转染外源性白细胞介素6基因增强乳腺癌细胞株MCF-7表达肿瘤相关抗原

目的 :探索白细胞介素 6 (interleukin 6 ,IL 6 )增强乳腺癌细胞表达乳腺癌抗原 (CA15 3)和癌胚抗原 (CEA)的作用。方法 :将含IL 6蛋白编码顺序1176bpcDNA插入Pci neo哺乳动物表达载体。将重组载体转染MCF 7乳腺癌细胞 ,采用ELASA方法测定细胞培养上清液内IL 6浓度 ,用MEIA(microp articalenzymeimmunoassay)方法测定上清液中肿瘤相关抗原CA15 3、CEA和CA12 5。结果 :含有外源IL 6基因的MCF 7细胞分泌IL 6浓度 (338.5±2 2 .6pg·10 -6细胞 )明显高于父本没有含外源基因的MCF 7细胞 (2 5 .4± 4 .6pg·10 -6细胞 )和仅含空载体Pci neo的MCF 7细胞 (19.6± 3.0pg·10 -6细胞 ) (P<0 .0 1)。细胞培养d 3后 ,带外源IL 6基因的MCF 7细胞培养上清液中CA15 3和CA12 5水平 (分别为 14 .9± 2 .3和 38.8± 5 .1μg·10 -6细胞 )明显高于父本 (分别为 6 .6± 1.5和 10 .0± 1.6 μg·10 -6细胞 )和空载体Pci neo的MCF 7细胞 (分别为 3.4±0 .7和 14 .6± 2 .2 μg·10 -6细胞 ,P <0 .0 5 )。而转染IL 6基因没有明显改变CEA表达 (P >0 .0 5 )。结论 :IL 6具有诱导肿瘤相关抗原的表达和增强肿瘤细胞的免疫原性的作用 ,提示IL 6可能增强机体对肿瘤的免疫反应性。     

股内侧肌穿支皮瓣的解剖学基础与膝周创伤修复

目的:探讨股内侧肌穿支皮瓣的解剖学特征,为临床膝周软组织缺损的修复提供一种新的术式。方法:实验于2005-01/10在解放军南京军区福州总医院军区临床解剖学研究中心实验室完成。30侧动脉内灌注红色乳胶的成人下肢标本由南京军区福州总医院军区临床解剖学研究中心提供。30侧成人下肢标本,以收肌结节、髌骨中点、腹股沟韧带中点为观测标志,重点解剖观测股内侧肌肌支的起源、走行、分支、分布及其远端吻合情况。一侧新鲜标本上,摹拟术式设计。结果:在股三角尖部自股动脉恒定发出的股内侧肌支,入肌门后在肌内沿肌束行向外下方,直至髌旁并与髌周动脉环相吻合。沿途除发出肌支营养股内侧肌外,还发出1～3支(77%为1支)、外径0.5～0.9mm肌皮穿支垂直穿过股内侧肌达深筋膜,并浅出至股内侧肌表面皮肤。皮瓣切取面积可达8.5cm×15.0cm。结论:可形成的以股内侧肌肌支-穿支逆行皮瓣转位可修复膝部软组织缺损。     

Cost-effectiveness of collaborative care for the treatment of major depressive disorder in primary care. A systematic review

Background  

The effectiveness of collaborative care for patients with major depressive disorder in primary care has been established. Assessing its cost-effectiveness is important for deciding on implementation. This review therefore evaluates the cost-effectiveness of collaborative care for major depressive disorder in primary care.     

阿司匹林锌的研制(Ⅰ)

研究阿司匹林锌的化学合成方法。方法：以阿司匹林和硫酸锌为原料,经两步反应合成阿司匹林锌。结果得到的样品经ＩＲ、ＮＭＲ、ＭＳ等波谱分析及元素分析、热分析证明该化合物为阿司匹林锌。总收率为８０％。结论该合成方法收率稳定,合成周期短,合成路线完全可行。     

Influence of two different resection techniques (conventional liver resection versus anterior approach) of liver metastases from colorectal cancer on hematogenous tumor cell dissemination – prospective randomized multicenter trial

Background

Preoperative radiotherapy of the pelvic abdomen presents with complications mostly affecting the small bowel. The aim of this study was to define the features of early radiation-induced injury on small bowel.

Methods

54 mice were divided into two groups (36 irradiated and 18 sham irradiated). Animals were placed on a special frame and (in the radiated group) the exteriorized segment of ileum was subjected to a single absorbed dose of 19 or 38 Gy radiation using 6 MV high energy photons. Specimens were collected for histology, immunohistochemistry (IHC) and ELISA analysis after 2, 24 and 48 hours. Venous blood was collected for systemic leucocyte count in a Burker chamber.

