Rapidcal. Immuno在骨髓脱钙中的应用

近年来血液病呈高发趋势,骨髓穿刺活检更广泛地应用于临床。骨髓活检标本制片的质量及免疫组化染色的准确表达,对骨髓病理诊断起着极其重要的作用。骨髓组织中含有丰富的钙盐、脂肪及大量造血细胞,成分复杂,软硬程度不一致。在去除不利于制片的钙盐成分的同时,又要保证造血细胞形态的完好及其免疫原性的完整保留。

Preliminary Study on Immuno-gold Chromatography Assay for Rapid Detection of Vibrio parahaemolyticus

[ Objective] Vibrio parahaemolyticus was chosen as the material to make rabbit anti- V. parahaemolyticus polyclonal antibody, the immuno-gold chromatography assay （IGCA） was used to develop immunogold labeling test strips for detecting V. parahaemolyticus. [Methods] Rabbit anti-V, parahaemolyticus IgG-2 was used to wrap nitrocellulose membrane detection line, goat anti-rabbit IgG was used to wrap control line, and im- muno-gold was used to label rabbit anti-V, parahaemolyticus IgG-1 to establish the immuno-gold chromatography assay （IGCA） for detection of V. parahaemolyticus. [Results] Test strip detection lines of positive results along with the control line were all red, test results could be got in only five to fifteen minutes, the minimum detectable amount of this method to V. parahaemolyticus was 3.60 x 104 cfu/ml, and cross reactions wouldnt occur when detecting common intestinal bacteria like Vibrio alginolyticus, vibrio cholerae, Vibrio damsela, Vibrio metschnikovii, Citrobacter freundii and salmonella etc. The test results were undifferentiated when the test strips were stored at 4 ~C for six months, stored at room temperature for three months or stored at 37 ~C for one month. [ Condusionl The established test strip assay was simple and rapid during operation, with high sensitivity, strong specificity and good stability, the test results were easy to be observed and judged, and this assay was very suitable for grassroots breeding department application.

Overview of cytokines and nitric oxide involvement in immuno-pathogenesis of inflammatory bowel diseases

Inflammatory bowel diseases(IBDs), including Crohn's disease and ulcerative colitis are complex disorders with undetermined etiology. Several hypotheses suggest that IBDs result from an abnormal immune response against endogenous flora and luminal antigens in genetically susceptible individuals. The dysfunction of the mucosal immune response is implicated in the pathogenesis of IBD. The balance between pro-inflammatory cytokines [tumor necrosis factor(TNF)-α, interleukin(IL)-1b, IL-8, and IL-17A], anti-inflammatory cytokines(IL-4 and IL-13), and immunoregulatory cytokines(IL-10 and transforming growth factors b) is disturbed. Moreover, evidence from animal and clinical studies demonstrate a positive correlation between an increased concentration of nitric oxide(NO) and the severity of the disease. Interestingly, proinflammatory cytokines are involved in the up-regulation of inducible oxide synthase(iN OS) expression in IBD. However, anti-inflammatory and immunoregulatory cytokines are responsible for the negative regulation of iN OS. A positive correlation between NO production and increased pro-inflammatory cytokine levels(TNF-α, IL-6, IL-17, IL-12, and interferon-γ) were reported in patients with IBD. This review focuses on the role of cytokines in intestinal inflammation and their relationship with NO in IBD.

Immunoregulatory Effects of Ethyl-acetate Fraction of Extracts from Tetrastigma Hemsleyanum Diels et. Gilg on Immune Functions of ICR Mice

