目的 通过观察血清胱抑素C水平的变化,探讨BDT新型化疗方案与VADT传统化疗方案在改善多发性骨髓瘤(MM)病人肾功能方面的差异.方法 回顾性分析84例确诊的MM病人,其中40例采用BDT化疗方案,44例采用VADT化疗方案.观察化疗前及4个周期化疗后血清胱抑素C、肌酐、尿素氮水平的变化,并进行统计学分析.结果 MM病人化疗前肾功能异常组较肾功能正常组血清胱抑素C水平明显升高(t=6.086,P＜0.05);根据ROC曲线下面积,血清胱抑素C水平在评价MM病人肾功能方面较肌酐、尿素氮更灵敏.Ⅰ、Ⅱ期MM病人血清胱抑素C水平比较差异无统计学意义(P＞0.05),Ⅲ期病人血清胱抑素C水平较Ⅰ、Ⅱ期显著升高(F=10.046,t =5.836、4.126,P＜0.05).MM病人采用BDT与VADT化疗方案治疗后血清胱抑素C水平均明显下降,且BDT化疗方案组化疗后血清胱抑素C下降程度较VADT化疗方案组更明显(t=6.242,P＜0.05).结论 血清胱抑素C为评价MM肾功能的一个灵敏指标;以硼替佐米为主的BDT化疗方案可明显降低MM病人血清胱抑素C水平,对改善肾功能更有优势,可作为合并肾功能不全MM病人的首选化疗方案.

目的 探讨骨髓涂片、活检并染色体检查对全血细胞减少症的诊断价值.方法 对206例全血细胞减少症病人的骨髓涂片、活检及染色体检查结果进行分析.结果 206例病人经骨髓涂片、活检并染色体检查确诊194例,确诊率为94.2％,骨髓涂片确诊率为51.0％,骨髓活检确诊率为74.3％,骨髓涂片并活检确诊率为85.9％,差异有显著性(x2 =7.83～23.89,P＜0.05).结论 骨髓活检诊断全血细胞减少的价值高于单纯骨髓涂片,骨髓涂片及活检并染色体检查能有效提高全血细胞减少症的诊断准确性.

目的 建立闭环式输血信息管理系统,提高临床输血的实用性、安全性和可控性,提高工作效率.方法 连接、整合输血信息管理系统与医院信息系统、实验室信息系统、医院不良事件报告系统,实现相关信息共享,将输血申请与审核、血清学相容性试验、血液发放、输注及输注后评价、输血不良事件报告等完整输血管理过程中的所有要素串联成一个环环相扣的工作流程,防止过程或要素遗漏,减少人为干预,使临床输血管理做到自动化、程序化、完整化.结果 输血科工作效率和临床合理用血水平大幅提高,规范了输血过程,差错发生率明显降低,杜绝了输血安全事故的发生,确保临床用血安全.结论 闭环式输血信息管理系统能有效提高医院的输血管理水平.

目的 探讨骨髓涂片、骨髓活组织检查(活检)、骨髓流式细胞术(FCM)联用检测初治非霍奇金淋巴瘤(NHL)骨髓侵犯的价值.方法 对96例初治NHL病人同时行骨髓涂片、骨髓活检、骨髓FCM检查,比较不同方法对NHL病人骨髓侵犯检测的阳性率,以及3种方法联合检测的价值.结果 骨髓涂片、骨髓活检、骨髓FCM检查及3种方法联用对NHL病人骨髓侵犯检测的阳性率分别为19.8％、22.9％、36.5％、45.8％.骨髓FCM检查对NHL骨髓侵犯检测的阳性率显著高于单用骨髓涂片和骨髓活检(x 2=7.76,P＜0.05);3种方法联用对NHL骨髓侵犯检测的阳性率最高(x 2=19.69,P＜0.05).弥漫大B细胞淋巴瘤骨髓侵犯构成比最高.结论 骨髓涂片、骨髓活检及骨髓FCM检查联用有助于提高初治NHL骨髓侵犯检测的阳性率,其优势互补,不可替代;对于部分不明原因发热病人进行骨髓检查有助于明确诊断.

