重组α干扰素-2b与异丙托溴铵治疗小儿毛细支气管炎的疗效对比研究

目的探讨重组α干扰素-2b和异丙托溴铵雾化吸入在小儿毛细支气管炎治疗中的临床效果。方法选取2020年4月至2022年8月湖州市中心医院诊治的112例毛细支气管炎患儿,根据信封法将其分为对照组和观察组,每组各56例。对照组患儿给予异丙托溴铵雾化吸入,观察组患儿给予重组α干扰素-2b雾化吸入,两组患儿均连续治疗7d。比较两组患儿的肺功能、γ干扰素(interferon-γ,IFN-γ)、白细胞介素-10(interleukin-10,IL-10)、免疫功能及不良反应。结果干预7d后,观察组患儿的喘息、咳嗽、肺部湿啰音及痰鸣音消失时间均显著短于对照组(P<0.05);两组患儿的潮气量(tidal volume,TV)、潮气呼气峰流量(peak tidal expiratory flow,PTEF)及达峰时间比(ratio of time to peak tidal expiratory flow to total expiratory time,TPTEF/TE)、IFN-γ及IL-10均显著高于本组干预前(P<0.05),观察组患儿的TV、PTEF、TPTEF/TE、IFN-γ及IL-10水平均显著高于对照组(P<0.05);两组患儿的CD3^(+)、CD4^(+)及CD4^(+)/CD8^(+)均显著高于本组干预前,CD8^(+)显著低于本组干预前(P<0.05);观察组患儿的CD3^(+)、CD4^(+)及CD4^(+)/CD8^(+)均显著高于对照组,CD8^(+)显著低于对照组(P<0.05);两组患儿的不良反应发生率比较差异无统计学意义(χ^(2)=0.704,P=0.401)。结论毛细支气管炎患儿采用重组α干扰素-2b和异丙托溴铵雾化吸入均可获得良好的治疗效果,但前者有助于缩短症状消失时间,提高患儿免疫水平,值得临床推广应用。

α干扰素-2bHis^58的分子构建、表达及纯化

目的通过定点突变,构建α干扰素-2bHis58(IFNα-2bHis58),以期获得高效的新型药物分子。方法采用PCR体外定点突变技术,使α干扰素-2b基因的第58位密码子由GAA突变为CAC。将扩增片段克隆入pET23b表达载体,重组质粒转化大肠杆菌BL21(DE3)。表达的IFNα-2bHis58经包涵体变复性、DEAE层析和CM层析纯化后,用WISH-VSV系统进行生物活性测定。结果IFNα-2bHis58主要以包涵体形式表达,表达量占菌体总蛋白的30%以上。纯化后,IFNα-2bHis58的纯度>95%,比活性>3.4×108IU/mg。结论构建了IFNα-2bHis58的表达载体,并成功在大肠杆菌中表达,获得了高活性突变分子IFNα-2bHis58,建立了IFNα-2bHis58的纯化工艺。

长效与短效α-干扰素治疗慢性丙型肝炎疗效观察

目的：对比长效干扰素联合利巴韦林与普通干扰素联合利巴韦林治疗慢性丙型肝炎的临床疗效及安全性。方法回顾性分析60例慢性丙型肝炎患者的临床资料，其中长效干扰素组采用聚乙二醇干扰素α-2a （PEG-INFα-2a）联合利巴韦林进行抗病毒治疗（n=30例），普通干扰素组采用普通IFNα-2a联合利巴韦林治疗（n=30例），均分别于治疗4、12、24、48w及治疗结束后24w评价疗效，并观察药物的不良反应。结果长效干扰素组持续病毒学应答率（SVR）为90.0%，显著高于普通干扰素组的56.7%（P〈0.01）；两组药物不良反应均有发热、肌肉酸痛、脱发及中性粒细胞、血小板下降等，但长效干扰素组发生发热和肌肉酸痛比率分别为36.7%和33.3%，显著低于普通干扰素组（90.0%和93.3%，P〈0.01）；长效干扰素组中性粒细胞下降明显高于普通干扰素组，在4～24w尤其明显。结论长效干扰素联合利巴韦林治疗方案疗效显著优于普通干扰素联合利巴韦林，且未增加不良反应。

Pegylated IFN-α2b added to ongoing lamivudine therapy in patients with lamivudine-resistant chronic hepatitis B