Objective To evaluate the effects of ethyl-acetate fraction （EAF） of extracts from Tetrastigma hemsleyanum Diels et. Gilg （TDG） on immune functions of ICR mice. Methods ICR mice were exposed to different doses of EAF for 15 or 30 days and then their immune functions were analyzed, including ConA-induced splenic lymphocyte transformation, SRBC- induced delayed type hypersensitivity response, serum hemolysin analysis, antibody-producing cells, peritoneal macrophage phagocytized chicken red blood cells, natural killer cell activity, and serum level of cytoldnes. Results EAF of extracts from TDG at different doses had various effects on immune functions of ICR mice. As compared with the controls, it increased the mouse spleen lymphocyte transformation induced by ConA, the left-hind voix pedis thickness and the number of plague forming cells （PFCs） at the dose of 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the ink clearance ability at the dose of 0.91 mg/mL, 1.82 mg/mL, 5.48 mg/mL, and 9.12 mg/mL, respectively; increased the phagocytosis index of mononuclear-macrophages and production of serum interferon-gamma （IFN-？） at the dose of 5.48 mg/mL; and could promote the production of serum tumor necrosis factor-alpha （TNF-α） at the dose of 9.12 mg/mL. Conclusion EAF of extracts from TDG can regulate mouse immune functions in vivo.

Rice bran hydrolysates induce immunomodulatory effects by suppression of chemotaxis, and modulation of cytokine release and cell-mediated cytotoxicity

Objective: To evaluate the immunomodulatory effects of rice bran hydrolysates on cultured immune cells and their underlying mechanism.Methods: Rice bran hydrolysates were prepared from pigmented rice(Oryza sativa L.) by hydrothermolysis and protease digestion. Rice bran hydrolysates were assayed for phenolic content and antioxidant activity. Cell proliferation of Jurkat, THP-1 and peripheral blood mononuclear cells(PBMC) was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Chemotaxis was evaluated by transwell chamber methods. Immunoadherence of THP-1 was performed on cultured human umbilical vein endothelial cells(HUVEC). Cytokine released from PBMC was measured by ELISA assay kits. Lymphocyte-mediated cytotoxicity was carried out on KKU-452 cells. Proteins associated with immunomodulation were analyzed by Western immunoblotting assay. Results: Rice bran hydrolysates were rich in phenolic compounds, such as ferulic acid, catechin, quercetin, and quercetin glycosides. Rice bran hydrolysates suppressed phytohemagglutinin(PHA)-stimulated proliferation of PBMC and Jurkat cells, chemotaxis of Jurkat and THP-1 cells, and immunoadherence of THP-1 on HUVEC cultured cells. The cellular mechanism of rice bran hydrolysates involved the activation of AMPK as well as suppression of m TOR, NF-κB and VCAM-1. Rice bran hydrolysates potentiated PBMC on the PHA-stimulated release of IL-2, TNF-α, and IL-4, and enhanced PHA-induced non-MHC-restricted cytotoxicity on KKU-452 cancer cells. Conclusions: The immunomodulatory effect of phytochemicals derived from rice bran hydrolysates suggests its therapeutic potential for further investigation.

Colorectal cancer screening: Comparison of transferrin and immuno fecal occult blood test

AIM: To evaluate the sensitivity and specificity of transfesrrin dipstick test (Tf) in colorectal cancer (CRC) screening and precancerous lesions screening. METHODS: Eight hundreds and sixty-one individuals at high-risk for CRC were recruited. Six hundreds and eleven subsequently received the three fecal occult blood tests and colonoscopy with biopsy performed as needed. Fecal samples were obtained on the day before colonoscopy. Tf, immuno fecal occult blood test (IFOBT) and guaiac fecal occult blood test (g-FOBT) were performed simultaneously on the same stool. To minimize false-negative cases, all subjects with negative samples were asked to provide an additional stool specimen for a second test even a third test. If the results were all negative after testing three repeated samples, the subject was considered a true negative. The performance characteristics of Tf for detecting CRC and precancerous lesions were examined and compared to those of IFOBT and the combination of Tf, IFOBT and g-FOBT. RESULTS: A total of six hundreds and eleven subjects met the study criteria including 25 with CRC and 60 with precancerous lesions. Sensitivity for detecting CRC was 92% for Tf and 96% for IFOBT, specificities of Tf and IFOBT were both 72.0% (95% CI: 68.2%-75.5%; χ2 = 0.4, P > 0.05); positive likelihood ratios of those were 3.3 (95% CI: 2.8-3.9) and 3.4 (95% CI: 2.9-4.0), respectively. In precancerous lesions, sensitivities for Tf and IFOBT were 50% and 58%, respectively (χ 2 = 0.8, P > 0.05); specificities of Tf and IFOBT were 71.5% (95% CI: 67.6%-75.1%) and 72.2% (95% CI: 68.4%-75.8%); positive likelihood ratios of those were 1.8 (95% CI: 1.3-2.3) and 2.1 (95% CI: 1.6-2.7), respectively; compared to IFOBT, g-FOBT+ Tf+ IFOBT had a significantly higher positive rate for precancerous lesions (83% vs 58%, respectively; χ 2 = 9.1, P < 0.05). In patients with CRC and precancerous lesions, the sensitivities of Tf and IFOBT were 62% and 69% (χ 2 = 0.9, P > 0.05); specificities of those were 74.5% (95% CI: 70.6%-78.1%) and 75.5% (95% CI: 71.6%-79.0%); positive likelihood ratios of those were 2.5 (95% CI: 2.0-3.1) and 2.8 (95% CI: 2.3-3.5). Compared to IF-OBT alone, combining g-FOBT, IFOBT and Tf led to significantly increased sensitivity for detecting CRC and cancerous lesions (69% vs 88%, respectively; χ 2 = 9.0, P < 0.05). CONCLUSION: Tf dipstick test might be used as an ad- ditional tool for CRC and precancerous lesions screening in a high-risk cohort.