目的 观察重组人血小板生成素(rhTPO)联合环孢素A(CsA)治疗难治性原发免疫性血小板减少症(R-ITP)的效果.方法 将41例R-ITP病人随机分为观察组(21例)和对照组(20例).观察组病人皮下注射rhTPO 300 U/(kg·d),疗程14d,同时分2次口服CsA 3～4 mg/(kg·d),疗程3个月.对照组病人皮下注射rhTPO 300 U/(kg·d),疗程14d.停药后定期观察病人血小板计数3个月.结果 治疗后第1、2周末观察组与对照组有效率分别为61.9％、55.5％和90.5％、90.0％,差异均无显著性(P＞0.05).治疗后第1个月末观察组与对照组复发率分别为15.8％、44.4％,差异无显著性(P＞0.05);第2、3个月末两组复发率分别为26.3％、61.1％和31.6％、83.3％,差异均有显著性(P=0.049、0.003).结论 TPO联合CsA治疗R-ITP,既能获得显著近期疗效,又能较长时间维持疗效.

自2013年开始,青岛大学医学院附属医院实施“无障碍就医”以来,医院在各个方面都取得了显著成效.文章通过对“无障碍就医”工程实施的动因、意义、主要做法和成效进行剖析,为公立医院探索内部管理方式改革,实现患者便捷就医、安全就医的目标,缓解群众“看病难、看病贵”问题提供了有益的借鉴.

Objective To compare the efficacy and safety of high-dose dexamethasone(HD-DXM)with conventional prednisone in the treatment of adults with primary immune thrombocytopenia(ITP).Methods A total of 73newly diagnosed ITP patients were divided into two groups in random.Dexamethasone group(DXM group,37patients):oral DXM 40mg/d,to be taken in two doses,for 4days and then discontinued,which was repeated for one more cycle on day 7after the discontinuation.Prednisone group(36patients),oral prednisone 1.0-1.5mg·kg-1·d-1 for 4weeks,and then the dose was gradually decreased to minimum maintenance dose or discontinued.The short-term and long-term efficacy and safety were compared between the two groups.Results For short-term efficacy,after 1-2-week treatment,the response in Dexamethasone group was significantly higher than that in Prednisone group(χ2=7.21,4.30;P<0.05),the efficacy on week 3remained higher,but the difference was not statistically significant(χ2=1.56,P>0.05).For long-term effect,a three-month follow-up showed that,on the first month,the recurrence rate between the two groups was not significant(χ2=0.11,P>0.05),and that on the second and the third month was much lower in Dexamethasone group than that in Prednisone group(χ2=4.49,4.80;P<0.05).In Dexamethasone group,little adverse reactions such as infections or Cushing syndrome were observed;In Prenisone group,Cushing syndrome manifestations were seen in more patients,and some complicated infections were also noted.Conclusion High-dose DXM therapy for ITP is superior than routine-dose Prednisone in terms of short-and long-term efficacy and safety.

Objective To analyze clinical features and risk factors of thrombosis in patients with hematologic malignancies(HM).Methods Clinical data of 874 HM patients(who hospitalized during 2007.4—2012.3) were reviewed retrospectively.The occurrence of thrombosis related to hematologic disease,location of thrombosis,its treatment and prognosis,and high risk factors was analyzed.Results The incidence of thrombosis accounted for 5.84%.Cerebral embolism and myocardial infarction were commonly seen,most of which were confirmed within six months after the diagnosis of HM,and 46.03% occurred after chemotherapy,18.60% died of thrombosis.Univariate analysis showed the high risks of thrombosis in HM were male,age ≥50 years,APL,ALL,unknown-type AL,MM,combined with other tumors,smoking index ≥400,high blood pressure,diabetes,coronary heart disease and varicose veins(χ2=2.763-34.984,P0.10).Multivariate Logistic regression analysis showed that male,advanced age,APL,unknown-type AL,MM,combined with other tumors,diabetes,coronary heart disease and varicose veins were independent risk factors.Conclusion Incidence of thrombosis is relatively high in patients with HM,male,advanced age,APL,unknown-type AL,MM,combined with other tumors,diabetes,coronary heart disease and varicose veins being independent risk factors.The thrombosis mostly occurs within six months after the diagnosis of HM,it is important to pay attention to its prevention and treatment,so as to improve its prognosis.

