Isolation and Identification of Cancer Stem Cells from Human Osteosarcom by Serum-free Three-dimensional Culture Combined with Anticancer Drugs

The cancer stem cells(CSCs)from human osteosarcoma by serum-free three-dimensional culture combined with anticancer drugs were isolated and identified.The primary cells derived from human osteosarcoma were digested by trypsin to prepare a single-cell suspension,and mixed homogeneously into 1.2% alginate gel.Single-cell alginate gel was cultured with serum-free DMEM/F12 medium.Epirubicin(0.8μg/mL)was added to the medium to enrich CSCs.After cultured conventionally for 7 to 10 days,most of cells suspended in ...

Serum-free culture of H pylori intensifies cytotoxicity

AIM： To perform a long culture passage of H pylori without serum, taking into account its cytotoxicity and the presence of the probable new cytotoxic factor. METHODS： One sample of H pylon 60190 （ATCC 49503） was grown on Brain Heart Infusion （BHI） agar containing 0.5% 2,6-di-O-methyl-β-cyclodextrin without any serum, being passaged 70-100 times every 3-4 d for approximately 2 h, while another sample of H pylori contained 70 mL/L fetal calf serum without 2,6-di-O- methyl-β-cyclodextrin. Their supernatant and extract after 16 h in culture were evaluated for changes in cell morphology and for cell viability using HeLa cells. Furthermore, the characteristics of the probable cytotoxic factor in the extract were examined on partial purification studies and its oytotoxicity was evaluated in various human cells. RESULTS： The supernatant and the extract of the bacterium grown on serum-free medium had strong cytotoxicity compared with those grown on serumcontaining medium. They irreversibly damaged HeLa cells without vacuolation that was altogether different from that of the bacterium when grown with serum. Their cytotoxicity was easily measured by cell viability assay. The probable cytotoxic factor partially purified and detected by chromatography had characteristics difference from that of vacuolating toxin and a broad cytotoxicity toward various cell lines. CONCLUSION： Serum-free long culture method of H pylorl makes its supernatant and its extract cytotoxic enough to be easily measured by cell viability assay. The probable cytotoxic factor has a unique characteristic and might be a new cytotoxin.

Novel sericin-based hepatocyte serum-free medium and sericin's effect on hepatocyte transcriptome

AIM To develop a novel hepatocyte serum-free medium based on sericin,and to explore the effect of sericin on the hepatocyte transcriptome.METHODS A controlled trial comparing novel serum-free medium and other media: C3 A cells were cultured in our novel serum-free medium,Hepato ZYME,complete medium(DMEM/F12 with 100 m L/L FBS),and DMEM/F12,andthen cell attachment,proliferation,and function as well as the biocompatibility of the media were assessed.A comparative study of serum-free media with or without 2 mg/m L sericin: the effect of sericin on C3 A growth was assessed by cell viability and proliferation,the effect of sericin on C3 A cell cycle distribution was determined by flow cytometry,and the effect of sericin on the C3 A transcriptome was assessed by gene-chip array and RT-q PCR.RESULTS More C3 A cells attached to the plate containing our serum-free medium than to those containing Hepato ZYME and DMEM/F12 at 24 h post-seeding.Both the viability and proliferation rate of C3 A cells in sericin-based serum-free medium were superior to those of cells in Hepato ZYME and DMEM/F12(P < 0.001).The content of albumin and urea in our serum-free medium was significantly higher than that in Hepato ZYME and DMEM/F12 throughout the whole culture period(P < 0.001) and was similar to that in complete medium at day 3,4,and 5.In part 2,cell viability and proliferation were greater in the presence of 2 mg/m L sericin(P < 0.001),as was the proportion of cells in S phase(16.21% ± 0.98% vs 12.61% ± 0.90%,P < 0.01).Gene-chip array analysis indicated that the expression of CCR6,EGFR,and FOS were up-regulated by 2 mg/m L sericin,and RT-q PCR revealed that the expression of CCR6,EGFR,FOS,AKT1,JNK1,NFk B1,MMP-9,MEK2,ERK1/2 and MYC was upregulated by 2 mg/m L sericin(P < 0.05).CONCLUSION We developed a novel hepatocyte serum-free medium.Sericin probably enhances cell attachment through the CCR6-Akt-JNK-NF-κB pathway and promotes cell proliferation through CCR6-mediated activation of the ERK1/2-MAPK pathway.

