Anti-VEGF treatment improves neurological function and augments radiation response in NF2 schwannoma model

Hearing loss is the main limitation of radiation therapy for vestibular schwannoma (VS), and identifying treatment options that minimize hearing loss are urgently needed. Treatment with bevacizumab is associated with tumor control and hearing improvement in neurofibromatosis type 2 (NF2) patients; however, its effect is not durable and its mechanism of action on nerve function is unknown. We modeled the effect anti-VEGF therapy on neurological function in the sciatic nerve model and found that it improves neurological function by alleviating tumor edema, which may further improve results by decreasing muscle atrophy and increasing nerve regeneration. Using a cranial window model, we showed that anti-VEGF treatment may achieve these effects via normalizing the tumor vasculature, improving vessel perfusion, and delivery of oxygenation. It is known that oxygen is a potent radiosensitizer; therefore, we further demonstrated that combining anti-VEGF with radiation therapy can achieve a better tumor control and help lower the radiation dose and, thus, minimize radiation-related neurological toxicity. Our results provide compelling rationale for testing combined therapy in human VS.Neurofibromatosis type 2 (NF2) is a dominantly inherited genetic condition with a birth prevalence of 1 in 25,000 (1). Bilateral vestibular schwannomas (VS), which are benign tumors composed of neoplastic Schwann cells that arise from the eighth cranial nerve, are the hallmark of NF2 (2). Standard approaches for treatment of growing VS include surgical removal and radiation therapy (RT). Hearing loss is the main limitation of radiation therapy for VS. For patients with sporadic VS who do not have NF2, RT is associated with long-term tumor control rates exceeding 95%. However, hearing preservation rates after radiation range from 50% to 80% (3, 4). Outcomes after radiation for patients with NF2 are inferior to those for sporadic patients, with short-term local tumor control rates approximately 80–85% and hearing preservation rates <50% (3). Thus, the identification of a novel adjunct therapy to enhance radiosensitivity while minimizing toxicity-related hearing loss in VS is urgently needed.Vascular endothelial growth factor (VEGF) and its receptors (VEGFRs) are expressed in VS, and its expression level positively correlates with schwannoma growth rate (5–7). In a retrospective review of 31 NF2 patients, treatment with bevacizumab, a humanized monoclonal antibody that specifically neutralizes VEGF-A, was associated with a reduction in the volume of most growing VS. More importantly, bevacizumab treatment improved hearing in 57% patients (7). Despite this progress, a number of challenges remain (8). First, not all NF2 patients respond to bevacizumab; second, the hearing response is not durable in all patients; and third, some patients are unable to tolerate long-term bevacizumab treatment. Studies to understand the mechanisms of anti-VEGF therapy-induced tumor growth inhibition and hearing improvement in schwannomas are urgently needed to optimize this therapy.In our study, first, we used the sciatic nerve model to characterize the effect and mechanisms of anti-VEGF treatment on neurological function. We revealed that anti-VEGF treatment alleviates tumor edema, which may further result in decreasing muscle atrophy and increasing nerve regeneration and, thus, improves neurological function. Second, we used the intracranial window model to monitor in real time the effect of anti-VEGF treatment on tumor vasculature. We showed that anti-VEGF treatment transiently normalizes the tumor vasculature, leading to improved perfusion and oxygen delivery. Using intravital microscopy imaging technique, we further defined the timing of this transient effect, termed the “normalization window,” in schwannoma models. Because oxygen is a potent radiosensitizer, finally, we showed that radiation therapy applied during the normalization window is most effective, and combined anti-VEGF and radiation therapy is superior to each monotherapy. Anti-VEGF and radiation combination therapy may thus help reduce the dose of each therapy and minimize treatment-associated adverse effect in NF2 patients.     

160例肺不张患者支气管镜检查分析

１６０例肺不张患者的支气管镜（下称纤支镜）检查结果表明：常见的主要病因中肿瘤（８２例，５１．３％），炎症（６１例，３８．１％），结核（８例，５．０％）。其诊断率依次为９２．０％，８７．０％，５０．０％；总诊断率９０．２％。对肺不张的病因与病变部位，支气管解剖特点，年龄关系以及ｘ线（包括ＣＴ）表现，纤支镜下特征等均作了分析讨论。     