Systemic study on the safety of immuno-deficient nude mice treated by atmospheric plasma-activated water

Cold atmospheric-pressure plasma is a new technology, widely used in many fields of biomedicine,especially in cancer treatment. Cold plasma can selectively kill a variety of tumor cells, and its biological safety in clinical trials is also very important. In many cases, the patient’s immune level is relatively low, so we first studied the safety assessment of plasma treatment in an immunocompromised animal model. In this study, we examined the safety of immuno-deficient nude mice by oral lavage treatment of plasma-activated water, and studied the growth status, main organs and blood biochemical indexes. Acute toxicity test results showed that the maximum dose of plasma treatment for 15 min had no lethal effect and other acute toxicity. There were no significant changes in body weight and survival status of mice after 2 min and 4 min of plasma-activated water(PAW)treatment for 2 weeks. After treatment, the major organs, including heart, liver, spleen, lung and kidney, were not significantly changed in organ coefficient and tissue structure. Blood biochemical markers showed that blood neutrophils and mononuclear cells were slightly increased, and the others remained unchanged. Liver function, renal function, electrolytes, glucose metabolism and lipid metabolism were not affected by different doses of PAW treatment. The above results indicate that PAW treatment can be used to treat immuno-deficient nude mice without significant safety problems.

Synthesis and evaluation of ^(64)Cu-radiolabeled NOTA-cetuximab(^(64)Cu-NOTA-C225) for immuno-PET imaging of EGFR expression

Objective: Epidermal growth factor receptor(EGFR) is overexpressed in a wide variety of solid tumors, serving as a well-characterized target for cancer imaging or therapy. In this study, we aimed to design and synthesize a radiotracer, ^(64) Cu-NOTA-C225, targeting EGFR for tumor positron emission tomography(PET) imaging.Methods: Cetuximab(C225) was conjugated to a bifunctional chelator, p-isothiocyanatobenzyl-1,4,7-triazacyclononane-1,4,7-triacetic acid(NOTA), and further radiolabeled with copper-^(64) for PET imaging. ^(64) CuNOTA-IgG and Cy5.5-C225 were also synthesized as control probes. A431 and A549 mouse models were established for micro-PET and/or near-infrared fluorescence(NIRF) imaging.Results: ^(64) Cu-NOTA-C225 exhibited stability in vivo and in vitro up to 24 h and 50 h post-injection,respectively. A431 tumors with average standard uptake values(SUVs) of 5.61±0.69, 6.68±1.14, 7.80±1.51 at 6, 18 and 36 h post-injection, respectively, which were significantly higher than that of moderate EGFR expressing tumors(A549), with SUVs of 0.89±0.16, 4.70±0.81, 2.01±0.50 at 6, 18 and 36 h post-injection, respectively. The expression levels of A431 and A549 were confirmed by western blotting. Additionally, the tracer uptake in A431 tumors can be blocked by unlabeled cetuximab, suggesting that tracer uptake by tumors was receptor-mediated.Furthermore, NIRF imaging using Cy5.5-C225 showed that the fluorescence intensity in tumors increased with time, with a maximal intensity of 8.17 E+10(p/s/cm^2/sr)/(μW/cm^2) at 48 h post-injection, which is consistent with the paradigm from micro-PET imaging in A431 tumor-bearing mice.Conclusions: The ^(64) Cu-NOTA-C225 PET imaging may be able to specifically and sensitively differentiate tumor models with different EGFR expression levels. It offers potentials as a PET radiotracer for imaging of tracer EGFR-positive tumors.