INFLUENCE OF MIXING DEVICE ON SERUM-FREE CULTIVATION OF INSECT CELLS IN SPINNER FLASKS

A culture system was developed and successfully employed for the serum-free cultivation of Tn-5B1-4 (Tn5 ) insect cells. With our adaptation procedure, it was possible to obtain cells fully adapted to serum-free media in stationary T-flasks and then enable these adapted cells to grow well in spinner flasks immediately. The spinner "ask with special stirring design proved tO provide favorable culture environment that made it desirable for use in the serum-free cultivation of Tn5 cells even at low seeding density.

Some biological characteristics of hybridomas and parental myelomas cultivated in serum-free medium for long-term period

It has been developed in this laboratory that a serum-free medium, designated asDMI,is adaptive for long-term culture,freezing and resurgence in liquid nitrogen,and cloningof hybridoma and parental myelomas as well as for cell fusions.In this report,it is describedthat the myeloma cells grown in DMI for more than 3 months maintained their biological charac-teristics such as induction of aseites and subcutaneous somatic tumor in BALB/c mice and toler-ance toward 8-azaguanine,ete..However,the ability of the tumor cells to attach to glass wallwas decreased.The selecting assay of these cells in HAT medium(medium containing hypoxan-thine,aminopterin and thymidine)showed that the death time in DMI was similar to that in theconventional 15％ newborn calf serum-supplemented medium(RPS15).The hybridoma cellsadapted in DMI secrete monoclonal antibodies in quantities comparable to those produced inRPS15.

Cholesterol requirement for growth of rodent parental myeloma cells in serum-free medium

Cholesterol,a major lipid component of plasma membrane,is thoughtto have profound effects on the structure and function of cells.Mostanimal tis-sues are capable of synthesizing cholesterol de novo from acetate;however,thereare relatively few mammalian cells in vitro expressing an absolute requirement foran exogenous source of cholesterol.In this paper,it was shown that IR983F(983)rat myeloma cells and P3X63-AgS-U1(P3U1)and P3X63-Ag8.653(653)mousemy eloma cells which had been cultivated in serum-free medium for more than 6months required cholesterol in vitro for growth in serum-free medium.Optimalgrowth of P3U1,653 and 983 occurred in cholesterol concentration of 5,10 and15μg/ml,respectively.Moreover,it was demonstrated that the cholesterol couldbe replaced by human low density lipoprotein in a concentration of 10μg/ml butnot by mevalonic acid lactone.In contrast to the parental myeloma cells,hybridoma cells derived from the mouse myeloma cells which had been cultivatedin serum-free medium for more than 6 months did not require cholesterol.

A PRIMARY OBSERVATION OF TPA EFFECT ON SSV-NIH3T3 CELLS IN SERUM-FREE MEDIUM

The effect of TPA, a potent tumor promoter, on SSV-NIH3T3 cells in serum-free medium was investigated. TPA stimulated DNA synthesis of SSV-NIH3T3 cells on the third day of culture in SFM. In SDS-PAGF of medium conditioned by TPA-treated SSV-NIH3T3 cells (in SFM+TPA), the amounts of four proteins of 31.0 Kd, 28.5 Kd, 25.5 Kd and 13.5 Kd strikingly increased over that of non-TPA-treated counterpart (in SFM). The PDGF-like activity was also detected in CM of SFM+TPA. When insulin and EGF were drown off the SFM+TPA (SFM-Ins-EGF+TPA), TPA lost its ability to stimulate DNA synthesis of SSV-NIH3T3 cells on the third day and SDS-PAGE of the conditioned medium showed that the amounts of the four proteins noted above grately reduced. However, cells in SFM-Ins-EGF+TPA were in almost the same growth condition as cells in complete SFM+TPA on the third day of culture. Results were discussed in the paper.

