抗伏马菌素B_1单克隆抗体的制备及鉴定

选择钥孔虫戚血蓝蛋白（ Ｋ Ｌ Ｈ）和牛血清白蛋白（ Ｂ Ｓ Ａ）作为半抗原伏马菌素 Ｂ１ （ Ｆ Ｂ１ ）的载体蛋白,以戊二醛一步法分别制备免疫原 Ｋ Ｌ Ｈ Ｆ Ｂ１ 和固相抗原 Ｂ Ｓ Ａ Ｆ Ｂ１ 蛋白结合物。用 Ｋ Ｌ Ｈ Ｆ Ｂ１ 免疫 Ｂａｌｂ／ｃ 小鼠后,应用细胞融合技术,筛选后获得了１株稳定分泌抗 Ｆ Ｂ１ 单克隆抗体的杂交瘤细胞株（ Ｆ Ｂ１ ４ Ｇ４ ）,分泌的抗体属于 Ｉｇ Ｇ１ 亚类。腹水的抗体效价为１ｖ１×１０７ ,抗体与其它３种真菌毒素、 Ｆ Ｂ３ 和２种载体蛋白均无交叉反应,与 Ｆ Ｂ２有轻微的交叉反应。     

抗A型产气荚膜梭菌α毒素单克隆抗体杂交瘤细胞系的建立与中和效应

用提取的Ａ型产气荚膜梭菌α毒素包涵体免疫ＢＡＬＢ／ｃ小鼠，取小鼠脾细胞与ＳＰ２／０骨髓瘤细胞进行融合和克隆化，经ＥＬＩＳＡ筛选，共获得７株稳定分泌单克隆抗体（ＭｃＡｂ）的杂交瘤细胞株，分别命名为１Ａ８、１Ｃ３、１Ｄ５、１Ｄ８、１Ｆ１、１Ｈ１和２Ｅ３。经鉴定，７株ＭｃＡｂ的Ｉｇ亚类有ＩｇＧ１（１Ｄ８）、ＩｇＧ３（１Ａ８、１Ｃ３和２Ｅ３）和ＩｇＭ（１Ｄ５、１Ｆ１和１Ｈ１）。细胞培养上清和腹水抗体效价分别为：１∶５１２～１∶１０２４和１∶１０６～１∶１０８。尤为重要的是，２Ｅ３杂交瘤细胞株分泌的ＭｃＡｂ不仅能够中和α毒素的磷脂酶Ｃ活性和溶血活性，而且能够对致死性腹腔感染小鼠产生良好的被动保护作用     

米酵菌酸单克隆抗体细胞株的建立

继制备米酵菌酸（ＢＡ）多克隆抗体以后，作者成功地运用免疫杂交瘤技术，获得了能分泌抗米酵菌酸的单克隆抗体细胞株１Ｆ７和２Ｈ９，其腹水效价可达１０－６，对ＢＡ的５０％有效抑制浓度为１０－４，两株细胞分泌抗体的亚型均为ＩｇＧ１型，纯化后获得ＢＡ－ＭｃＡｂ的ＩｇＧ浓度为１１ｇ／Ｌ     

异基因外周血造血干细胞移植治疗白血病

探讨用 Ｈ Ｌ Ａ 相合的同胞异基因外周血干细胞移植治疗急性髓细胞白血病及 Ａ Ｂ Ｏ 血型不合的 Ｐ Ｂ Ｓ Ｃ Ｔ 的情况,采用 Ｃ Ｔ Ｘ６０ｍｇ／ｋｇ ×２ ＋ Ｔ Ｂ Ｉ７０ ～７５ Ｇｙ 直线加速器照射＋ Ｃ Ｃ Ｎ Ｕ５０ｍｇ／ｋｇ ×１ ± Ｖ Ｐ１６３０ｍｇ／ｋｇ ×１ 对受者作预处理,供者经 Ｇ－ Ｃ Ｓ Ｆ６μｇ／ｋｇ ×５ｄ 动员后,用血细胞分离机（ Ｂａｘｔｅｒ Ｃ Ｓ３０００ｐｌｕｓ） 分离采集干细胞去除红细胞。对 Ａ Ｂ Ｏ 血型不合的 Ｐ Ｂ Ｓ Ｃ,强化移植物抗宿主病（ Ｇ Ｖ Ｈ Ｄ） 预防方案中 Ｍ Ｔ Ｘ 用量及次数;选输合适血制品。结果３ 例病人输注单个核细胞数分别为４０ 、１００ 、５０ ×１０８／ｋｇ , Ｃ Ｄ３４ ＋ 细胞分别为３６ 、１９０ 、２０ ×１０６／ｋｇ ;干细胞悬液中红细胞残存量分别为２３ｍｌ、１４ｍｌ、２６ ｍｌ,红细胞残存率分别为４ ％ 、３ ％ 、５３ ％ 。移植后造血重建迅速。例１ 于＋ ３４ｄ 染色体完全转变为供者型;例２ 于＋ ４０ｄ 血型完全转变为供者型,＋ ４５ｄ 染色体完全转变为供者型,例３ ＋ ５５ｄ 血型完全转变为供者型。没有输血溶血和免疫溶血发生。例２ 为主,次要 Ａ Ｂ Ｏ 均不合,则早期     

