中西医结合治疗慢性支气管炎急性发作30例

目的:在西医综合治疗基础上观察口服控涎丹与艾灸隐白穴联合治疗慢性支气管炎(慢支)急性发作的效果.方法:采用随机方法,将患者分为西医对照组共30例,中西医结合治疗组共30例.结果:治疗组痊愈率90%,总有效率为100%;对照组痊愈率为33.3%,总有效率为83.33%,痊愈率及总有效率治疗组明显高于西药对照组(P＜0.01和P＜0.05),有显著性差异.且治疗组住院时间较对照组明显缩短.结论:加用口服控涎丹与艾灸隐白穴治疗慢支急性发作疗效确切,未发现毒副作用,安全性好.     

伴NPM1基因突变的成年人急性髓系白血病临床研究

目的 探讨我国成年伴NPM1基因突变的急性髓系白血病患者(NPMc+AML)的临床特点,初步探讨定期定性检测该突变在早期判断AML复发中的意义.方法 采用聚合酶链反应(PCR)-毛细管电泳法对95例成年初治AML患者检测NPM1突变情况,并选取其中5例完全缓解患者定期检测该突变.结果 95例成年AML患者NPM1突变发生率为29.5%(28/95);≥40岁患者突变发生率[40.0%(22/55)]明显高于＜40岁患者[15.0%(6/40)](λ 2=6.963,P=0.012);正常核型AML患者突变发生率[51.1%(24/47)]明显高于异常核型患者[8.3%(4/48)](λ2=20.860,P=0.000).AML患者发生NPM1突变以M5[72.7%(16/22)]、M2[36.3%(8/22)]常见,在具有重现性染色体异常的AML中,未发现该突变.NPMc+AML患者白细胞、血小板计数及乳酸脱氢酶水平均明显高于NPMc-AML组(t值分别为4.132、4.603、4.069,均P＜0.05).NPMc+AML患者完全缓解率、无复发生存率及总生存率均明显高于NPMc-AML患者(λ2值分别为10.448、4.146、4.384,均P＜0.05).定期检测的患者血液学复发前1.5～2.0个月草新出现NPM1基因突变.结论 NPM1基因突变在成年AML患者中,尤其是正常核型AML患者中有较高的发生率,临床表现为患者年龄偏大,白细胞计数、血小板计数、乳酸脱氢酶均较高,NPM1基因突变是成年AML患者预后良好的指标.定期定性监测该突变可早期判断AML复发.     

多发性骨髓瘤微小RNA基因序列突变的检测及其意义

目的 了解多发性骨髓瘤(MM)患者微小RNA(miRNA)基因突变率及其临床意义.方法 采用聚介酶链反应单链构象多态性(PCR-SSCP)银染法检测4种人类MM细胞系(KM-3、ARH-77、U266、RPMI8226)、20例MM患者及20例血液学正常者12种miRNA基因突变,实验结果结合临床分析.结果 细胞系KM-3、RPMI8226检测出miRNA-335基因突变;MM患者中检出miR-19a、miR-19b和miRNA-335基因突变各1例,总突变率为15.00%(3/20);正常对照组未检出突变.病例组突变者中1例确诊4个月左右死亡,2例取材时已处于疾病终末期.结论 miRNA基因在MM中存在较高的突变率,miR-19a、miR-19b和miRNA-335基因突变可能与MM发病机制有关;发生突变的患者(表现)临床预后差,对miR-19a、miR-19b和miRNA-335基因突变的检测可能有助于对MM病程进展和预后的分析.     