CLONAL PROLIFERATION AND LONG-TERM CULTURE OF MALIGNANT LYMPHOMA CELLS UNDER SERUM-FREE CULTURE CONDITIONS

We developed a serum-free culture system that promoted the growth of B cell colonies in peripheral blood, bone marrow, lymph nodes and cerebrospinal fluid (CSF) from 7 out of 8 patients with non-Hodgkin's lymphomas of B cell type. The culture cells were pretreated with or without galactose oxi-dase (GO) prior to plating. Colony growth was best supported with BCGF. A moderate increment was observed with rIL-3, as well as rIL-1β and even to a lesser degree, by rlL-2, while B cell stimulating factor-2 (rBCSF-2) and rlL-1β did not show significant activity. rGM-CSF and rG-CSF had little effect, while rM-CSF enhanced the formation of lymphoma colonies. The cells from different patients had different requirements for Staphylococcus aureus protein A and GO pretreatment. It reflected the differences in activation and differentiation status and surface properties of lymphoma cells from different patients. The cells from CSF of one patient were successfully maintained in serum-free culture medium supplemented with 10% BCGF or 5% PHA-LCM for more than 4 months. The long-term culture cells were EBV negative, phenotypically consistent with B cells and gene rearrangements for JH, Kappa and myc. This serum-free culture system allowed extensive analysis of the growth requirements for clonogenic precursors.

Serum-free culture of dendritic cells from patients with chronic myeloid leukemia in vitro and estimation of their cytotoxicity

OBJECTIVE: To establish a serum-free culture system of dendritic cells (DCs) from chronic myeloid leukemia (CML) cells so that DCs vaccine may be applied to the adoptive immunotherapy of CML in the near future. METHODS: Fetal calf serum, serum-free medium and autologous serum were used for culture of DCs. The usage of cytokines was classified into two groups: group A (stem cell factor, granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4) and group B (granulocyte/macrophage colony-stimulating-factor, tumor necrosis factor-alpha and interleukin-4). The phenotypes of DCs were analyzed by using indirect immunofluorescence and flow cytometry. Mixed leukocyte responses were performed by methyl thiazolyl tetrazolium (MTT) assay. Chromosome analysis of DCs can be achieved by displaying G banding. T cells from CML patients were stimulated with autologous DCs and T-cell cytotoxicity was measured by (MTT) assay. RESULTS: CD34(+) cells or mononuclear cells were obtained from peripheral blood or bone marrow samples of eight patients of chronic-phase CML. Group A of serum-free medium was better than group B in expansion of total cell numbers and the rate of DCs. These results of serum-free medium were not significantly different from those of fetal calf serum medium, but the results of autologous serum medium were inferior to two groups above. The expression of major histocompatibility complex class II antigen on the surface of DCs was notable (> 50%), but the expression of CD83 and the costimulatory molecules CD86 was not noticeable (10% - 50%). Although CD1a(+)/CD14(-) DCs were potent stimulators of allogeneic lymphocytes, expansion of T cells from normal volunteers were not significant (average 27.2 fold at DCs: T cells ratio of 1:10). At day 12, CD1a(+) cells from three patients were studied by displaying G banding and Ph(+) cells in these populations were 100%, 98% and 60%, respectively. At an effector: target ratio of 40:1, 32% to 45% cytotoxicity was noted with DC-stimulated T cells against autologous leukemia cells. CONCLUSIONS: A stable serum-free culture system of CML-DCs was established. The expression of CD83 and CD86 on the surface of CML-DCs and DCs' potent stimulation of allogeneic lymphocytes were not notable. DCs in CML patients can be derived from the malignant clone and these malignant DCs could induce anti-leukemic reactivity in autologous T lymphocytes without the necessity for additional exogenous antigens.

