Objective To establish a adenovirus vector system to co-encode two shRNAs targeting miR-221 and miR-222,respectively,which has multilevel,semiquantitative and co-modulated abilities.Methods The co-modulated shRNA-encoding adenoviruses were constructed by restriction enzyme digestion method (mutant-shmiR-221/222 and class-shmiR-221/222).The two adenovirus vectors were transduced into LN229 and U251 cells (class-shmiR-221/222 transduction group and mut-shmiR-221/222 ransduction group);and blank control group was established.The expression of miR-221/222 in the three groups was examined by quantitative real time-PCR,increase of targeted protein expression was determined by Western blotting,and flow cytometry was used to detect the cell cycle kinetecs.The cellular damage was observed under a phase contrast microscope,whereas the cell apoptosis was measured by caspase-3/7 activating assay.Furthermore,the free energy states were analyzed after applying a NCM-based algorithm for predicting the accurate binding structures.Results The co-modulated shRNA-encoding adenoviruses were successfully constructed and transduced into glioblastoma cell lines.In the blank control group,mut-shmiR-221/222 ransduction group and class-shmiR-221/222 transduction group,the miR-221/222 expression level and rate of cells at stage S decreased accordingly;p27kip1 expression level,rate of cells at G0/G1 stage and rate of cells arrested at G 1 stage increased accordingly;degrees of cells being destroyed and loss their own morphology and cell apoptosis rate increased accordingly;number of cells decreased accordingly;the differences among them were significant (P＜0.05).The free energy states revealed that the mutated type was less stable in binding with miRNA than the classic type.Conclusions ShRNA encoding adenovirus to co-modulate the expressions of miR-221 and miR-222 in glioblastoma cells is successfully established.Together with the strategy of mutation to influence the stability of binding,the modulation could yield distinct reduction results in a semi-quantitative manner.

Neural tube defects (NTDs) are common, severe congenital malformations. The association between single nucleotide polymorphisms of the VANGL1 gene and NTDs in a Han population of Northern China was principally studied. Missense single nucleotide polymorphisms (rs4839469 c.346G > A p.Ala116Thr and rs34059106 c.1040A > C p.Glu347Ala) of the VANGL1 gene were analyzed by polymerase chain reaction (PCR) and sequencing methods in 135 NTD cases and 135 normal controls. Genotype and allele frequency distribution was calculated, and the spatial structure of the protein was predicted. The results showed that the VANGL1 gene sequence at the rs4839469 locus exhibited Ala116Thr and Ala116Pro polymorphisms, and allele and genotype distributions were significantly different (p = 0.036 and 0.010) between the case and control group. Genotype GC was newly discovered, and its odds ratio value versus GG genotype was 10.241; the a helix fragment of the Ala116Pro mutant was significantly shortened compared with wild type. The rs34059106 site showed alleles of A and did not display C alleles in the two groups. Therefore, the rs4839469 allele of VANGL1 was obviously associated with NTDs. And genotype GC increased the risk of NTDs, changes in the three-dimensional protein structure may have impacted its biological functions, and the rs34059106 polymorphism had no significant correlation with NTDs.

The transcriptional regulator PRDM1 controls cell-fate decisions and has been implicated in human tumorigenesis as a tumor suppressor. However, its pathological role in glioma remains elusive. In this study, we showed that PRDM1 protein levels were inversely correlated with the pathological grade of gliomas and were predictive of patient survival in a retrospective analysis. Restored expression of PRDM1 inhibited proliferation and suppressed invasion by glioma cells. Mechanistic investigation revealed that PRDM1 attenuated glioma malignancy by negatively modulating Wnt/β-catenin signaling and this modulation was dependent on the Wnt inhibitor Dkk1. Using bioinformatics and biological approaches, we found that PRDM1 was a direct target of miR-30a-5p, and PRDM1 dysfunction was attributable to miR-30a-5p-mediated repression. Our results provide evidence that PRDM1 deficiency contributes to the phenotype maintenance and pathogenesis of gliomas.