Preparation of Immuno-magnetic Beads and Their Separation & Detection to Ovary Cancer Cells

The organic monomer-molecule with nanometer magnetic powder by means of reforming the surface of nanometer magnetic powder have been synthesized. Magnetic beads in diameter of 2 mum or so am obtained by controlling conditions. Ovary cancer cells of ascites are separated and ovary cancer cells of blood arc detected by using immuno-magnetic beads linked with ovary cancer cell mono-antibodies. Results show that the specificity is 85% sensitivity is 87%, accuracy is 84%, cells acquiring purity is 90%, cells activity is 92% and detection sensitivity is 25 x 10(-7).

STUDY OF ENHANCED IMMUNOGENECITY OF B7-1 GENE TRANSFECTED HUMAN HELA CELL LINE

This work was supposed by CMB (No. 96—635) This is one of papers of the special issue on gene therapy research (Chin J Cancer Res Vol. 9 No. 4 December, 1997). Although cervical carcinoma cells may express the human papillomavirus protein E6 and E7, they fail to induce an effective specific cytotoxic T lymphocyte response. Recent studies suggest that expression of CD 80 (B7 1) on tumor cells is effective to induce antitumor immune responses. 1,2 In our study, CD 80 gene was transfected into human Hela cell line with a CD 80 expression plasmid (B7 1 +pcDNA 3) by electroporation, then the immunogenecity of the modified Hela cell was tested in TLMC (tumor lymphocyte mixed culture) system. Thymidine lymphocyte proliferation assays showed that the response of human peripheral blood lymphocytes (PBLS) to CD 80 positive Hela cells demonstrated a substantial increase in cell proliferation compared to the response to control cells. Cocultivation of allogeneic PBLs with CD 80 positive tumor cells for three days can induce an increased secretion of IL 2. Our results demonstrate an immunostimulatory effect of CD 80 expression on cervical cancer cells, which provides a basis for the development of a therapeutic tumor vaccine.

Lactobacillus isolates from healthy volunteers exert immunomodulatory effects on activated peripheral blood mononuclear cells

As probiotics in the gut, Lactobacilli are believed to play important roles in the development and maintenance of both the mucosal and systemic immune system of the host. This study was aimed to investigate the immunomodulatory function of candiate lactobacilli on T cells. Lactobacilli were isolated from healthy human feces and the microbiological characteristics were identified by API 50 CHL and randomly amplified polymorphic DNA （RAPD） assays. Anti-CD3 antibody activated peripheral blood mononuclear cells （PBMCs） were treated by viable, heat-killed lactobacilli and genomic DNA of lactobacilli, and cytokine profiles were tested by ELISA. Isolated lactobacilli C44 and C48 were identified as L. acidophilus and L. paracacei, which have properties of acid and bile tolerance and inhibitor effects on pathogens. Viable and heat-killed C44 and C48 induced low levels of proinflammatory cytokines （TNF-a, IL-6 and IL-8） and high levels of IFN-y and IL-12p70 in PBMCs. In anti- CD3 antibody activated PBMCs, viable and heat-killed C44 increased Th2 cytokine levels （IL-5, IL-6 and IL-10）, and simultaneously enhanced Thl responses by inducing IFN-y and IL-12p70 production. Different from that of lactabacillus strains, their genomic DNA induced low levels of IL-12p70, IFN-y and proinflammatory cytokines in PBMCs with or without anti-CD3 antibody activation. These results provided in vitro evidence that the genomic DNA of strains of C44 and C48, especially C44, induced weaker inflammation, and may be potentially applied for treating allergic diseases.