双胃镜粘膜切除法切除早期胃癌及其癌前病变

自１９９１年３月至１９９２年１２月，我院应用双胃镜对６例粘膜层早期胃癌及其癌前病变进行了内镜下粘膜切除术，其中Ⅱｃ型早期胃癌１例，Ⅱａ型早期胃癌２例，扁平隆起型重度异型增生２例，山田１型腺瘤１例，所有病变直径均小于２ｃｍ。一次操作病变完全切除３例，经第二次操作又完全切除２例，残留切除１例，后又补充外科手术切除。经追踪３～２１个月，尚未见复发征象。文中对本法适应症、完全切除的标准等也进行了讨论     

COMPARISON OF ENDOSCOPIC DETECTION RATE OF EARLY GASTRIC CANCER AND GASTRIC ADENOMA USING TRANSNASAL EGD WITH THAT OF TRANSORAL EGD

Background: To investigate the influence of the reduced image quality of transnasal esophagogastroduodenoscopy (EGD) with the ultrathin endoscope (transnasal EGD) on endoscopic diagnoses, we compared the detection rate (DR) of early gastric cancer and gastric adenoma by transnasal EGD with that of transoral EGD using a standard endoscope. Methods: Transnasal EGD was carried out in 2791 examinations for the purposes of screening or other reasons. Controls were examined by transoral EGD and numbered 3591 examinations. The transnasal endoscope used was an EG530N. Lesions graded C‐3 or higher by Kimura‐Takemoto's classification were regarded as endoscopic atrophy. Results: (i) DR in all subjects and those with atrophy were not different between transnasal and transoral EGD. (ii) Multivariate analysis of DR in subjects with atrophy was carried out using five variables: gender, age, purposes of endoscopy, endoscopic insertion route and the four endoscopists. DR was significantly higher in males or subjects ≥60 years. No difference was noted between the endoscopic insertion routes (transnasal vs transoral). (iii) The subjects analyzed in (ii) were divided into the transnasal and transoral groups, and multivariate analysis of DR was carried out using four variables. DR was not different among the endoscopists in the transoral group. However, in the transnasal group, DR increased as the years of endoscopic experience was prolonged. Conclusions: Multivariate analysis detected no significant difference in DR between transnasal and transoral EGD. However, a significant difference in DR by transnasal EGD among the endoscopists is detected. Transnasal EGD should be carefully carried out by experienced endoscopists.     

A CASE OF DIFFUSE‐TYPE EARLY GASTRIC CANCER WITH NODULAR GASTRITIS

A 39‐year‐old woman was referred to Osaka Police Hospital and admitted for surgical treatment of gastric cancer. Barium upper gastrointestinal study and endoscopic examination showed a 3.0 × 3.0 cm depressed lesion in the greater curvature of the middle corpus. An unusual miliary pattern resembling ‘goose flesh’ was observed endoscopically in the antrum. Biopsy specimens from the tumor showed poorly differentiated adenocarcinoma, and specimens from the antrum showed many lymphoid follicles with a germinal center. Rapid urease test and histological tests (Giemsa stain) for Helicobacter pylori were both positive. Early gastric cancer with nodular gastritis (NG) was diagnosed and a partial gastrectomy was performed. Histological examination of the resected specimen showed a stage I tumor consisting mainly of signet‐ring cell carcinoma restricted to the mucosa. Postoperatively H. pylori eradication therapy was performed and proved to be successful. One year after eradication therapy, endoscopy with biopsy showed no recurrence of gastric cancer and the remarkable regression of antral NG.     

FEASIBILITY OF ENDOSCOPIC SUBMUCOSAL DISSECTION FOR ELDERLY PATIENTS WITH EARLY GASTRIC CANCERS AND ADENOMAS

Background: Endoscopic submucosal dissection (ESD) has the advantage of permitting en bloc and histologically complete resection for early gastric cancer. Elderly patients often have surgical operative risks due to disease, and the feasibility of this treatment for such patients will improve the quality of life. The aim of the present study is to evaluate the efficacy and safety of ESD in elderly patients. Methods: We reviewed patients who underwent ESD for gastric lesions at Maebashi Red Cross Hospital. Among 251 gastric lesions treated by ESD from 2002 to 2006, 110 lesions were discovered in 93 elderly patients who were 75 years of age or older. The one‐piece resection with tumor‐free margin rate and the complications were assessed in comparison with younger patients under 75 years old. Results: The average age of the elderly patients was 79.8 years (range 75–92 years). The one‐piece resection with tumor‐free margin rate was 96.4% (106/110). Immediate bleeding occurred in one lesion (0.9%) and delayed bleeding requiring emergency endoscopy occurred in five lesions (4.5%). Perforation during ESD occurred in two patients (1.8%), and was immediately closed with endoclips and managed by conservative medical treatment. One case (0.9%) complicated with delayed perforation was managed by conservative medical treatment. The one‐piece resection with tumor‐free margin rate and the complication rate in elderly patients were not significantly different from those of younger patients. Conclusion: The present study shows the technical feasibility of ESD for gastric neoplasms in elderly patients.     

