Direct Infection of Colony Forming Unit-Megakaryocyte by Human Cytomegalovirus Contributes the Pathogenesis of Idiopathic Thrombocytopenic Purpura

Human cytomegalovirus （HCMV） late mRNA expression in megakaryoblast and in turn the pathogenesis of idiopathic thrombocytopenic purpura （ITP） patients with HCMV infection, and effectiveness of ganciclovir were investigated. Colony forming unit-megakaryocytes （CFU-MK） of 46 ITP patients with HCMV infection were incubated from patients＇ bone marrow mononuclear cells （MNC）. Reverse transcriptase-polymerase chain reaction （RT-PCR） was subsequently used for CFU-MK for HCMV-late mRNA detection, Ganciclovir therapy was given to both HCMV-late mRNA positive and negative groups for comparison of therapeutic effectiveness, The results in 19 of 46 CFU-MK culture cells specimens with positive HCMV-DNA by PCR or positive CMV-IgM by enzyme linked immunosorbent assay （ELISA） in the correspondent serum of peripheral blood were positive for HCMV-late mRNA, Sixteen out of 19 patients with positive HCMV-late mRNA CFU-MK had a positive response to ganciclovir. Amongst 27 patients with negative HCMV-late mRNA CFU-MK, only 4 positive responders to ganciclovir therapy were observed. Curative effectiveness of ganciclovir in HCMV-late mRNA positive group was significantly higher than that in HCMV-late mRNA negative group （P〈0.01）, It was suggested that HCMV could directly infect CFU-MK, which might be one of the mechanisms responsible for HCMV related ITE The ganci- clovir is an effective therapy in resulting in the increases in thrombocyte in the ITP patients whose HCMV- late mRNA was positive in their CFU-MK.

Hierarchy of mesenchymal stem cells: Comparison of multipotentmesenchymal stromal cells with fibroblast colony forming units

The organization of the compartment of mesenchymal stem cells is still obscure. Two types of human stromal precursor cells are known. Both of them are analyzed in in vitro system: mesenchymal multipotent stromal cells (MMSC) and fibroblast colony forming units (CFU-F). The aim of this study was to compare the main characteristics of MMSC and CFU-F derived from the bone marrow of 24 healthy donors. Growth and differentiation parameters, as well as relative expression levels of different genes were analyzed in MMSC and CFU-F. MMSC were cultivated for 5 passages. CFU-F concentration was determined for each bone marrow sample. The data obtained demonstrated the heterogeneity and hierarchical organization of both studied populations of stromal precursor cells-MMSC and CFU-F. These two types of stromal precursor cells turned to be different in most parameters studied. Altogether MMSC seemed to be more immature cells than CFU-F and took up the higher position in hierarchical tree of mesenchymal stem cells. The rate of differentiation and proliferative potential decreased with the donor’s age in both populations MMSC and CFU-F.

Machine learning for enumeration of cell colony forming units

As one of the most widely used assays in biological research,an enumeration of the bacterial cell colonies is an important but time-consuming and labor-intensive process.To speed up the colony counting,a machine learning method is presented for counting the colony forming units(CFUs),which is referred to as CFUCounter.This cellcounting program processes digital images and segments bacterial colonies.The algorithm combines unsupervised machine learning,iterative adaptive thresholding,and local-minima-based watershed segmentation to enable an accurate and robust cell counting.Compared to a manual counting method,CFUCounter supports color-based CFU classification,allows plates containing heterologous colonies to be counted individually,and demonstrates overall performance(slope 0.996,SD 0.013,95%CI:0.97–1.02,p value<1e-11,r=0.999)indistinguishable from the gold standard of point-and-click counting.This CFUCounter application is open-source and easy to use as a unique addition to the arsenal of colony-counting tools.