AIM： To investigate the role of pegylated-interferon （IFN）α-2b in the management of patients with lamivudineresistant chronic hepatitis B. METHODS： Twenty consecutive anti-HBe positive patients were treated with pegylated IFN α-2b （100 IJg sc once weekly） for 12 mo. There was no interruption in lamivudine therapy. Hematology, liver biochemistry, serum HBV DNA levels were detected by PCR, and vital signs were also assessed. Liver histology was assessed in some patients at entry and at wk 52 for comparison. RESULTS： Nine patients （45%） had a partial virological end-treatment response; seven patients （35%） showed complete virological end-treatment response. Eight patients （40%） showed biochemical end-treatment response. There was a trend for higher virological response rates in patients who had previously responded to IFN and relapsed compared to IFN non-responders （four out of seven patients vs none out of six patients, respectively; P=0.1）. Patients without virological endtreatment response showed significant worsening of fibrosis [median score 2 （range, 1 to 3） vs median score 3 （range, 1 to 4）], in the first and second biopsy respectively （P=0.014）, whereas necroinflammatory activity was not significantly affected. Patients with complete or partial virological end-treatment response did not show any significant changes in histological findings, possibly due to the small number of patients with paired biopsies （n = 5）. Nevertheless, after 12 mo of follow-up, only onepatient （5%） showed sustained virological response and only 2 patients （10%） showed sustained biochemical response. Two patients （10%） discontinued pegylated ]FN both after 6 mo of treatment due to flu-like symptoms. CONCLUSION： Pegylated IFNα-2b, when added to ongoing lamivudine therapy in patients with lamivudineresistant chronic hepatitis B, induces sustained responses only in a small minority of cases.

Psychiatric side effects of pegylated interferon alfa-2b as compared to conventional interferon alfa-2b in patients with chronic hepatitis C

AIM: To assess systematically the spectrum and extent of depressive symptoms comparing patient groups receiving peginterferon or conventional interferon.METHODS: Ninety-eight patients with chronic hepatitis C and interferon-based therapy (+ribavirin) were consecutively enrolled in a longitudinal study. Patients were treated with conventional interferon alfa-2b (48/98patients; 5 MIU interferon alfa-2b thrice weekly) or peginterferon alfa-2b (50/98 patients; 80-150 μgpeginterferon alfa-2b) in combination with weight-adapted ribavirin (800-1 200 mg/d). Repeated psychometric testing was performed before, three times during and once after antiviral therapy: Depression was evaluated by the Hospital Anxiety and Depression Scale (HADS), anger/hostility by the Symptom Checklist-90 Items Revised (SCL-90-R).RESULTS: Therapy with pegylated interferon alfa-2bproduces comparable scores for depression (ANOVA:P = 0.875) as compared to conventional interferon.Maximums of depression scores were even higher and cases of clinically relevant depression were frequent during therapy with peginterferon. Scores for anger/hostility were comparable for both therapy subgroups.CONCLUSION: Our findings suggest that the extent and frequency of depressive symptoms in total are not reduced by peginterferon. Monitoring and management of neuropsychiatric toxicity especially depression have to be considered as much as in antiviral therapy with unmodified interferon.

Risk factors for retinopathy associated with interferonα-2b and ribavirin combination therapy in patients with chronic hepatitis C

AIM： To elucidate the frequency and risk factors for retinopathy in patients with chronic hepatitis C who are treated by interferon-ribavirin combination therapy. METHODS： We prospectively analyzed 73 patients with histologically confirmed chronic hepatitis C, who underwent combination therapy for 24 wk. Optic fundi were examined before, and 2, 4, 12 and 24 wk alter the start of combination therapy. RESULTS： Fourteen patients （19%） developed retinopathy, which was initially diagnosed by the appearance of a cotton wool spot in 12 patients. Retinal hemorrhage was observed in 5 patients. No patient complained of visual disturbance. Retinopathy disappeared in 9 patients （64%） despite the continuation of combination therapy. However, retinopathy persisted in 5 patients with retinal hemorrhage. A comparison of the clinical background between the groups with and without retinopathy showed no significant differences in age, gender, viral genotype, RNA level, white blood cell count, platelet count, prothrombin time, complications by diabetes mellitus or hypertension, or pretreatment arteriosclerotic changes in the optic fundi. However, multiple logistic regression analysis revealed that complication by hypertension was observed with a high frequency in the group with retinopathy （P = 0.004, OR = 245.918, 95% CI = 5.6-10786.2）.CONCLUSION： Retinopathy associated with combination therapy of interferon α-2b and ribavirin tends to develop in patients with hypertension.

