B淋巴细胞缺乏影响小鼠心脏结构及心肌免疫细胞组成

目的探讨B淋巴细胞对心脏发育过程中心脏结构、功能及心肌免疫细胞的影响。方法使用心动超声、免疫荧光染色及流式细胞术分别评估野生型小鼠(WT小鼠)及B淋巴细胞缺陷小鼠(μMT小鼠)心脏结构、功能及心肌免疫细胞的组成。结果与WT小鼠相比,μMT小鼠心脏质量与小鼠体质量比及左心室质量显著减低(P<0.05),心肌细胞横截面积显著减小(P<0.001),且μMT小鼠的左心室射血分数明显升高(P<0.05)。mRNA测序结果显示,WT小鼠及μMT小鼠差异基因主要富集在心脏发育及肥厚型心肌病信号通路上。流式细胞术结果显示,与WT小鼠相比,μMT小鼠心肌具有更少的Ly6g^(+)中性粒细胞(P<0.001)、CD4^(+)阳性T淋巴细胞(P<0.01)及CD8^(+)T淋巴细胞(P<0.05)。结论B淋巴细胞缺失改变心肌中免疫细胞的组成,降低左心室质量,并增加心肌收缩力。

RBM3在卵巢上皮性癌中的表达及临床意义

目的 对RBM 3在卵巢上皮性癌中的表达及临床意义进行分析探讨,为今后的临床诊治工作提供有价值的参考信息。方法 选择手术切除的卵巢上皮性癌石蜡组织标本50例为实验组,选择同期良性卵巢上皮肿瘤20例、正常卵巢组织20例作为对照组,采取免疫组化法对RBM 3的表达水平进行检测,并对检测结果进行统计分析。结果 对比发现,在实验组中RBM 3表达阳性率较良性卵巢上皮肿瘤、正常卵巢组织检测结果发生显著升高（P〈0.05）;在FIGOⅢ~Ⅳ期中的表达阳性率显著高于Ⅰ期和Ⅱ期（P〈0.05）;低分化阳性率高于高、中分化（P〈0.05）;有腹水者表达阳性率高于无腹水组（P〈0.05）。结论 RBM 3对卵巢癌的发生以及发展产生了促进作用,在今后的临床工作中,应对其给予足够的重视。

Down-regulation of Wnt Antagonist,SFRP1 in Colorectal Tumorigenesis

OBJECTIVE To assess the functional change of SFRP1 （secreted frizzled-related protein1）, in colorectal tumorigenesis. METHODS Immunohistochemical investigation and the semiquantitative reverse transcription-polymerase chain reaction （RT-PCR） were used to assess the expression of SFRP1, β-catenin （β-cat） and E-caderin （E-cad） in matched samples of normal colorectal mucosa, adenomas and cancers. RESULTS SFRP1 mRNA expression was down-regulated in the neoplasms, and abnormal expressions of β-cat and E-cad were found in colorectal adenomas and colorectal cancers. CONCLUSION Down-regulation of SFRP1 observed is consistent with its acting as a tumor suppressor gene in colorectal tumorigenesis.

Recent advances in cytokines:Therapeutic implications for inflammatory bowel diseases

Inflammatory bowel diseases （IBDs） are complex and chronic disabling conditions resulting from a dysregulated dialogue between intestinal microbiota and components of both the innate and adaptive immune systems. Cytokines are essential mediators between activated immune and non-immune cells, including epithelial and mes- enchymal cells. They are immunomodulatory peptides released by numerous cells and these have significant effects on immune function leading to the differentiation and survival of T cells. The physiology of IBD is becom- ing a very attractive field of research for development of new therapeutic agents. These include cytokines involved in intestinal immune inflammation. This review will focus on mechanisms of action of oytokines involved in IBD and new therapeutic opportunities for these diseases.