Difference between resistant and susceptible maize to systematic colonization as revealed by DsRed-labeled Fusarium verticillioides

Fusarium verticillioides was labeled with DsRed via Agrobacterium tumefaciens-mediated transformation to examine differences in colonization and reactions of resistant and susceptible inbred lines of maize(Zea mays L.). The extent of systemic colonization of F. verticillioides in roots from maize lines either resistant or susceptible to the fungus was studied by visualizing the red fluorescence produced by the fungus expressing DsRed. The difference in quantities of colony forming units(CFU) in roots and basal stems, production of fumonisin B1, and pH of root were determined. Although F. verticillioides colonized both resistant and susceptible lines, differences were observed in the pattern and extent of fungal colonization in the two types of maize lines. The fungus colonized the susceptible lines producing mosaic patterns by filling the individual root cells with hyphae. Such a pattern of colonization was rarely observed in resistant lines, which were less colonized by the fungus than the susceptible lines in terms of CFUs. The production of mycotoxin fumonisin B1 in roots from different lines was closely correlated with the amount of F. verticillioides colonization, rather than the pH or amylopectin concentrations in the root. The findings from this study contribute to a better understanding of the defense mechanism in resistant maize lines to F. verticillioides.

Evaluation of the relationship between two different methods for enumeration fecal indicator bacteria: Colony-forming unit and most probable number

Most probable number （MPN） and colony-forming unit （CFU） estimates of fecal indicator bacteria （FIB） concentration are common measures of water quality in aquatic environments. Thus, FIB intensively monitored in Yeongsan Watershed in an attempt to compare two different methods and to develop a statistical model to convert from CFU to MPN estimates or vice versa. As a result, the significant difference was found in the MPN and CFU estimates. The enumerated Escherichia coli concentrations in MPN are greater than those in CFU, except for the measurement in winter. Especially in fall, E. coli concentrations in MPN are one order of magnitude greater than that in CFU. Contrarily, enterococci bacteria in MPN are lower than those in CFU. However, in general, a strongly positive relationship are found between MPN and CFU estimates. Therefore, the statistical models were developed, and showed the reasonable converting FIB concentrations from CFU estimates to MPN estimates. We expect this study will provide preliminary information towards future research on whether different analysis methods may result in different water quality standard violation frequencies for the same water sample.

基于肠道菌群平衡分析微生态制剂联合浙贝黄芩汤对急性淋巴细胞白血病大剂量化疗后患者的临床影响

目的探讨微生态制剂联合浙贝黄芩汤对急性淋巴细胞白血病(ALL)大剂量化疗后患者粒细胞集落刺激因子受体(G-CSFR)、粒单系集落形成单位(CFU-GM)、肠道菌群及红系爆式集落形成单位(BFU-E)的影响。方法选取延安大学附属医院2019年6月至2022年12月收治的ALL患者130例作为研究对象,根据治疗方法将患者分为A组、B组、C组,3组患者均接受大剂量化疗,化疗结束48 h后A组患者实施常规治疗,B组患者单纯浙贝黄芩汤治疗,C组给予微生态制剂联合浙贝黄芩汤治疗,治疗12 d后,对3组患者G-CSFR、CFU-GM、BFU-E表达情况及血细胞数量进行检测。结果治疗后,C组血红蛋白、白细胞、血小板[(79±6)g/L、(3.8±0.4)×10^(9)/L、(66.4±3.6)×10^(9)/L]与A组[(59±7)g/L、(3.2±0.4)×10^(9)/L、(52.6±2.8)×10^(9)/L]、B组[(61±7)g/L、(3.1±0.3)×10^(9)/L、(52.8±2.6)×10^(9)/L]对比,差异有统计学意义(P<0.05)。C组G-CSFR(5.35±0.16)pg/ml和白细胞介素-11受体(IL-11R)(6.38±0.54)μg/kg水平均高于A组[(2.23±0.13)pg/ml和(1.49±0.24)μg/kg]和B组[(2.31±0.16)pg/ml和(2.31±0.49)μg/kg]差异有统计学意义(P<0.05)。治疗后,C组患者7 d CFU-GM(18.5±6.0)个和14 d BFU-E(83.5±7.5)个高于A组[7 d CFU-GM(9.5±2.0)个和14 d BFU-E(59.5±6.5)个]和B组[7 d CFU-GM(12.0±6.5)个和14 d BFU-E(63.5±5.0)个],差异有统计学意义(P<0.05)。7 d后,C组双歧杆菌(12.56±3.25)lgCFU/g、乳酸杆菌(13.56±2.58)lgCFU/g、肠杆菌(5.12±1.45)lgCFU/g、肠球菌(5.14±0.58)lgCFU/g高于A组[(9.26±1.03)lg CFU/g、(8.65±0.84)lg CFU/g、(8.08±0.64)lgCFU/g、(8.15±0.46)lgCFU/g]和B组[(11.35±1.36)lg CFU/g、(12.43±1.14)lgCFU/g、(6.49±0.55)lgCFU/g、(6.66±0.43)lgCFU/g],差异有统计学意义(P<0.05)。结论微生态制剂联合浙贝黄芩汤治疗可以有效提高ALL大剂量化疗后患者的G-CSFR、CFU-GM、BFU-E水平,可能更好地改善化疗引起的患者骨髓抑制情况,改善肠道菌群,具有临床研究价值。