白细胞介素-2基因转导人胃癌细胞的实验研究

为研究白细胞介素－２（ＩＬ－２）基因转导人胃癌细胞的可行性和有效性，用真核表达载体，将人ＩＬ－２ｃＤＮＡ转导入人胃癌细胞株ＳＧＣ－７９０１细胞表达，并观察了基因转导和表达对胃癌细胞体外生长特性和ＬＡＫ细胞的肿瘤杀伤作用的影响，以及基因转导细胞的体内致瘤性。结果表明，人ＩＬ－２ｃＤＮＡ可以转导入人胃癌细胞，并表达有活性的ＩＬ－２，基因转导不改变胃癌细胞的体外生长特性，并能增强ＬＡＫ细胞的肿瘤杀伤作用，降低或消除胃癌细胞的体内致瘤性。本研究结果提示：ＩＬ－２基因转导胃癌细胞可能是防治胃癌的有效方法。     

弓形虫表面抗原SAG2的DNA疫苗载体构建及在Vero细胞中的表达

目的　构建弓形虫表面抗原SAG2的DNA疫苗载体 ,并在Vero细胞中表达。　方法　设计 1对引物 ,从弓形虫RH株速殖子基因组DNA中扩增SAG2全长编码基因 ,构建 pVAX1 SAG2真核表达重组质粒。以限制性内切酶KpnⅠ和EcoRⅠ进行双酶切、PCR鉴定 ,纯化后进行测序鉴定。脂质体介导法瞬时转染Vero细胞 ,同时以 pVAX1为对照 ,48h后收集细胞 ,Western blot鉴定。　结果　从弓形虫RH株DNA中扩增出了 5 77bp的SAG2基因 ,构建了真核表达载体 pVAX1 SAG2 ,在质脂体介导下转染Vero细胞 ,质粒DNA成功的转染到细胞中。通过Westen blot分析 ,细胞裂解液样品有 1条可被弓形虫免疫血清所识别的约 17ku大小的条带 ,与预计大小一致。　结论　真核表达载体pVAX1 SAG2在Vero细胞中有一定表达 ,且有一定的活性。     

The BTNL2 A allele variant is frequent in Danish patients with sarcoidosis

Background: The butyrophilin‐like 2 (BTNL2) gene is located on chromosome 6p21.3 close to the HLA‐class II genes. An association has been reported between sarcoidosis and a single nucleotide polymorphism in BTNL2, rs2076530, also termed the A allele. Objectives: To evaluate whether patients with sarcoidosis carry the A allele more frequently than healthy subjects. Methods: The series comprised 87 ethnic Danes with sarcoidosis and 113 healthy control subjects. Analysis of rs2076530 was performed by Taqman assay, polymerase chain reaction and sequencing of genomic DNA. Results: Sarcoidosis patients had a higher frequency of the A allele than controls (73.9% vs 55.8%) (P < 0.025). The frequency of GG, GA and AA genotype was 5.7%, 40.2% and 54.0% in patients vs 16.0%, 56.6% and 27.4% in controls (P < 0.001). The AA genotype was associated with increased risk of sarcoidosis in both a dominant [odds ratio (OR) 3.1; 95% confidence interval (CI) 1.1–8.7; P < 0.03] and a recessive model (OR 3.1; 95% CI 1.72–5.61; P < 0.001). Population attributable fraction for disease was 50% in a dominant model and 25% in a recessive model. Conclusions: The BTNL2 A allele variant occurs with a high frequency in Danish patients with sarcoidosis and the AA genotype is associated with a ～threefold higher risk of sarcoidosis than the GG genotype. Our results should encourage future studies on the interrelationship between the BTNL2 protein and granuloma formation in sarcoidosis. Please cite this paper as: Milman N, Svendsen CB, Nielsen FC and Hansen TvO. The BTNL2 A allele variant is frequent in Danish patients with sarcoidosis. Clin Respir J 2011; 5: 105–111.     

