Clinical relevance of soluble HLA class I molecules in Waldenstrom Macroglobulinemia

Objectives: Waldenstrom Macroglobulinemia (WM) is a B‐cell neoplasm characterised by secretion of IgM by lymphoplasmacytic bone marrow cells and by cytopenias and hypogammaglobulinemia in a subset of patients. Beta‐2 microglobulin (b2m) is a major prognostic factor in WM and the heavy chain of HLA class I molecules, which are known to have immunosuppressive properties and have been implicated in the pathogeny of several malignancies. Methods: We assessed the serum levels of the total soluble HLA‐I molecules and the HLA‐Gs molecules in 105 patients with IgM‐related disorders [WM (n = 42) and IgM MGUS (n = 63)], and compared the results to 41 healthy subjects. Results: We found higher levels of HLA‐Is in WM, compared to IgM MGUS and healthy donors. HLA‐Gs levels were similar in WM and in IgM MGUS, but higher than in healthy donors. The association between HLA‐Is at the cut‐off of 1.8 μg/mL and known markers of poor prognosis was then evaluated among WM patients using univariate and multivariate methods. Based on this, high HLA‐Is level was strongly associated with high serum β2M level >3 mg/L [OR = 2, (CI 95% 1.1–5.7); P = 0.04], age > 65 yrs [OR = 1.5, (CI 95% 0.5–4.1), P = 0.06] and haemoglobin ≤11.5 g/dL [OR = 3.3, (CI 95% 1.2–9.7); P = 0.03]. High levels of serum HLA‐Is were also found in patients with cryoglobulinemia, however irrespectively of WM or IgM‐MGUS status. Conclusion: Together our results suggest a possible role for soluble MHC class I molecules in WM disease. Further investigations are necessary to further demonstrate the prognostic impact of soluble MHC class I molecules in Waldenstrom Macroglobulinemia.     

Disseminated Nocardia asteroides Presenting as Pulmonary Non-caseating Granulomas in a Patient with Waldenstrom Macroglobulinemia

Disseminated nocardiosis has never been described before in a patient with Waldenstrom macroglobulinemia. We report an unusual case of disseminated nocardiosis in a patient with Waldenstrom macroglobulinemia who presented with pulmonary non-caseating granulomas. The patient was successfully treated with trimethoprim-sulfamethoxazole (TMP-SMX) for 1 year. Received: May 20, 2001 · Revision accepted: September 25, 2001     

Analysis of 6q deletion in Waldenstrom macroglobulinemia

The frequency and prognostic significance of chromosome 6q deletion were investigated in a cohort of patients with Waldenstrom macroglobulinemia (WM). By interphase cytoplasmic fluorescence in situ hybridization with probes for 6q21 and 6q25, we detected hemizygous 6q deletions in the clonal lymphoplasmacytic cells in 13 (38%) out of 34 patients. Patients with 6q deletions had a lower IgM paraprotein levels than non-deleted patients (P = 0.04). There was no correlation between 6q deletion and other clinical features of WM. There was no significant difference in overall survival between 6q deleted and non-6q deleted groups (P = 0.92). Our study confirms that 6q deletion is a frequent event, but it does not appear to be a prognostic marker for patients with WM.     

Serum concentrations of angiogenic cytokines in Waldenstrom macroglobulinaemia: the ration of angiopoietin-1 to angiopoietin-2 and angiogenin correlate with disease severity

Angiogenesis represents an essential step of disease progression in several haematological malignancies. Microvessel density is increased in 30% of patients with Waldenstrom macroglobulinaemia (WM), but there is very limited information regarding the role of angiogenic cytokines in this disease. Serum levels of vascular endothelial growth factor (VEGF), VEGF-A, angiogenin, angiopoietin (Ang)-1 and -2, and basic fibroblast growth factor (bFGF) were evaluated in 56 WM patients at different disease phases (24 untreated, 20 relapsed/refractory and 12 patients at remission) and 11 patients with immunoglobulin M type monoclonal gammopathy of undetermined significance (IgM-MGUS). All patients had increased levels of angiogenin, VEGF, VEGF-A, and bFGF compared with controls. The Ang-1/Ang-2 ratio was reduced in WM but not in IgM-MGUS patients. Angiogenin levels correlated with disease status: when compared with healthy subjects, patients with IgM-MGUS and untreated WM patients had increased angiogenin serum levels, which were higher in untreated WM patients than in MGUS. WM patients at remission had lower angiogenin serum levels compared with untreated patients, but these levels were increased again in active disease post-therapy. Angiogenin also correlated with albumin levels, while VEGF-A correlated with beta(2)-microglobulin (beta2M). Ang-1/Ang-2 ratio showed a strong, negative correlation with beta2M, and positive correlation with albumin, haemoglobin and lymphadenopathy. Our results indicate a potential use of angiogenin levels for follow-up in WM and angiogenic molecules as targets for the development of novel anti-WM agents.     