Clonogenic colony-forming ability of hepatic stem cells in the spleens of mice

To confirm the existence of hepatic stem cells (HSCs), fetal liver cells isolated from mice on embryonic day 13 (ED13) were long-term cultured in vitro. Growth of the cells was observed intensively and characteristics were iden-tified by immunocytochemistry. The results showed that some of the cells grew as colonies, in which some cells ex-pressed AFP, CD34 and Albumin. Then the cells were trans-planted intravenously into irradiated syngeneic mice. At day 12 a number of small hyperplasia nodules were seen in the apparently enlarged spleens of recipient mice. Moreover, some nodules were positive for AFP and CD34 and consisted of various types of cells, suggesting the very existence of he-patic stem cells in the mouse fetal liver.

地黄低聚糖对快速老化模型小鼠造血功能的影响

目的：研究地黄低聚糖（ＲＧＯＳ）对快速老化模型小鼠（ＳＡＭＰ８）造血功能的影响。方法：采用造血祖细胞体外克隆培养等实验血液学方法。结果：ＲＧＯＳ１０，２０ｍｇ·ｋｇ－１可使ＳＡＭＰ８小鼠的ＣＦＵ－Ｓ、ＣＦＵ－ＧＭ、ＣＦＵ－Ｅ和ＢＦＵ－Ｅ明显增多，并可使外周血中降低的ＷＢＣ数明显增加。提示ＲＧＯＳ可刺激ＳＡＭＰ８小鼠造血干细胞、祖细胞的增殖分化。集落刺激活性实验显示ＲＧＯＳ可使小鼠体内的集落刺激因子产生明显增多。小鼠骨髓组织学研究结果也进一步证实了ＲＧＯＳ增强ＳＡＭＰ８小鼠造血功能的作用。结论：ＲＧＯＳ可以增强ＳＡＭＰ８小鼠的造血功能。

系统性红斑狼疮血小板减少患者巨核祖细胞分化障碍

目的 明确系统性红斑狼疮 (SLE)血小板减少患者骨髓巨核祖细胞增殖分化情况 ,以及患者血清对巨核祖细胞增殖分化的影响。方法 分离 7例SLE血小板减少患者 (SLE血小板减少组 )、11例胸外科开胸手术切除肋骨患者 (正常对照组 )和 4例SLE血小板正常但病情活动患者 (SLE血小板正常组 )的骨髓单个核细胞 ,体外巨核祖细胞 (CFU MK)无血清半固体培养 ;并分别加入SLE血小板减少和SLE血小板正常患者的血清 ,观察血清对CFU MK集落形成的影响。结果 SLE血小板减少组CFU MK集落数为 (2 7.33± 9.30 )个 /片 ,明显低于正常对照组的 (6 1.2 2± 2 9.71)个 /片和SLE血小板正常组的 (6 0 .0 0± 2 9.71)个 /片 (P值均 <0 .0 5 ) ;加入SLE血小板减少患者血清后 ,SLE血小板减少组 (n =7)和正常对照组 (n =7)CFU MK集落数分别减少为(14 .2 9± 6 .73)和 (2 9.4 4± 2 3.35 )个 /片 (P <0 .0 5 )。加入SLE血小板正常患者血清后 ,正常对照组 (n =7)CFU MK增加到 (115 .6 0± 72 .99)个 /片 ,与未加血清者的差异无显著性 (P <0 .0 5 ) ;而SLE血小板减少组 (n =7)无显著增加 (P >0 .0 5 )。结论 SLE血小板减少患者骨髓巨核祖细胞分化障碍 。

