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.

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.

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.

Pegylated interferon α-2b up-regulates specific CD8+ T cells in patients with chronic hepatitis B

AIM:To investigate the effect of pegylated interferon (IFN) α-2b on specific CD8+ T lymphocytes in patients with chronic hepatitis B (CHB). METHODS:Twenty-one patients with CHB were treated with pegylated IFN α-2b. Periphery blood mononuclear cells were isolated from fresh heparinized blood by Ficoll-Hypaque density gradient centrifugation (density:1.077 g/L,Pharmingen) at weeks 0,4,8,12,and 24,respectively. Frequency of circulating hepatitis B virus (HBV) epitope-specific CD8 T cells was detected by flow cytometry. Cytokines were detected by cytometric bead assay. RESULTS:The frequency of circulating HBV core or env-specific CD8 T cells was higher (P < 0.05),the number of HBV core specific CD8 T cells was greater at week 24 (P < 0.05),the level of Th1-type cytokines [interleukin (IL)-12,tumor necrosis factor-α,and IFN-γ] was higher,while that of Th2-type cytokines (IL-4,IL-6,and IL-10) was lower in responders than in nonresponders (P < 0.05) after pegylated IFN α-2b treatment. The IL-6 level was correlated with HBV DNA (r = 0.597,P = 0.04),while the inducible protein-10 (IP-10) level was correlated with serum alanine aminotransferase (ALT) (r = 0.545,P = 0.005). The IP-10 level at week 8 after pegylated IFN α-2b treatment could predict the normalization of ALT in CHB patients (positive predict value = 56%,negative predict value = 92%). CONCLUSION:Pegylated IFN α-2b can enhance the immune response of CHB patients by increasing the frequency of HBV specific CD8+ T cells and regulating the Th1/Th2 cytokines.

Effects of interferon α-2b on liver function, complement level and oxidative stress in patients with hepatitis B

Objective:To observe the effect of interferonα-2b treatment on liver function,liver fibrosis,complement protein and oxidative stress in patients with hepatitis B.Methods:A total of 100 patients with hepatitis B in our hospital were randomly divided into the control group and the observation group,with 50 cases in each group.After admission,patients in the control group were treated with entecavir,while patients in the observation group were treated with interferonα-2b combined with entecavir.The levels of serum total bilirubin(TBil),aspartate aminotransferase(AST),alanine aminotransferase(ALT),type III procollagen(PCIII),type IV collagen(CIV),complement C3 protein(C3),complement C4 protein(C4),malondialdehyde(MDA),superoxide dismutase(SOD)and nitric oxide(NO)were compared between the two groups before and after treatment.Results:After treatment,the levels of TBil,AST,ALT,PCIII,CIV,MDA and NO in serum of patients with hepatitis B in both groups were significantly lower than those before treatment,and the levels of C3,C4 and SOD were significantly higher than those before treatment(P<0.05).After treatment,the levels of TBil,AST,ALT,PC III,C IV,MDA and NO in serum of patients in the observation group were significantly lower than those in the control group,while the levels of C3,C4 and SOD in serum of patients in the observation group were significantly higher than those in the control group(P<0.05).Conclusions:The combination of interferonα-2b and entecavir has a good curative effect on hepatitis B.It can significantly improve the liver function and immune function of patients,delay the process of liver fibrosis and reduce oxidative stress injury.It is worthy of clinical promotion.

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.

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。

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。

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。

Study of recombinant human IFN-α-2b bacilli Calmette–Guerin activated killer cells and against bladder cancer cell in vitro

Presently,one of the most potent immunothera-pies is the application of bacillus Calmette Guerin(BCG)to prevent recurrences of the superficial bladder cancer.Despite its successful use,nonresponders and certain side effects remain a major obstacle.Therefore,current studies aim at developing recombinant BCG(rBCG)strains secreting Th1-like cytokines to improve the effectiveness of the therapy.In this study,a new type of rBCG strain constructed by Tianjin institute of Urology was tested for its immunostimulatory capacity in vitro.Peripheral blood monocytes(PBMC)were stimulated by recombinant BCG and transformed into bacilli Calmette–Guerin activated killer(BAK)cells,and the effect of anticancer BAK cells was studied.Recombinant IFN-a-2b-BCG,wild-type BCG(wBCG),wild-type BCG and IFN-a-2b were coincubated with PBMCs respectively in vitro,and the proliferation of PBMC was detected with MTT in different time.BAK cells have the ability to kill bladder tumor cells,and the antitumor activity of effecter cells was determined by LDH release assay.The result of MTT showed that the proli-feration of PBMC in the recombinant BCG group was more powerful than in the other two groups(P<0.05).The result of LDH release assay showed that the antitumor activity of BAK cells stimulated by Recombinant BCG was the highest in all groups.We conclude that the recombinant BCG can activate more PBMCs to anti-bladder cancer in vitro than wild-type BCG does.

简易方法合成具有类过氧化物酶活性的聚乙烯亚胺包覆的四氧化三铁用于高灵敏检测干扰素α-2b

在病毒感染和癌症治疗中,干扰素α-2b(IFN-α2b)的灵敏检测至关重要,因此需要开发经济、稳定的灵敏检测IFN-α2b的方法.传统的酶联免疫吸附测定(ELISA)中使用的天然酶存在制备成本高和稳定性差等问题.为了提高其灵敏度并降低成本,我们合成了聚乙烯亚胺(PEI)修饰的四氧化三铁磁性纳米粒子(Fe_(3)O_(4)@PEI MNPs).在基于ELISA的IFN-α2b检测中,这些磁性纳米粒子作为辣根过氧化物酶的替代品,提供了比色谱和传统ELISA技术更高的灵敏度,并且能够实现IFN-α2b的可视化检测.该免疫分析方法的线性范围为0.075-25 ng mL^(-1),检测限为0.055 ng mL^(-1).基于Fe_(3)O_(4)@PEI MNPs优异的过氧化物酶活性,该方法在用于检测IFN-α2b和其他蛋白质生物标志物监测方面具有临床应用潜力.

