The influence of different chromosomal aberrations on molecular cytogenetic parameters in chronic lymphocytic leukemia

B-cell chronic lymphocytic leukemia (B-CLL) is the most common leukemia of adults in Western countries. The most frequent recurring chromosomal aberrations identified in B-CLL patients are trisomy 12 and deletions of 13q, 17p, and 11q. Cases with deletions of 11q and 17p have a poor prognosis, whereas cases with deletions in 13q have a favorable prognosis. It was previously shown that CLL patients with trisomy 12 and del(13)(q14) have a higher rate of asynchronous replication of normal structural genes when compared to those with normal karyotypes. We studied the replication pattern of the structural locus 21q22 and the imprinted gene SNRPN and its telomere (15qter) and the random aneuploidy of chromosomes 9 and 18 in CLL patients with trisomy 12 and deletions of 11q and 17p, and compared the results to those of CLL patients without these aberrations and to healthy controls. Random aneuploidy rate was higher in the group of patients with trisomy 12 as compared to all other groups. The replication pattern with higher asynchronous pattern was found in both aberration groups compared to the CLL patients without the aberrations and to the control group with involvement of 21q22 and 15qter, whereas the highest synchronous group was found in the 2 aberrations CLL patient groups compared to the other groups with the imprinted locus SNRPN. The existence and significance of chromosomal aberrations in CLL have a deleterious effect on the processes of cell cycle and gene replication and may have biological and prognostic implications.     

The effect of chlorambucil treatment on cytogenetic parameters in chronic lymphocytic leukemia patients

The most common treatment of chronic lymphocytic leukemia (CLL) is the alkylating agent chlorambucil (CLB), with or without prednisone. In the present study, our aim was to evaluate whether treatment with CLB for more than one year induced genetic changes manifested by comparative genomic hybridization (CGH) as new chromosomal aberrations. We also studied whether CLB affected the pattern of replication by using fluorescence in situ hybridization (FISH). We found a similar rate of asynchronous pattern of replication in both treated and untreated patients with CLL. Most of the aberrations found with CGH were previously reported in CLL. More prognostically unfavorable aberrations and more cases with genetic changes were found in the treated group. The changes found were not typical of the secondary genetic aberrations associated with alkylating agents. Thus, we conclude that treatment of CLL with CLB for at least a year does not affect the parameters analyzed in this study. Longer studies are needed to further explore the effects of alkylating agents on normal and malignant cells.     

Leg ulcer in hydroxyurea-treated patients

A cutaneous ulcer is a lesser known complication of hydroxyurea treatment. Out of 39 patients [18 polycythaemia vera (PV), 13 essential thrombocythaemia (ET), 4 chronic myeloid leukemia (CML), 4 undefined myeloproliferative diseases (MPD)] treated with hydroxyurea, 6 (4ET, 1PV, 1CML) developed a cutaneous ulcer during a period of less that 2 years' treatment. In all but one of the patients the ulcers were situated in the ankle region. At the time of onset of ulceration, none of them had extreme values in their peripheral blood counts. All had one or more of the predisposing factors such as minor trauma or mild varicosity. None of the patients had any alteration in arterial or venous circulation when examined by non-invasive means. No hyperviscosity was found as measured by capillary viscosimeter. The ulcers were cured in three patients without discontinuation of the drug. One patient later developed an ulcer on the other leg. The ulcers healed in two patients only after having stopped the hydroxyurea medication. One patient still had the ulcer when she succumbed to the underlying CML in transformation. In conclusion, cutaneous, ulceration of the leg is relatively common during hydroxyurea therapy. Predisposing factors are also involved in its development. Its healing does not necessarily require the discontinuation of the drug.     

