Serum CD 4 4 Variant 6 as an Indicator of Tumor Burden and Metastasis in Patients with Gastric Car cinoma

Objective: To study the expression of CD44 correlation with the ability ofmetastasis of tumor cells in gastric carcinoma, to find the correlation of the concentration ofsoluble CD 44 Variant 6 (sCD44v6) and the histologic expression of CD44 Variant 6 (CD44v6) in tumorswith the clinico- pathologic features, and to make serum concentration of the CD44v6 and theexpression of CD44v6 may be useful an indicator as an early diagnosis, invasion, metastasis, andprognosis. Methods: Serum samples were obtained from 70 patients with primary gastric carcinomabefore surgery and 20 patients after surgery. Serum levels of CD44v6 were determined with aquantitative- enzyme- link-immuno- adsorbent assay. The expression of CD44v6 in tumors was examinedby immuno-histo-chemical staining. Results: Both the serum concentration of CD44v6 and itsexpression in tumors were positively related to the depth of invasion of the tumor, lymph nodemetastasis, clinical stage, and diffuse type gastric carcinoma, but not to the tumor size. The serumlevel of CD44v6 in patients with gastric cancer was significantly higher than that in control. Theserum concentration of CD44v6 was markedly lowered after curative surgery (F<0. 001), and the serumlevel of CD44v6 was higher in patients with gastric cancer with CD44v6 positive tumor cells than inthose with CD44v6 negative tumor cells. The serum level of CD44v6 was a prognostic indicator inpatients with diffuse type gastric carcinoma, as was the histological expression of CD44v6.Conclusion: CD44v6 of gastric cancer and serum concentration of CD44v6 seems to be correlated to theprogression of diffuse type gastric carcinoma metas- tasis and clinical stage. An elevated level ofsCD44v6 may serve as an indicator of lymph node metastasis (especially early metastasis) and badprognosis in pa tients with gastric carcinoma.

The Characteristics of Immunophenotype in Acute Lymphoblastic Leukemia and Its Clinical Significance

Objective： To study the characteristics of immunophenotype in acute lymphoblastic leukemia （ALL） and its clinical significance. Methods： Immunophenotyping was performed on 81 ALL patients by three-color flow cytometry analysis using CD45/SSC gating, meanwhile the cytogenetic analysis was performed on 45 cases out of 81 ALL patients. Results： （1） CD19 was the most commonly expressed of all B-lineage antigens detected with the positive rate being 100%. In T-ALL, the positive expression rate of CD5 and CD7 was the highest, being 90%. Both B-ALL and T-ALL overlapped in expression of lineage antigens. There was no significant difference in the complete remission rate （CR rate） between T-ALL and B-ALL. （2） The incidence of ALL with rayeloid antigens expression （My＋ALL） was 39.5%. CD13 was most often seen among the myeloid markers. My＋ALL always involved in B-lineage antigens and the CR rate in children and adults was 72.2% and 78.6% respectively. （3） The incidence of HAL was 19.8%. Coexpression of B-lineage and myeloid-assoeiated antigens was the commonest subtype in HAL. The expression of CD34 was commonly seen in HAL patients （81.3%）. The CR rate was low in HAL, 50% for children and 40% for adults. （4） Compared to T-ALL, B-ALL, My＋ALL, and HAL had a higher positive rate of CD34 expression with the difference being significant （P〈0.025）. Conclusion： Immunophenotyping had remarkable predominance in diagnosing special category of ALL （such as HAL and My＋ALL）; CD19 and CD5 were highly sensitive in diagnosing B-ALL and T-ALL, but less special, and overlapping was found in expression. No significant association was found between the expression of CD34 or myeloid antigens and CR rate, while low CR rate was found in HAL patients, especially for those coexpressing CD34 antigen.