STUDY ON REPRODUCTIVE ENDOCRINOLOGY OF HUMAN PLACENTA--CULTURE OF HIGHLY PURIFIED CYTOTROPHOBLAST CELL IN SERUM-FREE HORMONE SUPPLEMENTED MEDIUM

A new method of long-term culture of cytotrophoblast cells in serum-free medium has been developed. Cytotrophoblast cells were isolated with cold trypsin and purified by unit gravity sedimentation through BSA density gradients. The cells were cultured in the FD medium with supplement of EGF, insulin, transferrin and sodium selenite. They could survive over three weeks. The results showed that both EGF and insulin stimulated hCG and progesterone secretion and that sodium selenite elevated hCG output but not progesterone secretion. Transferrin produced synergistic effect with EGF and insulin on hCG and progesterone secretion but it was ineffective when used alone. This study demonstrates that the four growth factors mentioned above are essential for the survival of cytotrophoblast cells in vitro. It is therefore suggested that EGF, insulin and selenium may possibly be involved in the regulation of hCG and progesterone secretion in the human placenta.

Enrichment of breast cancer stem cells using a keratinocyte serum-free medium

Background Keratinoyte serum-free medium （K-SFM） is a defined medium used to support the growth of primary keratinocytes and embryonic stem cell. The aim of this research was to optimize enrichment of breast cancer stem cells （CSCs） using K-SFM. Methods＇ A K-SFM was used to enrich CSCs from two breast cancer cell lines and a primary culture of breast cancer. RPMI-1640 supplemented with 10% fetal calf serum （FCS） was used as a control. CSCs were identified with flow cytometry using CD44＋/CD24-as molecular markers. The expression of a variety of CSC markers （Oct-4, ABCG2, Nanog, N-cadherin, and E-cadherin） was analyzed with real-time PCR. Results Much higher percentage of CSCs was achieved with K-SFM： 17.3% for MCF-7 cells, 17.4% for SKBR-3, and 20.0% for primary breast cancer culture. Less than 1% CSC was achieved using RPMI-1640 supplemented with 10% FCS. In comparison to the CSCs obtained with RPMI-1640, CSCs in the K-SFM expressed higher levels of Oct-4, ABCG2, Nanog and N-cadherin, and lower level of E-cadherin. Conclusion K-SFM is an optimal culture medium to maintain and to enrich breast CSCs.

Application of Serum-Free Culture Medium for Preparation of A-NK Cells

To compare the differences between proliferation and cytotoxicity of adherent natural killer （A-NK） cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also observe the assisting effect of IL-12 on the activation and the morphology character of IL-2-treated A-NK cells, cellular proliferation was evaluated by MTT method in vitro. The morphology of the target cells killed by A-NK cells was observed through electroscope. All of the A-NK cells cultured in serum-free medium AIMV could rapidly proliferate and keep high cytotoxicity compared with that in standard serum-containing medium. A-NK cells activated by both moderate-dose IL-2 and IL-12 were superior to the high-dose IL-2-treated A-NK cells. These data indicated that serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells, and moderate-dose IL-2 and IL-12 could reduce side effects caused by high-dose IL-2. The study provided a new experimental basis for experimental and clinical preparation of A-NK cells. Cellular ＆ Molecular Immunology.

