口服免疫药物后中国对虾某些血淋巴因子的测定及方法研究

于1991年4－9月，以海捕中国对虾亲虾及养殖中国对虾为材料，运用单向免疫扩散（SRID）方法测定对虾血淋巴免疫因子，并采用经改进后的Horowitz等人的方法测定对虾体内的酚氧化酶（PO）活性，目的在于运用该两种方法对口服不同免疫药物的养殖对虾的血淋巴免疫因子及PO活力进行测定。对亲虾和养殖对虾口服不同免疫药物前后的测定结果表明，几种情况下的对虾血淋巴中均存在类似IgM的因子，血细胞中存在酚氧化酶原（ProPO）并可被激活；正常对虾的血淋巴中存在PO活力；口服免疫药物可以刺激IgM，proPO，PO活力发生变化。研究初步表明，血淋巴因子的SRID测定及PO活力测定可作为衡量对虾免疫状态的定性指标。

Gfi1.1 regulates hematopoietic lineage differentiation during zebrafish embryogenesis

Growth factor independence 1 （GFI1） is important for maturation of mammalian lymphocytes and neutrophils and maintenance of adult hematopoietic stem cells （HSCs）. The role of GFI1 in embryonic hematopoiesis is less well characterized. Through an enhancer trap screen and bioinformatics analysis, we identified a zebrafish homolog of Gill （named grill） and analyzed its function during embryonic development. Expression of both an endogenous griLl gene and a GFP reporter gene inserted near its genomic locus was detected in hematopoietic cells of zebrafish embryos. Morpholino （MO） knockdown of gill.1 reduced expression of scl, Imo2, c-myb, mpo, ragl, gatal and hemoglobin alpha embryonic-1 （hbael）, as well as the total amount of embryonic hemoglobin, but increased expression ofpu.1 and l-plastin. Under the same conditions, MO injection did not affect the markers involved in vascular and pronephric development. Conversely, overexpression of gill.1 via mRNA injection enhanced expression ofgatal but inhibited expression ofpu.1. These findings suggest that Gill.1 plays a critical role in regulating the balance of embryonic erythroid and myeloid lineage determination, and is also required for the differentiation of lymphocytes and granulocytes during zebrafish embryogenesis.

Expression of mucosal addressin cell adhesion molecule 1 on vascular endothelium of gastric mucosa in patients with nodular gastritis

AIM:The interaction of mucosal addressin cell adhesion molecule 1 (MAdCAM-1) with integrin α4β7 mediates lymphocyte recruitment into mucosa-associated lymphoid tissue (MALT).Nodular gastritis is characterized by a unique military pattern on endoscopy representing increased numbers of lymphoid follicles with germinal center,strongly associated with H pylori infection.The purpose of this study was to address the implication of the MAdCAM-1/integrin β7 pathway in NG. METHODS:We studied 17 patients with NG and H pylori infection and 19 H pylori-positive and 14 H pylori-negative controls.A biopsy sample was taken from the antrum and snap-frozen for immunohistochemical analysis of MAdCAM- 1 and integrin β7.In simultaneous viewing of serial sections, the percentage of MAdCAM-1-positive to von Willebrand factor-positive vessels was calculated.We also performed immunostaining with anti-CD20,CD4,CD8 and CD68 antibodies to determine the lymphocyte subsets co- expressing integrin β7. RESULTS:Vascular endothelial MAdCAM-1 expression was more enhanced in gastric mucosa with than without H pylori infection.Of note,the percentages of MAdCAM-1-positive vessels were significantly higher in the lamina propria of NG patients than in H pylori-positive controls.Strong expression of MAdCAM-1 was identified adjacent to lymphoid follicles and dense lymphoid aggregates.Integrin β7-expressing mononuclear cells,mainly composed of CD20 and CD4 lymphocytes,were associated with vessels lined with MAdCAM-1-expressing endothelium.CONCLUSION: Our results suggest that the MAdCAM一1/ integrin a4p7 homing system may participate in gastric inflammation in response to H py/o}i-infection and contributes to MALT formation, typically leading to the development of NG.

