碘[^(131)I]爱克妥昔单抗注射液免疫反应活性的分析方法

研究建立了碘[^(131)I]爱克妥昔单抗注射液免疫反应活性检验分析方法。用活细胞结合百分数法,对细胞选择、反应时间、反应温度和分离方法进行研究,并应用该方法分析爱克妥昔单抗注射液免疫活性的稳定性及批间变异系数。结果表明:选择CEA表达阳性的LS180活细胞,反应时间为16h,反应温度为4℃,分离方法为直接分离法。该方法批间变异系数小于10%,重复性好,可以用于分析碘[^(131)I]爱克妥昔单抗注射液的免疫反应活性。

NMDA受体亚单位NR1在长期培养大鼠海马脑片中的免疫活性变化

目的 研究NMDA受体亚单位NR1在长期培养大鼠海马脑片各区中免疫组织化学反应活性变化规律。方法 将新生 6～ 9天大鼠海马和齿状回切成 4 0 0 μm厚脑片 (即海马脑片 ) ,分别体外培养 1、2、3、4、5周时再作 2 0 μm厚冰冻切片 ,行NR1免疫组织化学反应和图像分析。 结果 NR1在培养各周海马脑片各区锥体细胞和颗粒细胞胞体的免疫反应均呈阳性 ,胞膜深染 ,胞质淡染。培养 1周时 ,其反应强度为CA3区 >CA1区 >齿状回(DG) ;之后则为CA1区 >CA3区 >DG ;3周时各区反应均达高峰。结论 NR1在长期培养大鼠海马脑片各区的免疫活性有时空差异 。

豚鼠与大鼠促性腺激素释放激素的免疫与生物学特性比较

目的比较豚鼠促性腺激素释放激素(GnRH)与大鼠GnRH的免疫反应和生物学活性。方法将豚鼠和大鼠下丘脑匀浆,其GnRH含量用识别结构特异性和识别序列特异性的两种不同的抗体组成的放射免疫系统进行测量。生物学活性在大鼠和豚鼠活体内检测。结果用上述两种放射免疫系统测量,大鼠下丘脑内GnRH含量大致相当,而豚鼠下丘脑内GnRH含量有显著差别。大鼠GnRH的生物学活性在豚鼠和大鼠都比豚鼠GnRH的生物活性强。结论豚鼠GnRH与大鼠GnRH的免疫反应性和生物学活性都不一样。

人类端粒酶逆转录酶及c-kit蛋白在浆液性卵巢癌中的免疫组化表达:一项临床病理研究

. This study investigated the expression of human telomerase reverse transcriptase (hTERT) and c-kit in a cohort of serous ovarian carcinomas by immunohistochemistry with regard to outcome and clinicopathologic variables. Methods. Formalin-fixed, paraffin-embedded archival tissue sections of 10 benign serous cystadenomas, 10 serous neoplasms of low malignant potential (LMP), and 41 serous ovarian carcinomas were immunostained with antibodies to hTERT and c- kit. Immunostaining was scored with regard to quantity and intensity of positively stained cells as negative or weak, moderate, and strong. Mitotic activity was determined as mitotic figures per 10 high power fields. Results. hTERT expression was negative in serous cystadenomas; 70% of LMP showed strong nuclear immunoreactivity. In serous carcinomas, nuclear and sometimes cytoplasmic immunoreactivity was observed; 14% of cases were scored as negative, 42% as moderate, and 44% as strong. hTERT immunoreactivity increased with grade (P < 0.0192) and mitotic activity (P = 0.0018), but not with FIGO stage (P = 0.2752), and was related with outcome (P = 0.0477). No c-kit immunoreactivity was observed in serous cystadenomas and LMP; 27% of serous carcinomas were negative, 46% showed moderate, and 27% strong immunostaining. ckit immunoreactivity was positively correlated with grade (P = 0.0008) and FIGO stage (P = 0.0247), but not with mitotic activity (P = 0.1433) and outcome (P = 0.1145); however, c-kit expression was positively related with poor outcome in FIGO II and III stages (P = 0.0105). Conclusions. hTERT and ckit are frequently up-regulated in serous ovarian carcinomas; c-kit immunoexpression may serve as a marker of aggressive behavior in high stage tumors.