Mechanism of Immune Hyporesponsiveness Induced by Recipient-derived Immature Dendritic Cells in Liver Transplantation Rat

Objective To investigate the mechanism of immune hyporesponsiveness induced by donor-antigen- unloaded recipient-derived immature dendritic cell （imDC） of liver grafts in rats. Methods Forty Sprague-Dawley rats （donor） and forty male Wistar rats （recipient） were randomly divided into 4 groups： control, cyclosporine A （CsA）, mature DC （mDC）, and imDC groups respectively, with 10 donor rats and 10 recipient rats in each group. Recipient rats in CsA group were treated with 10 mg-kg-~＇d-I CsA starting day 2 after the transplantation. Recipients in the mDC or imDC groups were given Wistar rat derived mDCs （1 × 10^6/rat） or imDCs （1 × 10^6/rat） via dorsal vein of the penis respectively 1 day before the transplantation. In each group, 5 recipients were kept for determination of survival time and the other 5 rats were executed at day 10 after transplantation. Blood samples were collected for the measurement of serum alanine aminotransferase （ALT）, total bilirubin （TBIL）, interleukin 2 （IL-2）, interferon gamma （IFN-γ）, IL-4, and IL-10 levels. Liver tissue was harvested for HE staining and acute rejection evaluation. Expression levels of Fas-L/Fas in the grafts were detected by immunohistochemical staining; andWestern blot was used to detect the expression level of Scurfm. Results The survival time of CsA and imDC groups was significantly longer than that of control and mDC groups （all P〈0.05）. The levels of serum ALT and TBIL in the control group （2072.20±217.93 IU/L and 147.42±22.02pmol/L） and mDC group （2117.00±285.13 IU/L and 141.58±20.82 pmol/L） were significantly higher than those in the CsA group （59.68±13.48 IU/L and 15.40±2.13 pmol/L） or imDC group （50.80±9.63 IU/L and 14.44±3.49 pmol/L） （all P〈0.05）. In the CsA and imDC groups, the levels of IL-2 （22.52±3.75 pg/mL and 22.12±3.90 pg/mL） and IFN-γ （309.20±25.19 pg/mL and 321.00±21.64 pg/mL） were significantly lower, but the levels of IL-4 （297.60±25.07 pg/mL and 277.00±22.47 pg/mL） and IL-10 （1226.00±140.49 pg/mL and 1423.00±106.39 pg/mL） were higher than those of the control （IL-2：147.78±12.80 pg/mL, IFN-γ： 1758.60±106.22 pg/mL, IL-4：17.40±4.77pg/mL, IL-10： 81.00＋ 9.47 pg/mL） and mDC groups （IL-2：142.34±9.29 pg/mL, IFN-7：1835.00±82.63 pg/mL, IL-4： 15.60＋ 3.96 μg/mL, IL-10： 68.80± 11.23 pg/mL） （all P〈0.01）. The expression level of Scurfin protein on CD4＋CD25 ＋ T cells of the imDC group （1.34±0.29） was significantly higher than that in the control （0.72±0.13）, CsA （0.37±0.11）, and mDC groups （0.78±0.17） （all P〈0.05）.

Intestinal microbiota in inflammatory bowel disease:Friend of foe?

Inflammatory bowel disease （IBD） arises from disruption of immune tolerance to the gut commensal microbiota, leading to chronic intestinal inflammation and mucosal damage in genetically predisposed hosts. In healthy individuals the intestinal microbiota have a symbiotic relationship with the host organism and possess important and unique functions, including a metabolic function （i.e. digestion of dietary compounds and xenobiotics, fermentation of undigestible carbohydrates with production of short chain fatty acids）, a mucosal barrier function （i.e. by inhibiting pathogen invasion and strengthening epithelial barrier integrity）, and an immune modula- tory function （i.e. mucosal immune system priming and maintenance of intestinal epithelium homeostasis）. A fine balance regulates the mechanism that allows co- existence of mammals with their commensal bacteria. In IBD this mechanism of immune tolerance is impaired because of several potential causative factors. The gut microbiota composition and activity of IBD patients are abnormal, with a decreased prevalence of dominant members of the human commensal microbiota （i.e. Clostridium IXa and IV groups, Bacteroides, bifldobacteria） and a concomitant increase in detrimental bacteria （i.e. sulphate-reducing bacteria, Escherichia coll. The observed dysbiosis is concomitant with defectiveinnate immunity and bacterial killing （i.e. reduced mucosal defensins and IgA, malfunctioning phagocytosis） and overaggressive adaptive immune response （due to ineffective regulatory T cells and antigen presenting cells）, which are considered the basis of IBD pathogen- esis. However, we still do not know how the interplay between these parameters causes the disease. Studies looking at gut microbial composition, epithelial integrity and mucosal immune markers in genotyped IBD populations are therefore warranted to shed light on this obscure pathogenesis.