This study was purposed to investigate the difference of nucleated cell (NC) count, CD34(+) cell ratio and expansion multiple, cell cycle and colony formation capability in in vitro expanded human umbilical cord blood CD34(+) cells from HOXB4-transfecting directly and HOXB4-transfected human umbilical cord mesenchymal stem cells (HUCMSC) by means of prepared feeder layers of HUCMSC. The HUCMSC were divided into 2 groups:first group, in which HOXB4 gene was transfected into HUCMSC by using lentiviral vecfor, and feeder layers were set up; and second group in which feeder layers for HUCMSC of non-transfected HOXB4 gene were set up. The CD34(+) cells were separated from HUCB by magmatic activated cell sorting(MACS). After culture in medium with cytokines for 2 days, CD34(+) cells were divided into 5 groups, including control group and experimental group. The control groups included CD34(+) cells as group A (blank control group) and GFP-CD34(+) cells as group B (negative control group) and experimental groups included HOXB4-CD34(+) cells as group C, HUCMSC+CD34(+) cells as group D, HOXB4-HUCMSC+ CD34(+) cells as group E and cells in all groups were cultured in vitro. The number of nucleated cells were counted at day 6, 10, 14 of culture and CD34 immunophenotypes, cell cycle and colony forming capability were measured at day 10 of culture in different conditions. The results indicated that HOXB4 gene could be transfected into HUCMSC by lentiviral vector and feeder layers were set up successfully. After culture for 14 days, the nucleated cells in 5 groups could be amplified effectively, and the expansion levels in 5 groups were in order HOXB4-HUCMSC+CD34(+) cell group> HOXB4-CD34(+) cell group>HUCMSC+CD34(+) cell group> control groups (P < 0.05). At day 10 of in vitro expansion the CD34(+) cell percentage decreased significantly in all groups, while the number of CD34(+) cell increased in experiment groups, which were in order HOXB4-CD34(+) cells group> HOXB4-HUCMSC+CD34(+) cell group>HUCMSC+CD34(+) cell group>control groups (P < 0.05). The cell cycle detection showed that the percentage of cells in S+G2/M phase in experiment groups were higher than that in control groups (P < 0.05), and percentage of cells in HOXB4-HUCMSC+CD34(+) cells group was higher (41.57%) than that in HOXB4-CD34(+) cells group(37.87%) and HUCMSC+CD34(+) cell group (28.65%) (P < 0.05). There was no statistical difference in the CFU number between HOXB4-HUCMSC+CD34(+) cell group and HOXB4-CD34(+) cell group, which were both higher than that in HUCMSC+CD34(+) cell group and control groups (P < 0.05).It is concluded that the CD34(+) cells cultured on HOXB4-HUCMSC feeder layers can be amplified significantly and kept the characteristics of stem cells, The feeder lager of HOXB4-HUCMSC is relative safe for amplification of CD34(+) cells in vitro, it possesses the potential useful value.

