Development of novel-nanobody-based lateral-flow immunochromatographic strip test for rapid detection of recombinant human interferon a2b

Recombinant human interferon a2b(rhIFNa2b)is widely used as an antiviral therapy agent for the treatment of hepatitis B and hepatitis C.The current identification test for rhIFNa2b is complex.In this study,an anti-rhIFNa2b nanobody was discovered and used for the development of a rapid lateral flow strip for the identification of rhIFNa2b.RhIFNa2b was used to immunize an alpaca,which established a phage nanobody library.After five steps of enrichment,the nanobody I22,which specifically bound rhIFNa2b,was isolated and inserted into the prokaryotic expression vector pET28a.After subsequent purification,the physicochemical properties of the nanobody were determined.A semiquantitative detection and rapid identification assay of rhIFNa2b was developed using this novel nanobody.To develop a rapid test,the nanobody I22 was coupled with a colloidal gold to produce lateral-flow test strips.The developed rhIFNa2b detection assay had a limit of detection of 1 mg/mL.The isolation of I22 and successful construction of a lateral-flow immunochromatographic test strip demonstrated the feasibility of performing ligand-binding assays on a lateral-flow test strip using recombinant protein products.The principle of this novel assay is generally applicable for the rapid testing of other commercial products,with a great potential for routine use in detecting counterfeit recombinant protein products.

Study on pharmacodynamics of recombinant interferon α-2b suppository

Objective To investigate the antiviral activity of recombinant interferonα-2b suppository(IFNα-2b)in vivo and in vitro.Methods The cytopathic-effect inhibition assay was applied in this study to investigate the antiviral activity of this drug as well as yingtelong and axiluowei as positive control.The guinea pig model of vaginitis and skin infection caused by HSV-2 infection were established,treated with IFNα-2b suppository at dosages of 60000、180000、540000 IU,using IFNα-2b injection 180000 IU·kg-1 as controls.Score the pathological changes of appearance and skin,the virus activities of vaginal secretion and tissue sections of viginae were assayed after treatment.Results The TD50 of IFN α-2b and yingtelong for Vero cells was(>100)μg·mL-1 and(>100000)IU·mL-1,respectively.The IC50 of IFN α-2b and yingtelong and axiluowei for Herpes virus type 1 was(0.29±0.08)μg·mL-1 and(185.0±28.8)IU·mL-1 and(0.19±0.03)μg·mL-1,respectively.The mean scores for vaginal and skin lesion of the treated groups were lower than those of untreated group.Among these concentrations,the IFNα-2b suppository of 540000 IU·kg-1 group.Showed highest anti-viral activity.The virus activity in vaginal secretion of treated group was lower than that of untreated group too(P<0.01 or P<0.05).Tissue sections of viginae after treatment with IFNα-2b suppository showed significantly therapeutical effects on the degrees of vaginal lesion.At the same dosage,The anti-HSV activity of IFNα-2b suppository was also compared with IFNα-2b injection,the results showed that the activity of suppository of 540000 IU·kg-1 group was similar to that of the injection.Conclusions The IFNα-2b suppository has anti-viruses function both in vivo and in vitro.

SYNERGIC CYTOTOXICITY TO GASTRIC CANCER CELLS BY COMBINED USE OF TRICHOSANTHIN ANDRECOMBINANT INTERFERON α-2B

Objective To investigate a new approach of the combined use of trichosanthin (TCS) andrecombinant interferon alpha - 2b (rIFN α- 2b) against digestive system cancer cells. Methods Detect separatelythe cytotoxicity of TCS, rIFN α- 2b and their combination against digestive system cancer cell SGC- 7901.Results In the experiment in vitro, TCS, rIFN α- 2b both had direct, dose dependent cytotoxicity againstSGC - 7901. Their combined use demonstrated a toxicity signijicantly higher than that of the two drugs used alone,showing a signilicant synergic effect. This synergic cytotoxicity was confirmed in the animal experiment.Conclusion Combined use of TCS and rIFN α - 2b decreases the therapeutic dose of TCS and its toxic adverseellect, and this synergic effect is favorable to the clinical use of TCS protein against gastric cancer.

