荧光原位杂交技术检测多发性骨髓瘤染色体

目的研究荧光原位杂交技术(FISH)在多发性骨髓瘤(MM)染色体异常检测中的应用价值。方法选取从2012年1月至2014年1月该院收治的初治MM患者24例,同时采用常规G显带技术与FISH技术进行检测,比较两种检测方法的阳性检出率,分析染色体异常与MM的免疫学分型、临床分期与预后的相关性。结果常规G显带技术的染色体异常检出率比FISH检出率低(P0.01);常规G显带技术仅检出1例染色体异常者,MM患者的染色体异常以Ⅲ期和Ig G型为主。结论 FISH技术的染色体异常检出率与特异性较高,临床分期、分型与染色体异常存在相关性,Ⅲ期和Ig G型患者出现染色体异常的可能性更高。     

多发性骨髓瘤细胞遗传学变化及其蛋白酶体抑制剂的疗效观察

目的分析多发性骨髓瘤(MM)的细胞遗传学变化及其与蛋白酶体抑制剂治疗的关系。方法对2005年1月至2006年7月北京大学人民医院29例初诊的MM患者行染色体核型检查,采用骨髓细胞24h短期培养,用常规方法制备染色体,G显带进行核型分析。22例MM患者采用传统化疗即长春新碱联合阿霉素和地塞米松(VAD)方案或马法兰与泼尼松(MP)方案;7例患者应用蛋白酶体抑制剂(硼替佐米)为主的化疗方案。比较两组患者的有效率及缓解率。结果29例MM患者中异常核型检出率为37.9%,其中有复杂和高度复杂畸变的占81.8%。采用VAD或MP方案化疗的核型正常组的有效率为81.2%,核型异常组有效率为0,差异有显著性意义(P<0.05)。应用硼替佐米为主的化疗方案中核型异常组5例均有效,核型正常组2例均有效。核型异常组硼替佐米有效率与传统化疗组相比,差异有显著性意义(P<0.05)。结论染色体核型异常的患者,应首选蛋白酶体抑制剂硼替佐米等进行治疗。     

血液病150例染色体核型分析及其临床意义

目的 :探讨血液系统疾病的染色体核型异常及其与临床的关系。方法 :采用直接法和短期培养法制备骨髓及外周血细胞染色体 ,以R显带技术进行染色体核型分析。结果 :本组 15 0例 ,急性白血病 5 3例 ,其中急性淋巴细胞白血病 2 2例、急性非淋巴细胞白血病 30例 ,急性混合细胞白血病 1例 ,有核型异常者各为 10例、16例及 1例 ;慢性髓细胞白血病 (CML) 4 5例 ,4 1例有Ph染色体 ;CML急变期 9例中有 8例发现附加染色体异常 ;慢性淋巴细胞白血病 12例 ,仅 4例有核型异常 ;骨髓增生异常综合征 15例中 7例有核型异常 ;10例再生障碍性贫血 ,8例骨髓增殖性疾病 ,4例淋巴瘤 ,3例多发性骨髓瘤 ,核型均正常。结论 :染色体核型分析是血液系统疾病诊断、分型、预后判断和随访监测的一项重要指标     

92例多发性骨髓瘤的临床与细胞遗传学分析

目的探讨多发性骨髓瘤（MM）细胞染色体异常的临床意义。方法采用骨髓细胞直接法、24h短期培养法制备染色体标本,用R显带技术进行核型分析。结果92例MM中,常规细胞遗传学（CC）发现异常核型24例,发生率26．09％。部分病例在CC基础上应用荧光原位杂交（FISH）技术,异常检出率大为提高。结论FISH技术深入了MM的细胞遗传学研究,伴有14q32异常是MM患者最为常见的染色体核型异常,伴有多条染色体异常MM患者其临床预后差。     