黄芪注射液对人巨细胞病毒感染的粒-单系祖细胞体外增殖的影响

目的 探讨黄芪注射液对人巨细胞病毒 (HCMV)感染的粒—单系祖细胞 (CFU GM )体外增殖的影响。方法 采用造血祖细胞体外培养技术 ,用黄芪干预HCMV感染的CFU GM ,观察计数CFU GM的产率 ,大小 ,峰期及维持时间。结果 ①黄芪组每 2× 10 5个细胞中CFU GM簇和集落的产率分别为 333.33± 12 .6 9和81.2 3± 4 .93,病毒组每 2× 10 5个细胞中簇和集落的产率分别为 16 7.80± 16 .6 3,5 3.70± 1.6 7,两者相比差异有显著性 (P <0 .0 1)。②黄芪组大集落的比例占 (9.5± 0 .8) % ,明显高于病毒组 (2 .7± 1.0 ) % ,P <0 .0 1。③黄芪组大集落峰期出现早 ,集落和大集落维持时间长。结论 在体外黄芪注射液可以促进HCMV感染的CFU GM增殖。

鲜切菠萝片加工工艺的研究

以菌落总数和感官评价为指标,研究了鲜切菠萝片的合理工艺条件。结果表明:鲜切菠萝片经1.0%Vc-Na和1.0%柠檬酸处理,通过紫外灭菌(30W,照射高度40cm)30min,贮存于5℃,使货架期达到8d左右;菌落总数控制在105以内,符合鲜切果蔬的食品卫生要求,产品感官品质优良。

金龟子绿僵菌在森林土壤中的分布及对松墨天牛致病性测定

松墨天牛是重大森林植物检疫性病害——松材线虫病的主要媒介昆虫。本研究于2005年8月～2006年8月从福建、江西两省共110个林分样区（其中松林88个样区）采集土壤样品330份,采用选择性培养基分离土壤中的金龟子绿僵菌。从21个样区的26份土样中分离出的金龟子绿僵菌占采集样区的19．1％和样品的7．9％,成菌落数（CFU）500～72500CFU/g,表明金龟子绿僵菌在森林土壤中有较为广泛的分布。对分离到的9个产孢量高的菌株,采用浸渍法（1×10^7孢子/mL）接种3～4龄健康松墨天牛幼虫,采用跗节接种法接种2～15日龄健康成虫,测定其致病力。结果表明,MaYTTR-03、MaYTTR-04菌株对松墨天牛幼虫和成虫均有较高致病力,表现出良好的生防潜力。