Relationship between LYVE-1, VEGFR-3 and CD44 gene expressions and lymphatic metastasis in gastric cancer

AIM: To investigate the expression levels of lymphatic vessel endothelial hyaluronan receptor-1 (LYVE-1), vascular endothelial growth factor receptor-3 (VEGFR-3) and CD44 genes and the relationship between their lev- els and clinicopathological parameters in gastric cancer.METHODS: Tissue samples were obtained from 33 patients (8 females) with gastric cancer. mRNA levels of LYVE-1, VEGFR-3 and CD44 in normal and tumor tissues were quantitatively measured using real time polymerase chain reaction. The results were correlated with lymph node metastasis, histological type and differentiation of the tumor, T-stage, and presence of vascular, perineural and lymphatic invasions. The distribution of molecules in the tissue was evaluated using immunohistochemistry. RESULTS: LYVE-1, CD44 and VEGFR-3 gene expression levels were significantly higher in gastric cancer than in normal tissue. While there was no correlation between gene expressions and clinicopathologic fea- tures such as histologic type, differentiation and stage, gene expression levels were found to be increased in conjunction with positive lymph node/total lymph node ratio and the presence of perineural invasion. A significant correlation was also found between LYVE-1 and CD44 over-expressions and perineural invasion and lymph node positivity in gastric cancers. When the dis- tribution of LYVE-1 antibody-stained lymphatic vessels in tissue was evaluated, lymphatic vessels were located intra-tumorally in 13% and peri-tumorally in 27% of the patients. Moreover, lymph node metastases were also positive in all patients with LYVE-1-staining. CONCLUSION: LYVE-1, VEGFR-3 and CD44 all play an important role in lymphangiogenesis, invasion and metastasis. LYVE-1 is a perfectly reliable lymphatic vessel marker and useful for immunohistochemistry.

Association of serum vascular endothelial growth factor-C and lymphatic vessel density with lymph node metastasis and prognosis of patients with gastric cancer

AIM： To investigate whether serum vascular endothelial growth factor-C （SVEGF-C）, VEGF-C, and lymphatic vessel density （LVD） in tumor tissues are related to lymph node metastasis （LNM） and prognosis in gastric cancer. METHODS： SVEGF-C levels of 80 gastric cancer patients and 20 healthy donors were examined using ELISA. VEGF-C expression and LVD were examined using immunohistochemical staining. Kaplan-Meier survival analysis was performed to determine their influence on the prognosis of the patients. RESULTS： The SVEGF-C level in gastric cancer patients （595.9 ± 201.0 ng/L） was significantly higher （P = 0.000） than controls （360.0 ± 97.4 ng/L）. Both SVEGF-C and LVD were significantly higher in poorly differentiated adenocarcinomas, T3 and T4, LNM, distant metastasis, and pTNM groups Ⅲ and Ⅳ （P = 0.000）. The sensitivity and specificity of SVEGF-C for predicting LNM were 82.8% and 81.8%, respectively （cut-off = 542.5 ng/L）. The positive expression rate of VEGF-C was significantly higher in cancerous than in normal tissues （65% vs 20%; P = 0.001）. VEGF-C expression up-regulation was significantly related to differentiation, depth of invasion, LNM, distant metastasis, and pTNM stage （P = 0.000）. LVD was 10.7 ± 3.1/200 HP in the experimental group vs 4.9 ± 1.3/200 HP in controls （P = 0.000）; LVD in cancerous tissues with and without LNM was 12.0 ± 2.7/200 HP vs 7.6 ± 0.5/200 HP, respectively （P = 0.000）. SVEGF-C and LVD were significantly higher in VEGF-C positive than in negative patients （P = 0.000）; SVEGF-C level was related to LVD （P = 0.000）. Kaplan-Meier survival analysis factors predicating poor prognosis were： SVEGF-C level （P = 0.001）, VEGF-C expression and LVD （both P = 0.000）. CONCLUSION： SVEGF-C level, VEGF-C and LVD are related to LNI and poor prognosis of patients with gastric cancer, SVEGF-C may be a biomarker for LNI in gastric cancer,

Metastasis-associated protein 1 induces VEGF-C and facilitates lymphangiogenesis in colorectal cancer

AIM:To study the correlation between high metastasisassociated protein 1(MTA1)expression and lymphangiogenesis in colorectal cancer(CRC)and its role in production of vascular endothelial growth factor-C(VEGF-C). METHODS:Impact of high MTA1 and VEGF-C expression levels on disease progression and lymphovasculardensity(LVD,D2-40-immunolabeled)in 81 cases of human CRC was evaluated by immunohistochemistry. VEGF-C mRNA and protein expressions in human LoVo and HCT116 cell lines were detected by real-time polymerase chain reaction and Western blotting,respectively,with a stable expression vector or siRNA. RESULTS:The elevated MTA1 and VEGF-C expression levels were correlated with lymph node metastasis and Dukes stages(P<0.05).Additionally,high MTA1 expression level was correlated with a large tumor size(P< 0.05).A significant correlation was found between MTA1 and VEGF-C protein expressions in tumor cells(r=0.371, P<0.05).Similar to the VEGF-C expression level,high MTA1 expression level was correlated with high LVD in CRC(P<0.05).Furthermore,over-expression of MTA1 significantly enhanced the VEGF-C mRNA and protein expression levels,whereas siRNAs-knocked down MTA1 decreased the VEGF-C expression level. CONCLUSION:MTA1,as a regulator of tumor-associated lymphangiogenesis,promotes lymphangiogenesis in CRC by mediating the VEGF-C expression.