播散性黄色瘤(xanthoma disseminatum,XD)是一种罕见的良性非朗格汉斯细胞起源的组织细胞增生性疾病,以全身皮肤播散性橘黄色丘疹为特征[1].累及颅内的XD患者,国外仅19例[2],且多合并于全身系统病变,国内目前尚无报告.本文报道1例单纯发生于鞍区及松果体区而酷似生殖细胞瘤的XD,并结合文献进行讨论.

Brainstem glioma (BSG) is an entity which commonly occurs in pediatric patients and carries a dismal prognosis. However, the category of adult BSG has displayed considerably benign biological behavior, thus providing a unique perspective to comparatively understand the malignant features of pediatric BSG. microRNAs (miRNAs) have been associated with cancer development and progression. To identify miRNAs specifically involved in the acquisition of malignant progression of pediatric BSG, we analyzed the miRNA expression profiles in orthotopic models which could simulate the BSG heterogeneity by microarrays. Our research revealed that miR-20a and miR-106b (known to be closely related) were the two most robust upregulated miRNAs in pediatric BSG compared to adult subtype. Furthermore, the two types of human BSG tissue were utilized to verify the microarray data by qRT-PCR and in situ hybridization. The results indicated good consistency with that of the microarray method. In conclusion, our studies provide evidence that miR-20a and miR-106b may serve as putative causative involvement of malignant progression of pediatric BSG, thereby serving as likely novel targets for constraining the rapid fatal course of pediatric BSG.

OBJECTIVES To study the different expression of miRNA between pediatric and adult types of brainstem gliomas, and to provide the target miRNAs for explore the mechanism and miRNA interference of the malignant progression of pediatric BSG. METHODS miRNA expression profiles in orthotopic models which could simulate the BSG heterogeneity were examined by microarray and analyzed to obtain the aberrantly expressed miRNAs. The two types of human BSG tissue were utilized to verify the microarray data by qRT-PCR and in situ hybridization for the putative causative miRNAs. RESULTS There were 216 miRNAs detected in both the pediatric BSG group and the adult BSG group, 39 miRNAs to be differential expressed in the pediatric BSG group versus adult group, including 10 up-regulated and 29 down-regulated. qRT-PCR and in situ hybridization indicated good consistency with that of the microarray method. CONCLUSIONS Aberrantly expressed miRNA may serve as putative causative involvement of malignant progression of pediatric BSG, thereby might be potentially novel targets for therapy.

临床颅脑转移癌发生率约占15％～39％,主要以幕上占绝大多数,尤其是额、颞、顶叶皮质及皮质下,少见于颅骨和脑膜[1-2],而同时累及帽状腱膜下、颅骨、硬脑膜、海绵窦更为罕见,我科发现1例.现报告如下.病例报告患者 女,54岁.2010年11月主因左顶部头疼进行性加重1个月余.入院查体:昏睡,双眼睑水肿;左额顶较右侧隆起且皮肤呈紫色,约5 cm×7 cm,压痛;左侧额部痛觉减退,结膜水肿,眼球固定;右Babinski征阳性.腰穿初压为250mmH2O,血象、脑脊液化验无异常.头颅薄扫CT检查:左侧筛窦、上颌窦软组织影.头颅MRV检查:左侧横窦、乙状窦不显影.头颅MRI检查:左额顶帽状腱膜下软组织肿胀,左半球硬脑膜弥漫强化,右侧硬脑膜部分强化,左海绵窦明显受侵蚀.综上检查,高度怀疑感染,颅内压增高,给予抗感染,脱水治疗,2 d后体温下降(37℃).为明确病因请全院会诊,一致认为感染可能性大,确诊需活检。