A Study of the Relationship_Between Levels of SIgA and IFN-γ in the Female Genital Tract and the Clinical Course of NGU

Objective： To study the relationship of local immunity in the female genital tract with the clinical course of Nongonococcal Urethritis （NGU）. Methods： We collected cervical secretions from patients and examined levels of SIgA and IFN- γ. Results： Levels of SIgA and IFN- γ in the infected group were lower than those in the uninfected group, P1〈0.05 and P2〈0.05. The level of SIgA and IFN- γ in C.t,UU and C.t＋UU infected groups were not significantly different. Comparing the negative-changed group with thenonnegative-changed group, the level of SIgA and IFN-γ was 39.22±20.04mg/L and 103.19±29.94pg/ml, 26.00±10.56mg/I and 88.21±15.10pg/ml, P1〉0.05 and P2〉0.05. Conclusion： SIgA and IFN-γ secreted by genital tractmucosa may help prevent and resist local NGU infection.However, the effect is limited, and is insufficient to eliminate infection completely and prevent reinfection.

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.

Modulatory Effect of Rg_1 on the Activity of ProteinTyrosine Kinase of Lymphocytes in the Elderly

The effect of Rg1,a saponin extracted froin Panax ginseng, on the phenotype,receptor and the activity of protein tyrosine kinase (PTK) of lymphocytes isolated from 7 healthy oldpersons were studied. The CD25, CD45RA and CD45RO phenotypes of lymphocytes were 4eter-mined by indirect immunofluorescence technique. The percentage of CD25, CD45RA and CD45ROpositive lymphocytes was 38.3%±17.3%, 46.0% 15.1%, and 52.6%±14.1% respectively after incu-bation with PHA (5 μ±/ml) for 72 hours. However, there were 58.0%±12.5%, CD25, 64.1% ± 12.4%,CD45RA, and 74.0%±8.0%, CD45RO positive cells in the presence of Rg, ( 1μg/ml) along with PHA(5 μg/ml) over the sanie period of incubation. A significant increase was induced by Rgi (P<0.05).The activities of PTK in the cytoplasm and membrane of lymphocytes were measured by ELISAmcthod after incubation with PHA or PHA+Rg1. The absorbance value of PTK activity in cytoplasmafter 72 hr incubation was 0. 120±0.020 in PHA group, but 0. 1 38±0.015 in PHA+Rg1 group. In thelymphocyte membrane, it was 0.374± 0.060 in PHA group and 0.403 ± 0.008 in PHA+Rg1 group(P<0.001). These results showed that Rgi significantly arid simultaneously increased both the PT Kactivity and the expression of phenotype of lymphocytes.

Cellular immune function and liver damage in post hepatitic cirrhosis

AIM To study the cellular immune function in patients with post hepatitic cirrhosis (PHC) and its relation with different liver damages.

Effects of Echinacea Compound on the Immune Function of Weaned Piglets

[Objective] This study aimed to investigate effects of Echinacea compound on the immune function of weaned piglets. [Method] Eighty Duroc x Landrace x Yorkshire crossbred piglets were randomly divided into four groups： control group （drug-free group）, 1.5% close group, 1.0% dose group and 0.5% dose group. Blood samples of piglets were collected at 20, 35, 50, 60, 70 and 80 days old, respective- ly, to determine neutrophil leukocyte percentage in peripheral blood, lymphocyte trapsformation rate and the levels of antibodies against classical swine fever, porcine reproductive and respiratory syndrome disorder. [Result] Applying a certain dose of Echinacea compound could significantly increase neutrophil leukocyte per- centage in peripheral blood and lymphocyte transformation rate （P〈0.05）, and ex- tremely significantly improve the levels of antibodies against classical swine fever, porcine reproductive and respiratory syndrome disorder （P〈0.01）. [Conclusion] Echi- nacea compound has played a certain role in promoting nonspecific and specific im- mune function of piglets.

