L-精氨酸、L-硝基精氨酸对红藻氨酸诱导大鼠癫痫及其海马结构中iNOS mRNA表达的影响

目的　观察诱导型一氧化氮合酶 (iNOS)在红藻氨酸(KA)癫痫大鼠海马内的表达及L 精氨酸 (L Arg)和L 硝基精氨酸 (L NNA)慢性干预的影响。方法　采用惊厥剂量的KA(1 0mg·kg- 1 )诱导大鼠癫痫发作 ,以NOS抑制剂L NNA(50mg·kg- 1 )和NO前体L Arg(40mg·kg- 1 )进行干预 ,对大鼠的癫痫发作行为及KA后不同时间点的海马内i NOSmRNA ,通过RT PCR观察其表达。结果　KA可使动物发生时间相关性癫痫发作 ,L NNA预处理后使KA诱导的癫痫发作明显加重 ,而L Arg预处理后使KA诱导的癫痫发作减弱。iNOSmRNA在KA处理后 3h开始有微弱的表达 ,且随着时间的延长逐渐增加 ,2 4h达到最高水平 ,2d及3d时未见表达 ,但 7d时又出现高表达 ;经L NNA预处理的动物 ,KA后 1h其海马结构中未出现iNOSmRNA ,但L Arg预处理后再给予KA后 1h ,可见微弱的iNOSmRNA表达。结论　红藻氨酸给药后一定时间 ,癫痫大鼠海马结构中可出现iNOSmRNA表达 ,L Arg慢性干预也有一定影响     

Interleukin-12 enhances the sensitivity of human osteosarcoma cells to 4-hydroperoxycyclophosphamide by a mechanism involving the Fas/Fas-ligand pathway.

Cyclophosphamide (CY) and its derivative ifosfamide are alkylating agents used to treat osteosarcoma (OS). The purpose of these studies was to determine whether alkylating agents affect the expression of Fas ligand (FasL) and whether interleukin 12 enhances the sensitivity of human OS cells to alkylating agents. 4-Hydroperoxycyclophosphamide (4-HC), the preactivated CY compound, and 4-hydroperoxydidechlorocloclophosphamide (4-HDC), its nonalkylating analogue, human OS LM6 cells, and a clone of cells derived by transfection with the interleukin 12 gene (LM6-#6) were used for these studies. Incubation of LM6 and LM6-#6 with 10 micro M 4-HC increased the expression of FasL mRNA (2.5- and 3.0-fold, respectively). By contrast, 4-HDC, Adriamycin (ADR), cisplatin (CDP), and methotrexate (MTX) had no effect on FasL mRNA expression. Increased FasL expression after treatment with 4-HC was also demonstrated by immunohistochemistry and flow cytometry. Drug-induced FasL was functional and mediated cell death. We examined the effect of FasL up-regulation by 4-HC on LM6 and LM6-#6 cells. Flow cytometry showed that LM6-#6 cells expressed 2.2-fold more Fas than LM6 cells. Cytotoxicity of 4-HC, 4-HDC, ADR, CDP, and MTX on LM6, LM6-neo, and LM6-#6 were quantified. Colony-forming assay revealed an IC(50) of 2.10 micro M for 4-HC in LM6-neo cells compared with 0.41 micro M in LM6-#6 cells. The IC(50) for 4-HDC, ADR, CDP, and MTX were not significantly different between the two cell lines. We concluded that the increased expression of Fas enhanced LM6-#6 sensitivity to 4-HC. These data indicate that Fas/FasL may be involved in the cytotoxic pathway of CY. Combining biological agents with chemotherapeutic agents that have complementary Fas/FasL pathway actions may offer new therapeutic alternatives.     

Increased Fas expression reduces the metastatic potential of human osteosarcoma cells.