Asymptomatic lymphangioma involving the spleen and retroperitoneum in adults

Lymphangioma, a benign neoplasm of the lymphatic system, is common in children but rare in adults. Its clinical manifestations include abdominal pain, nausea, vomiting and a palpable mass. However, abdominal sonography or abdominal computed tomography （CT） scan can also incidentally reveal lymphangioma. A larger or symptomatic lymphangioma is treated with total resection to prevent recurrence, infection, torsion and enlargement. Although lymphangioma rarely becomes malignant, its prognosis is generally good. We report a cystic lymphangioma of the spleen and retroperitoneum, which was incidentally found in a 56-year-old man who was hospitalized due to a colon mass. Physical examination showed no specific findings. Abdominal CT revealed a 5.7 cm, non-enhanced multilobulated cystic mass with multiple sepia in the spleen and a 10 cm lobulated cystic mass in the paraaortic area. Splenectomy and retroperitoneal resection of the cystic mass were conducted. The endothelium of splenic and retroperitoneal cyst was immunohistochemically stained with D2-40 antibody. The patient was finally diagnosed with splenic cystic and retroperitoneal cavernous lymphangioma.

Isolated pancreatic granulocytic sarcoma: A case report and review of the literature

Granulocytic sarcoma (GS) is an extramedullary tumor mass consisting of immature myeloid cells. Isolated pancreatic granulocyte sarcoma is extremely rare. We report a very unusual pancreatic granulocytic sarcoma in a patient without acute myeloid leukemia. The patient presented with acute epigastric pain because of splenic infarction due to a mass consisting of myeloblasts in the pancreatic tail. The patients underwent splenectomy and distal pancreatectomy. Pathology and immunohistochemistry suggested a GS. Despite local surgery, an isolated tumor recurred 2 mo after operation and the patient died 3 mo after removal of the tumor. Only 7 reported cases of pancreatic GS were identified in the literature and the mass was located in the pancreatic head. This is the first report of GS in the pancreatic tail with splenic infarction.

Angiogenic markers endoglin and vascular endothelial growth factor in gastroenteropancreatic neuroendocrine tumors

AIM:To investigate the expression and potential prognostic role of vascular endothelial growth factor(VEGF) and endoglin in gastroenteropancreatic neuroendocrine tumors(GEP-NETs) . METHODS:Microvessel density(MVD) in GEP-NETs was evaluated using endoglin and CD31 immunohistochemistry.In addition,tissue levels of endoglin and VEGF were determined in homogenates by ELISA. RESULTS:Endoglin was highly expressed on tumor endothelial cells.CD31 MVD in GEP-NETs was significantly higher compared to endoglin MVD(P<0.01) .Two-tofour-fold higher tissue levels of endoglin and VEGF were seen in tumors compared to associated normal tissue. This increased endoglin tissue expression in tumors was significantly related to tumor size(P<0.01) ,presence of metastases(P=0.04) ,and a more advanced tumor stage(P=0.02) ,whereas expression of VEGF was not. CONCLUSION:We suggest that endoglin is a potential marker to indicate and predict metastases,which might be useful in the post-resection therapeutic approach of patients with GEP-NETs.

CONSTRUCTION AND IMMUNOGENICITY OF HUMAN PAPILLOMAVIRUS TYPE 6B L1 RECOMBINANT PLASMID

To construct a DNA vaccine as a prophylactic model to prevent condyloma acuminatum and detect its immu-nogenicity in mice. Methods The major capsid protein (L1) gene of human papillomavirus (HPV) 6b was inserted into an eukaryotic ex-pression plasmid (pcDNA3.1). The recombinant plasmid was transfected into COS-7 cells. Western blot were performed to detect whether L1 protein can be expressed in eukaryotic cells. Eighteen female BALB/c mice were tested for immunoge-nicity study. Results The recombinant plasmid (pcDNA3.1-HPV6bL1) was verified as HPV6b L1 gene by sequencing. Western blot showed specific strip. Anti-L1 protein antibodies could be detected in the mice’s sera inoculated with pcDNA3.1-HPV6bL1. Similarly, IL-4, IL-2, and IFN-γ were increased in the same mice. Conclusion HPV6b L1 recombinant plasmid was constructed successfully which had immunogenicity for BALB/c mice. It provided experimental evidence for the research of DNA vaccine of condyloma acuminata..-