Objective To compare the changes of biological characteristics and HOXB4 gene expression before and after expansion of human umbilical cord blood( HUCB) CD34+ cells in vitro. Methods CD34+ cells were separated from HUCB by MiniMACS and cultured in 10% FBS + IMDM including the cytokine combination of FL, SCF,TPO and IL-3. In the period,we observed cell morphology and biological properties,counted the number of nucleated cells ( NC) and colony-forming units( CFU) ,analysed CD34+ cells ratio with flow cytometry,and detected the HOXB4 gene expression changes by RT-PCR at different time points ( day 0,7,14 ) . Results The purity of selected CD34+ cell reached to ( 83. 17 ± 8. 67 ) % . The ratio of CD34+ cell decreased to ( 4. 78 ± 0. 64 ) % and ( 0. 82 ±0. 13) % on 7 days and 14 days culture. There was varying increase in the proportion of nucleated cells ( NC) and CD34+ cell,which the number of NC and CD34+ cell were respectively augmented( 76. 2 ± 14. 9) times, ( 254. 0 ±48. 85) times,( 3. 9 ± 0. 69) times and( 1. 8 ± 0. 64) times ( P < 0. 05) . The number of CFU on d7 was 3. 25 folds compared with day 0 culture. HOXB4 gene was highly expressed in separated CD34+ cells on day 0,but it declined rapidly in vitro expansion compared with its pre-culture level( P < 0. 05) . Conclusions The combination of FL + SCF + TPO + IL-3 enhances HUCB CD34+ cells effectively,and there is high amplification efficiency on 7 days culture compared with 14 days,but the self-renewal potential of CD34+ cells decrease in vitro culture.

目的:探讨地西他滨4d疗程方案治疗老年急性白血病的疗效及安全性。方法:应用地西他滨30mg/m2 4d疗程方案治疗4例老年急性髓系白血病,并评价其疗效和不良反应。结果:2例患者应用地西他滨4疗程后达完全缓解,1例在3疗程后获部分缓解,1例1疗程后因多器官功能衰竭死亡。4例患者治疗过程中均出现较明显的粒细胞和血小板减少。结论:地西他滨是治疗老年急性髓系白血病的有效选择,疗效较好,不良反应可以耐受。4d方案在疗效和耐受性上与3d和5d方案相比,无明显差别,应用方便。

This study was purposed to construct lentivirus vector containing human homeobox gene HOXB4 and explore changes of human umbilical cord mesenchymal stem cells (HUCMSC) after infected with HOXB4 mediated by lentivirus. PCR amplification was performed to obtain HOXB4, which was cloned in lenti-shuttle vector. Four-plasmid lentivirus packaging system was used to transfect HEK293T cells. After 48 h, lentivirus Lenti-HOXB4 was harvested and lentivirus titer was determined. Lenti-HOXB4 was used to infect HUCMSC. The infected cells were observed under inverted fluorescence microscope to determine the optimal multiplicity of infection (MOI). Meanwhile, RT-PCR, immune fluorescence staining, CCK-8 and flow cytometry (FCM) were used to determine the expression of HOXB4 and its effect on cell growth. The results indicated that lenti-HOXB4 was successfully obtained by co-transfecting the 293T cells with four plasmids. The determined virus titer was 3×10(8) TU/ml; when MOI was 20. Lenti-HOXB4 had a high transfection rate in HUCMSC, over 80%. In HUCMSC infected with lenti-HOXB4, the expression of target gene could be detected both at mRNA and protein levels. It could promote the proliferation of HUCMSC. FCM results indicated HOXB4 gene did not significantly influence the surface marker of HUCMSC. It is concluded that HOXB4 gene can promote the high proliferation of HUCMSC and does not significantly influence the expression of the surface marker of HUCMSC.

目的 探讨地西他滨4d方案治疗骨髓增生异常综合征(MDS)及老年人急性髓系白血病( AML)的疗效及安全性.方法 应用地西他滨每天30 mg/ m2,4 d方案治疗5例MDS及5例老年AML患者,观察其疗效及毒副作用.结果 3例患者获得了完全缓解,2例患者获得了部分缓解.不良反应主要为骨髓抑制和感染,较3d及5d方案毒副作用未增加.结论 地西他滨对于MDS及老年白血病患者有效,其不良反应可以耐受,4d方案经济、方便,适合在临床上使用.