急性白血病患者乳酸脱氢酶变化的临床意义

目的 观察急性白血病(AL)患者血清乳酸脱氢酶(LDH)的变化,探讨其对AL疗效和预后判断的临床意义.方法 采用全自动生化分析仪检测156例AL患者[包括初治组63例、完全缓解(CR)组46例、复发组47例]的LDH值,常规反带方法分析AL染色体核型,并根据核型分成预后好、预后中等及预后差3组,分析LDH值与上述分组、患者白细胞数及白血病亚型的相关性.结果 初治组和复发组的AL患者血清LDH值[其中位数(P50)分别为399和265 U/L]明显高于CR组(P50 153 U/L)(P＜0.05).LDH的增高与外周血白细胞数(rs=0.604)及骨髓中白血病细胞比例(rs=0.538)密切相关(均P＜0.01).LDH与不同白血病亚型有关(L2、M4明显增高)(P＜0.01).难治复发的AL患者初治时LDH值(P50 538 U/L)高于非难治复发组(P50 294 U/L).具有不良预后染色体核型的AL患者初治时LDH(P50 778 U/L)明显高于预后良好组(P50 306 U/L)和预后中等组(P50 405 U/L)(P＜0.01).结论 LDH在AL发病时明显升高,其升高程度与病期、亚型及恶性度有相关性,AL缓解期LDH可下降,LDH可作为AL诊治过程中判断病情、评估预后的参数.     

P53突变蛋白表达对弥漫大B细胞淋巴瘤预后的预测作用

目的 探讨p53突变蛋白表达对弥漫大B细胞淋巴瘤(DLBCL)预后的预测作用,指导个体化治疗.方法 随机选择初治DLBCL患者62例,应用免疫组织化学方法检测p53突变蛋白和CD10、bcl_6、MUM1的表达,分析p53突变蛋白表达与患者临床特征、分子分型以及预后的关系.结果 48.4%(30/62)的患者表达p53突变蛋白.p53突变蛋白表达与初始治疗反应有关(x2=20.365,P=0.040),阳性组的完全缓解率为33.3%(10/30),阴性组为59.4%(19/21);与分子分型有关(x2=31.023,P=0.021),阳性组非生发中心型比例显著高于阴性组,分别为83.3%和56.2%;与其他临床特征无关.多因素生存分析显示p53突变蛋白表达是独立的预后预测因子,阳性组的无进展生存期和中位生存期均短于阴性组(x2=36.784,P=0.005和x2=35.276,P=0.006).结论 p53突变蛋白表达是DLBCL独立的不良预后因子,能够用来指导个体化治疗.     

PTEN、MLL基因在T淋巴母细胞淋巴瘤/白血病的表达及其意义

目的 分析肿瘤抑制基因PTEN、混合系白血病(MLL)基因等在T淋巴母细胞淋巴瘤/白血病(T-LBL/ALL)的表达及意义.方法 选用76例T-LBL/ALl患者淋巴结存档蜡块,应用免疫组织化学EnVision法进行 PTEN标记,用20例反应性增生淋巴结标本作正常对照.并用荧光原位杂交(FISH)技术检测MLL基因所在1 1q23染色体的断裂和扩增情况.结果 76例T-LBL/ALL中,PTEN的表达率为64.47%(49/76),低于淋巴结反应性增生的100%(20/20)(λ2=19.220,P＜0.05).PTEN表达与临床分期、Ki-67、乳酸脱氢酶(LDH)呈负相关(P＜0.05).76例T-LBL/ALL中,MLL基因发生1 1q23染色体断裂13例(17.11%),扩增18例(23.68%).MLL基因断裂组总体生存率(25.0%)低于非断裂组(43.6%)(λ2=11.357,P＜0.05).MLL基因扩增组总体生存率(17.1%)低于非扩增组(42.7%)(λ2=4.533,P＜0.05).结论 抑癌基因PTEN表达降低在T-LBL/ALL的发生发展中可能具有重要作用.MLL基因发生染色体1 1q23断裂和扩增有助于对T-LBL/ALL预后的判断,发生MLL基因断裂或扩增的T-LBL/ALL预后较差,提示MLL基因断裂或扩增可能为T-LBL/ALL的一种分子亚型.     