Correlation of VEGF expression,angiogenesis and carcinomatous change in breast tumors

Objective: The aim of the study was to detect the expression of vascular endothelial growth factor (VEGF) and microvessel density (MVD) in breast benign tissues and malignant tumors to clarify the relationship between VEGF expression, angiogenesis and breast carcinoma occurrence. Methods: The expression of VEGF and MVD in 79 cases of invasive ductal breast carcinoma, 79 corresponding para-cancer normal tissues from primary invasive breast carcinomas, 35 breast carcinoma in situ, 23 breast atypical hyperplasia and 56 breast fibroid tumor were examined by immunohistochemistry staining (SP-method). Results: The positive rate of VEGF and MVD value increased significantly with the increase of the malignant degree of breast tissues (P = 0.000). In breast carcinoma group, the positive rate of VEGF and MVD value with lymph node metastasis were higher than those without lymph node metastasis (P = 0.011 and P = 0.023). A significant higher expression of VEGF and MVD value were observed as the clinical stage increased (P = 0.035 and P = 0.012). The MVD value was higher in VEGF positive group than negative group (P = 0.000). Conclusion: The combining detection of VEGF expression and MVD is helpful for evaluating malignant degree of breast carcinoma. Angiogenesis in breast tumors and occurrence of breast carcinoma might be correlated with the expression of VEGF.

Primary chondrosarcoma of the liver:case report and literature review

We present a rare case of primary chondrosarcoma of the liver in a 57-year-old man with pre-existing hepatitis B virus-related chronic hepatitis.The MRI scans showed a huge cystic-solid occupation of 18 cm×17 cm×11 cm in the right hepatic lobe.The tumor was completely resected,and the histological findings identified low-grade cartilaginous component with typical ring-and-arc chondroid matrix mineralization.Immunohistochemically,the neoplastic cells were positive for vimentin and S-100 protein.The patient received once postoperative adjuvant transcatheter arterial chemoembolization(TACE)at the 2nd month after discharge,and he is still alive for more than 13 months without recurrence and metastasis.To the best of our knowledge,this is the first case of primary chondrosarcoma of the liver.

Expression of caveolin-1 in clear cell renal cell carcinoma and its correlation with microvessel density

Objective:The aim of the study was to investigate the clinicopathologic significance of caveolin-1 expression in clear cell renal cell carcinoma (CCRCC) and its correlation with microvessel density (MVD).Methods:The expression of caveolin-1 was detected by the immunohistochemistry method,while the microvessel density was detected by the immunohistochemistry expression of CD34.Results:In the CCRCCs,the positive rate of caveolin-1 was 67.4%,the over expression of caveolin-1 was not related with sex and age,but related with clinicopathologic parameter,such as tumor sizes,clinical TMN stage,nuclear stage and survival time (P < 0.05).The MVD of positive caveolin-1 cases was significantly higher than that without caveolin-1 expression (P < 0.05).Conclusion:The expression of caveolin-1 is helpful in the prognostic evaluation of CCRCCs and it may be involved in the tumor angiogenesis.

Argyrophilic nucleolar organizer region in MIB-1 positive cells in non-small cell lung cancer: clinicopathological significance and survival

Objective： To evaluate the relation between argyrophilic nucleolar organizer region （AgNOR）-associated proteins and clinicopathological parameters and survival in non-small-cell lung cancer （NSCLC）. Methods： A total of 207 surgical specimens diagnosed as NSCLC were included in this study. Double-staining procedures were performed using antigen Ki-67 （clone MIB-1） and silver nitrate by immunohistochemical and AgNOR-staining methods. Results： The AgNOR area in MIB-l-positive cells of NSCLC is related to clinicopathological parameters under the TNM （tumor, node, and metastasis） system. The survival of patients with small AgNOR area in MIB-1-positive cells is better than that of patients with large AgNOR area. Molecular, biological （AgNOR area in MIB-l-positive cells）, and clinicopathological （greatest tumor dimension, metastases to regional lymph nodes, histology, and differentiation） parameters are independent prognostic factors of NSCLC.Conclusion： The AgNOR area in MIB- 1-positive cells is related to clinicopathological parameters and survival in NSCLC.