Development of novel interferon alpha2b muteins and study the pharmacokinetic and biodistribution profiles in animal model

Novel human interferon alpha 2b (hIFNα2b) muteins were developed by substituting cysteine residue (C) at positions 2 and 99 with aspartic acid residues (D). The mutein forms were then studied for pharmacokinetic profile. In addition, the influence of charge on the protein structure was tested in vivo for the biodistribution pattern. Codon substitutions were performed by Polymerase Chain Reaction (PCR)-based site-directed mutagenesis on a previously constructed synthetic hIFNα2b open reading frame (ORF) cloned in pET32b expression plasmid. The result of nucleotide sequencing analysis confirmed that all codons were replaced successfully without any additional mutation. Three mutant forms of hIFNα2b ORF were overexpressed in Escherichia coli BL21 (DE3) resulted in three muteins: hIFNα2b C2D, hIFNα2b C99D, hIFNα2b C2D C99D. To follow the kinetic and localization of the mutein interferon after intravenous administration, Tc99m was used to label the proteins. In particular of elimination half-life, it was shown that hIFNα2b C2D C99D > hIFNα2bC2D > hIFNα2bC99D > wild type. hIFNα2b C2D C99D mutein showed highest blood accumulation after 30 minutes administration. Taken together, the charge of hIFNα2b seems to be responsible for the fate of hIFNα2b in vivo.

Effects of anti-HPV bioprotein dressing combined with interferon α-2b therapy on the malignant molecule expression in patients with CINⅢ complicated by high-risk HPV positive

Objective: To study the effects of anti-HPV bioprotein dressing combined with interferon α-2b therapy on the malignant molecule expression in patients with cervical intraepithelial neoplasia (CIN) Ⅲ complicated by high-risk HPV positive. Methods: Patients who were diagnosed with CINⅢ and high-risk HPV positive and underwent conization in the 3201 Hospital Affiliated to Xi'an Jiaotong University between June 2014 and February 2017 were selected and randomly divided into the observation group who received preoperative anti-HPV bioprotein dressing combined with interferon α-2b therapy and the control group who received no special treatment. CIN lesion was collected to determine the expression of pro-proliferation molecules, pro-apoptosis molecules and epithelial-mesenchymal transition molecules. Results: Rsf1, Piwil2, TOPK, p38MAPK, ERK, Snail, Twist, N-cadherin and Vimentin mRNA expression in cervical intraepithelial neoplasia lesions of observation group were greatly lower than those of control group whereas LRIG3, SARI, IEX-1, FHIT and E-cadherin mRNA expression were greatly higher than those of control group. Conclusion: Anti-HPV bioprotein dressing combined with interferon α-2b therapy can inhibit the proliferation and invasive growth of tumor cells in patients with CINⅢ complicated by high-risk HPV positive.

重组人干扰素α-2b联合胸腺法新治疗慢性乙型肝炎分析

目的:探讨重组人干扰素α-2b联合胸腺法新治疗慢性乙型肝炎临床疗效。方法:分析我院收治的80例慢性乙型肝炎临床资料,依据治疗方式不同分为常规治疗组40例和联合治疗组(重组人干扰素α-2b联合胸腺法新治疗组)40例。结果:联合治疗组ALT、TBIL、ALB明显优于常规治疗组,同时联合治疗组HBVDNA病毒学转换率和HBeAg血清学转换率均高于常规治疗组,P<0.05,差异均有统计学意义。结论:重组人干扰素α-2b联合胸腺法新治疗慢性乙型肝炎患者临床效果明显,预后良好。

柴芍六君子汤联合重组人干扰素α-2b治疗慢性乙型病毒性肝炎47例

目的:观察柴芍六君子汤联合重组人干扰素α-2b治疗慢性乙型病毒性肝炎(Chronic Hepatitis B,CHB)的临床疗效。方法:将90例CHB患者随机分为试验组47例和对照组43例。对照组在常规西医治疗的基础上给予重组人干扰素α-2b,试验组在对照组基础上加用柴芍六君子汤。观察两组患者的临床疗效、应答率及不良反应发生率,检测两组患者治疗前后血清丙氨酸氨基转移酶(alanine transaminase,ALT)、天冬氨酸氨基转移酶(aspartateaminotransferase,AST)水平,比较两组患者治疗后中医证候积分。结果:对照组有效率为83.72%,试验组有效率为91.48%,两组患者临床疗效比较,差异具有统计学意义(P<0.05)。试验组治疗后中医证候积分高于对照组,差异具有统计学意义(P<0.05)。两组患者治疗后血清AST、ALT水平均低于治疗前,且试验组低于对照组,差异具有统计学意义(P<0.05)。试验组完全应答率为53.19%,对照组完全应答率为46.51%,两组患者完全应答率比较,差异具有统计学意义(P<0.05)。对照组不良反应发生率低于试验组,差异具有统计学意义(P<0.05)。结论:柴芍六君子汤可提高重组人干扰素α-2b治疗CHB抗病毒疗效显著,可改善患者生存质量,减少不良反应的发生。