近年来,陆续有研究者报道一类特殊的骨髓瘤,其形态表现为淋巴样浆细胞,并有较独特的生物学特点.此型骨髓瘤较为少见,并且因其临床表现、细胞病理学与某些淋巴瘤存在部分相似性,因此不易诊断.现对我院收治的2例患者资料进行回顾性分析,并对相关文献进行复习.

The purpose of this study was to detect the expression levels of geminin and cdt1 in peripheral blood and bone marrow from patients with newly diagnosed acute leukemia (AL), and further explore effects of them in the pathogenesis of AL. mRNA expression of geminin and cdt1 in peripheral blood and bone marrow of newly diagnosed AL patients was detected by SYBR Green real-time quantitative reverse transcription polymerase chain reaction(SYBR-RT-PCR). The results showed that mRNA expressions of both geminin and cdt1 in peripheral blood were positive in 10 out of 13 newly diagnosed ALL patients (76.92%) and in 9 out of 14 newly diagnosed AML patients (64.29%), while no positive expression of these 2 genes was detected in 10 normal controls; mRNA expression levels of geminin and cdt1 in bone marrow of newly diagnosed ALL and AML patients were 108.06 ± 67.34 and 52.37 ± 35.16, 62.66 ± 58.69 and 26.68 ± 22.29, respectively, which were higher than those in normal controls (11.81 ± 2.83 and 7.32 ± 5.77), there were significant differences (p < 0.01 and p < 0.05). mRNA expression of geminin was significantly positive related to mRNA expression of cdt1 in bone marrow of 34 newly diagnosed AL patients (r = 0.55, p < 0.01). It is concluded that mRNA expressions of geminin and cdt1 are enhanced and significantly positively related between them in bone marrow of AL patients. The over-expression of geminin and cdt1 mRNA may play an important role in pathogenesis of AL.

Objective To detect the expression levels of geminin and cdt1 in bone marrow of patients with acute myeloid leukemia(AML),and investigate their relationship with the pathogenesis and prognosis of this condition. Methods Using SYBR Green real-time quantitative reverse transcription polymerase chain reaction(SYBR-RT-PCR),the expressions of geminin and cdt1 mRNA were detected in 47 AML patients at different phases,including 25 initially-treated,16 complete-remission and six non-remission or relapsed.A follow-up of six months was done in those initially treated. Results The expression levels of geminin and cdt1 in initially-treated,non-remission or relapsed patients were higher than that in the controls and complete-remission,the differences being significant(H=4.95,3.22;Z=2.67-4.95;P0.01);the differences of geminin and cdt1 between the complete-remission and the controls were not significant(P0.05).In AML,the expressions of geminin and cdt1 in bone marrow cells were positively correlated(r=0.59,P0.01),and a positive correlation was noted of both the two expressions in initially-treated,remission and relapsed groups(r=0.49-0.87,P0.05).A follow-up of six months was conducted in 25 initially-treated patients,six of them died.The expression of geminin in those who died was higher than that who survived,the difference being significant(t=2.16,P0.05),but the difference of cdt1 expression between the two groups was not significant(t=0.61,P0.05). Conclusion Geminin and cdt1 expressions may be correlated with the course of the disease,and the high expression of geminin in initially-treated patients is likely to be a poorly prognostic prediction of AML.

In order to investigate the special role of HOXB4 in expansion and self renewal of hematopoietic stem cells, the cDNA of HOXB4 was extracted and cloned from umbilical cord blood mononuclear cells by using RT-PCR. Then the eukaryotic expression bicistronic plasmid vector pIRES2-EGFP/HOXB4 was designed and constructed after cutting HOXB4 and pIRES2-EGFP respectively by restriction enzyme EcoRI and BamHI. The recombinant plasmid was delivered into competent cells of Escherichia coli. The successful construction of plasmid was confirmed by the identification of endonuclease cutting and sequencing. The results showed that the HOXB4 cDNA was cloned successfully from umbilical cord blood mononuclear cells and the recombinant eukaryotic expression bicistronic plasmid vector was constructed, and then introduced it into 293T cells successfully. It is concluded that a pIRES2-EGFP/HoxB1 eukaryotic expression bicistronic plasmid vector has been constructed successfully, which results provide a useful material basis for exploration of HoxB4 function in the proliferation and differentiation of hematopoietic cells.