肝卵圆细胞分化调控机制的研究进展

卵圆细胞是肝脏的干细胞，具备多向分化潜能，体内外实验证实其可以分化为肝细胞、胆管细胞、胰腺及肠型上皮细胞。在胚胎发育过程中，肝脏干细胞以肝细胞的形式存在，而在成年哺乳动物的肝组织中则以卵圆细胞的形式存在。对卵圆细胞分化调控机制的研究在基础理论和临床应用等方面均有重要意义，一方面，卯圆细胞可能参与肝脏损伤的修复与重建，研究其分化机制有助于阐明肝脏的发育机制；另一方面，卵圆细胞可分化为具有功能的成熟肝细胞，将为肝细胞移植和生物型人工肝提供重要的细胞来源，可能缓解供体肝脏严重缺乏的矛盾。但卵圆细胞的分化过程和机制非常复杂，分化异常可能导致肝细胞癌的发生。     

Expression of hENTl and ERCC1 genes in tumor tissues non-small cell lung cancer

ObjectiveTo investigate the expression of hENTl and ERCC1 genes in tumor tissues non–small cell lung cancer (NSCLC).MethodsFresh non–small lung cancer specimens were transplanted into nude mice. Twenty mice were randomized into two groups: experimental group receiving gemcitabine plus cisplatin and control group receiving 0.9% physiological saline. The expressions of hENTl and ERCC1 mRNA in tumor tissue were detected by real–time fluorescent quantitative PCR. The volume of tumor, the weight of nude mice and tumor volume were respectively measured and calculated 2–3 times per week. Tissue samples were collected from NSCLC mice treated with gemcitabine plus carboplatin.ResultsThe histological examination showed that many tumor cells were well preserved in nude mice. The rate of transplanted tumor cells was 86.7%. The concomitant treatment study showed that the rate of TV, RTV, T/C in GEM + DDP group was the lowest. LBP + DOC, DDP + DOC obviously influenced the body weight. Compared with NS group, DDP group, GEM group, the survival period and the level of hENTl of DDP+GEM group increased obviously, the level of ERCC1 decreased significantly (P<0.05).ConclusionsThe expression of hENT1 and ERCC1 genes in tumor tissues were closely correlated with the response to chemotherapy and prognosis of patients with NSCLC treated with gemcitabine plus cisplatin.     

哈萨克族食管鳞癌组织中CyclinD1、survivin的表达

目的研究Cyc linD1和survivin基因在哈萨克族食管鳞状上皮细胞癌组织中的表达。方法采用RT-PCR技术检测Cyc linD1和survivin基因在30例哈萨克族食管癌组织及30例非癌组织中的表达。结果食管癌组织、非典型增生组织、正常组织中Cyc linD1阳性表达率分别为63.33%、70.00%、15.00%,正常组织低于非典型增生组织及食管癌组织（P〈0.05）;survivin阳性表达率分别为93.33%、60.00%、15.00%,三者比较均有统计学差异（P〈0.05）。Cyc linD1、survivin mRNA表达与食管鳞状上皮细胞癌病理分化程度无相关关系,两者在食管鳞状细胞癌组织中表达无相关关系。结论在哈萨克族食管鳞状细胞癌形成过程中,Cyc linD1、survivin过表达是两个相对独立的分子事件;检测survivin可能更有助于新疆食管癌高发地区进行早期食管癌筛查。     

Attainment of complete/very good partial response following rituximab-based therapy is an important determinant to progression-free survival, and is impacted by polymorphisms in FCGR3A in Waldenstrom macroglobulinaemia

The incorporation of rituximab into various regimens has improved depth of response in Waldenstrom macroglobulinaemia (WM), though the impact of achieving better responses remains to be determined. We examined response depth on progression‐free survival (PFS) in 159 rituximab‐naïve WM patients who received rituximab‐based therapy. The median follow‐up was 33·5 months, and categorical responses were as follows: complete response (CR, 8·8%); very good partial response (VGPR, 13·2%); partial response (50%); minor response (18·9%); Non‐Responders (8·8%). Sequencing for polymorphic variants of FCGR2A, FCGR2B, and FCGR3A was performed, and impact on response depth determined. Achievement of better categorical responses was incrementally associated with improved PFS (P < 0·0001). No separation was observed between CR and VGPR, and attainment of at least a VGPR was associated with improved time‐to‐progression. Neither age, serum IgM, haematocrit, platelet count, serum β₂microglobulin, WM International Prognostic Scoring System score, and treatment group predicted for CR/VGPR. Polymorphisms at FCGR3A‐48 and ‐158 were associated with improved categorical responses, particularly attainment of CR/VGPR (P ≤ 0·03). The attainment of CR/VGPR was associated with significantly longer PFS in rituximab‐naïve WM patients undergoing rituximab‐based therapy, and was predicted by polymorphisms in FCGR3A.     