胃癌活组织c—erb—B2过度表达与预后的关系

探讨胃癌活检组织ｃ－ｅｒｂ－Ｂ２过度表达与预后的关系。用免疫组化法检测１０３例胃癌胃镜活检组织及术后１５１个转移淋巴结ｃ－ｅｒｂ－Ｂ２表达。结果：１８．４％胃癌活检组织出现阳性表达，进展期胃癌、乳头状腺癌及伴淋巴与肝转移者阳性率增高（Ｐ均＜０．０５）；阳性表达更多见于高中分化胃癌（但Ｐ＞０．０５）；转移淋巴结表达阳性率高于胃癌原发病灶（Ｐ＜０．０５）。高中分化胃癌ｃ－ｅｒｂ－Ｂ２过度表达者５年生存率低于阴性者（Ｐ＜０．０１）。以上结果提示ｃ－ｅｒｂ－Ｂ２过度表达与否有助于胃癌预后的估计。     

重组钩体基因疫苗免疫豚鼠脾细胞 LTT，IL-2，IL-6的活性测定

目的为从多方面证实赖型钩体重组基因疫苗的免疫活性。方法将赖型钩体重组基因多肽疫苗(分子量68kDa)多点皮下注射免疫豚鼠。(以质粒载体pT7-7为阴性对照,全钩死疫苗WCV为阳性对照)取其脾细胞,用3H-TdR法和MTT比色法分别测定特异性淋巴细胞转化试验(LTT)的相对转化指数(RPI)及IL-2,IL-6活性。结果1)重组基因疫苗免疫组特异性LTT的RPI为2.19±0.18明显高于pT7-7阴性对照1.42±0.27(P＜0.005),与阳性对照WCV无统计学差异(P＞0.05)。2)基因疫苗免疫组IL-2,IL-6活性分别为34.8±3.11,94.6±6.02测定值明显高于pT7-7阴性对照20.4±3.05,61±6.28(P＜0.005),与WCV阳性对照无统计学差异(P＞0.05)。结论1)钩体基因重组疫苗能使免疫豚鼠体内Th1、Th2活化增强,有胸腺依赖性抗原性质(TDAg),可激起体内强有力T-B细胞协同效应的特异性体液免疫反应,确有增强免疫活性作用,是良好的免疫原。2)重组钩体基因疫苗与阳性对照WCV各实验值无统计学差异,提示二者免疫活动可能相当,但重组基因疫苗有毒副作用小的优势,应用前景良好。     

Rapid gastric emptying of a liquid meal in long-term Type 2 diabetes mellitus

Both delayed and accelerated gastric emptying rate (GER) have been reported in patients with diabetes mellitus. Delayed GER has been attributed to autonomic neuropathy in established diabetes but rapid GER was demonstrated in early Type 2 diabetes. The aim of the study was to investigate rapid gastric emptying in a group of people with long-duration Type 2 diabetes. GER of a radiolabelled liquid meal was studied scintigraphically in 20 Type 2 patients with a mean (± SEM) duration of diabetes 13 (±1) years. The 50 % emptying time (t₅₀) for the liquid meal was shorter in diabetic patients (29.6 ± 2.1 min) than in controls (39.2 ± 1.9 min; p<0.0005). Accelerated emptying (t₅₀ value below the shortest t₅₀ of controls) was evidenced in 14/ 20 patients and delayed emptying (t₅₀ value exceeding the upper t₅₀ of controls) in none. Patients with accelerated GER were comparable for BMI, diabetes duration, HbA_1c and fasting glycaemia to those with normal GER. Rapid GER for liquids was found in the presence or absence of autonomic neuropathy. Seven of the patients with rapid emptying of the liquid meal were reassessed using a solid meal. Only one patient demonstrated rapid emptying of the solid meal, which was normal in 3 and delayed in 3 patients. In conclusion, accelerated GER can be found in long-term Type 2 diabetes but there is no concordance between GER of a liquid and a solid meal. Copyright © 1998 John Wiley & Sons, Ltd.     