Serum-free media for articular chondrocytes in vitro expansion

Background In vitro chondrocyte expansion is a major challenge in cell-based therapy for human articular cartilage repair. Classical culture conditions usually use animal serum as a medium supplement, which raises a number of undesirable questions. In the present study, two kinds of defined, serum-free media were developed to expand chondrocytes in monolayer culture for the purpose of cartilage tissue engineering. Methods Bovine chondrocytes were expanded in serum-free media supplemented with fibroblast growth factor-2 and platelet-derived growth factor or fibroblast growth factor-2 and insulin-like growth factor. Expansion culture in a conventional 10% fetal bovine serum （FBS） medium served as control. Fibronectin coating was used to help cell adhesion in serum-free medium. Next, in vitro three-dimensional pellet culture was used to evaluate the chondrocyte capacity. Cell pellets were expanded in different media to re-express the differentiated phenotype （re-differentiation） and to form cartilaginous tissue. The pellets were assessed by glycosaminoglycans contents, collagen II, collagen I and collagen X immunohistological staining. Results Chondrocytes cultured in serum-free media showed no proliferation difference than cells grown with 10% FBS medium. In addition, chondrocytes expanded in both serum-free media expressed more differentiated phenotypes at the end of monolayer culture, as indicated by higher gene expression ratios of collagen type II to collagen type I. Pellets derived from chondrocytes cultured in both serum-free media displayed comparable chondrogenic capacities to pellets from cells expanded in 10% FBS medium. Conclusion These findings provide alternative culture approaches for chondrocytes in vitro expansion, which may benefit the clinical use of autologous chondrocytes implantation.

接种密度对牛颗粒细胞形态及相关基因表达的影响

牛卵巢颗粒细胞是雌激素(Estrogen,E_(2))合成的主要来源,具有E_(2)活性的牛颗粒细胞体外模型的建立是研究E_(2)合成与分泌过程中潜在调控机制的重要工具,该细胞模型的建立可为E_(2)合成分子调控机制及牛卵巢卵泡发育机制的研究提供理论与技术支持。细胞接种密度是颗粒细胞体外培养模型的关键因素,高密度可引起细胞生理及分子的显著变化。研究通过不同接种密度下的形态变化和基因表达筛选牛颗粒细胞体外培养最优接种密度,采用长期无血清法培养牛原代颗粒细胞,培养液中添加FSH及IGF-1以诱导E_(2)的合成。培养7 d后,采集6孔板中高(3.0×10^(6)个细胞/孔)、中(2.0×10^(6)个细胞/孔)、低(1.0×10^(6)个细胞/孔)3种不同接种密度的细胞图像进行观测,并利用qRT-PCR技术检测3种不同接种密度中相关基因的表达情况。结果表明,低密度(1.0×10^(6)个细胞/孔)接种细胞呈现成纤维细胞样外观,细胞无聚集倾向;中密度组(2.0×10^(6)个细胞/孔)可观察到少量聚集的细胞团;高密度(3.0×10^(6)个细胞/孔)培养条件下,大多数细胞聚集成细胞团。低密度组(1.0×10^(6)个细胞/孔)中,CYP19A1及FSHR高表达,RGS2及VNN2低表达;高密度组(3.0×10^(6)个细胞/孔)表达量则相反。综上所述,低密度组(1.0×10^(6)个细胞/孔)细胞可作为具有E_(2)活性的牛颗粒细胞体外模型。