重组人干扰素α-2b凝胶联合微波治疗慢性宫颈炎临床体会

目的探讨重组人干扰素α-2 b凝胶联合微波治疗慢性宫颈炎的疗效。方法按照数字随机表法将我院妇科门诊收治的282例慢性宫颈炎患者分为微波治疗组(对照组)和重组人干扰素α-2 b凝胶联合微波治疗组(观察组),每组各141例,比较两组患者的临床疗效。结果观察组患者治疗总有效率为94.3%高于对照组患者的85.1%,对比两组患者在治疗总有效率,P<0.05,差异具有统计学意义。结论重组人干扰素α-2 b凝胶联合微波治疗慢性宫颈炎的疗效较为理想,是一种理想的治疗方案。

人重组干扰素-α-2b治疗子宫内膜异位症的临床分析

目的观察分析人重组干扰素-α-2b治疗子宫内膜异位症的临床疗效。方法选取2010年6月-2012年6月子宫内膜异位症的患者56例,按照随机数字法分成观察组和对照组,各28例,观察组使用人重组干扰素-α-2b治疗,对照组使用戈舍瑞林治疗,观察对比两组治疗效果。结果两组术后血清CA125、EMAb水平术后1、2、3个月与术前对比存在明显差异(P<0.05),具有统计学意义;两组治疗期间药物不良反应发生率进行比较存在一定差异(P<0.05),具有统计学意义。结论人重组干扰素-α-2b治疗子宫内膜异位症的临床疗效显著,与常规使用戈舍瑞林治疗作用类似,药物不良反应表现在使用早期较多,但持续时间短,停药后皆能自行消失,安全性较高。

注射用重组人干扰素α2b联合胸腺肽肠溶片治疗慢性乙型肝炎48例

目的探讨注射用重组人干扰素α2b联合胸腺肽肠溶片治疗慢性乙型肝炎的临床疗效。方法 96例慢性乙型肝炎患者,分成观察组和对照组。对照组给予重组人干扰素α2b,观察组给予重组人干扰素α2b联合胸腺肽肠溶片。结果观察组总有效率79.17%高于对照组60.42%,差异有统计学意义(P<0.05)。结论注射用重组人干扰素α2b联合胸腺肽肠溶片治疗慢性乙型肝炎具有较佳疗效。

Lys-2-rhTNFα单用或联合其它细胞因子抗肿瘤作用的研究

研究了 L ys- 2 - rh TNFα(1)单用或联合 1 2 5Ser- rh IL - 2 (2 )和 /或 rh INFα- 2 a(3)的体内外抗肿瘤作用。实验结果表明 1对人结肠癌细胞株 (M76 0 9)、小鼠黑色素瘤细胞株 (B16 )、小鼠白血病细胞株 (Yac- 1)均有抑制细胞增殖的作用。 1合用 2及 3较它们各自单用及任意两者合用的细胞毒作用明显增强 (P<0 .0 1)。 1与 2、3联合用药组较它们各自单独用药对小鼠结肠癌 Colon2 6细胞移植性肿瘤的抑瘤作用明显提高 (P<0 .0 5 )。

重组人干扰素α-1b联合消旋卡多曲治疗婴幼儿病毒性腹泻疗效观察

目的:分析和探讨重组人干扰素α-1b联合消旋卡多曲治疗婴幼儿病毒性腹泻临床效果.方法:选取我院2013-01/2014-01收治的110例婴幼儿病毒性腹泻患者,通过随机数字表法分为对照组(n=55)和观察组(n=55),对照组患儿采用消旋卡多曲治疗,观察组患儿采用重组人干扰素α-1b联合消旋卡多曲治疗,对两组患儿临床治疗效果作比较分析.结果:观察组患儿临床治疗有效率显著高于对照组,住院时间较对照组患儿更低(P<0.05).结论:在婴幼儿病毒性腹泻治疗中,采用消旋卡多曲与重组人干扰素α-1b联合治疗,能明显改善临床症状,缩短住院时间,减轻患儿痛苦,疗效显著,值得临床推广应用.