Evaluate the Expression of uPA,PAI-1 in Human Gastric Cancer and its Correlation with the Angiogenesis by the Application of Tissue Microarray

OBJECTIVE To investigate the expression of urokinase-type plasminogen （uPA）, its inhibitor-1 （PAI-1） mRNA and its protein in human gastric cancer and to find out the relationship among the tumor differentiation, angiogenesis, and other clinical pathologic factors. METHODS In situ hybridization （ISH） was used to get the uPA, PAI-lmRNA in 110 cases with human gastric cancer in 2-tissue microarray （TMA）. Immunohistochemical staining （S-P method） for uPA, PAI-1 protein and CD34 were performed in the 110 cases in 2 TMA. RESULTS The expression of the uPA, PAI-lmRNA and their protein happened in the cytoplasm of gastric cancer cells were induced by the poor differentiation of the GC, and the expression of uPA had an increasing trend while the expression of the PAI-1 had a decreasing trend. The microvessel density （MVD） had a positive correlation with the clinical stages and the significant relationship with the lymph node metastasis （P 〈 0.05）. The MVD in uPA positive group was significantly higher than those in uPA negative group （P 〈 0.05）. The expression of PAI-1 has no correlation neither with the clinical stages nor the lymph node metastasis. CONCLUSION The uPA play an important role in invasion and metastasis of GC through promoting angiogenesis. Interdicting the secretion and function of the uPA may allow the target therapy against the tumor invasion. As a new high-throughput technology, the tissue microarray is a valuable way to be used in clinical treatment.

Synthesis of Nanostructured Erbium Oxides and Investigation of Influence Oxides on the Immune System and of Some Animal Tissues

Nanostructured erbium oxides （Er2O3） with coherent scattering length 26, 31, 62 and 65 nm were obtained using as a precursor of erbium chloride and erbium oleate. The influences of Er203 on the immune system and some animal tissues were carried out. The experiments have been made on white mouse＇s and outbred rats. Complex pharmaco-toxicological research presented erbium oxide nanostructure size of coherent scattering regions 26, 31, 56 and 65 nm showed that when administered orally no acute toxicity, no effect on the immune system of the body, has no effect on blood cells. But, long-term （30 day） intragastric administration shows toxicities on the internal organs of experimental animals, which lead to structural changes and functional impairment due to tissue accumulation of nanoparticles.

IMMUNOHISTOCHEMICAL DISTRIBUTION OF ERYTHROPOIETIN AND ITS RECEPTOR EXPRESSION IN POSTNATAL RAT RETINA DEVELOPMENT

Objective To investigate the distribution of erythropoietin （EPO） and erythropoietin receptor （ EPOR ） expression in the postnatal rat retina development. Methods Forty-two male Sprague-Dawley rats were divided into 7 groups according to their various postnatal days： postnatal 1 d （D1 group）, 3 d （D3 group）, I week （W1 group）, 2 weeks （W2 group）, 3 weeks （W3 group）, 4 weeks （W4 group） and8 weeks （W8 group） （ n = 6 ）. Single eye was randomly chosen from each rat for the study. The retinal sections were stained with hematoxylin and eosin （HE） and used for the retina development observation. Immunohistochemical staining was used to localize EPO and EPOR expressions in retinas.of differentstages of development, and the expression intensities were determined by an image plus 4 program~~ Results The retinal inner nuclear layer （INL） and outer nuclear layer （ONL） were mixed together and had not yet fully differentiated in D1 and D3 groups. The INL and ONL formed their own independent regions and the outer plexiform layer （OPL） appeared between two layers in W1 group. With the postnatal retinal development, the inner plexiform layer （ IPL ） , rods and cones layer （ RCL ）, and OPL were gradually widened and stabilized in W2 to W3 groups. EPO/EPOR expressions located prominently in the inner part of the postnatal rat developing retinas. The expression of EPO in GCL and INL gradually increased from DI to W4, then the expression decreased in W8. Expression of EPOR in GCL gradually increased from DI to WI , then decreased in W2 ; and it gradually increased again from W3 to W8. Expression of EPOR in INL gradually increased from D1 to W1, then decreased in W2 ; and it continued to decrease from W3 to W8. Expression of EPOR in the external segment of RCL gradually increased from D1 to W8. However, expression in the internal segment of RCL gradually decreased from D1 to W3 , then no obvious expression was seen in the internal segment of RCL in W4 and W8. Conclusion EPO/EPOR expressions locate prominently in the inner part of the postnatal rat developing retina. And EPO/EPOR expressions in the rat retinas exist the dynamic changes during the postnatal retina development period.