微乳液-8-羟基喹啉体系测定Fe(Ⅲ)

研究了 Ｏ／ Ｗ 阴离子微乳液十二烷基硫酸钠（ Ｓ Ｄ Ｓ）／正丁醇（ｎ－ Ｃ４ Ｈ９ Ｏ Ｈ）／正庚烷（ｎ－ Ｇ７ Ｈ１６ ）／ Ｈ２ Ｏ 对 Ｆｅ 与８－ 羟基喹啉（ Ｈ Ｏｘ ）显色体系的增敏增稳作用,实验结果表明其灵敏度为ε４６０ ＝ ６５６×１０３ Ｌ·ｍ ｏｌ－ １ ·ｃｍ － １ ,ε５８０ ＝ ５５９×１０３ Ｌ·ｍ ｏｌ－ １ ·ｃｍ － １ ,与相同含水量的 Ｓ Ｄ Ｓ胶束体系（ε４６０ ＝ ４７２×１０３ Ｌ·ｍ ｏｌ－ １ ·ｃｍ － １ ,ε５８０ ＝ ４０５×１０３ Ｌ·ｍ ｏｌ－ １ ·ｃｍ － １ ）比较,发现其测定灵敏度显著增加,增加百分比分别为３８９％ 、４６９％ ,某些实验条件得以改善。     

HCl—NaBH4—K3Fe（CN）6体系气态原子吸收光谱法测定饮料中铅

采用ＶＡ－９０气态原子化装置与ＷＦＸ－１Ｄ原子吸收分光光度计联用测定饮料中铅，采用ＨＣｌ－ＮａＢＨ４－Ｋ３Ｆｅ（ＣＮ）６体系，经Ｌ９（３４）正交试验选择最佳实验条件为：ＮａＢＨ４２０ｇ／Ｌ，Ｋ３Ｆｅ（ＣＮ）６１００ｇ／Ｌ，ＨＣｌ１．５ｍｌ，氩气流量为０．８Ｌ／ｍｉｎ。在０～１００μｇ／Ｌ范围内，相关系数ｒ＝０．９９９６，回收率为９４．１％～１０７．５％，Ｒ·Ｓ·Ｄ％＝３．３～５．２特征浓度和检出限分别为１．２９μｇ／Ｌ／１％和０．０４９μｇ／Ｌ。干扰试验表明，Ｓｎ、Ｈｇ、Ｃｒ、Ｃｕ、Ｍｎ在５００μｇ／Ｌ以下，Ｚｎ、Ａｓ、Ｆｅ、Ｎｉ、Ａｇ在１０００μｇ／Ｌ以下，不干扰测定     

新疆出血热北疆株单克隆抗体杂交瘤细胞株的建立

作者应用新疆出血热病毒北疆株９０－９株鼠脑免疫ＢＡＬＢ／Ｃ小鼠，取脾细胞与ＳＰ２／０骨髓瘤细胞融合，共获得４株分泌抗ＸＨＦＶ北疆株单克隆抗体（ＭｃＡｂ）杂交瘤细胞系。核型分析表明符合杂交瘤细胞的标准。用间接免疫荧光法及间接酶联免疫吸附试验检测表明，ＭｃＡｂ的腹水具有很高滴度，其中２株可达５×１０５（ＥＬＩＳＡＰ／Ｎ≥２．１）。ＭｃＡｂ免疫球蛋白亚型鉴定均属ＩｇＧ，其中３株为ＩｓＧ１，１株为ＩｇＧ２ａ。对ＭｃＡｂ的初步应用表明，在反向被动血凝抑制试验（ＲＰＨＩ）中，分别采用南、北疆ＭｃＡｂ致敏血球检测小鼠腹水，其结果有明显差异。     