Objective To investigate the clinical significance of the level of serum soluble human leukocyte antigens Ⅰ(sHLA-Ⅰ) in patients with acute leukemia,lymphoma or multiple myeloma.Methods Concentrations of serum sHLA-Ⅰ in 112 patients with acute leukemia,lymphoma multiple myeloma were determined by enzyme linked immunosorbent assay.Results Concentrations of serum sHLA-Ⅰ in acute leukemia,lymphoma multiple myeloma patients were significantly increased compared with normal control(all P<0.01).The levels of sHLA-Ⅰ of Lymphoma and multiple myeloma patients were significantly higher than those of acute leukemia subjects(all P<0.05),but no difference was detected between lymphoma and multiple myeloma(P>0.05).The sHLA-Ⅰ levels in untreated and relapsed patients were significantly higher than those in the complete remission subjects(all P<0.01),while there was no difference between the untreated and relapsed patients significantly(P>0.05).The serum level of sHLA-Ⅰ was closely correlated with the peripheral leukocytes and contents of serum LDH as well as β2 macroglobulin.Conclusions Serum sHLA-Ⅰ can be as a prognosis and relapse index for malignant hemopathy.

OBJECTIVE:To explore the expression and correlation of MK and VEGF gene in acute leukemia(AL),and evaluate their relationship with angiogenesis and prognosis.METHODS:The expression level of MK and VEGF in 95 AL patients in different phase were detected by Taqman real-time fluorescent quantitative RT-PCR.RESULTS: The expressions level of MK and VEGF in acute myelogenous leukemia(AML) and acute lymphoblastic leukemia(ALL) were significantly higher than those in control group(P=0.000 0);The expression level of MK and VEGF in untreated/relapsed groups was significantly higher than that in control/remission groups(P0.01);The expression level of MK and VEGF in remission group was also significantly higher than that in control group(P0.01).Linear correlation was observed between the level of MK and VEGF in relapsed group(r=0.526,P0.05).In remission group,MK had linear relationship with VEGF and the ratio of the blast cell in bone marrow(r=0.673,P0.01;r=0.468,P0.01),but in untreated/control groups,there was no linear relationship among MK,VEGF level and the ratio of the blast cell in bone marrow.CONCLUSIONS: The high expressions of MK and VEGF are correlated with angiogenesis and prognosis in AL,and there is synergistic effect between MK and VEGF.Monitoring MK and VEGF can be used to guide the treatment and to estimate prognosis.

Objective To investigate the features of Ph chromosome and its significance in chronic myelogenous leukemia(CML).Methods Karyotyping was performed in 124 CML patients by R-banding,and dynamic detection of chromosomal changes was conducted in some of them in the course of progress.Results Of the 124 cases,93.55% was Ph chromosome positive(Ph+) and 6.45% negative(Ph-).In the Ph+ patients,94.83% was classical Ph and 5.17% variant rearrangement.Extra chromosome changes were demonstrated in 11.21% of classical Ph cases.The rate of extra chromosome was found in 7.48% of patients in chronic phase(CP),33.33% in accelerated phase(AP),and 66.67% in blast crisis phase(BP),of which,the most common ones were +8,2Ph,-21 and i(17q).The detection of extra chromosome in AP and BP was higher than that in CP(χ2=19.285,P<0.01),many abnormal clones were seen simultaneously in BP patients.Conclusion There is a close correlation between extra chromosome changes and the progress of CML.Chromosome karyotype analysis is important in the diagnosis,treatment and prediction the prognosis of CML.