Clinical characteristics of a group of adults with nodular lymphoid hyperplasia:A single center experience

AIM： To describe the clinical and histological characteristics of a group of adults with small-bowel nodular lymphoid hyperplasia （NLH）. METHODS： Patients were searched for five years in pathology records of our institution. The biopsy material was reassessed using strict histopathological criteria. Clinical data were obtained from medical records. RESULTS： Small-bowel NLH was diagnosed in 18 cases. The female： male ratio was 2 ： 1. The most frequent symptoms were diarrhea （72%）, involuntary weight loss （72%） and abdominal pain （61%）. Nine patients （50%） had immunodeficiency. Small-bowel bacterial overgrowth was found in three （17%） cases. At small-bowel NLH diagnosis, three （17%） had associated lymphoma： two intestinal and one extra-intestinal lymphomas. In two patients with villous atrophy and anti-endomysial antibodies the diagnosis of celiac disease was established. Giardia larnblia infection was found in only one patient with hypogammaglobulinemia （Herman＇s syndrome）. CONCLUSIONS： NLH is uncommon in adult patients. Associated diseases are immunodeficiency and lymphoid tissue malignancies.

Acute liver failure due to natural killer-like T-cell leukemia/ lymphoma: A case report and review of the Literature

Acute liver failure （ALF） is a medical emergency requiring immediate evaluation for liver transplantation. We describe an unusual case of a patient who presented with ascites, jaundice, and encephalopathy and was found to have ALF due to natural killer （NK）-Iike T cell leukemia/lymphoma. The key immunophenotype was CD2＋, CD3＋, CD7＋, CD56＋. This diagnosis, which was based on findings in the peripheral blood and ascitic fluid, was confirmed with liver biopsy, and was a contraindication to liver transplantation. A review of the literature shows that hematologic malignancies are an uncommon cause of fulminant hepatic failure, and that NK-like T-cell leukemia/lymphoma is a relatively recently recognized entity which is characteristically CD3＋ and CD56＋. This case demonstrates that liver biopsy is essential in diagnosing unusual causes of acute liver failure, and that infiltration of the liver with NK-like T-cell lymphoma/leukemia can cause acute liver failure.

PROPERTIES OF PROTEOLYTIC TOXIN OF VIBRIO ANGUILLARUM FROM DISEASED FLOUNDER

Extracellular products (ECP) produced by Vibrio anguillarum strain M3 originally isolated from diseased flounder ( Paralichthys olivaceus ) were prepared. ECP of M3 showed gelatinase, casinase, amylase and haemolytic activity on agarose plates. High protease activity against azocasin was detected. Bacterium M3 showed highest growth and protease activity at 25℃. The protease present in ECP showed maximal activity at pH 8 and 55℃; was completely inactivated by application of 80℃ heat for 30 min; was completely inhibited by EDTA and HgCl 2, and was partially inhibited by PMSF, SDS, MnCl 2 and iodoacetic acid; but not inhibited by CaCl 2 and MgCl 2. The ECP was toxic to flounder fish at LD 50 values of 3.1 μg protein /g body weight. The addition of HgCl 2 and application of heat at 50℃ decreased the lethal toxicity of ECP. When heated at 100℃, ECP lethality to flounder was completely inhibited. After intramuscular injection of ECP into flounder, it showed evident histopathological changes including necrosis of muscle, extensive deposition of haemosiderin in the spleen, dilated blood vessels congested with numerous lymphocytes in the liver. These results showed that ECP protease was a lethal factor produced by the bacterium V. anguillarum M3.

Adult T-cell leukemia/lymphoma presenting multiple lymphomatous polyposis

Multiple lymphomatous polyposis (MLP) is an unusual form of non-Hodgkin's lymphoma characterized by polyps throughout the gastrointestinal tract. It has been reported that most MLP are observed in cases with mantle cell lymphoma of B-cell type. We herein present a case of a 66-year-old man with adult T-cell leukemia/lymphoma (ATLL). Colonoscopy revealed MLP throughout the colon and histopathological findings of ATLL cell infiltration. The patient died despite combination of chemotherapy. The literature of manifestations of colonic involvement of ATLL isreviewed and the importance of endoscopic evaluation to differentiate ATLL intestinal lesions from opportunistic infectious enterocolitis is discussed.