Baseline survey of pregnant women's knowledge of treatment to prevent mother- to-child human immunodeficiency virus transmission in a resource limited setting

Despite continuing advances in scientific knowledge about treatments to prevent mother-to-child(MTCT)of the human immune deficiency virus(HIV),there is a paucity of data on pregnant women’s knowledge of treatments to prevent MTCT of HIV in Aba,South Eastern Nigeria.We investigated pregnant women’s knowledge of the availability of treatments to prevent MTCT of HIV in the low resourced setting of Aba town in South Eastern Nigeria.A descriptive study involving 100 consecutive and consenting patients at the antenatal clinic of Abia State University Teaching Hospital(ABSUTH),Aba,South Eastern Nigeria over the period 1st November, 2007 to 30th November,2007.Using a structured questionnaire,the respondents’sociodemographic data were obtained as well as their knowledge of the availability of treatments to prevent mother to child HIV transmission in pregnant women and self-reported data on their having ever tested for HIV.Knowledge regarding availability of treatment to prevent mother to child transmission of HIV was good as 95%of the respondents were aware that such treatment was available.Knowledge that treatment is available to help someone infected by HIV to live longer was also good as 96% of the respondents were aware of the availability of such treatment.Seventy one percent of the respondents had ever tested for HIV.The levels of knowledge regarding the availability of treatment to prevent mother to child HIV transmission and to help someone infected with HIV are good.Thus, utilization of anti-retroviral prophylaxis amongst HIV infected pregnant women in this community would be expected to be high if the drugs were available.This will reduce the impact of HIV,especially in children.

Immunological protection of small bowel by simultaneously transplanted liver graft in pigs

BACKGROUND: A simultaneously transplanted liver shields a bowel graft from immunologic attack in small animals, while the possible immuno-tolerance induced by the liver in liver and small bowel transplantation (LSBT) is uncertain in large animal models. To investigate the clinically suspected beneficial effect of the liver on small bowel allograft, we developed a new model of composite LSBT in the pig. METHODS: Seventy outbred long-white pigs were randomized into four groups. LSBT without immunosuppressive treatment (n=10, group A); LSBT with routine immunosuppressive treatment (n=10, group B); LSBT with a lower dose of immunosuppressive treatment (n=10, group C); and small bowel segment allotransplantation without immunosuppressive treatment (n=10, group D). RESULTS: There was no remarkable difference in survival time between groups A and D (10.33 vs. 12.89 days, P>0.05), but the initial time of acute rejection of the intestinal graft in group A was clearly delayed when compared to group D (8.22 vs. 4.33 days, P<0.05), and the rejection scores in group A were remarkably lower than those in group D at each postoperative time point (0 vs. 0.44 on day 3, P<0.05; 0.22 vs. 1.78 on day 5, P<0.05; 1.11 vs. 2.56 on day 7, P<0.05). There were evident differences in postoperative survival time, initial time of acute rejection and postoperative rejection scores between groups A, B and C. Postoperative survival time (30.00 vs. 28.13 days, P>0.05), initial acute rejection time (25.40 vs. 22.13 days, P>0.05) or rejection score did not differ between groups B and C within one postoperative month. CONCLUSIONS: Compared to isolated segment small bowel allotransplantation, the intestinal graft in LSBT (group A) had a delayed initial time of acute rejection and a lower postoperative acute rejection score, and a lower dose of immunosuppressive treatment led to persistent graft immuno-tolerance in LSBT. Thus the simultaneously transplanted liver graft may reduce the risk of intestinal rejection and protect the bowel graft from severe acute rejection.