某院儿科注射用重组人干扰素α_(2)b超说明书用药分析

目的分析儿科住院患儿注射用重组人干扰素α_(2)b(rhIFNα_(2)b)的使用情况,为临床合理用药和儿童安全用药提供参考。方法采用回顾性调查方法,从医院信息系统调取2021年1月~6月儿科住院患儿使用注射用rhIFNα_(2)b的所有病历。用Excel软件统计患者情况、临床诊断、用药剂量、联合用药和治疗效果等信息,以相关说明书、共识为参考,并采用倾向性评分匹配法选取对照组患者使用rhIFNα_(2)b的治疗效果并进行对比分析。结果注射用rhIFNα_(2)b可以作为雾化制剂使用,有效率高、不良反应低。但需要注意儿童的年龄、感染情况以及使用方法。结论该院注射用rhIFNα_(2)b超说明书用药现象普遍,虽然大部分适应证有较为充分的证据支持,但仍需进一步规范和管理,以促进临床合理用药。

不同剂量重组人干扰素α_2b注射液雾化吸入治疗小儿病毒性肺炎临床疗效的对比研究

目的比较不同剂量重组人干扰素α2b注射液雾化吸入治疗小儿病毒性肺炎的临床疗效。方法选取2013年9月—2014年8月保定市第一中心医院儿科收治的病毒性肺炎患儿120例,其中中途退出19例,最终纳入患儿101例,采用随机数字表法分为对照组35例、低剂量组36例、高剂量组30例。3组患儿均给予常规治疗,低剂量组患儿给予低剂量重组人干扰素α2b注射液(10万U·kg^(-1)·次^(-1))雾化吸入,高剂量组患儿给予高剂量重组人干扰素α_2b注射液(20万U·kg^(-1)·次^(-1))雾化吸入;3组患儿均连续治疗1周。比较3组患儿临床疗效及咳嗽、喘憋、肺部啰音消失时间,治疗前后血清炎性因子[肿瘤坏死因子α(TNF-α)、白介素6(IL-6)]水平,并记录低剂量组、高剂量患儿治疗期间不良反应发生情况。结果低剂量组、高剂量组患儿临床疗效优于对照组(P<0.05),高剂量组患儿临床疗效优于低剂量组(P<0.05)。低剂量组、高剂量组患儿咳嗽、喘憋、肺部啰音消失时间短于对照组(P<0.05),高剂量组患儿咳嗽、喘憋及肺部啰音消失时间短于低剂量组(P<0.05)。治疗前3组患儿TNF-α、IL-6水平比较,差异无统计学意义(P>0.05);治疗后低剂量组、高剂量组患儿血清TNF-α、IL-6水平低于对照组(P<0.05),高剂量组患儿血清TNF-α、IL-6水平低于低剂量组(P<0.05)。低剂量组患儿治疗期间不良反应发生率低于高剂量组(P<0.05)。结论与低剂量重组人干扰素α_2b注射液(10万U·kg^(-1)·次^(-1))相比,高剂量重组人干扰素α_2b注射液(20万U·kg^(-1)·次^(-1))雾化吸入治疗小儿病毒性肺炎的临床疗效更佳,能更有效地降低患儿血清炎性因子水平,但高剂量重组人干扰素α_2b注射液所致不良反应风险发生升高,临床应根据患儿个体情况及耐受性等选择应用。

重组人干扰素α组人干对慢性乙肝患者进行不同疗程治疗的临床效果评价

目的探讨重组人干扰素α讨重组对慢性乙肝患者进行不同疗程治疗的临床效果。方法 2014年6月～2016年5月,选取本院收治的78例慢性乙肝患者作为研究对象,参照患者入院序号奇偶数按照1:1比例划分为对照组与观察组,每组患者39例。两组患者均行重组人干扰素α组患者治疗,对照组持续治疗6个月,观察组持续治疗12个月,观察对比两组患者治疗结束后3个月、6个月丙氨酸氨基转移酶(ALT)复常率、HBeAg阴转率、HBV-DNA阴转率和不良反应发生率。结果治疗结束后3个月观察组患者ALT复常率、HBeAg阴转率和HBVDNA阴转高于对照组,治疗结束后6个月两组患者上述指标均明显好转,且观察组高于对照组(P<0.05);治疗结束后,观察组不良反应发生率为7.68%,与对照组5.12%相比差异无统计学意义。结论针对慢性乙肝患者,重组人干扰素α对慢性效果显著,且长疗程治疗较短疗程治疗效果更为理想,安全性较高,值得进一步推广。