小麦内生细菌及其对根茎部主要病原真菌的抑制作用

对小麦植株不同生育期、不同器官的内生细菌进行了分离和数量变化分析.结果表明,根、茎、叶及未成熟籽粒等器官中存在大量的内生细菌,鲜组织中平均约含内生细菌5.0×105CFU.g-1,其中根系中内生细菌数量达7.8×105CFU.g-1,而茎秆、叶片和未成熟籽粒中内生细菌数量分别为4.8×105、3.2×105和2.8×105CFU.g-1.内生细菌数量在不同生育期也存在差异,幼苗期平均约为3.1×105CFU.g-1、拔节期和灌浆期分别为5.7×105和7.0×105CFU.g-1.不同小麦田块之间存在明显差异,长武县一田块植物鲜组织中内生细菌的数量为6.1×105CFU.g-1,而大荔县一田块约为3.9×105CFU.g-1.试验结果发现,对小麦全蚀病菌具有拮抗作用的内生细菌有51株、对小麦纹枯病菌具有抑制作用的内生细菌有45株.用平板对峙法测定,有71株对两种病原真菌均有拮抗作用,对小麦全蚀病菌抑菌圈直径大于10mm的有23株,其中来源于根系、茎秆、叶片和籽粒的分别为6株、7株、9株和1株;对小麦纹枯病菌抑菌圈超过10 mm的有20株,其中来源于根系、茎秆、叶片和籽粒的分别为7株、5株、7株和1株,说明从小麦叶片诱捕分离的内生细菌中,对小麦全蚀病菌和纹枯病菌抑菌作用较强的分离株比率最高,其次为茎秆,而根部和未成熟籽粒中比例明显较低.

响应面法优化海洋枯草芽孢杆菌HS-A38增殖发酵培养基

在摇瓶培养条件下,优化提高海洋枯草芽孢杆菌菌体浓度的发酵培养基组分。采用Plackett-Burman法确定了影响菌体浓度的显著性因子,即葡萄糖和ZnSO4;通过最陡爬坡实验逼近这两个重要因子的最大响应稳定区域,采用中心组合实验确定显著性因子的最佳水平,并用Design expert 7.13软件进行回归分析。优化后培养基的组成为:葡萄糖8.49g/L,豆粕粉12.0g/L,尿素0.625g/L,酵母膏4.0g/L,K2HPO44.0g/L,ZnSO40.053g/L。优化后的活菌浓度达到1.64×109 cfu/mL,与优化前菌体浓度(7.22×108 cfu/mL)相比,提高了1倍多。通过统计优化培养基组分可有效提高海洋枯草芽孢杆菌HS-A38的发酵液菌体浓度。

扶正祛邪丹治疗骨髓增生异常综合征的临床及实验研究

目的:研究扶正祛邪丹对骨髓增生异常综合征(MDS)的疗效与作用机理。方法:用扶正祛邪丹为主治疗MDS患者33例,观察血象、骨髓象、骨髓病理切片及免疫学的变化;用干细胞培养法观察扶正祛邪丹对BALB/C小鼠骨髓造血干细胞增殖的影响。结果:临床患者基本缓解率2424%,总有效率7272%。实验研究发现扶正祛邪丹对骨髓红系造血干细胞有明显促进作用。结论:扶正祛邪丹对MDS的难治性贫血疗效较好,可能是通过促进红系造血干细胞增殖实现的。

蝎毒对B16黑色素瘤细胞生长的抑制作用

目的 :观察蝎毒 (scorpion venom,SV)及其分离组分对小鼠 B1 6黑色素瘤细胞生长的抑制作用。方法 :采用 MTT法检测肿瘤细胞存活率 ,集落形成法检测肿瘤细胞增殖 ,采用流式细胞术检测细胞周期的变化。结果 :4 0 0和 80 0 mg· L- 1 的 SV可明显抑制 B1 6黑色素瘤细胞生长 (P<0 .0 1或 P<0 .0 5 )。蝎毒组分 (SV- 、SV- 和 SV- )在浓度为 2 0 0、4 0 0及 80 0 mg· L- 1 时 ,对 B1 6细胞生长有明显的抑制作用 (P<0 .0 5～ P<0 .0 1 ) ;在 4 0 0和 80 0 mg· L- 1 时 ,可使 G0 / G1 期细胞数明显减少 (P<0 .0 5～ P<0 .0 1 ) ,G2 +M期细胞明显增多 (P<0 .0 5～ P<0 .0 1 ) ,S期细胞呈降低趋势 ,而 SV对细胞周期进程未见明显影响。当 SV、SV- 、SV- 及 SV- 的浓度为 4 0 0和 80 0 mg· L- 1 时 ,对 B1 6细胞集落形成均有明显的抑制作用 (P<0 .0 1 )。结论 :在一定浓度范围内蝎毒及其组分对 B1 6黑色素瘤细胞的存活与增殖均有明显的抑制作用 ,纯化后各组分的抑瘤作用优于蝎毒 ,但其抑制作用低于阳性对照丝裂霉素 (Mitom ycin- c,MMC)组。提示蝎毒引起细胞周期进程改变可能是其抑瘤作用的机制之一。