在自定义高危群体中M蛋白定量筛查多发性骨髓瘤及相关预后研究

目的探讨M蛋白定量检测在符合自定义高危群体中对多发性骨髓瘤的诊断及预后分层价值。方法纳入2020年6月至2022年12月就诊于本院所筛查出的200例符合自定义多发性骨髓瘤高危群体进行血清蛋白电泳M蛋白检测及血清游离轻链定量测定,记录入组患者血清蛋白电泳结果、血清游离轻链定量及κ/λ比值;阳性患者进一步完善检测确诊为多发性骨髓瘤,确诊患者再进一步行Fish检测评估预后危险分层,记录相关检测结果。结果200例高危群体中血清游离轻链比值异常共检出58例,血清蛋白电泳阳性检出39例,进一步确诊多发性骨髓瘤共34例;确诊患者血清游离轻链比值异常为34例,而血清蛋白电泳M蛋白阳性为27例,7例漏诊;免疫原位杂交(Fish)检测结果提示确诊患者血清游离轻链高比值13例中(>100或<0.01)有8例存在高危遗传学改变,其他21例有3例存在高危遗传学改变。而多发性骨髓瘤高危影响因素的多因素分析显示:女性、贫血、球蛋白异常、血肌酐升高是高危影响因素,符合本研究中自定义多发性骨髓瘤高危群体的诊断标准。结论血清游离轻链检测相较于血清蛋白电泳检测可以明显降低多发性骨髓瘤漏诊率,是高危人群检测筛查的有效手段,并且血清游离轻链高比值跟基因风险分层存在一定的正相关性,能在一定程度上反应其预后风险。

血清游离轻链在肺癌中的表达及诊断价值

目的:血清游离轻链(free light chain,FLC)在多种疾病中表达异常,但其在肺癌中的表达尚不清楚,本研究旨在探讨血清FLC在肺癌中的表达及诊断价值。方法:选取2021年1至12月湖南师范大学附属湘东医院收治的80例肺癌患者作为肺癌组,另选取同时期的80例健康体检人员作为对照组。收集所有参与者的一般资料、血清κFLC和λFLC水平;收集肺癌组患者住院期间的相关临床指标[血清癌胚抗原(carcinoembryonic antigen,CEA)、细胞角蛋白-19片段抗原(cytokeratin fragment antigen 21-1,CYFRA21-1)水平,以及肿瘤直径、组织学分型、TNM分期、是否有淋巴结转移]。比较肺癌组和对照组血清FLC[κFLC、λFLC、FLC(κ+λ)]的表达水平。将80例肺癌患者按性别、年龄、吸烟史、肿瘤直径、TNM分期、组织学分型、淋巴结转移进行分组,比较组间血清κFLC、λFLC表达水平的差异。采用受试者操作特征(receiver operating characteristic,ROC)曲线评价血清FLC水平单独及联合其他指标在肺癌中的诊断价值。结果:肺癌组血清FLC(κ+λ)、κFLC表达水平均显著高于对照组,差异均有统计学意义(均P<0.001);而2组间血清λFLC表达水平的差异无统计学意义(P>0.05)。不同肿瘤直径、组织学分型、TNM分期的肺癌血清κFLC表达水平差异均无统计学意义(均P>0.05);但是,有淋巴结转移的肺癌患者血清κFLC水平高于无淋巴结转移的肺癌患者,且差异有统计学意义(P=0.033)。不同肿瘤直径、组织学分型肺癌患者的血清λFLC表达水平差异均无统计学意义(均P>0.05);但是,TNM分期III期+IV期、有淋巴结转移的肺癌患者血清λFLC表达水平分别高于TNM分期I期+II期、无淋巴结转移的肺癌患者,差异均有统计学意义(分别P=0.033,P=0.019)。κFLC、CEA诊断肺癌的曲线下面积(area under the curve,AUC)差异无统计学意义(P=0.333)。在2项联合诊断肺癌的指标中,κFLC+CYFRA21-1的诊断效能(AUC=0.875)及敏感性(71.3%)最高。κFLC+λFLC+CEA+CYFRA21-1联合诊断肺癌的AUC为0.915(95%CI 0.860~0.953,P<0.001)。结论:血清FLC在肺癌中高表达,并且与肺癌的浸润和转移有关。血清FLC尤其是κFLC对肺癌的诊断具有价值,FLC、CEA、CYFRA21-1联合检测的诊断效能最佳。