The objective of this study was to develop a combination chemotherapy of implantation of a 3-bis(2-chloroethyl)-1-nitrosourea (BCNU)-loaded wafer and intracarotid perfusion of BCNU-loaded nanoparticles for glioma treatment in vivo. BCNU-loaded poly(D,L-lactic acid) (PLA) nanoparticles coated with transferrin (Tf) were prepared by a solvent evaporation/diffusion method using Tf as the emulsifier. X-ray photoelectron spectroscopy, Bratton-Marshall colorimetric assay and zeta-potential analysis confirmed the existence of Tf on the nanoparticles and their functional activities. BCNU-loaded PLA wafers were made of BCNU-loaded PLA microspheres. In vitro drug release behavior demonstrated that BCNU was released from the Tf-PLA nanoparticles and wafers in two distinct phases. The biodistribution of Tf-coated nanoparticles investigated by 99mTc-labeled single-photon emission computed tomography (SPECT) showed that the surface-containing Tf-PLA nanoparticles were concentrated in the brain. Inhibition of tumor growth in the C6 glioma-bearing animal model showed that combinational chemotherapy of BCNU-loaded wafer and BCNU-loaded PLA nanoparticles had a stronger inhibitory effect and prolonged the average survival time of rats (164%) compared with that of the control group. Furthermore, the tumors of this treatment group were not visible by examination at 4 weeks. The results of this study demonstrate for the first time that combination therapy of implantation of a BCNU-loaded wafer and intracarotid perfusion of BCNU-loaded nanoparticles may be a new strategy for glioma gene therapy.

颅脑创伤合并肢体骨折是颅脑创伤中常见的多发伤类型,具有易漏诊、伤情复杂、病情危重、并发症多等特点.治疗中易因关注颅脑损伤而忽视对骨折的诊疗,严重骨折如果漏诊,可导致患者骨折愈合延期、骨不连、重度残疾甚至死亡.本研究对我院2005年1月-2008年12月收治的颅脑创伤合并四肢骨折患者的临床情况进行分析,报告如下.

Aberrantly expressed miRNAs are linked to the regulation of oncogenes and/or tumor suppression genes within the cell signal transduction pathway network, thereby contributing to carcinogenesis. miRNA function can be antagonized, thus representing a novel anti-tumor approach for integrated cancer therapy. In this study, we designed adenovirally-expressed shRNAs that functionally co-repressed the expression of miR-221 and miR-222, which are related to glioblastoma, to overcome the low efficiency of gene therapy. In addition, we generated novel shRNAs whose 3' ends were mutated in the region complementary to the target miRNA's 5' seed region to reduce the stability of binding with the miRNA. Various inhibition levels of miRNA were achieved: classic shRNAs yielded the greatest reduction in miRNA levels, followed by mutated shRNAs and the blank control, as determined by qRT-PCR. These results were confirmed by the protein expression of p27kip1, the validated target of miR-221/222, the effect on cell cycle arrest in G1 phase, and the impact on cell apoptosis. These results suggested that we could produce shRNAs encoded by adenovirus that co-repressed multiple tumor-related miRNAs simultaneously, and that the level of repression and the effect on the function of a specific miRNA could be achieved in a semi-quantitative manner.

Objective: To obtain more information about frizzled6 gene and Frizzled6 protein,for further study on Wnt-Fz signaling pathway and the correlation between FZ gene and neural tube defects.Methods: Bioinformatics methods were used to analyze FZ6 gene structure,single nucleotide polymorphisms,protein secondary structure,protein interaction network,and multiple sequence alignment of Fz protein family.Results: FZ6 gene locates on the 8 chromosome of human (8q22.3-q23.1) and has full length of 33995 base pairs,encoding 706 amino acids;there are 8 SNPs in coding region,including 4 missense;the 10 members of Fz protein family are divided into four clusters,Fz3 and Fz6 in the same cluster;Fz6 protein has two conserved domains,but unlike other family members,it does not contain a C-terminal S/T-X-V motif;its secondary structure mainly consists of α-helix and coil.Conclusion we get more biological information about FZ6 gene and Fz6 protein by bioinformatic analysis,which is very useful for further research.