Interleukin-24 is correlated with differentiation and lymph node numbers in rectal cancer

AIM:To assess the significance of interleukin(IL)-24 and vascular endothelial growth factor(VEGF)expression in lymph-node-positive rectal cancer. METHODS:Between 1998 and 2005,90 rectal adenocarcinoma patients with lymph node involvement were enrolled.All patients received radical surgery and postoperative pelvic chemoradiotherapy of 50.4-54.0 Gy.Chemotherapy of 5-fluorouracil and leucovorin or levamisole was given intravenously during the first and last week of radiotherapy,and then monthly for about 6 mo.Expression of IL-24 and VEGF was evaluated by immunohistochemical staining of surgical specimens, and their relations with patient characteristics and survival were analyzed.The median follow-up of surviving patients was 73 mo(range:52-122 mo). RESULTS:IL-24 expression was found in 81 out of 90 patients;31 showed weak intensity and 50 showedstrong intensity.VEGF expression was found in 64 out of 90 patients.Negative and weak intensities of IL-24 expression were classified as negative expression for analysis.IL-24 expression was significantly reduced in poorly differentiated tumors in comparison with well or moderately differentiated tumors(P=0.004),N2b to earlier N stages(P=0.016),and stageⅢc to stageⅢ a orⅢb(P=0.028).The number of involved lymph nodes was also significantly reduced in IL-24-positive patients in comparison with IL-24-negative ones. There was no correlation between VEGF expression and patient characteristics.Expression of IL-24 and VEGF was not correlated with survival,but N stage and stages were significantly correlated with survival. CONCLUSION:IL-24 expression was significantly correlated with histological differentiation,and inversely correlated with the degree of lymph node involvement in stageⅢrectal cancer.

Colitis associated with biological agents

In the past,there has been considerable focus on a host of drugs and chemicals that may produce colonic toxicity.Now,a variety of new biological monoclonal antibody agents,usually administered by infusion,have appeared in the clinical realm over the last decade or so to treat different chronic inflammatory or malignant disorders.For some of these agents,adverse effects have been documented,including apparently new forms of immune-mediated inflammatory bowel disease.In some,only limited symptoms have been recorded,but in others,severe colitis with serious complications,such as bowel perforation has been recorded.In others,adverse effects may have a direct vascular or ischemic basis,while other intestinal effects may be related to a superimposed infection.Some new onset cases of ulcerative colitis or Crohn's disease may also be attributed to the same agents used to treat these diseases,or be responsible for disease exacerbation.Dramatic and well documented side effects have been observed with ipilimumab,a humanized monoclonal antibody developed to reduce and overcome cytotoxic T-lymphocyte antigen 4,a key negative feedback regulator of the T-cell anti-tumor response.This agent has frequently been used in the treatment of different malignancies,notably,malignant melanoma.Side effects with this agent occur in up to 40% and these are believed to be largely immune-mediated.One of these is a form of enterocolitis that may be severe,and occa-sionally,fatal.Other agents include rituximab(an antiCD20 monoclonal antibody),bevacizumab(a monoclonal antibody against the vascular endothelial growth factor) and anti-tumor necrosis factor agents,including infliximab,adalimumab and etanercept.

Retinoic acid regulated insulin-like growth factor gene expression in cord blood lymphocytes

Objective To explore the immune promotion mechanism of retinoic acid (RA).Methods Reverse Transcription-Polymerase chain reaction (RT-PCR) was used to detect the gene expression of insulin like growth factor Ⅰ (IGF-Ⅰ) and IGF receptors (IGF-IR and IGF-IIR) in vitro, and their regulation by RA in human cord blood lymphocyte (CBL). Results Significant upregulation of IGF-I, IGF-IR and IGF-IIR mRNA was found at 6-24 hours in CBL incubated with physiologic concentrations of RA as compared to those without RA. Conclusion The enhancement of IGF expression may be an important pathway for vitamin A to promote immune cellular functions.