PURPOSE: The process of metastasis requires the single tumor cell that seeds the metastatic clone to complete a complex series of steps. Identifying factors responsible for these steps is essential in developing and improving targeted therapy for metastasis. Resistance to receptor-mediated cell death, such as the Fas/Fas ligand pathway, is one mechanism commonly exploited by metastatic cell populations. EXPERIMENTAL DESIGN AND RESULTS: LM7, a subline of the SAOS human osteosarcoma cell line with low Fas expression, was selected for its high metastatic potential in an experimental nude mouse model. When transfected with the full-length Fas gene (LM7-Fas), these cells expressed higher levels of Fas than the parental LM7 cells or LM7-neo control-transfected cells. These cells were also more sensitive to Fas-induced cell death than controls. When injected intravenously into nude mice, the LM7-Fas cell line produced a significantly lower incidence of tumor nodules than control cell lines. Lung weight and tumor nodule size were also decreased in those mice injected with LM7-Fas. Levels of Fas were quantified in osteosarcoma lung nodules from 17 patients. Eight samples were Fas negative, whereas the remaining 9 were only weakly positive compared with normal human liver (positive control). CONCLUSIONS: Our results demonstrate that altering Fas expression can impact the metastatic potential of osteosarcoma cells. We conclude that the increase of Fas on the surface of the LM7 osteosarcoma cells increased their sensitivity to Fas-induced cell death in the microenvironment of the lung, where Fas ligand is constitutively expressed. Thus, loss of Fas expression is one mechanism by which osteosarcoma cells may evade host resistance mechanisms in the lung, increasing metastatic potential. Fas may therefore be a new therapeutic target for osteosarcoma.     

人子宫内膜异位症在位和异位内膜细胞原代培养及形态学观察

目的:探讨子宫内膜异位症(endometriosis,EM)异位子宫内膜细胞原代培养方法,并比较EM在位及异位子宫内膜细胞与正常对照在位子宫内膜细胞形态的差异。方法:采用改良EM细胞原代培养方法,免疫细胞化学法鉴定细胞类型,在光学显微镜及电子显微镜下观察细胞形态差异。结果:正常对照子宫内膜细胞及EM在位、异位子宫内膜细胞分离培养成功率分别为91.67%、93.75%和75.00%。EM异位、在位子宫内膜腺上皮细胞与正常在位子宫内膜腺上皮细胞大小相似,但EM异位子宫内膜腺上皮细胞染色质增多,核增大。EM异位子宫内膜间质细胞较EM在位及正常在位子宫内膜间质细胞小,且细胞膜表面有较多的微绒毛和胞浆突起。结论:注意取材方式,采用改良原代培养方法,可以提高EM异位子宫内膜细胞培养成功率。EM异位子宫内膜腺上皮细胞和间质细胞超微结构与正常妇女及EM在位子宫内膜细胞有显著不同。     

COL5A2 as a potential clinical biomarker for gastric cancer and renal metastasis

Background:Gastric cancer, characterized by insidious onset and multiple metastasis, is almost incurable and has poor prognosis, and also one of the leading causes of treatment failure and death in patients with gastric cancer (GC). However, the prognosis of collagen type V alpha2 chain (COL5A2) in GC and renal metastasis is unknown.Methods:Recruited 148 patients who underwent GC. The diagnosis of GC was confirmed by ultrasound imaging and pathological examination. Immunohistochemistry and RT-qPCR were performed to exam the expression level of COL5A2. The statistical methods included Pearson chi-square test, Spearman-rho correlation test, univariate and multivariate cox regression analysis. Finally, this research constructed receiver operating characteristic (ROC) curves and applied the area under the curve (AUC).Results:Based on Pearson''s chi-square test, Spearman-rho test, and univariate/multivariate cox regression, pathologic grade (P < .001), renal metastasis (P < .001) and staging (P < .001) were significantly related to COL5A2. And COL5A2 expression (hazard ratio [HR]: 18.834, P < .001) is an independent risk factor of GC. The AUC was used as the degree of confidence in judging each factor: COL5A2 (AUC = 0.878, P < .001), COL1A1 (AUC = 0.636, P = .006), COL1A2 (AUC = 0.545, P = .368), and COL3A1 (AUC = 0.617, P = .019). Through the ROC result, COL5A2 had more advantage as a biomarker for GC than other collagens.Conclusions:COL5A2 gene expression level might be a risk factor for GC. COL5A2 has a strong correlation with the prognosis of the disease.     