β-catenin and PPAR-γ levels in bone marrow of myeloproliferative neoplasm: an immunohistochemical and ultrastructural study

In accordance with increased proliferation in myeloproliferative neoplasm (MPN), the goal is to evaluate the immunoexpression of: β-catenin, PPAR-γ and Ki67 protein, to compare them with bone marrow ultrastructural characteristics in patients with MPN. Immunoexpression and electron microscopy of bone marrow was analyzed in 30 Ph-negative MPN patients, including per 10 patients with polycythemia vera (PV), essential thrombocythemia (ET) and primary myelofibrosis (PMF). The quantity of β-catenin immunoreactive cells was significantly higher in PV then in ET (p < 0.01) or PMF group of patients (p < 0.01) and also in ET versus PMF group of patients (p < 0.01). Erythroid lineage showed absent β-catenin staining without immunoreactivity in nucleus. In contrast, immunoreactivity for PPAR-γ was localized mostly in megakaryocytes and the highest number of PPAR-γ immunopositive cells was detected in PMF group of patients. In addition, the proliferative Ki67 index was significantly increased in the PMF and PV patients compared to patients with ET. Also, the megakaryocytes showed abnormal maturation in PMF group of patients as determined by ultrastructural analysis. These results indicated that PV dominantly expressed β-catenin and proliferation marker Ki67 in bone marrow, while PMF is linked preferentially to PPAR-γ immunopositive megakaryocytes characterized by abnormal maturation.     

Modified order of allelic replication in lymphoma patients at different disease stages

Asynchronous replication of homologous loci was reported in lymphocytes of patients with lymphoma, ovarian and renal cancer as well as in lymphocytes of patients with premalignant conditions, for example, essential mixed cryoglobulinemia associated with hepatitis C virus and in monoclonal gammopathy of unknown significance. In the present study we evaluated the replication pattern in lymphocytes of four groups of patients with intermediate grade of non-Hodgkin lymphoma at various stages of their disease: 1) at diagnosis; 2) during cytotoxic treatment; 3) in remission; and 4) in relapse. A significantly higher proportion of the asynchronous pattern of replication at diagnosis, during cytotoxic treatment, and in relapse was noted as compared to healthy controls and to patients who achieved remission of their lymphoma. Also, the frequency of the two doublets (DD) pattern in every group studied was significantly lower than in the controls. If our findings can be confirmed in larger, long-term prospective studies, it may allow the use of a simple and inexpensive tool to closely observe patients with lymphoma who are at high risk for relapse.     

The JAK2V617F allele burden and STAT3- and STAT5 phosphorylation in myeloproliferative neoplasms: early prefibrotic myelofibrosis compared with essential thrombocythemia, polycythemia vera and myelofibrosis

Early prefibrotic myelofibrosis (early PMF) is a diagnosis that clinically and histologically mimic essential thrombocythemia (ET), but is important to distinguish from ET, polycythemia vera (PV) and primary myelofibrosis (PMF) due to its different prognosis and clinical evolution. In this study, we assessed the allele burden of JAK2V617F in bone marrow biopsies from patients with these chronic myeloproliferative neoplasms. We correlated our findings with the amount of phosphorylated STAT3 (P-STAT3) and STAT5 (P-STAT5) in megakaryocyte nuclei in the bone marrow. The JAK2V617F allele burden was significantly higher in patients with PV (median: 50.99, range: 23.08-97.29, p < 0.01 and p < 0.01) and PMF (median: 44.13, range: 33.61-92.17, p < 0.05 and p < 0.01) compared with a low allele burden in ET (median: 23.465, range: 8.67-47.92) and early PMF (median: 25.68, range: 0.61-49.13) respectively. In addition, we found a significantly higher phosphorylation of STAT5 and STAT3 in the JAK2V617F positive group than in the negative group. There was no positive correlation between increasing JAK2V617F allele burden and the amount of P-STAT3 and P-STAT5. However, we found low values of P-STAT5 in bone marrow biopsies from patients with ETJAK2V617F+ as compared with patients with early PMFJAK2V617F+. Although this difference was statistically significant, larger studies are needed to firmly support this conclusion.     

The influence of cytogenetic aberrations on gene replication in chronic lymphocytic leukemia patients

Chronic lymphocytic leukemia (CLL) is the most common leukemia in humans, with the major cytogenetic aberrations of trisomy 12 and deletion of 13q14. This study examined the influence of these aberrations on general gene replication. The study group included three subgroups: (1) 15 CLL patients, (2) 4 CLL patients with trisomy 12, (3) 3 CLL patients with deletions in 13q14. Five healthy individuals served as a control group. Monocolor fluorescence in situ hybridization (FISH) with probes for c-myc, HER-2/neu, and p53 was applied to lymphocyte nuclei for the evaluation of replication timing. Asynchronous replication (SD) rate was significantly higher in all CLL patients (P < 0.01) when compared to the control group and was even higher in the group of CLL patients with trisomy 12 and 13q14 deletion (P < 0.01). The asynchrony rate was significantly higher in cells with trisomy 12 for all three probes analyzed, compared to "healthy" cells in the same patients (P < 0.001). To conclude, in CLL patients with a chromosomal aberration such as trisomy 12 and 13q14 deletion we were able to demonstrate a high rate of asynchrony of replication. The high correlation between cells with trisomy 12 and SD pattern could reflect direct influence of the aberration on gene replication and cell cycle control.     