Costimulation of resting B lymphocytes alters the IL-4-activated IRS2 signaling pathway in a STAT6 independent manner: implications for cell survival and proliferation

IL-4 is an important B cell survival and growth factor. IL-4 induced the tyrosine phosphorylation of IRS2 in resting B lymphocytes and in LPS- or CD40L-activated blasts. Phosphorylated IRS2 coprecipitated with the p85 subunit of PI 3’ kinase in both resting and activated cells. By contrast, association of phosphorylated IRS2 with GRB2 was not detected in resting B cells after IL-4 treatment although both proteins were expressed. However, IL-4 induced association of IRS2 with GRB2 in B cell blasts. The pattern of IL-4- induced recruitment of p85 and GRB2 to IRS2 observed in B cells derived from STAT6 null mice was identical to that observed for normal mice. While IL-4 alone does not induce activation of MEK, a MEKI inhibitor suppressed the IL-4-induced proliferative response of LPS-activated B cell blasts. These results demonstrate that costimulation of splenic B cells alters IL-4-induced signal transduction independent of STAT6 leading to proliferation. Furthermore, proliferation induced by IL-4 in LPS-activated blasts is dependent upon the MAP kinase pathway.

Malignant lymphoma of spleen presenting as acute pancreatitis: A case report

This is a case report of a patient who presented with acute pancreatitis without the common causes. A pancreatic biopsy revealed large B cell lymphoma. Spleen lymphoma with pancreatic involvement inducing acute pancreatitis, which is a rare disorder, was diagnosed. Here we also review the few similar cases reported in the literature.

Gastric cancer-derived heat shock protein-gp96 peptide complex enhances dendritic cell activation

AIM To investigate the role of heat shock protein (HSP)glycoprotein (gp) 96 in dendritic cells (DCs) and lymphocytes induction in gastric cancer (GC). METHODS Human GC cell lines KATOIII, MKN-28 and SGC-7901 were infected with adenovirus gp96 at a multiplicity of infection of 100. gp96-GC antigen peptide complexes were purified. MTT (3-(4,5-dimethylthiazol-2-yl)2,5- diphenyltetrazolium bromide) assay, lactate dehydrogenase (LDH) release assay and enzyme-linked immunosorbent assay were used to determine allo-reactive T cell stimulation, natural killer (NK) cell activity and expression of cytokines (such as interleukin (IL)-10, IL-12, interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha), respectively. Effect of cytotoxic T lymphocyte (CTL) on DCs incubated with HSP-gp96 was also evaluated by LDH release. All assays were performed in triplicate and the average values were reported. Comparison between groups was conducted using Student's t test. RESULTS T cells incubated with HSP-gp96 exhibited a marked increase in proliferation in a dose-dependent manner (P < 0.05). NK cell activity after gp96-GC peptide complex treatment was significantly higher than that after antigen peptide treatment (P < 0.05). The activity of CTLs incubated with DCs from three GC cells lines was obviously higher than that stimulated by GC antigen at ratios of 50: 1, 25: 1, 10: 1, and 5: 1 (P < 0.05). Furthermore, the secretion of TNF-alpha, IL-10, IL-12 (P70) and IFN-alpha markedly increased after incubation with HSP-gp96 (P < 0.05). CONCLUSION HSP-gp96 promotes T cell response, enhances DC antigen presentation and induces cytokine secretion, as well. HSP-gp96 has potential as immunotherapy for elimination of residual GC cells.

Alpha-fetoprotein-producing colon cancer with atypical bulky lymph node metastasis

Alpha-fetoprotein (AFP)-producing colorectal cancer is extremely rarely reported until now. All of the reported cases harboring synchronous hematogenous spread including liver and/or lung metastasis had a poor prognosis and died within 12 mo. We here describe a 71-year old man with AFP-producing colon cancer who presented with an unusual bulky lymph node metastasis instead of hematogenous spread. He underwent adjuvant chemotherapy in addition to curative surgical resection, which prolonged his survival.