Expression of iron regulatory genes in a rat model of hepatocellular carcinoma.

BACKGROUND/AIMS: The altered iron metabolism in hepatocellular carcinomas (HCCs), characterized by the iron-deficient phenotype, is suggested to be of importance for tumour growth. However, the underlying molecular mechanisms remain poorly understood. We asked whether these iron perturbations would involve altered expression of genes controlling iron homeostasis. METHODS: HCCs were induced in rats by the Solt and Farber protocol of chemical hepatocarcinogenesis, and to evaluate the effects of iron loading, one group of animals were supplemented with dietary iron during tumour progression. Tissue iron contents were determined, labelling indices of S-phase nuclei were calculated, and mRNA levels of iron-regulatory genes were quantitated. Protein levels of ferroportin1 were determined with Western blot. RESULTS: HCCs displayed reduced amount of tissue iron and lack of histologically stainable iron. HCCs expressed significantly higher mRNA levels of genes involved in iron uptake (transferrin receptor-1, divalent metal ion transporter-1), ferroxidase activity (Ferritin-H), and iron extrusion (ferroportin1). The protein levels of ferroportin1 in iron-deficient HCCs were similar as in control livers, and did not increase in HCCs exposed to iron. Hepcidin mRNA levels were decreased in iron-deficient HCCs, rose in response to iron loading and correlated to the tissue iron content. CONCLUSIONS: Taken together, the altered expressions of iron-regulatory genes in HCCs possibly reflect an increased demand for bioavailable iron and a high iron turnover in neoplastic cells.     

黄芪等12味中药在体外培养中对人调节性T细胞分化的影响

目的:探讨黄芪等12味中药对人调节性T细胞的分化的影响,为溃疡性结肠炎的中药组方研究提供实验依据.方法:分离正常人外周血淋巴细胞和单核细胞,在体外培养中用人肠道厌氧茵抗原刺激单核细胞,然后将激活的单核细胞与淋巴细胞混合培养,分别加入黄芪等12种不同的中药煎液,最后用流式细胞术检测其对CD4 CD25 FOXP3 调节性T细胞分化的影响.结果:与PBS组比较,CD4 细胞中CD25及FOXP3双阳细胞的百分比在黄芪及丹参组明显升高(5.6%±0.6%,5.5%±0.8%vs 4.3%±0.8%;均P＜0.01),同时CD4 CD25 细胞中FOXP3的表达率也显著升高(50.0%±3.8%,45.1%±3.3%vs 30.9%±4.5%;均P＜0.01).结论:黄芪和丹参可能通过调节性T淋巴细胞的分化平衡来发挥其治疗作用,可作为溃疡性结肠炎组方的首选中药.     

Expression of the preoptic regulatory factor-1 and ?2 genes in rat testis

Hormone-responsive peptides play a vital role in development and regulation of testicular function. The preoptic regulatory factors, porf-1 and porf-2, were originally discovered in the rat brain, but are also expressed in the rat and human testis. In the brain expression is age-related, hormone-responsive, region-specific, and gender-related, suggesting that porf-1 and porf-2 are involved in gender-specific brain development and function. Tissue-specific porf-1 and porf-2 mRNAs are also found in the testis and hypophysectomy may alter testicular porf-2 expression. It was thus of interest to further examine porf-1 and porf-2 expression in the testis to evaluate their potential as hormone-responsive peptides that regulate testicular development and function. Testicular expression of both porf-1 and −2 was analyzed as a function of maturational stage, aging and hypophysectomy by the solution hybridization/nuclease protection assay, and cellular location determined byin situ hybridization histochemistry. Expression was quantitatively compared in normal male rats at 15,30, and 60 d (n=4) and at 2, 6, 12, and 24 mo of age (n=5). During development porf-1 is expressed at a constant level at 15, 30, and 60 d, then declines significantly with advancing age; levels at 24 mo are only 20% of those seen at 2 mo (p<0.05). In contrast, porf-2 expression is highest at 15 d of age and steadily declines at 30 and 60 d, plateaus in the mature adult (6 and 12 mo), then exhibits an additional significant decline in the aged 24 mo animals (6 vs 24 mo,p<0.05). Hypophysectomy of young adult rats at day 42 results in increased testicular expression 12 d later of both porf-1 (p<0.05) and porf-2 (p<0.005) compared to intact 54-d-old rats (n=5).In situ hybridization histochemistry confirms that both porf-1 and porf-2 are expressed in the mature testis at 60 d of age. Porf-2 mRNA is localized to immature germ cells including spermatogonia and primary spermatocytes. Porf-1 mRNA is associated with mature sperm and at low levels in the Sertoli cell cytoplasm surrounding spermatocytes. These data suggest that porf-2 is a pituitary hormone-responsive factor in the developing testis and that both porf-1 and porf-2 have cell-type specific functions in the germ cell compartment of the mature testis.     