Primary Purification of Co-expressed Soluble and Insoluble Alpha-interferon 2b from Recombinant E. coli

Alpha-interferon 2b (IFN 2b) was produced both in soluble and insoluble forms from recombinant E. coli. The dissolution of the expressed IFN 2b in inclusion body was carried out and it was found that the optimal condition to dissolve the expressed protein was 7 mol稬1guanidinium salt solution at pH 3.0. The resultant solution was diluted 20 times using pH 6.0 buffer to refold the protein correctly. The cation exchange column was employed to purify both refolded and soluble IFN 2b. For soluble IFN sample, high IFN 2b recovery yield (92.1%) with 91.7% purity was obtained in the eluate. However, for refolded IFN sample, only 72.7% of IFN 2b was recovered with relatively low purity (56.8%) by cation exchange chromatography. Although the expression level of insoluble IFN was higher than that of co-expressed soluble IFN in this recombinant E. coli cells, the productivity of bioactive IFN 2b was higher with soluble expressed IFN after primary purification process. Soluble expression of foreign proteins in recombinant bacteria might be an alternative strategy for efficient production of heterogeneous pro-teins due to high bioactivity and simple downstream protein purification process.

An extremely rare case of delusional parasitosis in a chronic hepatitis C patient during pegylated interferon alpha-2b and ribavirin treatment

During treatment of chronic hepatitis C patients with interferon and ribavirin, a lot of side effects are described. Twenty-three percent to 44% of patients develop depression. A minority of patients evolve to psychosis. To the best of our knowledge, no cases of psychogenic parasitosis occurring during interferon therapy have been described in the literature. We present a 49-year-old woman who developed a delusional parasitosis during treatment with pegylated interferon alpha-2b weekly and ribavirin. She complained of seeing parasites and the larvae of fleas in her stools. This could not be confirmed by any technical examination. All the complaints disappeared after stopping pegylated interferon alpha-2b and reappeared after restarting it. She had a complete sustained viral response.

大剂量干扰素辅助治疗国人口腔粘膜恶性黑色素瘤的临床研究

目的：探讨大剂量干扰素辅助治疗口腔粘膜恶性黑色素瘤的疗效和安全性。方法回顾性分析2004年5月至2012年11月我科经治的Ⅲ～ⅣA期口腔粘膜恶性黑色素瘤患者的治疗及预后情况。在经治的117例患者中，73例术后接受了大剂量干扰素α-2b治疗，其中有58例完成了治疗计划，设为治疗组，另15例中途停止；其余44例未采用大剂量干扰素治疗者作为对照组。比较两组患者的总生存期（ OS）和无复发生存期（ RFS）以及相关不良反应的发生情况。结果117例患者的中位OS为40个月（95％CI：33～62个月）。治疗组与对照组中Ⅲ期患者的中位OS（69个月vs．65个月）和RFS（50个月vs．34个月）的差异均无统计学意义（P＞0.05），治疗组与对照组中ⅣA期患者的中位OS（37个月 vs．20个月）和中位RFS（33个月 vs．10个月）的差异均有统计学意义（ P＜0.05），治疗组与对照组中Ⅳ期伴颈淋巴结转移患者的中位OS（40个月vs．20个月）和中位RFS（33个月 vs．10个月）的差异均有统计学意义（P＜0.05）。大剂量干扰素治疗的常见血液学毒性为骨髓抑制，包括白细胞减少和血小板减少；非血液性不良反应包括流感样症状、恶心呕吐、肝功能损害等。全组患者不良反应以1～2级为主，仅有7例出现3～4级毒副反应，给予对症治疗后均能缓解，无治疗相关性死亡。结论术后大剂量干扰素辅助治疗能显著提高ⅣA期口腔粘膜恶性黑色素瘤患者的OS和RFS，且治疗的不良反应可耐受。

67例慢性丙型肝炎治疗过程及随访中血清ALT、病毒载量及肝纤维化指标的变化

目的 观察聚乙二醇干扰素α-2b联合利巴韦林治疗慢性丙型肝炎过程中肝功能、病毒复制及肝纤维化指标的改变情况.方法 检测67例慢性丙型肝炎患者在干扰素联合利巴韦林治疗开始（0周）、结束（48周）和停药12周（60周）时血清丙氨酸转氨酶（ALT）、丙肝病毒核糖核酸（HCV RNA）、透明质酸（HA）、Ⅲ型前胶原肽（PCⅢ）、Ⅳ型胶原（Ⅳ-C）和层粘连蛋白（LN）水平.结果 治疗后完全应答组（CR-S,43/67） ALT、HCV RNA及血清4项纤维化指标均显著下降（P ＜0.05或P＜0.01）,部分应答组（CR-R,13/67）和无应答组（NR,11/67） ALT、HCV RNA及血清4项纤维化指标变化不明显,反跳甚至更高.结论 聚乙二醇干扰素α-2b联合利巴韦林治疗慢性丙型肝炎约65％患者完全应答,随着肝细胞炎症的改善,病毒RNA滴度、肝纤维化指标水平明显下降,表明干扰素联合利巴韦林能抑制HCV RNA复制,调节机体免疫功能,减轻肝脏炎症反应,改善肝功能,减少肝纤维化.