ESTABLISHMENT OF IMMUNO－PCR TECHNIQUE FOR THE DETECTION OF TUMOR ASSOCIATED ANTIGEN MG＿7－Ag ON THE GASTRIC CANCER

The gastric cancer associated antigen MG_7-Ag was detected by means of a newly established method,termed Immuno-PCR A McAb-recombinant DNA chimeric molecule was made which possesses bispecific binding affinity for antigen that had been immobilized on microtiter wells and the segment of the attached DNA was amplified by PCR. The antigen of gastric cancer cell line KATO III was monitored by this method. Analysis of PCR products by agarose gel electrophoresis after staining with ethidium bromide allowed as few as 20 cells to be detected readily and reproducibly. Immuno-PCR showed a 104 enhancement in detection sensitivity compared with ELISA assay. When the same numbers of cells ( 2×10e/ml ) were immobilized and then the serial diluted chimeric molecule was added. 3.8×10-14 moles and 3.0 × 10-11 moles were needed to give positive results with the Immuno-PCR and ELISA assay. respectively.Therefore, Immuno-PCR could give an enomous amplification capability with good specificity, and has a sensitivity much higher than any existing techniques for antigen detection.

A Novel Immunological Technique for Detection of Gramicidin (S) Producing Bacteria

A serological kit was prepared for the first time to detect bacteria that produce Gramicidin (S). Since, an immunocomplex of antibiotic Gramicidin (S) was prepared followed by immunonization of laboratory animales to obtain specific antibodies in their serum. Serial dilutions of the prepared antibodies were tested directly against samples of bacteria and compared with control samples, which include bacteria that produce Gramicidin (S) (as a positive control) and the bacteria that does not produce Gramicidin (S) (as a negative control) depending slide method after the incubation of bacteria in a suitable media for 3 - 4 hours at 37°C. The prepared kites were evaluated in reference Laboratories and they proved to be highly sensitive (100%), specific (100%), more economic and reliable procedure, in addition to its stability for more than one year. It was the first time to prepare a kit for detection of bacteria which produce Gramicidin (S) directly within few minutes.

Usefulness of Immuno-Magnetic Beads Conjugated with Anti-EpCAM Antibody for Detecting Endometrial Cancer Cells

A simple and non-invasive method for detecting endometrial cancer in women with abnormal uterine bleeding is required. For this purpose, we prepared immuno-magnetic beads conjugated with anti-human EpCAM rat monoclonal antibody (mAb) for isolating exfoliated endometrial cells including endometrial cancer cells in vaginal discharge. The affinities of the anti-human EpCAM rat mAbs were analyzed by flow cytometry and immunocytochemistry and then magnetic beads were conjugated with the mAbs. The rate of retrieval of endometrial cells using the immuno-magnetic beads was calculated. Endometrial cells were isolated using the immuno-magnetic beads from the vaginal discharges of 22 patients with endometrial cancer and 16 non-malignant controls. The isolated cells were stained using endometrial cancer specific-mAbs and analyzed by flow cytometry and imaging cytometry. The immuno-magnetic beads conjugated with high-affinity mAb (clone 1456) appeared to have very low auto-fluorescence. Sufficient enrichment of Ep-CAMpositive cells using immuno-magnetic beads was observed in both simulation and clinical samples. The overall sensitivities of flow cytometry and imaging cytometry to detect endometrial cancer cells were 72.7% and 45.5%, respectively. Meanwhile, the overall specificities of flow cytometry and imaging cytometry for healthy controls were 75.0% and 81.3%, respectively. Our immuno-magnetic beads have very low auto-fluorescence, so they could be useful for fluorescent analysis, such as fluorescent immunochemical staining. In the future, these novel immuno-magnetic beads could be used for cytological study.

Immuno-Chemotherapy Using Repeated Vaccine Treatment Can Produce Successful Clinical Responses in Advanced Metastatic Melanoma