Aberrant expression of caspase cascade regulatory genes in adult T-cell leukaemia: survivin is an important determinant for prognosis

Derangement of either apoptosis or cell division is known to play an important role in tumorigenesis. Fas-mediated apoptosis on normal and leukaemic T cells is finely tuned by inhibitory proteins, such as FAP-1, FLIP and survivin, and defective caspase isoform which can attenuate the function of its intact caspase as a decoy molecule. However, complex involvement of such inhibitors in tumour biology relating to apoptotic pathology remains unclear in the neoplasms. We report the aberrant expression of FAP-1, FLIP and survivin mRNAs on leukaemic T cells from adult T-cell leukaemia (ATL) patients. Among these inhibitors, only survivin was aberrantly expressed in all ATL cases, but not in any normal peripheral blood mononuclear cells (PBMCs). Furthermore, survivin mRNA expression level was characteristic in each subtype of ATL and represented an important determinant for ATL prognosis. However, the apoptotic effector of casp-8, which is essential in Fas-mediated signal transduction, was dominant in defective casp-8 rather than intact casp-8 in ATL cells, suggesting a favourable biological situation for escape from apoptosis. Taken together, ATL cells probably possess many different regulatory mechanisms in order to attenuate Fas-mediated signalling and subsequently expand their populations under escape from apoptosis. Among these inhibitors, survivin is a useful bio-marker to assess tumour biology and may be a potential new target for apoptosis-based selective therapy in neoplasms as the expression is a general feature of neoplasia, but not normal tissues.     

肝癌组织中Muc-1和Survivin表达与癌细胞分化程度的关系研究

目的探讨Muc-1和Survivin在肝细胞癌组织中表达及其与癌细胞分化程度的关系。方法采用免疫组化法检测50例肝细胞癌患者癌组织 Muc-1和Survivin的表达,分析两者与肝癌细胞分化程度的关系。结果 Muc-1和Survivin在肝癌组织中的表达阳性率分别为86%和88%,且其表达程度越强,肝癌细胞的分化程度越低;肝癌细胞越趋向于低分化时,Muc-1和Survivin双表达率越高。结论 Muc-1和Survivin可能作为肝癌恶性程度及预后的分析指标。     

Expression and significance of tumor-related genes in HCC

AIM: To investigate the expression and clinical significance of DEK, cyclin D1, insulin-like growth factor Ⅱ (IGF-Ⅱ), glypican 3 (GPC3), ribosomal phosphoprotein 0 (rpPO) mRNA in hepatocellular carcinoma (HCC) and its paraneoplastic tissues. METHODS: The expression of mRNAs of DEK, cyclin D1, IGF-Ⅱ, GPC3 and rpP0 mRNA was detected in HCC and its paraneoplastic tissues by multiplex RT-PCR. RESULTS: By the simplex RT-PCR, the overexpression of mRNAs of DEK, cyclin D1, IGF-Ⅱ, GPC3, rpP0 mRNA in HCC and its paraneoplastic tissues was 78.1%, 87.5%, 87.5%, 75.0%, 81.3% and 15.6%, 40.6%, 37.5%, 21.9%, 31.3% respectively (P<0.05). By the multiplex RT-PCR, at least one of the mRNAs was detected in all HCC samples and in 75.0% of paraneoplastic samples (P>0.05). However, all these five mRNAs were found in 68.8% of HCC samples, but only in 9.4% of paraneoplastic tissues (P<0.05). The positive expression of mRNAs of DEK, cyclin D1, IGF-Ⅱ, GPC3, rpP0 in well- and poorly-differentiated HCC was 89.0%, 66.7%, 66.7%, 66.7%, 77.8% and 73.9%, 95.7%, 95.7%, 95.7%, 82.6%, respectively (P>0.05). The expression of these genes in HCCs with α-feto protein (AFP) negative and positive was 90.0%, 80.0%, 90.0%, 90.0%, 90.0% and 72.7%, 86.3%, 77.3%, 90.9%, 68.2% respectively (P>0.05). CONCLUSION: The expression of DEK, cyclin D1, IGF-Ⅱ, GPC3, rpP0 mRNA in HCC is much higher in HCC than in its paraneoplastic tissues. Multiplex RT-PCR assay is an effective, sensitive, accurate, and cost-effective diagnostic method of HCC.