MT方案治疗急性单核细胞白血病的疗效及其与染色体核型的关系

目的 研究米托蒽醌联合替尼泊苷(MT)方案在急性单核细胞白血病(M5)诱导缓解中的疗效及患者不良反应,并观察疗效与白血病染色体核型的关系.方法 将33例M5患者按治疗史分两组:初治组23例(A组)、DA(柔红霉素联合阿糖胞苷)或HDA(三尖杉酯碱、柔红霉素和阿糖胞苷)1个疗程无效组10例(B组).按核型预后分两组:预后中等组29例(C组),预后不良组4例(D组),均采用MT方案2个疗程诱导缓解,分别统计4组的临床疗效及患者不良反应.结果 MT方案对A、B组的M5诱导完全缓解(CR)率分别为83%(19/23)及60%(6/10),有效率达91%(21/23)及70%(7/10).C、D组CR率分别为83%(24/29)及25%(1/4),有效率为88%(26/29)及50%(2/4),其中复杂核型CR率为0(0/3),非复杂核型的11q23染色体异常患者一次化疗达CR率100%(4/4).MT方案对M5化疗后白细胞最低点在第(7±3)天出现,为(0.4±0.2)×109/L,白细胞<1×109/L时间达(8±5)d,未见化疗相关死亡病例.结论 MT方案简单有效、较安全,是治疗M5的较佳化疗方案,对1个疗程DA、HDA方案无效者亦可试用.MT方案化疗疗效与核型预后分组有关,对11q23染色体异常的M5患者疗效较好,对复杂核型患者疗效欠佳.     

沙利度胺治疗急性白血病的疗效及对血管调控因子水平的影响

目的 观察沙利度胺联合化疗治疗急性白血病的临床疗效及其对血浆血管内皮生长因子(VEGF)、血管内皮生长因子受体(VEGFR)、碱性成纤维细胞生长因子(bFGF)水平的影响.方法 急性白血病患者36例,随机分为试验组及对照组各18例.每组均予以常规化疗方案标准剂量化疗,试验组同时口服沙利度胺100 mg/d.治疗前及治疗后8周分别采集外周血,双抗体夹心酶联免疫吸附法(ELISA)检测血浆VEGF、VEGFR、bFGF含量.以15位健康体检者为健康对照组.结果 试验组与对照组有效率分别为88.9%(16/18)和77.8%(14/18),差异有统计学意义(x2=4.103,P＜0.05).试验组与对照组治疗前血浆VEGF水平分别为(389.78±249.94)和(318.54±125.78)pg/ml,高于健康组的(132.91±26.66)pg/ml(t=3.141、3.024,均P＜0.01);治疗后分别为(211.74±36.72)和(288.02±31.77)pg/ml,高于健康组(t=2.413、2.324,均P＜0.05);试验组与对照组治疗前VEGF差异无统计学意义(t=1.384,P＞0.05),治疗后差异有统计学意义(t=2.793,P＜0.05).试验组与对照组治疗前血浆VEGFR水平分别为(2490.75±1695.9)和(2322.78±1105.87)pg/ml,高于健康组的(1134.98±378.45)pg/ml(t=2.914、2.783,均P＜0.01);治疗后分别为(1359.71±390.24)和(1753.89±337.04)pg/ml,与健康组相比差异有统计学意义(t=2.572、2.447,均P＜0.05);试验组与对照组治疗前VEGFR差异无统计学意义(t=1.276,P＞0.05),治疗后差异有统计学意义(t=2.486,P＜0.05).试验组与对照组治疗前血浆bFGF水平分别为(2.43±0.27)和(2.4l±0.33)ng/ml,高于健康组的(1.83±0.44)ng/ml(t=4.982、4.171,均P＜0.05);治疗后分别为(2.09±0.17)和(2.11±0.31)ng/ml,与健康组相比差异有统计学意义(t=3.01l、2.773,均P＜0.05);试验组与对照组治疗前及治疗后相比差异无统计学意义(t=0.953、1.282,均P＞0.05).结论 沙利度胺联合化疗可提高急性白血病患者的缓解率,有可能成为一种通过抗血管新生从而抑制白血病细胞生长及浸润的有效治疗方法.     