Advanced Stage IV and IIIc melanoma has a dismal survival, with or without, standard chemotherapy. New therapies are required to improve survival and reduce morbidity. Repeated vaccine dosing does not appear to have been explored, so Vaccinia Melanoma Cell Lysate (VMCL) vaccine repetitive therapy was tested, either alone, or combined with chemotherapy. 37 patients (31 Stage IV [M1a(6), b(7), c(18)] and 6 Stage IIIc) were studied using intra-dermal VMCL vaccine therapy. If disease progressed, vaccine was continued with standard chemotherapy (DTIC and/or Fotemustine). Overall survival was assessed and clinical responses were also recorded. From vaccine commencement, median overall follow-up was 10 months. Survivals ranged from 4 to 73 months. Median (mean) overall survival was 10 (23.5) months;overall survival at 1, 2 and 3 years was 40.5%, 21.6% and 10.8% respectively. CR and PR occurred in 18.9% (7) and 18.9% (7) of patients;these were durable for up to 6.1 years in 4 patients. Stable disease was noted in a further 17 patients (45.9%). In 6 patients (16.2%) no response to therapy was apparent. Repeated vaccinations with or without chemotherapy produced strong, durable clinical responses with overall survival > 23 months occurring in nearly 25% of advanced melanoma patients. The overall disease control rate (CR, PR and SD) was 83.7%, including CR in very advanced cases. These results, in a largely unselected population of advanced metastatic melanoma patients, compare very favourably with other regimens, and notably were associated with minimal, if any, toxicity. Further analysis of this approach appears warranted.

The immunoreactivity of Cra a 4 decreased after Maillard reaction with xylose and glucose

Cra a 4 is a heat stable allergen in oyster,the effect of Maillard reaction on the immunoreactivity and structure of Cra a 4 was investigated.Firstly,Cra a 4 was cloned,expressed and identified.The purified Cra a 4 was incubated with xylose at 100℃,pH 8.5 for 30 min or with glucose for 60 min.After Maillard reaction,the IgE-binding activity of Cra a 4 and in vitro digested samples was reduced by about 50%.The unfoldα-helix and higher surface hydrophobicity of glycated Cra a 4 were identified as the reason for hypo-immunoreactivity.The higher frequency of lysine(13%)in the primary structure and exposure on the surface of the spatial structure was attributed to the hypo-immunoreactivity of Cra a 4 after Maillard reaction.It comprehensively evaluated the effects of Maillard reaction on Cra a 4 and provids new methods for the industrial production of hypoallergenic oyster.The modified Cra a 4 by Maillard reaction could be a hypoallergenic therapeutic agent for allergy to oyster.

Clinical remission from treatment dilemma of CMV colitis with ganciclovir use in a critical ill elderly immunocompetent patient

INTRODUCTION Cytomegalovirus(CMV)infects more than half of all adults by 40 years of age.It is estimated that 40~100%of immunocompetent adults worldwide are CMV seropositive,CMV remains in a latency state and allows reactivation in immune insufficiency,such as acquired immunodeficiency syndrome(AIDS),inflammatory bowel disease(IBD),organ transplantation,and active malignancy[1,2].CMV diseases in immunocompromised patients invade the broad tropism of the human body,such as the lung,retina,and kidney.The gastrointestinal(GI)tract is one of the most common organs of CMV disease.

Isolation,Purification and Immunomodifying Activity of Polysaccharides from Phellinus igniarius

Phellinus igniarius polysaccharides were obtained by hot water extraction of mushroom fruit bodies followed by ethanol precipitation,and further purified by anion exchange and gel filtration chromatography.Immuno-enhancing activity was evaluated by determining the effect of different polysaccharide fractions on mouse spleen lymphocyte proliferation in vitro.Three crude polysaccharide preparations,designated PIP30,PIP60 and PIP80 according to the percentage concentration of ethanol required for precipitation from hot aqueous extracts,promoted lymphocyte proliferation to varying degrees.Purification of polysaccharide present in PIP30 and PIP60 was carried out using anion exchange chromatography.Proliferation rates in samples treated with 200 μg/mL of fractions P30U,P30W and P30S1(obtained following anion exchange chromatography of PIP30) were approximately 5-,4-,and 3.5-fold higher,respectively compared with negative controls.Similarly,marked increases(3.4-and 2.8-fold) in cell proliferation rates compared with negative controls were observed with 200 μg/mL concentrations of P60W and P60S2(obtained following anion exchange chromatography of PIP60),respectively.Two purified polysaccharide preparations,P60W1-1 and P60S1-1,were obtained following gel filtation chromatography of fractions P60W1 and P0S1,respectively.Fraction P60W1-1(10-100 g/mL) enhanced splenocyte proliferation 0.5-1.4-fold compared with negative controls,whereas no effects were observed with P60S1-1.