乙肝病毒基因免疫小鼠初步研究

将经过基因修饰的乙肝病毒核心区基因定向克隆于质粒ｐｃＤＮＡ３巨细胞病毒启动子下游，构建成真核表达载体，称为ｐｃＤＮＡ３－ＨＢＶ。将ｐｃＤＮＡ３－ＨＢＶ通过多点肌肉注射，给ＢＡＬＢ／ｃ小鼠免疫接种１００μｇ／只，２周后追加免疫１次，追加免疫１周后８只免疫鼠中７只出现抗－ＨＢｃＩｇＧ血清转化，而接种空载质粒ｐｃＤＮＡ３的对照组及未经任何免疫接种的ＢＡＬＢ／ｃ小鼠无一只抗－ＨＢｃＩｇＧ阳性。     

不稳定心绞痛与急性心肌梗死血脂变化比较

目的：探讨不稳定心绞痛（ＵＡＰ）和急性心肌梗死（ＡＭＩ）之间血脂变化。方法：对１０９例ＵＡＰ和１４３例ＡＭＩ病人血脂值进行统计学处理分析。结果：ＵＡＰ组ＴＧ（１７１．２±１４６．９）ｍｇ／ｄｌ高于ＡＭＩ组（１３６．９±８４）ｍｇ／ｄｌ（Ｐ＜０．０１）及ＴＧ／ＨＤＬ－Ｃ（５．５±６．２）比（４．１±３．３）和ＬＤＬ－Ｃ／ＨＤＬ－Ｃ（４．０±１．９）比（３．６±１．６）高于ＡＭＩ组（两者均Ｐ＜０．０５）。结论：ＵＡＰ病人ＬＤＬ－Ｃ／ＨＤＬ－Ｃ比值增高可能与其血管病变严重性相关     

肾病综合征患儿输血后疟疾1例

１ 病例患儿陈某 ,男 ,４岁。因受凉“感冒”后反复尿少、全身浮肿３月余于１９９４年１月３１日入本院。曾在当地医院检查 :尿蛋白 ,血胆固醇１５．９ｍｍｏｌ／Ｌ,白蛋白１５ｇ／Ｌ,Ａ／Ｇ倒置 ,拟诊“肾病综合征”给予强的松治疗６周无效。入本院时检查 :Ｔ３７．２℃ ,Ｐ１２０次／分 ,ＢＰ１４／１０．６ｋＰａ ,柯兴面容 ,全身高度水肿 ,咽部充血 ,心尖部Ⅰ级ＳＭ ,肝下缘于右肋下３ｃｍ ,脾未触及。血常规ＷＢＣ１３．６×１０９／Ｌ ,ＲＢＣ４．５５×１０１２／Ｌ ,血胆固醇９．６２ｍｍｏｌ／Ｌ ,ＡＬＢ１ｇ／Ｌ ,ＧＬＢ２…     

抗黄曲霉毒素M1的单克隆抗体细胞株的建立

本研究建立了抗黄曲霉素素ＡＦＭ１的单克隆抗体细胞株,用ＡＦＭ１－肟＝朱血清白蛋白（ＡＦＭ１－ｏｘｉｍｅ－ＢＳＡ）ＷＪＭ ＢＵ ＴＲ ＱＫＵＭ本研究建立     

NanH and PknG putative virulence factors as a recombinant subunit immunogen against Corynebacterium pseudotuberculosis infection in mice

Despite the economic and zoonotic relevance of caseous lymphadenitis, a competent immunoprophylaxis tool is still necessary. Here, we evaluated two putative virulence factors of Corynebacterium pseudotuberculosis, rNanH, and rPknG, as recombinant subunit vaccines in a murine model against the infection by C. pseudotuberculosis. Three groups of ten Balb/c mice each were inoculated with a sterile 0.9% saline solution (G1), rNanH (G2), or rPknG (G3) in formulations containing saponin as an adjuvant. The mice received two vaccine doses intercalated by a 21-day interval and were challenged with 2 × 10⁴ CFU/mL of the C. pseudotuberculosis MIC–6 strain 21 days after the last immunization. The total IgG, IgG1, and IgG2a production levels increased significantly in the experimental groups (G2 and G3) on day 42. The highest levels of IgG2a antibodies in G2 and G3 were observed compared to IgG1 levels. G3 showed a significant (p < 0.05) humoral response through higher production of total IgG at day 42 when compared to G2. A significant increase of mRNA expression levels of interleukin (IL)-17, tumor necrosis factor, and interferon-γ was observed only in G2, while IL-4 was significantly produced only by G3. The levels of IL-10 and IL-12 obtained were not significant in any group. The survival rates after the challenge were 20% for G3 and 60% for G2 (p < 0.05). Our findings suggest that the formulation containing rNanH and saponin (G2) resulted in the best protection against the challenge and was able to elicit a Th1 immune response in mice, and can be considered as a promising antigen in the development of an effective vaccine against caseous lymphadenitis.     