人参皂甙对肾衰大鼠的肾保护作用与延髓腹外侧部TH-IR变化的相关性

[目的]探讨人参皂甙对急性肾衰大鼠的肾保护作用与延髓腹外侧部TH-IR的变化的相关性。[方法]选用52只健康雄性SD大鼠,随机均分为4组:实验对照组(ARF+NS组)、实验给药组(ARF+GS组)、空白对照组(NS+NS组)和空白给药组(NS+GS组)。采用甘油致急性肾衰的动物模型,通过整体实验和免疫组织化学的方法,观察急性肾衰大鼠口服人参皂甙(25 mg/2 mL)48 h后,血清BUN,Cre,GSH水平的变化及肾组织病理学的改变;同时观察延髓腹外侧部TH免疫反应活性的变化。[结果]ARF+NS组血清BUN和Cre水平明显升高,GSH水平明显降低(P<0.05),肾小管坏死程度严重。ARF+GS组血清BUN和Cre水平降低,GSH水平升高,肾小管坏死程度明显减轻。免疫组织化学的结果显示ARF+NS组延髓腹外侧部TH免疫反应阳性神经元数目和免疫染色强度显著增强(P<0.05)。ARF+GS组延髓腹外侧部TH免疫反应活性进一步增强,明显高于ARF+NS组(P<0.05)。[结论]急性肾衰大鼠口服人参皂甙48 h后,能明显增强机体抗氧化损伤的能力和肾保护的作用;延髓腹外侧部儿茶酚胺能神经元活性增强可能是人参皂甙抗急性肾衰和肾保护的作用的中枢机制之一。

Analysis of the Antimicrobial Activity of Antimicrobial Peptides Extracted from Immunized Tenebrio molitor Induced by Different Microbial Treatments Against 26 Pathogens

[Objective] The study was to explore whether antimicrobial activity of the antimicrobial peptides extracted from immunized Tenebrio molitor varied with different pathogens as inducers.[Method]By inducing T.molitor hungry larvaes to generate immune response via feeding with bacteria and with fungi or actinomycete post to pricking,the antimicrobial peptides extracts were obtained by grinding and centrifuging the cultures.Its antimicrobial activity against 26 pathogens was measured by bacteriostatic ring,and evaluated by trisection to four types and inhibitory spectrum.[Result]Both the antimicrobial activity and antimicrobial spectrum of the antimicrobial peptides extracts varied remarkably among different pathogens as inducers.[Conclusion]Bio-control used strains have obvious advantage in inducing the insect to express body fluid immunity material-antimicrobial peptides.

外用激素药膏六不宜

外用激素药膏六不宜是明启学用的外用激素类药膏药主要有肤轻松软膏、地塞米松软膏、氢化可的松软膏、氯氟舒松软膏等，使用时有以下六不宜：霉菌性感染的皮肤病不宜使用。这是由于激素可阻碍巨噬细胞对病原菌的吞噬，改变白细胞的数量和趋向性，破坏有免疫反应活性的淋巴...

Advances in immunotherapy for treatment of lung cancer

Different approaches for treating lung cancer have been developed over time, including chemotherapy, radiotherapy and targeted therapies against activating mutations. Lately, better understanding of the role of the immunological system in tumor control has opened multiple doors to implement different strategies to enhance immune response against cancer cells. It is known that tumor cells elude immune response by several mechanisms. The development of monoclonal antibodies against the checkpoint inhibitor programmed cell death protein 1 （PD-1） and its ligand （PD-L1）, on T cells, has led to high activity in cancer patients with long lasting responses. Nivolumab, an anti PD-1 inhibitor, has been recently approved for the treatment of squamous cell lung cancer patients, given the survival advantage demonstrated in a phase III trial. Pembrolizumab~ another anti PD-1 antibod）5 has received FDA breakthrough therapy designation for treatment of non-small cell lung cancer （NSCLC）, supported by data from a phase I trial. Clinical trials with anti PD-1/PD-L1 antibodies in NSCLC have demonstrated very good tolerability and activity, with response rates around 20% and a median duration of response of 18 months.