国产人α＿1型基因工程干扰素治疗慢性肝炎Ⅲ期临床试验

３９例慢性肝炎患者，其中乙型肝炎１２例，丙型肝炎２７例，接受人α＿１型基因工程干扰素治疗（ｒＨＩＦＮα＿１）。１０例丙型肝炎病人ｒＨＩＦＮα＿１每日肌注１×１０￣６Ｕ，连用９０天，１７例丙型肝炎病人和乙型肝炎ｒＨＩＦＮα＿１每日肌注３×１０￣６Ｕ，连用９０天。治疗后乙型肝炎组ＨＢｅＡｇ和ＨＢＶＤＮＡ的阴转率均为４１．６７％；丙型肝炎用１×１０￣６Ｕ／ｄ和３×１０￣６Ｕ／ｄ组ＨＣＶＲＮＡ阴转率分别为５０％和６４．７％，两个剂量组阴转率比较差异无显著性（Ｐ＞０．０５），治疗期间不良反应发生率较低，远期疗效尚待观察。

国产基因工程干扰素α1治疗慢性乙型肝炎的临床研究

２２５例慢性乙型肝炎分三组治疗，Ａ和Ｂ组７４例配对随机用干扰素α１及安慰剂双盲对照观察，Ｃ组１５１例为干扰素α１４０微克连用三个月，ＨＢｅＡｇ、ＨＢＶ－ＤＮＡ、ＨＢｅＡｇ和ＨＢＶ－ＤＮＡ双转阴率及抗ＨＢｅ转阳率分别为４０．５％、５７．１％、３９．３％及２９．７％，与对照组差异有非常显著性（Ｐ＜０．０１）。扩大治疗组结果与Ａ组相似，均明显优于对照组。治疗组随访半年和一年ＨＢｅＡｇ及ＨＢＶ－ＤＮＡ转阴率与治疗结束时相似，表明有较持久的效果。

重组人干扰素联合薄芝糖肽治疗高甲胎蛋白慢性乙型肝炎的临床观察

目的:观察重组人干扰素联合薄芝糖肽治疗高甲胎蛋白(AFP)慢性乙型肝炎的疗效及安全性。方法:选择我院82例高AFP慢性乙型肝炎患者,随机均分为观察组和对照组。观察组患者采用重组人干扰素α-2b注射液300万U肌肉注射、每日1次,连用4周后改为500万U、隔日1次;并以薄芝糖肽4 ml加入5%葡萄糖注射液200 ml中静脉滴注,每日1次。对照组患者仅采用重组人干扰素α-2b注射液治疗,用法用量同观察组。两组疗程均为16周。比较两组患者治疗前后丙氨酸氨基转移酶(ALT)、乙型肝炎e抗原(HBe Ag)、乙型肝炎病毒脱氧核糖核酸(HBV-DNA)拷贝数和AFP的变化,并观察不良反应发生情况。结果:治疗后,观察组患者ALT复常率和HBe Ag转阴率均显著高于对照组,差异有统计学意义(P<0.05);两组患者HBV-DNA拷贝数和AFP水平均显著低于本组治疗前,且观察组低于对照组,差异有统计学意义(P<0.05)。观察组患者不良反应发生率显著低于对照组,差异有统计学意义(P<0.05)。结论:重组人干扰素联合薄芝糖肽治疗高AFP慢性乙型肝炎较单用重组人干扰素疗效更好,可有效改善肝功能、抑制病毒增殖,且安全性较好。

重组人干扰素联合阿德福韦酯治疗慢性乙型肝炎的疗效观察

目的探讨重组干扰素联合阿德福韦酯治疗慢性乙型肝炎的临床疗效。方法将2011年12月—2012年12月72例慢性乙肝患者随机分成二组:对照组36例单纯采用重组人干扰素α-2b治疗;治疗组36例采用重组人干扰素α-2b+阿德福韦酯治疗;比较二组临床疗效、乙肝病毒脱氧核糖核酸(HBV DNA)转阴率、乙肝病毒e抗原(HBeAg)转阴率、HBeAg血清转换率和患者肝功能指标水平的变化情况。结果治疗组的总有效率明显高于对照组(P<0.05);治疗组治疗后的HBV DNA转阴率、HBeAg转阴率、HBeAg血清转换率均显著高于对照组(P<0.05);治疗组患者治疗后的丙氨酸转氨酶(ALT)、冬氨酸转氨酶(AST),与对照组差异无统计学意义。结论重组人干扰素联合阿德福韦酯治疗慢性乙型肝炎,提高乙肝病毒标志物转阴率,改善肝功能水平。