三氧化二砷联合全反式维甲酸治疗初发急性早幼粒细胞白血病的临床观察

目的 观察全反式维甲酸(ATRA)联合三氧化二砷(As2O3)治疗初诊急性早幼粒细胞白血病(APL)的疗效及不良反应.方法 对ATRA每天25mg/m2联合As2O310mg/d(联合组)治疗的35例APL患者达完全缓解(CR)时间、CR率、早期病死率及不良反应进行观察,并与单用As2O3 10 mg/d(单药组)治疗的33例进行比较.结果 联合组CR率为94.3%(33/35),与单药组[90.9%(30/33)]比较差异无统计学意义(P>0.05);联合组获得CR时间为26.1 d,短于单药组的30.5 d,差异有统计学意义(P<0.05);联合组与单药组APL分化综合征及不良反应发生率、早期病死率比较,差异均无统计学意义(均P>0.05);高WBC组比中、低WBC组CR率低,死亡率高,差异均有统计学意义(均P<0.05),低WBC组与中WBC组差异无统计学意义(P>0.05).结论 As2O3联合ATRA较单用As2O3治疗初诊APL获得CR时间短,WBC>10×109/L为预后不良的因素,APL分化综合征应尽早发现,及时处理.     

抑肝扶脾法治疗腹泻型肠易激综合征临床观察

为探讨抑肝扶脾法治疗腹泻型肠易激综合征(IBS)的疗效,将120例随机分为治疗组和对照组各60例,治疗组采用抑肝扶脾法治疗,对照组则以思密达加谷维素治疗,治疗组总有效率为90%,对照组总有效率为80%,两组差异有统计学意义(p＜0.05),结果表明,抑肝扶脾法治疗腹泻型肠易激综合征有较好的临床效果.     

High mutation rate of a long microsatellite allele in Drosophila melanogaster provides evidence for allele-specific mutation rates

Within recent years, microsatellite have become one of the most powerful genetic markers in biology. For several mammalian species, microsatellite mutation rates have been estimated on the order of 10(-3)-10(-5). A recent study, however, demonstrated mutation rates in Drosophila melanogaster of at least one order of magnitude lower than those in mammals. To further test this result, we examined mutation rates of different microsatellite loci using a larger sample size. We screened 24 microsatellite loci in 119 D. melanogaster lines maintained for approximately 250 generations and detected 9 microsatellite mutations. The average mutation rate of 6.3 x 10(-6) is identical to the mutation rate from a previous study. Most interestingly, all nine mutations occurred at the same allele of one locus (DROYANETSB). This hypermutable allele has 28 dinucleotide repeats and is among the longest microsatellite reported in D. melanogaster. The allele-specific mutation rate of 3.0 x 10(-4) per generation is within the range of mammalian mutation rates. Future microsatellite analyses will have to account for the dramatic differences in allele-specific mutation rates.     

Detection of loss of heterozygosityat microsatellite loci in esophageal squamous-cell carcinoma

Microsatellite alterations at 3 genetic loci (chromosomes 2p, 3p and 17p) were analyzed in 25 tumors (20 primary tumors and 5 metastatic lymph nodes) from 20 patients after surgical treatment for esophageal cancer. DNA samples from tumors were compared with control DNA from lymphocytes obtained from the peripheral blood of the individual patients. Microsatellite alterations [microsatellite instability (MSI) and loss of heterozygosity (LOH)] were detected in 15% of 20 primary tumors with marker D2S123 (chromosome 2p), 55% with marker D3S1067 (chromosome 3p) and 50% with marker TP53 (chromosome 17p). The 3-year disease-free survival rate of the 10 patients who had tumors without alterations or with an alteration at only 1 of 3 microsatellite loci was 75% and it was better than that of the 10 patients who had tumors with alterations at 2 or 3 microsatellite loci (48%, p = 0.049). This finding suggests that esophageal cancer with alterations at multiple microsatellite loci might have strong malignant potential. However, MSI was only detected in one of 20 patients, which suggests that MSI might not play an important role in the development of this cancer. Three of 5 metastatic lymph nodes showed no LOH even though primary tumors of these patients exhibited LOH with 1 or 2 markers, and 1 metastatic lymph node had LOH that was detected with D3S1067 even though the primary tumor of this patient had no LOH with all markers. Thus, clonal heterogeneity might exist in esophageal squamous-cell carcinomas.     