Epitope mapping of a common 57 kDa antigen of Leishmania species by monoclonal antibodies

BALB/c mice were immunized with freeze-thawed promastigote of Leishmania infantum. Five monoclonal antibodies (mAb) were selected, four IgM (designated as P1A9, P2G8, P5E3 and P6B3) and one IgG1 (P3D2). ELISA and Western blot analysis suggested that all monoclonal antibodies are specific to a band of 57 kDa antigen of L. infantum as well as other three Leishmania species (L. tropica, L. major and L. donovani). ELISA additivity tests revealed four epitopes on 57 kDa antigen as defined by four IgM monoclonal antibodies. Three distinct epitopes were recognized by P1A9, P2G8, and P6B3 antibodies and one epitope recognized by P5E3 antibody that shared with P2G8, and P6B3 epitopes. The 57 kDa protein was purified with affinity column and was shown to possess proteolytic activity. It seems that 57 kDa protein is the major surface Leishmania antigen (gp63) that has been used as subunit vaccine with appropriate adjuvant and induced protection against L. major infection in BALB/c mice.     

Partial protection to respiratory syncytial virus (RSV) elicited in mice by intranasal immunization using live staphylococci with surface-displayed RSV-peptides

A live bacterial vaccine-delivery system based on the food-grade bacterium Staphylococcus carnosus was used for delivery of peptides from the G glycoprotein of human respiratory syncytial virus, subtype A (RSV-A). Three peptides, corresponding to the G protein amino acids, 144-159 (denoted G5), 190-203 (G9) and 171-188 (G4 S), the latter with four cysteine residues substituted for serines, were expressed by recombinant means as surface-exposed on three different bacteria, and their surface accessibility on the bacteria was verified by fluorescence-activated cell sorting (FACS). Intranasal immunization of mice with the live recombinant staphylococci elicited significant anti-peptide as well as anti-virus serum IgG responses of balanced IgG1/IgG2a isotype profiles, and upon viral challenge with 10(5) tissue culture infectious doses(50) (TCID(50)), lung protection was demonstrated for approximately half of the mice in the G9 and G4 S immunization groups. To our knowledge, this is the first study in which protective immunity to a viral pathogen has been evoked using food-grade bacteria as vaccine-delivery vehicles.     

东海加工用河鲀鱼种毒性研究

应用酶联免疫方法，对分布于中国东海，应用地加工河豚鱼干制品的五种主要河鱼种毒性进行了调查研究。毒素含量范围为＜４．０×１０－２＞１．０×１０４ＭＵ／ｇ。即使处于繁殖期，暗鳍、淡鳍腹刺仍均属无毒或弱毒级。头纹宽吻和横纹东方则具有相当大的毒性。毒素含量随其生活的环境、海域及季节变化。建议在监督控制下，批准部分具备条件的企业进行河鱼干制品的加工生产，但应明确限制允许加工的腹刺属鱼种     

TAM和MPA联合用药对卵巢癌顺铂耐药细胞株SKOV-3/DDP生长的影响

目的:探讨三苯氧胺(tamoxifen,TAM)和甲孕酮(medroxyprogesterone,MPA)联合用药对顺铂耐药的卵巢癌细胞株SKOV-3／DDP生长影响的机制。方法:用四甲基偶氮唑蓝(MTT)法、电子显微镜及流式细胞仪测定用药前后SKOV-3／DDP细胞生长、形态学、周期及凋亡率的变化。结果:低浓度的TAM和MPA联合用药组对SKOV-3／DDP细胞生长无影响。高浓度用药组可抑制卵巢癌细胞生长,电子显微镜可观察到细胞凋亡的形态学变化,1×10-6~1×10-4mol／L的TAM和MPA联合用药组在抑制细胞增殖的同时尚可诱导细胞凋亡,其细胞凋亡率分别为4.06％、11.77％、19.3％,两两比较,差异均有显著性(P<0.05)。结论:TAM和MPA联合用药既对顺铂耐药的卵巢癌细胞有杀灭作用,又因其较小的副作用及肯定的临床意义有可能成为卵巢癌有效的化疗辅助用药。     