The significance of bone marrow biopsy and JAK2V617F mutation in the differential diagnosis between the "early" prepolycythemic phase of polycythemia vera and essential thrombocythemia

It has been suggested that polycythemia vera (PV) could be preceded by an "early" phase of the disease (e-PV), in which the increase in the red cell parameters is lower than required for a PV diagnosis. In this study, we compared the clinicopathologic and molecular features of 17 patients with e-PV with those of 14 patients with essential thrombocythemia (ET) and 19 with PV.The results for e-PV were more similar to those for PV than for ET. In fact, patients with e-PV were characterized by an increase in the red cell parameters, splenomegaly (P<.05), and hepatomegaly (P=.038), together with hypercellular bone marrow due to increased erythropoiesis and granulopoiesis, associated with megakaryocytic hyperplasia, with pleomorphic aggregates (P<.001). The frequency of the JAK2V617F mutation was similar in e-PV (16 cases tested [100%]) and PV (18/19 [95%]) but was significantly lower (7/13 [54%]) in ET (P=.0007). We propose a diagnostic algorithm helpful to distinguish ET from the early prepolycythemic phase of PV.     

甲亢和甲减患者血浆心钠素和内皮素的检测及其临床意义

采用放免法检测58例甲亢和47例甲减患者血浆心钠素(ANF)和内皮素(ET)水平.甲亢未治组ANF和ET水平显著高于甲亢缓解组、 甲减未治组、甲减治疗组及对照组(P＜0.01), ANF和ET水平与血清FT3、FT4存在正相关.甲亢缓解组、 甲减未治组和治疗组ANF浓度与对照组比较无显著性差异(P＞0.05),但甲减未治组ANF明显低于治疗组(P＜0.05).甲亢缓解组、 甲减治疗组ET与对照组比较无显著性差异(P＞0.05),但甲减未治疗组ET均明显低于其它组(P＜0.01,P＜0.05).测定ANF和ET对甲亢和甲减的辅助诊断和疗效观察具有一定临床价值.     

Concurrent cisplatin-based radiotherapy and chemotherapy for locally advanced cervical cancer

BACKGROUND AND METHODS: On behalf of the Gynecologic Oncology Group, we performed a randomized trial of radiotherapy in combination with three concurrent chemotherapy regimens -- cisplatin alone; cisplatin, fluorouracil, and hydroxyurea; and hydroxyurea alone -- in patients with locally advanced cervical cancer. Women with primary untreated invasive squamous-cell carcinoma, adenosquamous carcinoma, or adenocarcinoma of the cervix of stage IIB, III, or IVA, without involvement of the para-aortic lymph nodes, were enrolled. The patients had to have a leukocyte count of at least 3000 per cubic millimeter, a platelet count of at least 100,000 per cubic millimeter, a serum creatinine level no higher than 2 mg per deciliter (177 micromol per liter), and adequate hepatic function. All patients received external-beam radiotherapy according to a strict protocol. Patients were randomly assigned to receive one of three chemotherapy regimens: 40 mg of cisplatin per square meter of body-surface area per week for six weeks (group 1); 50 mg of cisplatin per square meter on days 1 and 29, followed by 4 g of fluorouracil per square meter given as a 96-hour infusion on days 1 and 29, and 2 g of oral hydroxyurea per square meter twice weekly for six weeks (group 2); or 3 g of oral hydroxyurea per square meter twice weekly for six weeks (group 3). RESULTS: The analysis included 526 women. The median duration of follow-up was 35 months. Both groups that received cisplatin had a higher rate of progression-free survival than the group that received hydroxyurea alone (P<0.001 for both comparisons). The relative risks of progression of disease or death were 0.57 (95 percent confidence interval, 0.42 to 0.78) in group 1 and 0.55 (95 percent confidence interval, 0.40 to 0.75) in group 2, as compared with group 3. The overall survival rate was significantly higher in groups 1 and 2 than in group 3, with relative risks of death of 0.61 (95 percent confidence interval, 0.44 to 0.85) and 0.58 (95 percent confidence interval, 0.41 to 0.81), respectively. CONCLUSIONS: Regimens of radiotherapy and chemotherapy that contain cisplatin improve the rates of survival and progression-free survival among women with locally advanced cervical cancer.     