Results

Histology demonstrated progressive infiltration of inflammatory cells with cryptitis and increased apoptosis. MIP-2 (macrophage inflammatory protein) concentration was significantly increased in irradiated animals up to 48 hours. No significant differences were observed in IL-10 (interleukin) and TNF-α (tumour necrosis factor) levels. IHC with CD45 showed a significant increase at 2 hours of infiltrating leucocytes and lymphocytes after irradiation followed by progressive decrease with time. Caspase-3 expression increased significantly in a dose dependent trend in both irradiated groups up to 48 hours.

Conclusion

Acute small bowel injury caused by local irradiation is characterised by increased apoptosis of crypt epithelial cells and by lymphocyte infiltration of the underlying tissue. The severity of histological changes tends to be dose dependent and may affect the course of tissue damage.     

Binding kinetics differentiates functional antagonism of orexin-2 receptor ligands

Orexin receptor antagonism represents a novel approach for the treatment of insomnia that directly targets sleep/wake regulation. Several such compounds have entered into clinical development, including the dual orexin receptor antagonists, suvorexant and almorexant. In this study, we have used equilibrium and kinetic binding studies with the orexin-2 (OX₂) selective antagonist radioligand, [³H]-EMPA, to profile several orexin receptor antagonists. Furthermore, selected compounds were studied in cell-based assays of inositol phosphate accumulation and ERK-1/2 phosphorylation in CHO cells stably expressing the OX₂ receptor that employ different agonist incubation times (30 and 5 min, respectively). EMPA, suvorexant, almorexant and TCS-OX-29 all bind to the OX₂ receptor with moderate to high affinity (pK_I values ≥ 7.5), whereas the primarily OX₁ selective antagonists SB-334867 and SB-408124 displayed low affinity (pK_I values ca. 6). Competition kinetic analysis showed that the compounds displayed a range of dissociation rates from very fast (TCS-OX2-29, k_off = 0.22 min⁻¹) to very slow (almorexant, k_off = 0.005 min⁻¹). Notably, there was a clear correlation between association rate and affinity. In the cell-based assays, fast-offset antagonists EMPA and TCS-OX2-29 displayed surmountable antagonism of orexin-A agonist activity. However, both suvorexant and particularly almorexant cause concentration-dependent depression in the maximal orexin-A response, a profile that is more evident with a shorter agonist incubation time. Analysis according to a hemi-equilibrium model suggests that antagonist dissociation is slower in a cellular system than in membrane binding; under these conditions, almorexant effectively acts as a pseudo-irreversible antagonist.Linked ArticlesThis article is part of a themed section on Orexin Receptors. To view the other articles in this section visit http://dx.doi.org/10.1111/bph.2014.171.issue-2     

Bone tumors: magnetic resonance imaging versus computed tomography

The magnetic resonance (MR) imaging characteristics of bone tumors are described and the clinical utility of MR imaging in patient evaluation is reported. Fifty-two patients with skeletal lesions were examined with a Picker MR imager (0.15-T resistive magnet). Twenty-five patients had primary malignancies, seven had benign bone neoplasms, 15 had skeletal metastases, and five had neoplasm simulators. Forty-five patients had CT scans available for comparison. For demonstrating the extent of tumor in marrow, MR was superior to CT in 33% of cases, about equal to CT in 64%, and inferior to CT in 2%. For delineating the extent of tumor in soft tissue, MR was superior to CT in 38% of cases and about equal to CT in 62%. CT was superior in all cases for demonstrating calcific deposits and pathologic fractures. In four patients with metal prostheses or surgical clips, MR was superior to CT in documenting recurrent tumor because of artifactual degradation of the CT image. Direct sagittal and coronal images from MR permit accurate assessment of the relationship of tumor to adjacent normal structures, including the physis, joints, and neurovascular structures. MR is useful in the evaluation of bone tumors: it is of greatest value in evaluations of the peripheral skeleton, the medullary canal, soft tissues, and postoperative tumor recurrence. With a 0.15-T magnet, MR is less useful in the evaluation of the axial skeleton and cortical bone.     