Persistence of Lactobacillus reuteri DSM17938 in the Human Intestinal Tract: Response to Consecutive and Alternate-Day Supplementation

Background: Probiotics may enhance gastrointestinal health and immune function. The efficacy of different probiotic dosing strategies on colonization and persistence of probiotics is undefined.

Objective: The authors assessed colonization and persistence of Lactobacillus reuteri (L. reuteri) DSM17938 (BioGaia AB, Stockholm, Sweden) after daily or alternate-day dosing.

Methods: Volunteers ate pudding with L. reuteri (10⁹ CFU) daily (n = 9) or on alternate days (n = 9) over 7 days. Fecal samples were collected on dosing days (D1–7) and after dosing ended (D13–15 and D20–22) and were analyzed for the presence of L. reuteri. Results are reported in 3-day increments (D2–4, D5–7, D13–15, and D20–22).

Results: L. reuteri count rose in response to daily supplementation ([mean ± SD] D2–4: 4 × 10⁴ ± 2 × 10⁴ CFU, p < 0.01; D5–7: 10 × 10⁴ ± 9 × 10⁴ CFU, p < 0.01) and alternate-day supplementation (D2–4: 21 × 10⁴ ± 20 × 10⁴ CFU, p < 0.01; D5–7: 11 × 10⁴ ± 15 × 10⁴ CFU, p = 0.06) and fell in both groups 1 week after dosing ended (p < 0.01). Total volunteers with detectable L. reuteri 1 and 2 weeks after dosing ended was similar in response to daily feeding (4/9 and 2/9, respectively) and alternate-day feeding (3/9 and 2/9, respectively). L. reuteri count was higher D2–4 in response to alternate-day vs daily feeding (p < 0.05) but similar thereafter.

Conclusions: Alternate-day probiotic intake achieves equivalent colonization to daily intake, but colonization declines rapidly once dosing stops. It is possible that, initially, responsiveness to probiotics may differ between individuals, but those differences do not persist with longer consumption.     

Human IgG subclass responses in relation to serum bactericidal and opsonic activities after immunization with three doses of the Norwegian serogroup B meningococcal outer membrane vesicle vaccine

Ten adult volunteers, with low prevaccination levels of serum IgG antibodies against meningococcal antigens (< 1 microg ml(-1)), received three doses of the Norwegian group B meningococcal outer membrane vesicle (OMV) vaccine intramuscularly at weeks 0, 6 and 46. Anti-OMV IgG subclass responses were measured and compared with serum bactericidal activity (SBA) and opsonic activity against the vaccine strain 44/76. All vaccinees showed an IgG1 antibody response after each vaccine dose. The vaccine-induced median serum IgG1 antibody levels were 16, 17 and 18 microg ml(-1) 2-6 weeks after the first, second and third dose, respectively. Three vaccinees showed a weak IgG3 response after the first dose, whereas 8 and 9 showed a response after the second (median = 10 microg ml(-1)) and third dose (median = 10 microg ml(-1)), respectively. Low levels of anti-OMV IgG2 antibodies were found, whilst specific IgG4 antibodies were only detected for one vaccinee. The vaccine induced at least a fourfold increase in SBA titre in 8 vaccinees after the first dose, in 9 vaccinees after 2 doses and in all vaccinees after 3 doses. A positive correlation was found between IgG1 subclass antibody levels and SBA (r = 0.62, P < 0.0001). Elevated opsonophagocytic activity, measured as respiratory burst (RB), was observed in all vaccinees after one vaccine dose and usually increased after 2 and 3 doses. A strong positive correlation was found between IgG1 antibody levels and RB (r = 0.76, P < 0.0001). In conclusion, we have shown that systemic meningococcal OMV vaccination in adult vaccinees mainly induced IgG1 antibodies which correlated with bactericidal and opsonic activity, but also a considerable amount of IgG3 antibodies, which, in contrast to the IgG1 response, was induced only after 2 or 3 vaccine doses and declined more rapidly.