以pcD-IL-2为基因佐剂的含日本血吸虫SjFABPc基因真核表达重组质粒裸DNA免疫小鼠的研究

目的　在小鼠模型中评价含日本血吸虫脂肪酸结合蛋白分子基因的真核表达质粒pcD -SjFABPc与含IL - 2基因佐剂的质粒构建体 pCD -IL - 2经肌注、皮内途径导入机体后所诱生的免疫反应。 方法　碱裂解法大量制备重组质粒 pcD-SjFABPc、pCD -IL - 2重组质粒及空质粒 pcDNA3,经肌肉及皮内注射免疫BALB/c小鼠 ,每只鼠注射 10 0 μg ,两周后同量加强免疫一次。分别于末次免疫后的第 2 0d、5 0d及 65d用MTT法检测脾脏T淋巴细胞增殖与NK细胞活性 ,并用双夹心ELISA测定血清细胞因子IL - 2、IFN -γ及IL - 10含量 ;ELISA法测定免疫鼠血清IgG抗体。 结果　NK细胞杀伤活性及脾淋巴细胞增殖测定 ,加佐剂组 ( pcD -SjFABPc联合 pCD -IL - 2组 )较不加佐剂组 ( pcD -SjFABPc组 )NK细胞杀伤及脾淋巴细胞增殖活性皆增加 (P <0 .0 5 )。两途径三次检测IL - 2及IFN -γ值均明显高于NS对照组和空质粒组 ,且加佐剂组的明显高于不加佐剂组的 (P <0 .0 5 ) ;不同途径各组IL - 10值与NS及空质粒对照组的比较则无显著性差别 (P >0 .0 5 )。注射质粒后 2 0d和 5 0d ,未测出抗体 ;免疫后 65d检测 ,pcD -SjFABPc和pcD -SjFABPc联合 pcD -IL - 2免疫组的OD值明显高于对照组 (P <0 .0 1) ,而该两组OD490 值之间无显著性差异 (P >0 .0 5     

日本血吸虫中国大陆株基因重组抗原Sj22.6kDa基因克隆与表达的研究

目的 :分离 Sj2 2 .6抗原基因 ,构建 Sj2 2 .6表达克隆。方法 :抗体筛选 Sj成虫 c DNA库 ,PCR扩增目的基因片段 ,亚克隆入表达载体 p GEX- 1λt。对重组体进行诱导表达并对表达产物进行 SDS- PAGE和 Western blot分析。结果 :PCR扩增产物为约 930 bp的 DNA条带。亚克隆入p GEX- 1λt,获得两个表达克隆 p GSj6和 p GSj2 4 ,特异性表达产物为 2 2 .6k Da抗原。重组体的表达方式为分离表达。结论 :从日本血吸虫成虫 c DNA库筛选到 Sj2 2 .6k Da克隆化基因 ,并构建了表达克隆 p GSj2 4和 p GSj6。     

Acid Instillation Increases Gastric Luminal Prostaglandin E2 Output in Man

ABSTRACT. The capacity of the gastroduodenal mucosa to maintain integrity when exposed to acid and pepsin may require formation of endogenous prostaglandins (PG). The gastric mucosa is capable of PG biosynthesis, and PGE₂ is present in the gastric contents of man. The purpose of this study was to examine if acidification of the human stomach affects the output of PGE₂. Gastric perfusion was made with 150 mM HC1 in seven healthy subjects pretreated with a histamine-2-receptor blocker (ranitidine). Gastric luminal PGE₂ was measured by gas chromatography-mass spectrometry. Basal output of PGE₂ was 1.42 ± 0.24 pmol/min (mean+SEM), which increased to 5.37 ± 0.91 pmol/min (p < 0.02) during acid perfusion. Gastric acidification did not cause mucosal damage as judged by luminal DNA. We conclude that PGE₂ is synthesized in the gastric mucosa even during nearly complete inhibition of parietal cell secretion. Luminal acid, a likely physiological stimulator of mucosal defense, induces a fivefold increase in PGE₂ output from the intact mucosa.     

锁骨颅骨发育不全1个家系2例患者RUNX2基因突变检测报告及文献复习

目的 探讨RUNX2基因突变在锁骨颅骨发育不全(CCD)发病中的意义及中国家族性CCD的发病机制.方法 一个中国家系2例CCD患者均表现为锁骨发育不良、头颅矢状缝明显增宽、巨大骨质缺损和坐骨支缺失.本研究提取该家系中2例患者、6位健康成员的外周血及100名健康志愿者进行RUNX2基因测序.结果 发现2例患者均携带RUN...     

T载体的构建及在恶性疟原虫RAP2基因克隆中的应用研究

目的　构建PCR产物高效克隆载体T载体 ,并应用于克隆恶性疟原虫RAP2基因。方法　PUC18用SmaI酶切纯化后 ,与dTTP在 70℃孵育 3h ,构建T载体。另设计一对特异引物 ,采用PCR方法从恶性疟原虫FCC1/HN株基因组DNA中特异扩增RAP2基因 ,并将其克隆入T载体 ,重组克隆经蓝白斑初筛后 ,再用双酶切及PCR法进行鉴定。结果　从恶性疟原虫基因组DNA中特异扩增出大小为 12 15bp基因片段 ,与预期长度相符。克隆入T载体后的重组克隆经双酶切及PCR鉴定均正确无误。结论　成功构建T载体 ,并获得阳性重组克隆T -RAP2 ,为进行该RAP2基因的序列测定及研究该基因的结构与功能奠定基础