Objective To investigate the inhibition effects on tumor invasion of malignant glioma U251 cells by small hairpin RNA targeting AKT1 and COX-2 in vitro.Methods The recombinant adenovirus vector plasmid expression vector which contained short hairpin RNA (shRNA) expression construct of AKT1 and COX-2 was transfected into human malignant glioma U251 cells.AKT1 and COX-2 expressions were assessed using Real-time PCR and western blot analysis and the expressions of MMP-2,MMP-9 and TIMP-2 were detected by Western blot.The ectocytic density changes of MMP-2 and MMP-9 were detected using ELESA.The invasion ability of the tumor cells were measured by using Scarification and Transwell.Results Recombinant adenovirus vector rAd5-A-C mediated shRNA targeting AKT1 and COX-2 dramatically down-regulated the expressions of AKT1,COX-2,MMP-2 and MMP-9 in U251 cells and up-regulated the expression of,TIMP-2.The extracellular concentrations of MMP-2 and MMP-9 decreased in cell cultures transfected with shRNA targeting AKT1 and COX-2.Scarification indicated that the ability of invasion were decreased in cells transfected with shRNA targeting AKT1 and COX-2 compared with control group and nonsense sequence group.Transwell showed that the number of cells invading through the matrigel was significantly lower in rAd5-A-C transfection group (35.2±7.3) compared with control group (87±4.7)and nonsense sequence group (82±3.4).Conclusions Knockdown of AKT1 and COX-2 by small interference RNA can significantly downregulate AKT1 and COX-2r expressions and inhibits the invasion ability of U251 cells in vitro.

The dorsal cutaneous appendage, or so-called human tail, is often considered to be a cutaneous marker of underlying occult dysraphism. The authors present a case of human tail occurring in a 9-month-old infant with multiple abnormalities of the spinal cord and spine. Examination revealed unremarkable except for a caudal appendage and a dark pigmentation area in the low back. Neuroradiological scans revealed cleft vertebrae and bifid ribbon, split cord malformations, block vertebrae, and hemivertebra. Surgical excision of the tail and untethering the spinal cord by removal of the septum were performed. The infant had an uneventful postoperative period and was unchanged neurologically for 18 months of followup. To our knowledge, no similar case reports exist in the literature. The specific features in a rare case with a human tail treated surgically are discussed in light of the available literature.

To gain a broader appreciation of the clinical presentation, operative treatment, and outcome of fibrous dysplasia involving the calvarium in children, we retrospectively reviewed a series of cases of fibrous dysplasia involving the calvarium (4 males and 2 females) with patients’ age ranging from 5 to 12 years old. The clinical manifestation, radiographic findings, surgical treatment, outcome and follow-up were evaluated on the basis of medical records. Fibrous dysplasia in the series was monostotic, involving frontal bone (2 cases), temporal bone (1 case), parietal bone (2 cases) and occipital bone (1 case). The patients most commonly presented with enlarging mass and cosmetic complaints. The treatment given, depending on clinical presentation, was simple biopsy with conservative follow-up (2 cases) to cranial resection (4 cases). All the cases were histopathologically confirmed as fibrous dysplasia. It was demonstrated that fibrous dysplasia involving the calvarium is a typically benign but slowly progressive disorder of bone. Modern imaging modalities and histopathologic analysis have made diagnosis relatively straightforward. Surgery should be reserved for patients with functional impairment or cosmetic deformity. Because of the benign nature of the condition, the surgery itself should be contemplated with great caution in children.

目前,许多医院相继开展癫痫病的外科手术治疗,为难治性癫痫病患者的治疗带来了新的希望,但癫痫外科手术治疗应具备相应的条件.首先开展癫痫外科治疗工作的医院应拥有受过专业训练的癫痫内、外科医生,并有临床神经电生理、神经心理学、神经影像学等多学科人员参与的团队.

The spinal epidural space is an uncommon presenting site in primary non-Hodgkin’s lymphomas, especially for children. A boy suffered spinal cord compression syndrome caused by primary spinal epidural non-Hodgkin’s lymphoma. Thoracolumbar magnetic resonance imaging (MRI) demonstrated an intraspinal mass. An operation was performed with gross total tumor removal. Histological examination revealed a non-Hodgkin’s B-cell lymphoma. Bone marrow aspiration was negative for lymphoma involvement. No other therapies (chemotherapy and/or radiotherapy) were performed according to the parents’ opinion. The patient died approximately one year after the operation due to brain metastases. The clinical course and imaging features were discussed with a review of literatures.

松果体区肿瘤是少见的中枢神经系统肿瘤,在颅内肿瘤中不到1%.肿瘤类型以生殖细胞肿瘤、松果体实质肿瘤、其他神经上皮肿瘤、脑膜瘤等为主.