Constitutive in vivo cytokine and hematopoietic growth factor gene expression in the bone marrow and peripheral blood of healthy individuals

We investigated hematopoietic growth factor (HGF) and cytokine gene expression in the bone marrow (BM) and peripheral blood (PB) of healthy individuals as a starting point for delineating the physiologic role of cytokines in steady state hematopoiesis. BM biopsy specimens and PB samples from 7 healthy individuals were analyzed by polymerase chain reaction amplification of reverse-transcribed RNA using gene-specific primer sets. Consistent gene expression in the BM of all 7 individuals was detected for macrophage colony-stimulating factor (CSF), stem cell factor, interleukin-6 (IL-6), IL-7, erythroid-potentiating factor, erythroid-differentiating factor, and insulinlike growth factor 1, all cytokines with reported direct stimulatory effects on in vitro hematopoiesis. Of these, erythroid-potentiating factor and erythroid- differentiating factor appeared to be the only stimulating factors that were also expressed in the PB. Among the cytokines with inhibitory effects on in vitro hematopoiesis IL-4, tumor necrosis factor-alpha (TNF-alpha), TNF-beta, transforming growth factor-beta, and macrophage inflammatory protein-1 alpha were expressed in the BM of the 7 individuals. Except for TNF-alpha, the latter cytokines were also expressed in the PB. Consistent expression in the BM and PB of all tested individuals was also observed for IL-1 beta, IL-1 receptor antagonist, and IL-1 beta converting enzyme, which are all members of the IL-1 family with a possible indirect effect on hematopoiesis. Remarkably, no expression of granulocyte CSF, granulocyte-macrophage CSF, and IL-3 was found in the BM or PB of all investigated individuals (n = 15). This was also the case for IL-1 alpha, IL-2, IL-5, IL-9, IL- 12, IL-13, leukemia-inhibiting factor, interferon-gamma, and inhibin. Weak IL-8 and IL-10 expression was found in the BM and/or PB of a minority of investigated individuals. These findings provide insight into which cytokines or HGFs potentially are involved in the autocrine or paracrine regulation of in vivo steady state hematopoiesis. The absence of expression of granulocyte CSF, granulocyte-macrophage CSF, and IL-3 in the BM of healthy individuals implicates that it is highly unlikely that these HGFs are involved in the autocrine or paracrine regulation of constitutive hematopoiesis.     

Effects of Treatments by Calcium and Sex Hormones on Vertebral Fracturing in Osteoporosis

Lateral radiographs of the thoracic and lumbar spine were takenperiodically in 49 patients with osteoporosis. Thirty patientswere postmenopausal, and 19 nonmenopausal with osteoporosisdue to steroids, male hypogonadism, alcoholism, thyrotoxicosisor unknown cause. Patients were studied before, during and aftertreatment with high calcium alone, or with combined calciumand sex steroids. Calcium was given as effervescent calciumlactate gluconate, and sex hormones as oestradiol valerate,testosterone oenanthate, or methenolone oenanthate. A totalof 964 films covering 409 patient-years were available for measurement.On each vertebra, deformity due to loss of anterior height wasmeasured and assigned to one of four grades. For the time intervalbetween each consecutive pair of films, a patient's vertebralfracture rate score was calculated and expressed per thousandpatient-years. In comparison with the corresponding pretreatment fracture ratescore, both the postmenopausal and the nonmenopausal groupswho had not received sex hormones previously, failed to showsignificant changes (p=0.144; p=0.017) on high calcium aloneduring mean periods of 4.3 and 2.8 years respectively. If thefirst 2 years on high calcium were excluded for the postmenopausalgroup, they still failed to show a reduction in fracture ratescore (observed for a mean period of 5.0 years; p=0.04). When treated with combined calcium and sex hormones, both postmenopausaland nonmenopausal groups showed a lower fracture rate scoreof 20 and 207 respectively when compared with the pretreatmentlevels of 1500 and 1697 (in mean treatment periods of 3.2 and4.4 years; p<0.001 in each case). When given high-dose calciumalone, but after treatment with sex hormones as well, the postmenopausalgroup showed no change in fracture rate score from pretreatment(in a mean of 3.1 years; p=0.069); however the nonmenopausalgroup still showed a significant reduction in fracture ratescore from 1697 to 42 over a mean period of 2.3 years (p=0.001).The postmenopausal group, after stopping all treatment, showeda higher fracture rate score of 1286 (in a mean of 2.6 years)than did those on combined calcium and sex hormones, in whomthe fracture rate score was 20 (in a mean of 3.2 years; p=0.008).A subgroup of 11 patients with osteoporosis of both the menopausaland nonmenopausal types, had data both before (in a mean of5.5 years) and during (for a mean of 2.5 years) treatment withcalcium alone; the fracture rate scores were 1473 and 918 (p=0.247).Data were available for nine patients both before (for meanof 5.5 years) and during (for a mean of 5.5 years) treatmentwith calcium and sex hormones; the fracture rate score fellfrom 1397 to 100 (p=0.001). It is concluded that in groups with both menopausal and nonmenopausalosteoporosis, vertebral fracturing was reduced by treatmentwith combined calcium and sex hormones, but no significant effectfrom calcium alone was shown. In both groups, cessation of therapywas associated with a return to near the pretreatment fracturerate score, strongly suggesting the need for lifelong treatment.