Effect of bovine parvovirus replication on DNA,RNA, and protein synthesis in S phase cells

The kinetics of replication of bovine parvovirus (BPV) and the effect of viral replication on cellular macromolecular synthesis were examined in hydroxyurea (HU)-synchronized fetal bovine spleen cells. Immediately after release of cells from HU block, 80–85% of the cells began to synthesize DNA. The production of infectious progeny BPV proceeded at a faster rate in synchronized cells infected at the beginning of S phase than in asynchronous cultures. In synchronized cells, titers of infectious virus increased at 8 hr p.i. and the maximum titer was achieved by 20 hr. BPV DNA synthesis preceded the production of progeny virus by 2 hr. Although the rates of RNA and protein synthesis in infected cells were severely reduced after 8 hr p.i., BPV replication did not affect the rate of progression of cells through S phase.     

甲泼尼龙联合羟基脲治疗原发性膜性肾病的疗效

目的 探讨甲泼尼龙联合羟基脲治疗原发性膜性肾病的临床效果。方法 选取2016年1月~2018年12月我院经肾穿刺活检病理确诊为原发性膜性肾病患者42例,按照随机数字表法分成观察组（20例）和对照组（22例）。观察组给予甲泼尼龙联合羟基脲治疗,对照组接受甲泼尼龙联合环磷酰胺治疗,比较两组24h尿总蛋白、血浆白蛋白、临床疗效及并发症发生率。结果 两组治疗前后24h尿总蛋白、血浆白蛋白比较,差异无统计学意义（P＞0.05）;观察组总有效率为80.00%,低于对照组的86.36%,但差异无统计学意义（P＞0.05）。观察组并发症总发生率为35.00%,低于对照组的40.91%,但差异无统计学意义（P＞0.05）。结论 甲泼尼龙联合环磷酰胺治疗原发性膜性肾病较甲泼尼龙联合羟基脲降尿蛋白效果具有非劣效性,且无严重并发症发生,可作为治疗的备选方案。     

Impact of JAK2V617F Mutation Burden on Disease Phenotype in Chinese Patients with JAK2V617F-positive Polycythemia Vera (PV) and Essential thrombocythemia (ET)

Most patients with polycythemia vera (PV) and half of essential thrombocythemia (ET) possess an activating JAK2V617F mutation. The objective of this study was to better define the effect of JAK2V617F mutant allele burden on clinical phenotypes in Chinese patients, especially thrombosis. By real-time polymerase chain reaction (RT-PCR), the JAK2V617F mutation burden was detected in 170 JAK2V617F-positive patients, including 54 PV and 116 ET. The results showed that JAK2V617F allele burden was higher in PV than in ET (P< 0.001). Higher percentage of patients had JAK2V617F allele burden over 20% in PV than in ET (68.5% VS 26.7%) (P< 0.001). In PV patients, higher JAK2V617F allele burden was observed in female (P< 0.05) and leukocytosis patients (WBC above 10×10⁹/L) (P< 0.001). Meanwhile, ET patients showed increased JAK2V617F allele burden in the group with higher hemoglobin (HGB above 150g/L) (P< 0.05), leukocytosis (WBC above 10×10⁹/L) (P< 0.001), splenomegaly (P< 0.05) and thrombosis (P< 0.05). In conclusion, the JAK2V617F mutation allele burden is higher in Chinese patients with PV than ET. In PV patients, JAK2V617F mutation burden had influence on WBC counts. And the clinical characteristics of ET patients, such as WBC counts, hemoglobin level, splenomegaly and thrombosis, were influenced by JAK2V617F mutation burden. Male, high hemoglobin (HGB above 150g/L), and increased JAK2V617F mutation burden (JAK2V617F allele burden ≥16.5%) were risks of thrombosis (P< 0.05) for ET patients by Logistic Regression.     

