SH3GL2在无病灶性难治性癫(癎)患者脑组织中的表达

目的:分析无病灶性难治性癫患者脑组织中SH3GL2 mRNA与蛋白的表达水平,探讨可能的发病机制。方法:运用逆转录-聚合酶链式反应(RT-PCR)和Westernblot分别检测10例无病灶性难治性癫患者及10例对照组脑组织SH3GL2 mRNA及蛋白的表达。结果:RT-PCR检测显示,无病灶性难治性癫患者脑组织SH3GL2 mRNA表达的相对水平为(102.23±54.68),对照组为(44.59±17.12),两者差异有显著统计学意义(P<0.01);Westernblot分析表明,无病灶性难治性癫患者脑组织的SH3GL2蛋白表达相对水平(229.54±89.82)高于对照组(143.88±59.69),两者差异也有显著统计学意义(P<0.05)。结论:SH3GL2 mRNA与蛋白表达水平在无病灶性难治性癫患者脑组织中明显增强,提示表达于中枢神经系统的SH3GL2可能在人类难治性癫的发病机制中起了一定作用。     

两株鼠抗人GL50单克隆抗体的制备及其生物学特性的初步研究

目的　研制鼠抗人GL5 0分子单克隆抗体并对其生物学特性进行初步研究。方法　以天然高表达人GL5 0分子的Daudi细胞为免疫原 ,人GL5 0 L92 9转基因细胞为目的单克隆抗体(McAb)的筛选细胞 ,采用B淋巴细胞杂交瘤技术 ,获得分泌特异性鼠抗人GL5 0分子McAb的杂交瘤细胞株 ;免疫荧光标记和流式细胞术分析McAb识别GL5 0的表达 ;Westernblot检测抗体对特异性细胞膜蛋白的识别 ;台盼蓝 (Trypanblue)染色细胞计数法和MTT法检测McAb对Daudi细胞体外生长的抑制作用 ;3 H TdR法检测抗体对GL5 0 L转基因细胞介导的活化T细胞体外增殖的效应。结果　成功制备 2株分泌特异性抗人GL5 0抗体的杂交瘤细胞株 12B11和 11C4 ;两者皆为IgG2a亚型 ;腹水效价皆在 1∶10 0 0以上 ;12B11、11C4特异性地识别GL5 0分子。 11C4McAb可以明显抑制Daudi细胞在体外的生长 ,并可抑制GL5 0 L转基因细胞介导的活化T淋巴细胞的体外增殖效应。结论　成功获得了 2株特异性鼠抗人GL5 0抗体 ,其中 11C4McAb具有诱导表达GL5 0分子的B淋巴瘤细胞Daudi的体外生长抑制作用 ;在T淋巴细胞体外增殖反应中发挥了重要的调节作用。     

Audit of minimally-invasive surgery for submandibular sialolithiasis

Sialolithiasis is one of most common diseases to affect major salivary glands, with a symptomatic incidence of 27 cases per million per annum. The majority form within the submandibular gland where minimally-invasive treatments have all but eliminated adenectomy. All records of patients presenting with submandibular stones between 1997 and 2015 were reviewed. Stones <5 mm were retrieved through endoscopic or radiographic techniques, 5-7 mm stones were initially considered for extra-corporeal shock wave lithotripsy, but after poor results were treated through intraoral surgical removal with those >7 mm. Follow up was performed at 1 week and 3 months with current status performed with postal and telephone questionnaires. 378 patients had 424 stones removed, successful retrieval in 94% (n = 356), with 50 having had previous failures. Median number of stones per patient was 1 (range 1-4), with a mean size of 8.6 mm (SD 4.5 mm) mainly located at the hilum (50.5%), anterior duct (30%) and Genu (17%). 256 patients (65%) treated through intraoral surgical extraction, 92 (24%) endoscopic alone. Inpatient stay was 1.4 days in first third and 0.5 days in final third. Adenectomy occurred in 14 patients, due to failure to retrieve the sialolith or unresolved symptoms. Complications involved 11 patients with permanent paraesthesia, 7 ranulas and 14 strictures. Patients with preoperative strictures were more likely to develop complications (p = 0.002) with paraesthesia being most common. Intraoral minimally-invasive surgery is aesthetic, curative and spares the risk to marginal mandibular nerve and submandibular gland. Length of inpatient stay improved and ranula risk reduced throughout the study.     