High effectiveness and safety of one-week antibiotic regimen in Helicobacter pylori eradication

Forty-three sporadic gastric cancers were analyzed with regard to whether mutations of simple repeated sequences in the transforming growth factor beta type II receptor (TbetaR-II) gene are associated with microsatellite instability (MSI) and gastric carcinogenesis. In 12 of the 43 cancers (28%), MSI was observed at least at 1 of the 2 microsatellite loci. Frameshift mutations of the TbetaR-II gene, all of which were 1 base deletion of 10 adenine repeats, were detected in 3 of 6 cancers, with MSI at 2 loci. However, mutations were not detected in 6 cancers, with MSI only at 1 locus and 31 cancers without MSI. Moreover, microanalysis in these cases revealed that the mutant-type alleles of TbetaR-II were invariably common in different areas within the tumor, in contrast to the markedly variable alleles of microsatellite loci. Our results suggest that frameshift mutation of the TbetaR-II gene may be a critical event associated with MSI and may contribute to carcinogenesis of the stomach. One of the possible mechanisms of escape from growth control by TGFbeta during gastric carcinogenesis could involve frameshift mutations of the TbetaR-II gene caused by DNA replication errors.     

The mutation rates of di-, tri- and tetranucleotide repeats in Drosophila melanogaster

In a recent study, we reported that the combined average mutation rate of 10 di-, 6 tri-, and 8 tetranucleotide repeats in Drosophila melanogaster was 6.3 x 10(-6) mutations per locus per generation, a rate substantially below that of microsatellite repeat units in mammals studied to date (range = 10(-2)-10(-5) per locus per generation). To obtain a more precise estimate of mutation rate for dinucleotide repeat motifs alone, we assayed 39 new dinucleotide repeat microsatellite loci in the mutation accumulation lines from our earlier study. Our estimate of mutation rate for a total of 49 dinucleotide repeats is 9.3 x 10(-6) per locus per generation, only slightly higher than the estimate from our earlier study. We also estimated the relative difference in microsatellite mutation rate among di-, tri-, and tetranucleotide repeats in the genome of D. melanogaster using a method based on population variation, and we found that tri- and tetranucleotide repeats mutate at rates 6.4 and 8.4 times slower than that of dinucleotide repeats, respectively. The slower mutation rates of tri- and tetranucleotide repeats appear to be associated with a relatively short repeat unit length of these repeat motifs in the genome of D. melanogaster. A positive correlation between repeat unit length and allelic variation suggests that mutation rate increases as the repeat unit lengths of microsatellites increase.     

Measuring quality of life in cardiac rehabilitation clients

OBJECTIVE: To clarify the origin of defective mismatch repair (MMR) in sporadic endometrial cancers with microsatellite instability (MSI), a thorough mutation analysis was performed on the human mismatch repair gene MSH3. METHODS: Twenty-eight MSI-positive endometrial cancers were investigated for mutations in the human mismatch repair gene MSH3 using single-strand conformation variant (SSCV) analysis of all 24 exons. All variants were sequenced. Loss of heterozygosity was investigated at all MSH3 polymorphisms discovered. A subset of tumors were investigated for methylation of the 5' promoter region of MSH3 using Southern blot hybridization. RESULTS: An identical single-base deletion (delta A) predicted to result in a truncated proteins was discovered in six tumors (21.4%). This deletion occurs in a string of eight consecutive adenosine residues (A8). Because simple repeat sequences are unstable in cells with defective MMR, the observed mutation may be an effect, rather than a cause, of MSI. Evidence of inactivation of the second MSH3 allele in tumors with the delta A mutation would strongly support a causal role for these MSH3 mutations. However, there was no evidence of a second mutation, loss of sequences, or methylation of the promoter region in any of the tumors with the delta A mutation. CONCLUSION: Although the delta A mutation is a frequent event in sporadic MSI-positive endometrial cancers, it may not be causally associated with defective DNA MMR.     

Mutation of the type II transforming growth factor-beta receptor is coincident with the transformation of human colon adenomas to malignant carcinomas

The transforming growth factor-beta (TGF-beta) type II receptor (RII) is a colon cancer suppressor gene that is inactivated by mutation in 90% of human colon cancers arising via the microsatellite instability (MSI) pathway of carcinogenesis. To determine the pathophysiological consequence of RII mutations, we have determined the timing of their onset among 22 MSI human colon adenomas of varying stages. No RII mutations were detected in any early MSI adenoma, including all those with simple tubular or villous histology. The earliest RII mutation detected was in a region of high-grade dysplasia but was absent from the surrounding simple adenoma. Six additional RII mutations were all found in highly progressed adenomas that contained regions of frankly invasive adenocarcinoma. These RII mutations were detected in both the advanced adenomas and their adjacent regions of carcinoma. RII mutation is a late event in MSI adenomas and correlates tightly with progression of these adenomas to cancer.     