Partial suppression by pyridinolcarbamate of growth and necrosis of atherosclerotic lesions in swine subjected to an atherogenic regimen that produces advanced lesions.

Conflicting results have been reported in the literature on the effects of pyridinol-carbamate on the prevention of experimentally induced atherosclerosis in rabbits. In the current study the effects of pyridinolcarbamate have been tested in a swine model in which advanced atherosclerosis is produced by a combination of balloon-intimal trauma and a hyperlipidemic diet. Results were compared with those in two reference groups of swine subjected to the same atherogenic regimen—one left untreated by drugs and the other treated with hydroxyurea which is one of the antimetabolites used in cancer chemotherapy.In both drug-treated groups the intimal surface involved by atherosclerotic lesions, and the lesion area expressed as a ratio to the total medial area were significantly less than those of the untreated control group. Both drugs also significantly reduced the number of necrotic atheromatous lesions as compared to the untreated group. The mechanism of action of pyridinolcarbamate in our experimental condition is not clear at present. The demonstration of a beneficial effect of a relatively non-toxic drug such as pyridinolcarbamate seems to us to warrant carrying out further “in depth” study of this drug.     

聚乙二醇干扰素联合利巴韦林治疗慢性丙型肝炎的疗效研究

目的 观察聚乙二醇干扰素联合利巴韦林对慢性丙型肝炎初治与复发患者的疗效,并分析可能影响疗效的因素.方法 对64例慢性丙型肝炎的初治和复发患者应用聚乙二醇干扰素联合利巴韦林抗病毒治疗.对上述病例进行回顾性研究,分析两组患者的快速病毒学应答、完全早期病毒学应答、治疗结束时病毒学应答及持续病毒学应答,并探讨影响病毒学应答的相关因素.结果 初治组患者获得快速病毒学应答、完全早期病毒学应答、治疗结束应答、持续应答的概率明显高于复发组,差异具有统计学意义（P＜0.05）;初治组与复发组复发率不具有统计学意义（P＞0.05）,但复发组慢性丙型肝炎患者治疗结束后复发率明显高于初治组（33.33％与6.38％）.发生持续应答患者的快速病毒学应答、完全早期病毒学应答率明显高于未发生持续应答患者,且差异有统计学意义（P＜0.05）.结论 初治CHC患者较复发患者可获得较高病毒学应答率,复发率较低,获得快速病毒学应答和完全早期病毒学应答是疗效的阳性预测指标.     

Nucleolar organizer regions of megakaryocytes in chronic myeloproliferative disorders

Summary To study megakaryocyte activation, the argyrophilic staining method of nucleolar organizer regions (AgNOR) has been applied to decalcified bone marrow biopsies of 16 individuals with no haematopoietic disorders and 59 patients with chronic myeloproliferative disease. Of the 59 patients, 18 had chronic myeloid leukaemia (CML), 21 chronic megakaryocytic granulocytic myelosis (CMGM), 13 polycythaemia vera (PV) and 7 essential thrombocythaemia (ET). The AgNOR number of megakaryocytes in CML was significantly lower, and in CMGM, PV and ET significantly higher than in healthy individuals. The high number and the clusters of fine-grained AgNORs of megakaryocytes in CMGM, PV and ET are suggestive of active, proliferating cells. The AgNOR number of megakaryocytes and the platelet counts of the patients did not show a convincing correlation. In CMGM, PV and ET the pyknotic, heterochromatinized megakaryocytes with narrow rims of cytoplasm called bare (nude) nuclei, possessed few, large AgNOR granules. The AgNOR staining of bare nuclei and the roughly identical number of granules found in CMGM, PV and ET indicate a common, active mechanism of apoptosis.     