Anti-tumor effect of the extract from Qingyihuaji formula on pancreatic cancer by down-regulating Notch-4 and Jagged-1

OBJECTIVE： To investigate, in terms of Notch signaling pathway, the effect on pancreatic cancer of the extract of an anti-tumor prescription -- Qingyihuaji formula （QYHJ） -- from Traditional Chinese Medicine （TCM）.METHODS： Nude mice were implanted subcutaneously with human pancreatic cancer cell line SW1990 and then randomly divided into four groups： Control, QYHJ extract, Gemcitabine, and Combination of QYHJ extract and gemcitabine. Treatments were given for 21 days and tumor growth was evaluated simultaneously. Then, expression of Notch receptors （Notch-I, Notch-2, Notch-3, and Notch-4） and their Jagged ligands （Jagged-1 and Jagged-2） in dissected tumor tissue were detected by real-time quantitative reverse transcription-polymerase chain reaction and Western blot. Finally, immunohistochemistry was performed to detect CD133, a marker of pancreatic cancer stem cells （CSCs）, to evaluate the impact of QYHJ extract on pancreatic CSCs.RESULTS： QYHJ extract treatment effectively inhib- ited the tumor growth in nude mice. The expression of both Notch-4 and Jagged-1 were decreased significantly in QYHJ treatment groups （P 〈 0.05）, while gemcitabine alone had no significant effect in down-regulating Jagged-1 （P 〉 0.05）. No significant difference was observed in the ex- pression of Notch-1, Notch-2, Notch-3, and Jagged-2 between three treatment groups and control group （P 〉 0.05）. Moreover, immunohistochemical analysis showed that the number of CD133 positive cells was significantly reduced by QYHJ treatment （P 〈 0.05）, and the combined treatment was more effective than gemcitabine alone （P 〈 0.05）.CONCLUSION： The role of the extract in pancreatic cancer treatment was associated with down-regulation of Notch-4 and Jagged-1 in Notch signaling pathway. The extract could enhance the antitumor activity of gemcitabine and was more effective than gemcitabine in regulating Notch signaling pathway to some extent.

Applications of multiplexed immunohistochemistry/immunofluorescence and multispectral imaging technology in the field of tumor immunotherapy

Multiplexed immunohistochemistry/fluorescence(mIHC/IF)in combination with multispectral unmixing is a novel multitarget histopathological staining and imaging technique.By simultaneously revealing expression level and spatial information for up to eight biomarkers in situ,in addition to a nuclear stain within a single formalin-fixed paraffin-embedded(FFPE)tissue section,this technology can analyze the phenotype,abundance,morphology and intercellular relationship of cells while providing statistically significant results.In recent years,technical improvements have brought new insight into mIHC/IF and multispectral imaging approaches,which have been successfully applied in the field of cancer immunotherapy,specifically in regard to tumor microenvironment research,immunotherapy drug discovery,and prognostic and metastatic risk evaluation.This review highlights the principle,workflow,advantages and disadvantages of the technology,and discusses the latest applications of mIHC/IF-based imaging technology in the field of TME-related research and immunotherapy drug discovery.

STIM1 and Orai1:novel targets for vascular diseases?

The past five years have witnessed the discovery of the endoplasmic reticulum calcium(Ca2+) sensor STIM1 and the plasma membrane Ca2+channel Orai1 as the bona fide molecular components of the store-operated Ca2+ entry(SOCE) and the Ca2+ release-activated Ca2+current(I CRAC) .It has been known for two decades that SOCE and ICRAC are required for lymphocyte activation as evidenced by severe immunodeficient phenotypes in patients lacking ICRAC.In recent years however,studies have uncovered expression of STIM1 and Orai1 proteins in various tissues and described additional roles for these proteins in physiological functions and pathophysiological conditions.Here,we will summarize novel findings pertaining to the role of STIM1 and Orai1 in the vascular system and discuss their potential use as targets in the therapy of vascular disease.