Increased proliferation activity measured by immunoreactive Ki67 is associated with survival improvement in rectal/recto sigmoid cancer

AIM: To assess the expression of Ki67 as prognosticator in rectal/recto sigmoid cancer.METHODS: Samples from 146 patients with rectal and recto sigmoid cancer were studied for expression of Ki67 and its prognostic significance in comparison with clinicopathological predictors of survival. Formalin-fixed, paraffin-embedded tissues from 6 (4.1%) patients with T1, 26 (17.8%) with T2, 94 (64.4%) with T3, and 20 (13.7%) with T4 tumors were studied. Ki67 expression was determined immunohistochemically. Samples were divided according to mean value into high (＞40%) and low (≤40%) expression. Areas of extensive proliferation (＞50%) were defined as 'hot spot' areas. RESULTS: Hot spot areas were present in samples regardless of histopathological grade. Lower TNM and Dukes stage and higher expression of Ki67 and presence of Ki67 hot spot areas in histopathological samples were associated with better survival, whereas no association was observed with histopathological grade (P = 0.78). In Cox multivariate regression analysis, significant prognostic factors were Dukes stage (P＜0.001), presence of lymph node metastases (P = 0.015), age (P = 0.035) andpresence of Ki67 hot spot areas (P = 0.044). CONCLUSION: Proliferative activity as measured by Ki67 in rectal cancer is associated with survival improvement compared with patients with low Ki67. Areas of prognostically significant increased proliferation were found independently of histopathological tumor grade.

Inhibition of PMA-induced endothelial cell activation and adhesion by over-expression of domain negative IκBα protein

AIM: NF-κB, regulate the expression of cytokine-inducible genes involving immune and inflammatory responses, will be potential therapy approach for allograft from rejection. In this study, we use pCMV-IκBαM vector to inhibit NF-κB activation and investigate the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells. METHODS: The NF-κB activity was detected with pNF-κB reporter gene and electrophoretic mobility shift assay. Expression of cell surface molecules was detected by RT-PCR and flow cytometer. The cell-cell adhesion assay was performed to determine the effect of pCMV-IκBαM in inhibition of T cells adhesion to endothelial cells. RESULTS: We could find that NF-κB activity is inhibited by over-expression of non-degraded IκBα protein. Expression of adhesion molecules like ICAM-1, VCAM-1, and P-selectin as well as cell-cell adhesion were inhibited significantly by transfection of the pCMV-IκBαM vector. CONCLUSION: Our results indicate that the pCMVIκBαM, which inhibit the activity of NF-κB through over-expression of non-degraded IκBα protein, can be used for gene therapy in diseases involving NF-κB activation abnormally like organ transplantation via inhibiting cell adhesion.

In Vitro and in Vivo Study of the Antitumor Effects of a THANK Modified Hepatoma Cell Line

Objective THANK, known as a member of TNF superfamily, is a potent costimulator of both B and T lymphocytes and can promote astrong immune response. To investigate its role in liver immunotherapy, the anti-tumor effects of the THANK-transduced hepatoma cellline SMMU-7721 in vitro and in vivo were studied.Methods THANK full-length cDNA was transfected into SMMU-7721 cell line. The transfectant with stable expression of THANK wasobtained by clone selection and THANK s effects on hepatoma cells were analyzed, further the tumorigenicity of THANK-transduced7721 cells was examined in nude mice.Results THANK's expression in 7721 cells inhibited the growth of hepatoma cells and induced a strong CTL response in vitro. The cellcycle analysis showed that THANK transfected 7721 cells were arrested in the S phase. The expression of THANK in SMMU-7721 cellline not only inhibited the tumorigenicity of 7721 cells, but also induced a systemic immune response against re-challenge of parental7721 tumors.Conclusion THANK transduction in SMMU-7721 cells can induce an effective immune response in nude mice and may be useful for theimmunotherapy of hepatomas.