一氧化氮和内皮素对肝硬化大鼠离体内脏血管条舒缩的影响

目的:探讨一氧化氮(NO)和内皮素(ET)血管活性作用的动静脉差异在门脉高压形成机制中的意义。方法:以四氯化碳皮下注射复制大鼠肝硬化门脉高压模型,取动物门静脉(PV)和肠系膜动脉(MA)制成螺旋血管条,观察NO和ET的最大舒缩反应(Rmax,Cmax),并计算得到EC50值。结果:模型组(n=8)大鼠PV和MA对NO的Rmax显著高于对照组(n=7),EC50显著低于对照组(P<0.05,P<0.01);而对ET的Cmax则显著低于对照组,EC50显著高于对照组(P<0.05,P<0.01)。两组大鼠对NO和ET的最大舒缩反应和EC50,在PV和MA之间均存在显著差异,但模型组相差的幅度显著大于对照组(P<0.05或P<0.01)。结论:NO和ET在肝硬化大鼠内脏血管的舒缩作用存在动静脉差异,这可能是它们参与门脉高压形成的重要因素。     

核苷类似物和抗结核药治疗携带HBV肺结核患者的临床观察

目的观察肺结核并HBV携带初治患者抗结核治疗的同时核苷类似物抗病毒治疗的临床效果。方法将肺结核并HBV携带者随机分为A、B、C组,每组30例。A组抗结核治疗（2HREZ／4HR）的同时予拉米夫定或者恩替卡韦抗病毒及甘草酸二胺肠溶胶囊护肝治疗,B组抗结核治疗（2HREZ／4HR）的同时予甘草酸二胺肠溶胶囊护肝治疗,C组仅给予抗结核治疗（2HREZ／4HR）,比较3组患者肝功能受损情况、HBV—DNA变化情况。结果A组肝损率较B组和C组低,差异均具有统计学意义（P〈0．05）;B组肝损率较C组低,差异有统计学意义（P〈0．05）;A组治疗后2月、6月HBV-DNA水平较治疗前明显下降,差异有统计学意义（P=0．000）;B、C组治疗后6月HBV-DNA水平较治疗前无明显下降,差异无统计学意义（P=0．476、0．941）。结论对肺结核并HBV携带者早期予核苷类似物抗病毒治疗,能有效抑制HBV-DNA复制,减少肝功能损伤发生率,从而顺利完成抗结核治疗疗程,可广泛应用于临床。     

口服避孕药预治疗对多囊卵巢综合征患者体外受精-胚胎移植结果的影响

目的探讨多囊卵巢综合征(PCOS)患者应用口服避孕药预治疗对体外受精-胚胎移植(IVF-ET)结局的影响。方法 192例PCOS患者IVF-ET周期随机分成两组:研究组IVF-ET前先用口服避孕药(妈富隆或达英35)预治疗93例,对照组99例。比较两组年龄、促性腺激素(Gn)用量、获卵数、受精率、植入率、妊娠率、流产率和卵巢过度刺激综合征(OHSS)的发生率。结果两组年龄、Gn用量、获卵数、受精率无明显差异,预治疗组的植入率、妊娠率显著高于对照组(P0.05)。流产率和卵巢过度刺激综合征(OHSS)的发生率对照组(9.1%)显著高于研究组(5.2%)。结论 PCOS患者IVF-ET前先用口服避孕药顸治疗可提高植入率和妊娠率,降低OHSS的发生率。     

Allogeneic hematopoietic cell transplantation for advanced polycythemia vera and essential thrombocythemia

Allogeneic hematopoietic cell transplantation (HCT) is curative for selected patients with advanced essential thrombocythemia (ET) or polycythemia vera (PV). From 1990 to 2007, 75 patients with ET (median age 49 years) and 42 patients with PV (median age 53 years) underwent transplantations at the Fred Hutchinson Cancer Research Center (FHCRC; n = 43) or at other Center for International Blood and Marrow Transplant Research (CIBMTR) centers (n = 74). Thirty-eight percent of the patients had splenomegaly and 28% had a prior splenectomy. Most patients (69% for ET and 67% for PV) received a myeloablative (MA) conditioning regimen. Cumulative incidence of neutrophil engraftment at 28 days was 88% for ET patients and 90% for PV patients. Acute graft-versus-host disease (aGVHD) grades II to IV occurred in 57% and 50% of ET and PV patients, respectively. The 1-year treatment-related mortality (TRM) was 27% for ET and 22% for PV. The 5-year cumulative incidence of relapse was 13% for ET and 30% for PV. Five-year survival/progression-free survival (PFS) was 55%/47% and 71%/48% for ET and PV, respectively. Patients without splenomegaly had faster neutrophil and platelet engraftment, but there were no differences in TRM, survival, or PFS. Presence of myelofibrosis (MF) did not affect engraftment or TRM. Over 45% of the patients who undergo transplantations for ET and PV experience long-term PFS.