A clinical study of Weining Granules in the treatment of gastric precancerous lesions

OBJECTIVE:To investigate the clinical effects of Weining Granules on gastric precancerous lesions(GPLs).METHODS:120 patients with GPLs were randomly assigned in a 1:1 ratio to receive Weining Granules(trial group) or the comparator Weifuchun tablets(control group) for 6 months.Outcomes were compared between the two groups including:overall response;gastroscopically-determined response;pathologically-confirmed response;eradication of Helicobacter pylori(HP);microvessel density(MVD) in the gastric mucosa;expression of vascular endothelial growth factor(VEGF);interleukin 2(IL-2);interleukin 6(IL-6);T lymphocyte subsets;immunoglobulins;symptom scores;quality of life(QOL);and adverse reactions.RESULTS:Patients in the trial group had a significantly higher(P<0.05) overall response rate(81.7%) as compared with those in the control group(63.3%).Relative to treatment with Weifuchun tablets treatment with Weining Granules resulted in a significant improvement(P<0.05) in the scores for gastric pain,distension and stuffiness in the hypochondrium,and anorexia.As compared with the tablets the Granules were associated with a significantly higher overall gastroscopically-determined response rate(78.3%;P<0.05).Pathological examination of tissue samples indicated that 61.7% of patients receiving the granules were cured with an overall response rate of 75.5%;these rates were significantly higher than in the control group(P<0.05).In comparison with patients receiving the tablets,those given the granules were significantly more likely to have their HP eradicated(75.0% vs.51.4%;P<0.05).Improvements in MVD,VEGF,CD4+,CD4+/CD8+,IL-2,IL-6 and IgG were significantly greater with the Weining Granules than with the Weifuchun tablets(P<0.05 or P<0.01).After follow-up of 1 year,17.5% of patients in the trial group relapsed as compared with 39.5% in the control group(P<0.05).Relative to the control group,the trial group showed significantly greater improvements in physical,psychological and social relationships,and in environmental domains(P<0.05 or P<0.01).No significant adverse reactions were observed during treatment.CONCLUSION:The Weining Granules appear to be effective in improving the gastric precancerous state and the main symptoms,in inhibiting angiogenesis,enhancing immune function and QOL,and in reducing 1-year relapses.In addition,this preparation seems to be associated with a low risk of adverse events,making it a safe and efficacious option for the treatment of GPLs.

Efficacy and effect on serum VEGF-C of mild moxibustion plus functional exercise for upper-limb lymphedema after breast cancer surgery

Objective To observe the efficacy of mild moxibustion combined with functional exercise in the treatment of upper-limb lymphedema after breast cancer surgery and its effect on serum vascular endothelial growth factor C(VEGF-C).Methods Seventy-eight patients were divided into a control group and an observation group by the random number table method,with 39 cases in each group.The control group received functional exercise and the observation group received mild moxibustion plus functional exercise.The differences in circumference between the two upper limbs,the lymphatic flow of the affected upper limb,the disability of arm,shoulder and hand(DASH)score,the functional assessment of cancer therapy-breast(FACT-B)score and serum VEGF-C level between the two groups were compared before and after treatment.Efficacy was evaluated after treatment.Results The total effective rate was significantly higher in the observation group than in the control group(P<0.05).The difference in circumference between the two upper limbs of the two groups decreased significantly after treatment(both P<0.05),but it was significantly lower in the observation group than in the control group(P<0.05).The lymphatic flow of the affected upper limb of the two groups increased significantly after treatment(both P<0.05),but it was significantly greater in the observation group than in the control group(P<0.05).The DASH scores of the two groups decreased significantly after treatment(both P<0.05),but it was significantly lower in the observation group than in the control group(P<0.05).The FACT-B scores of the two groups increased significantly after treatment(both P<0.05),but it was significantly higher in the observation group than in the control group(P<0.05).After treatment,the serum VEGF-C level increased significantly in the observation group(P<0.05),whereas the control group did not show significant change(P>0.05).The post-treatment serum VEGF-C level was significantly higher in the observation group than in the control group(P<0.05).Conclusion The efficacy of mild moxibustion combined with functional exercise for upper-limb lymphedema after breast cancer surgery is certain,which can reduce the difference in circumference between the two upper limbs,increase the lymphatic flow of the affected upper limb,improve the limb function and the quality of life,and regulate the serum VEGF-C level.