豆酱微波杀菌工艺

以微波功率、辐照时间和装填量为工艺参数,以菌落总数、大肠菌群为检测指标,研究了豆酱的微波杀菌工艺条件,并与传统巴氏杀菌工艺进行比较,分析了杀菌前后豆酱感官品质和色差值的变化。结果表明:微波功率、杀菌时间与装填量对杀菌效果均有明显影响,豆酱微波杀菌最佳工艺参数为:3 400 W,120 s,100 g/袋。菌落总数减少99.9%,大肠菌群数<3.0 MPN/g,产品符合国家标准。与巴氏杀菌工艺相比,豆酱感官品质尤其是色泽变化更小。

骨髓源性肝干细胞的确认及定向分化的实验研究

目的 确定骨髓源性肝干细胞的表面标记 ,体外培养诱导其分化为肝细胞。方法 采用流式细胞仪检测多种大鼠肝损伤模型骨髓中干细胞标记的细胞群体的数量变化 ,寻找可能的肝干细胞 ,免疫磁珠分离各骨髓干细胞标记的细胞群体 ,进行体外培养诱导分化 ,观察细胞形态的变化 ,免疫组化技术检测清蛋白 (白蛋白 ) ,AFP ,CK 8/ 18等肝细胞标记的表达。结果 各模型大鼠骨髓内 β2微球蛋白阴性 (β2 m- )细胞显著增高 ,体外培养经诱导后呈多角形细胞表现 ,清蛋白 ,AFP ,CK 8/ 18表达阳性。其他干细胞类型细胞数量变化小 ,体外培养未见肝细胞样变化。结论 β2 m- 细胞的数量随肝损伤而变化 ,在体外具有向肝细胞分化的能力 。

乌贼墨对小鼠造血干细胞、粒-单系祖细胞及外周白细胞的影响

目的探讨乌贼墨对小鼠造血干细胞、粒-单系祖细胞及外周血WBC的影响。方法用不同剂量的乌贼墨灌胃正常小鼠、环磷酰胺(Cy)和辐射骨髓损伤模型小鼠,采用造血祖细胞体外培养方法和实验血液学技术,检测小鼠骨髓造血干细胞生成数(CFU-S)、粒-单系祖细胞集落生成数(CFU-GM)和外周血WBC数量。结果乌贼墨能够显著提高正常小鼠CFU-S,CFU-GM和外周血WBC数量,有效拮抗模型小鼠体内CFU-S,CFU-GM及外周WBC的降低,并显著促进模型小鼠上述各指标的恢复。结论乌贼墨具有显著的促进小鼠骨髓粒系造血作用。其作用机制可能通过调节机体的免疫机能,诱导机体产生粒/单核细胞集落刺激因子和多种细胞因子,促进造血细胞的增殖,并诱导造血细胞向粒单系细胞分化。

股骨头缺血性坏死骨髓活性的研究

目的 :股骨头缺血性坏死的病因及发病机制不明。本研究旨在探讨因非创伤所致股骨头缺血性坏死骨髓活性有无异常 ,揭示骨坏死发生机理。方法 :对 1999年 9月～ 2 0 0 0年 4月间骨坏死病人骨髓成纤维样细胞集落形成单位 (FCFUs)进行培养 ,观察其生长特点及集落数目 ,观察骨坏死病人与对照组之间有无差异。结果 :骨坏死组骨髓成纤维样细胞集落形成单位的贴壁生长增殖速度慢于对照组 ,FCFUs形成集落数目与对照组相比存在明显差异。结论 :非创伤性股骨头缺血性坏死存在骨髓活性下降 。