378例白血病患者染色体核型分析及其临床意义

目的探讨白血病患者染色体改变及其在诊断、治疗及预后判断中的意义。方法采用短期培养法制备骨髓细胞染色体标本,以R显带技术用显微镜对240例急性白血病(AL)患者、138例慢性粒细胞白血病患者(CML)进行常规染色体核型分析,每例患者分析10～20个中期分裂细胞,并动态观察部分患者病程进展中染色体核型的变化。结果 240例AL中核型异常182例,占75.83%,其中单纯数目异常占20.33%(37/182),以多倍体和非整倍体为主;结构异常113例,占62.08%,主要为特异性染色体重排且与分型有关。138例CML患者中Ph染色体阳性(Ph+)127例,占92.03%;其中90.55%(115/127)为典型易位,9.45%(12/127)为变异易位。127例Ph+患者中22例(13.39%)出现额外染色体,加速期和急变期额外染色体的检出率明显高于慢性期(P<0.05)。结论染色体核型分析对白血病的诊断、鉴别诊断、治疗及预后判断具有重要意义。     

骨髓增生异常综合征99例临床分析

目的探讨骨髓增生异常综合征(MDS)患者WHO亚型分布、细胞遗传学特点及其与MDS诊断分型、疾病进展和预后的关系。方法回顾性分析2001年1月至2007年12月安徽医科大学附属安徽省立医院血液科收治的99例成人原发MDS患者的染色体核型、WHO分型及预后情况,随访观察并进行相关性研究。结果99例MDS患者难治性贫血(RA)型26例(26.26%);难治性贫血伴环形铁幼粒细胞增多(RAS)型6例(6.06%);难治性贫血伴多系发育异常(RCMD)型23例(23.23%);难治性贫血伴原始细胞增多(RAEB)型44例(44.44%)。按IPSS预后分组,中危Ⅱ和高危组的染色体核型异常检出率明显高于低危和中危Ⅰ组(χ2=17.88,P<0.01);中危Ⅱ和高危组患者进展为急性白血病的发生率明显高于低危和中危Ⅰ组(χ2=40.22,P<0.01)。按IPSS染色体核型分组,预后好、中、差的患者中位存活期分别为45(95%CI:39~51)、37(95%CI:25~49)和23(95%CI:13~31)个月,Log-rank检验三组总体生存(OS)率差异有统计学意义(P=0.010)。结论中国有别于西方国家MDS患者的WHO亚型分布,染色体核型分析是MDS诊断分型及预后评估的重要指标。     

24例多发性骨髓瘤患者T辅助细胞亚群的检测及遗传学分析

目的:分析多发性骨髓瘤(MM)患者外周血Th1、Th2细胞数量和比例及骨髓细胞分子遗传学变化,探讨其相关关系及临床意义。方法:流式细胞术(FCM)检测分析24例MM患者外周静脉血Th1、Th2细胞的比例并采用荧光原位杂交技术(FISH)对其骨髓细胞13q14缺失、p53缺失、IgH易位重排进行检测。结果:①Ⅲ期MM患者Th1细胞比例较对照组显著降低,Ⅱ、Ⅲ期MM患者Th2细胞比例较对照组及Ⅰ期降低,Ⅲ期MM患者Th1/Th2比值较对照组、Ⅰ及Ⅱ期增高(P0.05或P0.01);②24例MM患者中检出9例13q14缺失,其中5例D13S319异常(20.83%),6例RB1异常(25.00%),8例IGH易位重排(33.33%),3例P53缺失(12.50%);③按FISH预后风险分组,高危组Th1、Th2细胞较标危组明显降低,Th1/Th2比值较之增高。结论:MM患者外周血Th1、Th2细胞比例降低及Th1/Th2比值增高,且在del(13q14)和(或)del(17p13)的高危组更为明显,与临床分期及预后相关。13q14缺失及IgH基因重排在MM患者中的发生率较高,与临床分期的关系有待进一步研究。     