GL50分子在活化T细胞表面的表达及对细胞增殖的影响

目的探讨GL50分子在活化T细胞表面的表达及其生物学意义。方法分别取不同时相（24-144h）激发型CD3单抗联合CD28单抗活化的T细胞，采用流式细胞术分析T细胞表面GL50分子表达阳性率，RT-PCR法检测T细胞内GL50分子mRNA转录水平；^3H-TdR掺入法分析阻断GL50信号对T细胞增殖的影响。结果激发型CD3单抗联合CD28单抗活化T细胞后24h GL50分子阳性表达率为17．2％，其后逐渐升高至活化120h时达最大阳性率95．7％，并维持至活化144h时基本不变；T细胞活化24—144h均可检测到GL50分子mRNA表达；阻断性GL50分子单抗对活化T细胞增殖具有显著抑制作用。结论GL50分子在活化T细胞表面呈诱导性表达，且可与T细胞表面自身受体结合在免疫应答放大效应中发挥重要作用。     

Aberrant promoter methylation of SH3GL2 gene in vulvar squamous cell carcinoma correlates with clinicopathological characteristics and HPV infection status

Objective: This study attempted to examine the methylation status of SH3GL2 gene in different types of human vulvar lesions and its correlation with clinicopathological parameters. Methods: Immunohistochemical analysis was used to identify the expression status of SH3GL2 in vulvar squamous cell carcinoma (VSCC), vulvar intraepithelial neoplasia (VIN) and benign vulvar squamous epithelium tissues. Bisulfite genomic sequencing method was used to detect methylation status of the SH3GL2 gene. Clinicopathological correlation of the alterations was analysed by the chi-square tests. Results: Immunohistochemical analysis showed expression of SH3GL2 in VSCC was significantly downregulated than that in VIN and normal vulvar tissues. In accordance with higher frequency of methylation status in SH3GL2, statistical analysis showed methylation status of SH3GL2 was closely related to tumor TNM stage (P=0.003), but not related to age, tumor volume, tumor differentiation, lymph node metastasis and VIN grade. High-methylation status of SH3GL2 showed significant association with HPV infection status. Conclusions: Our results indicated that the methylation status of SH3GL2 gene was associated with the TNM staging and HPV infection status of VSCC, suggesting that it might play a synergistic role in the development of VSCC.     

Pre-existing CD19-independent GL7− Breg cells are expanded during inflammation and in mice with lupus-like disease

Interleukin 10 (IL-10)-producing regulatory B-cells (Bregs) suppress inflammatory responses that mediate autoimmune diseases. However, it is unknown whether Bregs derive from a pre-existing dedicated B-cell lineage or if any B-cell can differentiate into Bregs in response to BCR or TLR activation. GL7⁺ B-cells are antigen-experienced differentiated B-cells while GL7^−/lo are at an early stage of B-cell differentiation. While both GL7^−/lo and GL7⁺ B cells can produce IL-10, differentiation of GL7⁻ B-cells into Bregs does not require CD19- or Bcl6-induced signals, suggesting that BCR-induced proliferation or Ig class-switching is not necessary for generation of Breg cells. Of particular importance, we show that GL7⁻ Breg cells are dramatically expanded in lupus-like mice and GL7⁻ Bregs suppressed inflammatory responses in lupus-like mice by inducing expansion of Foxp3⁺Treg cells. Taken together, these results suggest that pre-existing GL7⁻IL-10⁺ cells are expanded during inflammation, differentiate into GL7⁺ Bregs and contribute to immune-regulation in lupus-like mice.     

Evidences of endocytosis via caveolae following blood–brain barrier breakdown by Phoneutria nigriventer spider venom

Spider venoms contain neurotoxic peptides aimed at paralyzing prey or for defense against predators; that is why they represent valuable tools for studies in neuroscience field. The present study aimed at identifying the process of internalization that occurs during the increased trafficking of vesicles caused by Phoneutria nigriventer spider venom (PNV)-induced blood–brain barrier (BBB) breakdown. Herein, we found that caveolin-1α is up-regulated in the cerebellar capillaries and Purkinje neurons of PNV-administered P14 (neonate) and 8- to 10-week-old (adult) rats. The white matter and granular layers were regions where caveolin-1α showed major upregulation. The variable age played a role in this effect. Caveolin-1 is the central protein that controls caveolae formation. Caveolar-specialized cholesterol- and sphingolipid-rich membrane sub-domains are involved in endocytosis, transcytosis, mechano-sensing, synapse formation and stabilization, signal transduction, intercellular communication, apoptosis, and various signaling events, including those related to calcium handling. PNV is extremely rich in neurotoxic peptides that affect glutamate handling and interferes with ion channels physiology. We suggest that the PNV-induced BBB opening is associated with a high expression of caveolae frame-forming caveolin-1α, and therefore in the process of internalization and enhanced transcytosis. Caveolin-1α up-regulation in Purkinje neurons could be related to a way of neurons to preserve, restore, and enhance function following PNV-induced excitotoxicity. The findings disclose interesting perspectives for further molecular studies of the interaction between PNV and caveolar specialized membrane domains. It proves PNV to be excellent tool for studies of transcytosis, the most common form of BBB-enhanced permeability.