,2 but percutaneous management of CAF can obviate median sternotomy and cardiopulmonary bypass, and there may be less morbidity with transcatheter embolization techniques,3,4 including the use of occluders or microcoils. We report a successful percutaneous closure of a giant CAF from the left coronary artery to the right ventricle using patent duct occluder.…… Biography：Correspondence to: Dr. ZHOU Sheng-hua, Department of Cardiology, Second Xiangya Hospital, Central South University,Changsha 410011, China (Tel: 86-731-2233716. Email:zhougqin @ 21 cn.com)