Lamprey buccal gland secretory protein-2 (BGSP-2) inhibits human T lymphocyte proliferation

Lamprey is a representative of the agnathans, the most ancient class of vertebrates. Parasitic lampreys secrete anticoagulant from their buccal glands and prevent blood coagulation of host fishes. We identified a buccal gland secretory protein-2 （BGSP-2） from a buccal gland cDNA library of Larnpetrajaponica. The full-length BGSP-2 gene was cloned and the recombinant BGSP-2 protein was generated. The role of BGSP-2 on lymphocyte proliferation was studied by examining its effects on human T lymphocytes. We found that lamprey BGSP-2 was able to effectively block the proliferation of T cells in vitro by inducing G1/S cell cycle arrest. Furthermore, it inhibited the proliferation of hmnan T lymphocytes stimulated by phytohemagglutinin （PHA） at a minimum concentration of 0.1μg/ml. Our data suggest that lamprey BGSP-2 is able to block the mitosis of human T lymphocytes at the G1/S point, and has the potential of anti-proliferative effect on PHA-activated T lymphocytes.

The expression of cyclooxygenase-2 （COX-2） and p16 in non-Hodgkin＇s lymphomas and its clinical significance

Objective： To investigate the expression of cyclooxygenase-2 （COX-2） and p16 proteins in non-Hodgkin＇s lymphomas （NHL） and their relationship with the genesis and progress of it. Methods： The expression of COX-2 and p16 protein were studied in the lymph nodes tissue from 60 NHL patients and 10 control patients with non-malignant diseases by flow cytometry. Results： Positive rate of COX-2 protein expression in NHL tissues （63.3%, 38/60） was higher than that in normal lymphaden tissues （0, 0/10）. The difference was significant between the two groups （P 〈 0.01）. Expression of COX-2 protein was related with the clinical stage of NHL. In stage Ⅰ ＋ Ⅱ patients, it was significantly lower （35.0% ± 54.6%） than that in stage Ⅲ ＋ Ⅳ patients （84.6% ±87.5%） （P 〈 0.01）. In different sex, age, tumor malignant degree, IPI grade, extranodal involvement and B symptoms groups, the differences of COX-2 expression were not statistically significant （P 〉 0.05）. Positive rate of p16 protein expression （41.7%, 25/60） in NHL＇ was statistically lower than that in normal lymphomas （100%, 10/10） （P 〈 0.01）. Expression of p16 protein was related to malignant degree of NHL. The positive rates of p16 protein in low malignant degree tissues （64.7%, 11/17） was higher than that in high malignant degree tissues （14.3%, 2/14） （P 〈 0.05）. Positive rates of p16 protein of NHL tissues in different sex, age, IPI grade, extranodal involvement, clinical stages and B symptoms were not statistically significant （P 〉 0.05）. The p16 protein expression in COX-2 positive patients was 47.4% （18/38）, and in negative patients it was 31.8% （7/22）. There was no statistically difference between them （P 〉 0.05）. Correlation analysis revealed there was no correlation between expression of COX-2 and p16 protein. Conclusion： Both COX-2 and p16 protein may all have relationship with the genesis and progress of NHL. The expression of COX-2 protein in NHL may be a poor prognostic indicator. COX-2 and p16 protein probably have different mechanisms in the genesis and progress of NHL. Their relationship is firstly put forward in this article and needed further studying.