sFLC、血清钙离子在多发性骨髓瘤患者诊断及预后判断中的应用价值

目的:探讨血清游离轻链(sFLC)、血清钙离子在多发性骨髓瘤(MM)患者诊断及预后中的临床应用价值。方法:选取2018年1月至2022年1月在河南省人民医院治疗的MM患者40例作为观察组,同时选取健康志愿者40例作为对照组,比较两组患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子等差异性,同时分析观察组不同国际分期系统(ISS)、不同化疗疗效及不同预后患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子等差异性。结果:观察组sFLC-κ水平[(98.39±21.19)对(12.01±4.45)mg/L]、sFLC-λ水平[(210.20±45.54)对(14.10±5.11)mg/L]、低钙血症比例(65%对0)均明显高于对照组(P<0.05),而sFLC-κ/λ比值[(0.44±0.10)对(0.87±0.12)]、血清钙离子[(1.98±0.46)对(2.42±0.40)mmol/L]明显低于对照组(P<0.05)。观察组ISS分期Ⅲ期患者sFLC-κ、sFLC-λ浓度、低钙血症比例和低钙血症病程均明显高于Ⅰ期和Ⅱ期患者(P<0.05),而sFLC-κ/λ、血清钙离子均明显低于Ⅰ期和Ⅱ期患者(P<0.05)。观察组有低钙血症患者sFLC-κ水平[(107.76±21.22)对(94.67±20.11)mg/L]、sFLC-λ水平[(245.54±41.12)对(205.54±50.22)mg/L]明显高于无低钙血症患者(P<0.05),而sFLC-κ/λ比值明显低于无低钙血症患者[(0.42±0.04)对(0.47±0.06);P<0.05]。化疗无效患者sFLC-κ水平[(107.29±20.14)对(91.11±18.92)mg/L]、sFLC-λ水平[(247.98±42.26)对(179.29±39.32)mg/L]明显高于化疗有效患者(P<0.05),而sFLC-κ/λ比值明显低于化疗有效患者[(0.43±0.10)对(0.50±0.09);P<0.05]。sFLC-κ、sFLC-λ、sFLC-κ/λ预测化疗无效的ROC曲线下面积分别为0.803、0.793和0.699,P<0.05。存活和死亡患者的sFLC-κ、sFLC-λ、sFLC-κ/λ、血清钙离子、低钙血症比例和低钙血症病程比较差异无统计学意义(均P>0.05)。结论:sFLC、血清钙离子与MM患者ISS分期有关,sFLC水平在预测MM患者化疗疗效方面有一定应用价值,sFLC与血清钙离子在判断MM患者预后方面暂未发现有应用价值。

河北省免费NIPT联合PAPP-A、F-β-HCG 及超声检查诊断孕中期胎儿染色体异常的意义

目的 评价河北行免费的无创产前DNA检测(NIPT),联合血清妊娠相关血浆蛋白-A(PAPP-A)、游离β-人绒毛膜促性腺激素(F-β-HCG)检测及超声对孕中期染色体异常的诊断价值。方法 选取衡水市第二人民医院2019年1月至2023年1月接收的3 591例拟行染色体检测的孕中期妇女,均行免费NIPT联合血清学指标(PAPP-A、F-β-HCG)及超声检查。将羊水穿刺染色体核型分析结果记为金标准,采用受试者工作特征曲线(ROC)评价不同方法诊断孕中期染色体异常的价值。结果 经随访,在3 591例中,共有39例染色体异常,其中,32例经超声检查有染色体异常,34例经NIPT检测有染色体异常。在3 552例染色体正常的胎儿中,34例经超声检查有染色体异常,36例经NIPT检测有染色体异常;染色体异常胎儿的母体PAPP-A、F-β-HCG水平均高于染色体正常胎儿的母体(P<0.05)。PAPP-A、F-β-HCG共同诊断胎儿染色体异常的敏感度及ROC曲线下方的面积(AUC)均高于母体PAPP-A、F-β-HCG单独诊断(P<0.05);NIPT,PAPP-A、F-β-HCG及超声的联合诊断的敏感度及AUC均高于NIPT,PAPP-A、F-β-HCG,超声的单独诊断(P<0.05),且不同方法诊断染色体异常的特异度对比差异均无统计学意义(P>0.05)。结论 孕中期胎儿染色体异常的母体PAPP-A、F-β-HCG水平高,且NIPT联合PAPP-A、F-β-HCG及超声诊断染色体异常有较好的价值。