α-2-HS-glycoprotein is a potential marker predicting hepatitis B e antigen seroconversion in patients with chronic hepatitis B during treatment with pegylated interferon alfa-2b

The efficacy of interferon (IFN) is limited in about 1/3 of patients with chronic hepatitis B (CHB). We used two-dimensional electrophoresis (2-DE)-based proteomic strategies to identify potential serum markers predicting hepatitis B e antigen (HBeAg) seroconversion in these patients during IFN therapy. Two groups of patients were enrolled: training and validation. In the training group, 2-DE experiments and subsequent identification of altered levels of proteins showed that α-2-HS-glycoprotein, leucine-rich α-2-glycoprotein, and haptoglobin were significantly upregulated as compared with baseline levels in the HBeAg seroconversion group, whereas apolipoprotein C-III precursor, leucine-rich α-2-glycoprotein, and α-albumin were downregulated in the non-seroconversion group. For patients with HBeAg seroconversion in the training group, Western blot analyses showed that α-2-HS-glycoprotein levels in 75% of patients were significantly upregulated at the end of the treatment as compared with baseline levels. Subsequent experiments in the validation group showed that α-2-HS-glycoprotein levels were significantly increased at week 4 in 83.33% of patients in the HBeAg seroconversion group. Dynamic changes in the serum level of α-2-HS-glycoprotein may be a potential early marker for predicting HBeAg seroconversion during IFN treatment for CHB.

Frequency ofT-cell FoxP3＋ Treg and CD4＋/CD8＋ PD-1 expression is related to HBeAg seroconversion in hepatitis B patients on pegylated interferon

Background Host immune responses against hepatitis B virus （HBV） induced by antiviral therapy play a crucial role in viral clearance. To further investigate the immune mechanisms underlying the differences between respondents and non-respondents, we analyzed myeloid dendritic cells （mDCs）, plasmacytoid dendritic cells （pDCs）, FoxP3＋ regulatory T cells （FoxP3＋ Treg） and programmed death 1 （PD-1） expression in CD4＋/CD8＋ T cells in chronic hepatitis B patients undergoing pegylated interferon （PeglFN）α-2b treatment. Methods Patients received PeglFNα-2b for 24 or 48 weeks, with follow-up at 24 weeks. The frequencies of mDCs, pDCs, FoxP3＋ Treg, and PD-1 expression by CD4＋/CD8＋ T cells were evaluated by flow cytometry at baseline, weeks 4 and 12, end of treatment, and follow-up （12/24 weeks）. Results In HBeAg seroconverters （respondents）, the mDC relative frequency decreased at week 4 and then rebounded at week 12. The pDC relative frequency decreased consistently. In non-HBeAg seroconverters （non-respondents）, both mDC and pDC frequencies decreased slightly. The FoxP3＋ Treg relative frequency decreased during treatment and remained low during follow-up in respondents, while in non-respondents it decreased slightly during therapy but rebounded after discontinuation. In patients with HBeAg 〈17.55 PEI-U/ml at week 12 and 〈8.52 PEI-U/ml at week 24, the FoxP3＋ Treg frequency decreased during treatment and at follow-up. In respondents, CD4~PD-1 and CD8＋PD-1 levels decreased at week 4 and remained low at week 12. In non-respondents, PD-1 expression decreased at week 4 but rebounded at week 12. Conclusions The results indicate that the dynamic changes in DCs, FoxP3＋ Treg frequency, and PD-1 expression by CD4＋ and CD8＋ T cells exhibit different trends in HBeAg and non-HBeAg seroconversion patients. During PeglFNa-2b treatment of chronic hepatitis B patients, these changes may be of predictive value for HBeAg seroconversion. HBsAg and HBeAg levels are related to FoxP3＋ Treg frequency.

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。

重组干扰素α治疗重症急性胰腺炎效果观察

目的探讨重组干扰素α对重症急性胰腺炎(SAP)的治疗作用及机制。方法将84例SAP患者随机分为临床干预组与对照组,对照组予以胃肠减压、预防感染、纠正水电解质平衡紊乱等常规治疗,干预组在常规治疗措施基础上肌注重组干扰素α40μg/d,7～14 d。观察治愈天数、发生并发症例数和病死例数,采用急性生理学及慢性健康状况评分-Ⅱ评分和CT评分评价病情。酶联免疫吸附法检测患者血清中肿瘤坏死因子-α(TNF-α)、白细胞介素-8(IL-8)、白细胞介素-2(IL-2)、白细胞介素-10(IL-10)和核因子кB浓度的变化。结果干预组较对照组治愈天数、发生并发症例数和病死例数有所降低,急性生理学及慢性健康状况评分-Ⅱ评分和CT评分显著降低。干预组TNF-α、IL-8和核因子кB较干预前下降,IL-2升高,IL-10升高后又回降。结论重组干扰素α在治疗SAP过程中,可减轻炎性反应、减少严重并发症发生、促进治愈。