Hypoxic Lung-Cancer-Derived Extracellular Vesicle MicroRNA-103a Increases the Oncogenic Effects of Macrophages by Targeting PTEN

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Essential role of Rap signal in pre-TCR-mediated beta-selection checkpoint in alphabeta T-cell development

We demonstrate that lck promoter-driven conditional expression of transgenic SPA-1, a Rap GTPase-activation protein, causes a profound defect of alphabeta T-cell development at the CD4/CD8 double-negative (DN) stage due to enhanced cell death without affecting gammadelta T-cell development. The effect was specific to the DN stage, because CD4 promoter-driven SPA-1 expression hardly affected T-cell development. Rap1A17, a dominant-negative Rap mutant, interfered with the generation of double-positive (DP) cells from Rag2(-/-) fetal thymocytes in vitro in the presence of anti-CD3epsilon antibody and Notch ligand. Rap GTPases were activated in a DN cell line by the expression of self-oligomerizing CD3 (CD8:CD3epsilon chimera), which substituted autonomous pre-T-cell receptor (TCR) signal, inducing CD69 expression and CD25 down-regulation. Reciprocally, expression of C3G, a Rap guanine nucleotide exchange factor, in both normal and Rag2(-/-) DN cells markedly enhanced Notch-dependent generation and expansion of DP cells without additional anti-CD3epsilon antibody, thus bypassing pre-TCR. Defective alphabeta T-cell development in the conditional SPA-1-transgenic mice was restored completely by introducing a p53(-/-) mutation. These results suggest that endogenous Rap GTPases downstream of pre-TCR play an essential role in rescuing pre-T cells from the p53-mediated checkpoint response, thus allowing Notch-mediated expansion and differentiation.     

先天性单眼上斜肌麻痹不同手术方式疗效观察

目的:探讨不同手术方式治疗先天性单眼上斜肌麻痹的疗效以及术后双眼视觉功能的恢复重建情况。方法:回顾性病例研究。选择2016-05/2019-05郑州市第二人民医院斜视与小儿眼科收治的82例先天性上斜肌麻痹患儿作为研究对象,根据患者第一眼位垂直斜视度、患眼下斜肌功能亢进程度、单眼及双眼运动情况等术前检查结果,设计相应的手术方式。包括下斜肌断腱术(3例)、下斜肌部分切除术(63例)、下斜肌徙后术(6例)、健眼下直肌徒后术(4例)、下斜肌减弱+对侧/同侧直肌术(5例)、上斜肌折叠术(1例)。结果:和手术前比较,手术后同时知觉、融合功能、远立体视、近立体视、矫正视力、代偿头位均得到明显改善(P<0.05);有无代偿头位患儿手术后立体视无差异(P>0.05)。结论:根据先天性单眼上斜肌麻痹病情严重程度选择不同的手术方式,在改善患儿视力、代偿头位方面具有积极意义,有助于重建双眼视觉功能。     

Inhibitory effects of cultured Dendrobium tosaense on atopic dermatitis murine model

Dendrobium tosaense is one of the most valuable Chinese medicines and well developed health food. Atopic dermatitis (AD) is a chronic skin disease that occurs mainly in childhood. The pathogenesis of atopic dermatitis had been studied in BALB/c mice modeling by skin-inoculated ovalbumin (OVA) with 2,4,6-trinitro-1-chrolobenzene (TNCB). These mice exhibit features of chronic dermatitis, including skin rash, mast cells infiltration, and elevated serum anti-OVA specific IgE and cytokines modulation. In this study, a standardized ethyl acetate extract of D. tosaense (DtE) was used to protect these mice from the OVA/TNCB-induced skin lesions of atopic dermatitis. The results indicated an increased population of natural T regulatory cell was accompanied by immunosuppression in cytokine profiles and anti-OVA IgE level to significantly reduce Th2 polarization. Finally, toluidine blue staining indicated mast cell infiltration and degranulation was reduced in skin lesion. Our results were shed light on the usage of D. tosaense in AD.