Microsatellite instability analysis: a multicenter study for reliability and quality control

The molecular biology section of the Hereditary Non-Polyposis Colorectal Cancer study group-Germany, instituted a multicenter study to test the reliability and quality of microsatellite instability (MSI) analysis. Eight laboratories compared MSI analyses performed on 10 matched pairs of normal and tumor DNA from patients with colorectal carcinomas. A variety of techniques were applied to the detection of microsatellite changes: (a) silver and ethidium bromide staining of polyacrylamide gels; (b) radioactive labeling; and (c) automated fluorescence detection. The identification of highly unstable tumors and tumors without MSI was achieved in high concordance. However, the interpretation of the band patterns resulted in divergent classifications at several microsatellite marker loci for a large fraction of this tumor/normal panel. The data on more than 30 primers per case suggest that the enlargement of the microsatellite panel to more than 10 loci does not influence the results. In this study, cases with MSI in less than 10% of loci were classified as microsatellite stable, whereas MSI was diagnosed in cases with more than 40% of all markers unstable. We propose that a panel of five microsatellite loci consisting of repeats with different lengths should be analyzed in an initial analysis. When less than two marker loci display shifts in the microsatellite bands from tumor DNA, the panel should be enlarged to include an additional set of five marker loci. The number of marker loci analyzed as well as the number of unstable marker loci found should always be identified. These criteria should result in reports of MSI that are more comparable between studies.     

Mutation and evolution of microsatellites in Drosophila melanogaster

Levels of nucleotide polymorphism in the Drosophila melanogaster genome are correlated with rates of recombination. This relationship may be due to hitchhiking of advantageous mutations (selective sweeps) or to continual removal of deleterious mutations from the genome (background selection). One test of the relative contributions of selective sweeps and background selection to the observed levels of variation in the genome of D. melanogaster is to compare levels of nucleotide variability (with a mutation rate on the order of 10(-9) per nucleotide per generation) with more rapidly evolving DNA loci such as microsatellites. This test depends critically on details of the mutational process of microsatellites. In this paper, we summarize our studies of microsatellite characteristics and mutation rates in D. melanogaster. We find that D. melanogaster microsatellites are short and have a mutation rate (6.5 x 10(-6) per locus per generation) several orders of magnitude lower than mammals studied to date. We further show that genetic variation at 18 dinucleotide repeat microsatellites in a population of D. melanogaster from Maryland is correlated with regional rates of recombination. These and other microsatellite data suggest that both background selection and selective sweeps may contribute to the correlation between DNA sequence variation and recombination in Drosophila.     

Mutation rate of a microsatellite sequence in normal human fibroblasts

Dinucleotide repeats, because of their repetitive nature, are prone to frameshift mutations, most likely via a DNA-polymerase slippage mechanism. Mutation rates in microsatellite DNA sequences are high in mismatch repair-defective cells. In normal cells, only estimates of maximal rates of mutation in microsatellites have been possible previously, because of the low sensitivity of screening assays for mutations in endogenous sequences. We have measured the spontaneous mutation rate of a dinucleotide repeat in diploid human foreskin fibroblasts. In our system, the mutation target is a (CA)17 repeat contained within a stably integrated plasmid. The repeat disrupts the reading frame of a neomycin (neo) resistance gene within the plasmid. Cells containing frameshift mutations in the CA repeat that correct the reading frame of the neo gene are selected using the neo analogue G418. This system of measuring microsatellite mutation rates is highly sensitive, because there is a specific target within which mutations can be selected. Fluctuation analysis of cells containing the target DNA yielded mutation rates of <3.1 x 10(-8) to 44.8 x 10(-8) mutations/cell/generation. This is the first report of a direct measurement of a spontaneous mutation rate of a microsatellite sequence in normal human cells.