多发性骨髓瘤中SOX2和KLF5的表达及临床意义

目的:检测多发性骨髓瘤(MM)组织中SOX2和Krüppel样因子5(KLF5)的表达情况,并探讨其临床意义。方法:收集100例MM患者的骨髓标本作为研究对象,同期选择72例正常人的骨髓标本作为对照组,采用免疫组织化学法检测MM及正常骨髓细胞中SOX2和KLF5蛋白表达情况,分析SOX2、KLF5蛋白表达与MM临床病理特征的关系,并对所有患者随访3年,分析SOX2和KLF5表达对MM患者预后的影响。结果:MM组细胞中SOX2的阳性表达率为69.00%,显著高于对照组的20.83%(P0.05);MM组细胞中KLF5的阳性表达率为66.00%,显著高于对照组的25.00%(P0.05);MM细胞中SOX2表达情况与患者D-S分期、ISS分期有关(P0.05);MM细胞中KLF5表达情况与患者ISS分期、髓外转移有关(P0.05);术后随访3年,共有46例患者死亡,其中SOX2高表达MM患者3年总存活率显著低于低表达患者(P0.05),KLF5高表达MM患者3年总存活率显著低于低表达患者(P0.05);Cox模型多因素分析结果显示,ISS分期、SOX2、KLF5是影响MM预后的独立危险因素。结论:MM患者细胞中SOX2和KLF5表达显著升高,与患者ISS分期等临床病理特征有关,可能成为MM患者不良预后的生物标志物。     

年轻多发性骨髓瘤患者24例临床分析

目的:探讨年轻多发性骨髓瘤(MM)患者(50岁)的临床表现、实验室检查特点、疗效及生存情况。方法:回顾性分析24例(50岁)MM患者的临床资料。结果:年轻MM患者占同期MM患者的10.1%(24/238),IgG型和轻链型为主,DS分期均为Ⅲ期,ISS分期Ⅰ期6例,Ⅱ期14例,Ⅲ期4例;8例出现髓外浸润;染色体核型异常(G带染色)4/12例,FISH异常2/12例。15例使用万珂,5例使用来那度胺,22例使用反应停,6例参加临床试验,2例使用脂质体阿霉素,2例使用砷剂,1例异基因造血干细胞移植。疗效:完全缓解10例,非常好的部分缓解4例,部分缓解8例,疾病稳定1例,疾病进展1例;总有效率91.7%,完全缓解率41.7%。达到完全缓解的时间为2~27个月(中位时间6个月),达到完全缓解后维持的时间为8~48个月(中位时间18个月)。1年总生存率100%,3年预计总生存率95.45%,5年预计总生存率70.67%。结论:MM多见于老年人,但发病年龄有年轻化趋势,年轻患者骨骼损害严重,髓外浸润多见,万珂及来那度胺靶向药物疗效良好,中位生存时间比老年患者长。     

修订的国际分期系统对初诊多发性骨髓瘤的预后意义

目的:探讨修订的国际分期系统(R-ISS)对初诊多发性骨髓瘤(MM)的预后价值。方法:对189例初诊的MM患者进行FISH或cIg-FISH检测细胞遗传学异常,并统计疾病分期及生存情况。结果:t(4;14)阳性者24例(13.5%,24/178),del(17p)阳性者20例(10.7%,20/187)。对比阴性患者,t(4;14)及del(17p)阳性患者具有较短的中位无进展生存(PFS),差异有统计学意义(P0.05),而中位总生存(OS)差异无统计学意义,但t(4;14)阳性患者与阴性患者生存曲线有分离趋势。t(14;16)阳性患者仅5例,未单独分析。多因素分析显示,年龄、乳酸脱氢酶(LDH)和del(17p)是评估OS的独立预后因素(P0.05);LDH、del(17p)和t(4;14)是评估PFS的独立预后因素(P0.05)。R-ISS分期Ⅰ、Ⅱ和Ⅲ期患者分别为16、142和31例,1年OS率分别为100%、89%和65%(P0.001),1年PFS率分别为86%、77%和52%(P=0.002);ISS分期Ⅰ、Ⅱ、Ⅲ期患者分别为23、77和89例,1年OS率分别为90%、88%和83%(P=0.239),1年PFS率分别为72%、73%和74%(P=0.672)。结论:t(4;14)、del(17p)核型异常及LDH均是预后不良因素;基于LDH和细胞遗传学异常基础上的R-ISS分期在初诊MM患者的预后评估中优于ISS分期系统。     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.