MDCK细胞无血清悬浮培养技术在流感疫苗研究与生产中的应用进展

流感是一种常见的呼吸道疾病,由流感病毒引起,接种疫苗是预防流感的有效手段。MDCK细胞(Madin-Darby Canine Kidney Cells)具有易于培养和高产量的特点,可以支持流感病毒的复制和增殖,被广泛用于流感疫苗的研究和生产。随着对流感病毒研究的不断深入,为了进一步拓展MDCK细胞用于流感病毒研究和工业应用的能力,研究人员开始发展MDCK细胞无血清全悬浮培养技术。通过长期的培养和优化,驯化的MDCK悬浮细胞株可以更好地适应流感病毒的生长环境,提高流感病毒的产量和感染性。总之,MDCK细胞无血清悬浮培养技术在流感病毒研究和工业应用中发挥着重要作用,为流感疫苗生产和抗流感药物研发提供更好的工具。同时也必须重视MDCK细胞的安全性,采取合适的措施来确保其应用的安全性和可靠性。

饲粮代谢能水平对14~17周龄北京油鸡血清抗氧化、免疫指标及肌肉游离氨基酸含量的影响

试验旨在研究饲粮代谢能水平对14~17周龄北京油鸡血清抗氧化和免疫指标及肌肉游离氨基酸含量的影响。选取13周龄北京油鸡600只(公母各半),随机分为3组,每组200只。组内各性别分别设置5个重复。3个处理组分别饲喂低代谢能(12.20 MJ/kg)、中代谢能(12.64 MJ/kg)、高代谢能(13.08 MJ/kg)3种不同的日粮。预试期1周,正试期4周(14~17周龄)。结果显示:①饲粮代谢能水平和性别对血清过氧化物酶活性有显著交互作用(P<0.05),中代谢能组母鸡过氧化物酶活性显著高于中代谢能组公鸡(P<0.05);性别对血清超氧化物歧化酶、谷胱甘肽过氧化物酶活性有显著影响(P<0.05),母鸡血清超氧化物歧化酶活性显著高于公鸡(P<0.05),谷胱甘肽过氧化物酶活性显著低于公鸡(P<0.05);②代谢能水平和性别对血清IgA、IgG、IgM、γ-干扰素含量有显著交互作用(P<0.05),饲粮代谢能水平对血清IgA、IgM、白细胞介素-6含量有显著影响(P<0.05);中代谢能组IgA含量显著低于其他两组(P<0.05);低代谢能组IgM含量显著低于其他两组(P<0.05);高代谢能组白细胞介素-6含量显著高于其他两组(P<0.05)。性别对血清IgA、IgG、α-干扰素含量有显著影响(P<0.05)。母鸡IgA和α-干扰素含量显著高于公鸡(P<0.05)。公鸡IgG含量显著高于母鸡(P<0.05)。③饲粮代谢能水平和性别对14~17周龄北京油鸡肌肉中部分游离氨基酸含量有显著影响(P<0.05),且在中代谢能水平下含量最高;公鸡总氨基酸、甜味氨基酸、鲜味氨基酸含量显著高于母鸡(P<0.05)。研究表明:代谢能水平对14~17周龄北京油鸡血清抗氧化指标无显著影响;代谢能为13.08 MJ/kg时,免疫性能较好;代谢能为12.64 MJ/kg时,肌肉游离氨基酸含量最高,甜味氨基酸、鲜味氨基酸、必需氨基酸、总氨基酸含量最高,味道更鲜,品质最优;从游离氨基酸含量分析,公鸡较母鸡风味更佳。