Hepato-biliary profile of potential candidate liver progenitor cells from healthy rat liver

AIM： To evaluate the presence of progenitor cells in healthy adult rat liver displaying the equivalent ad- vanced hepatogenic profile as that obtained in humans. METHODS： Rat fibroblastic-like liver derived cells （rFLDC） were obtained from collagenase-isolated liver cell suspensions and characterized and their phenotype profile determined using flow cytometry, immunocyto- chemistry, reverse transcription polymerase chain reac- tion and functional assays. RESULTS： rFLDC exhibit fibroblastoid morphology, ex- press mesenchymal （CD73, CD90, vimentin, m-smooth muscle actin）, hepatocyte （UGTIA1, CK8） and biliary （CK19） markers. Moreover, these cells are able to store glycogen, and have glucose 6 phosphatase activity, but not UGTIA1 activity. Under the hepatogenic differentia- tion protocol, rFLDC display an up-regulation of hepatocyte markers expression （albumin, tryptophan 2,3-di- oxygenase, G6Pase） correlated to a down-regulation of the expression of the biliary marker CK19. CONCLUSION： Advanced hepatic features observed in human liver progenitor cells could not be demonstrated in rFLDC. However, we demonstrated the presence of an original rodent hepato-biliary cell type.

肝动脉结扎微波固化治疗肝癌的研究

原发性肝癌在我国发病率较高,但手术切除率一般不到50％。我院1990年始对术中不能切除的肝癌,采用肝功动脉结扎微波固化治疗,收到较好的效果。 肝动脉结扎方法,根据病变范围和肝门显露程度,选择以下3种途径:①结扎肝动脉支的近端后,经肝动脉支的远端注入丝裂霉素4mg。

Mechanical Stress Induces Neuroendocrine and Immune Responses of Sea Cucumber(Apostichopus japonicus)

Grading procedure in routine sea cucumber hatchery production is thought to affect juvenile sea cucumber immunologi- cal response. The present study investigated the impact of a 3-min mechanical perturbation mimicking the grading procedure on neuroendocrine and immune parameters of the sea cucumber Apostichopusjaponicus. During the application of stress, concentrations of noradrenaline and dopamine in coelomic fluid increased significantly, indicating that the mechanical perturbation resulted in a transient state of stress in sea cucumbers. Coelomocytes concentration in coelomic fluid increased transiently after the beginning of stressing, and reached the maximum in 1 h. Whereas, coelomocytes phagocytosis at 3 min, superoxide anion production from 3 min to 0.5 h, acid phosphatase activity at 0.5 h, and phenoloxidase activity from 3 min to 0.5 h were all significantly down-regulated. All of the immune parameters recovered to baseline levels after the experiment was conducted for 8 h, and an immunostimulation occurred after the stress considering the phagocytosis and acid phosphatase activity. The results suggested that, as in other marine invertebrates, neuroendocrine/immnne connections exist in sea cucumber A. japonicus. Mechanical stress can elicit a profound influence on sea cucumber neuroendocrine system. Neuroendocrine messengers act in turn to modulate the immunity fimctions. Therefore, these ef- fects should be considered for developing better husbandry procedures.

电针对抑郁大鼠行为学、海马生长抑素及其mRNA表达的影响

目的:观察电针对抑郁大鼠行为学、海马生长抑素(somatostatin,SS)及SS mRNA表达的影响,探讨电针治疗抑郁症的可能机制。方法:30只健康Wistar大鼠,随机分为正常组、模型组和电针组,每组10只。慢性应激法刺激造抑郁大鼠模型,电针"百会""三阴交",每日1次,每次20 min,连续14 d。使用开野试验和糖水试验检测大鼠行为学变化,使用免疫组化法和逆转录多聚酶链反应法测定SS及SS mRNA的表达。结果:造模后模型组大鼠水平和垂直运动次数、糖水摄入量与正常组比较均降低(P<0.01),治疗后电针组的水平、垂直运动次数较治疗前及模型组均有升高(P<0.01)。模型组与正常组相比,SS、SS mRNA表达均降低(P<0.05);电针组与模型组比较,SS、SS mRNA表达均升高(P<0.05)。结论:电针可明显改善抑郁大鼠行为学,提高抑郁大鼠海马SS及SS mRNA的表达,电针的抗抑郁作用可能与之有关。