DETECTION OF B LYMPHOMA CELLS UNDERGOING APOPTOSIS BY ANNEXIN-V ASSAY

To quantitatively analyze apoptotic and secondary necrotic cells unde r apoptosis conditions. Methods. The cells of Burkitt lymphoma (BL) cell line Raji were incubated with 1 .0 μmol/L dexamethasone (DEX) for 2, 4 and 8 h respectively, then stained with Annexin V FITC (fluorescein isothiocyanate conjugated) which was used to detec t the exposed phosphatidylserine (PS) on the epimembrane resulting from a loss o f phospholipid asymmetry in the early stage of apoptosis, and also stained with propidium iodide (PI) which allows analysis of secondary necrotic cells related with cell membrane and DNA damage that probably represent late stage of apoptosi s, then apoptotic cells were quantified by flow cytometry (FCM). Furthermore, An nexin+/PI and Annexin+/PI+ cells were sorted by fluoresence activated cell sorter (FACS), and identified by electron microscopy (EM) and DNA gel electroph oresis. Results. The percentage of apoptotic cells was found to increase with the incuba tion time (r=0.97). This method was sensitive with low detection limit (0.02%), and was reproducible with low coefficient variance (CV)(4.2%). Meanwhile, the Annexin+/PI and Annexin+/PI+ cells were identified as apoptotic and necroti c cells under EM, and DNA extracted from the Annexin+/PI cells was characteri zed by “ladder pattern”. Conclusions. Annexin V assay is a specific, sensitive, accurate, reproductive and quantitative method for analyzing apoptotic cells.

EFFECTS OF ACUMOXI ON IL2-IFN-NKC IMMUNOREGULATORY NETWORK AND ITS RELATED MACROPHAGE-IL1-Th NETWORK AND ON TUMOR——Observation on the Effect of Acumoxi on the Macrophage-IL1-Th Network and on the Metastasis of Hepatoma

Objective: To observe the effect of acumoxi (acupuncture and moxibustion) on macrophage (Mφ)-lL1-Th net-work and hydroperitoneum hepatoma (H 22) metastasis in mice. Methods: A total of 36 BALB/ c male mice bearing H 22 are randomly divided into control, acupuncture and acumoxi groups with 12 cases in each group. In the later 2 groups, Dazhui (GV 14) and Guanyuan (CV 4) are punctured once daily, continuously for 18 days, and in acumoxi group, the two acupoints were also moxibustioned alternatively with moxa stick once every day. After killing the mice, the tissue samples of the 3 groups are treated routinely step by step and analyzed by means of colorimetric analysis for determining the phagocytic function of the macrophages; and the content of IL1 of the Mφ supernatant is assayed with serum plate agglutination (SPA)-Ig floral hoop method of T helper cell (Th) monoclonal antibody; the weight of the reniportal lymph node, the kidney and the lung, and the number of the cancerous nodes on the pulmonary surface are calculated. Results: After acupuncture and moxibustion treatment, the immunoregulatory network indices of acumoxi group increase obviously compared with those of control group(P<0.01), showing an anti-metastasis effect of acumoxi on H 22. Conclusion: Results of the present study and those of our former research prove that acupuncture and moxibustion can suppress the tumor growth and H 22 metastasis by the enhancement of the immunoregulatory network.

PRAME Gene Expression in Acute Leukemia and Its Clinical Significance

Objective To investigate the expression of the preferentially expressed antigen of melanoma （PRAME） gene in acute leukemia and its clinical significance. Methods The level of expressed PRAME mRNA in bone marrow mononuclear cells from 34 patients with acute leukemia （AL） and in 12 bone marrow samples from healthy volunteers was measured via RT-PCR. Correlation analyses between PRAME gene expression and the clinical characteristics （gender, age, white blood count, immunophenotype of leukemia, percentage of blast cells, and karyotype） of the patients were performed. Results The PRAME gene was expressed in 38.2% of all 34 patients, in 40.7% of the patients with acute myelogenous leukemia （AML, n=27）, and in 28.6% of the patients with acute lymphoblastic leukemia （ALL, n=7）, but was not expressed in the healthy volunteers. The difference in the expression levels between AML and ALL patients was statistically significant. The rate of gene expression was 80% in M~, 33.3% in M2, and 28.6% in M~. Gene expression was also found to be correlated with CDl5 and CD33 expression and abnormal karyotype, but not with age, gender; white blood count or percentage of blast cells. Conclusions The PRAME gene is highly expressed in acute leukemia and could be a useful marker to monitor minimal residual disease. This gene is also a candidate target for the immunotherapy of acute leukemia.