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.

舒肝益脾颗粒联合注射用重组人干扰素α2b治疗慢性乙型肝炎患者的效果

目的:观察舒肝益脾颗粒联合注射用重组人干扰素α2b治疗慢性乙型肝炎患者的效果。方法:选取2020年6月至2023年6月该院收治的86例慢性乙型肝炎患者进行前瞻性研究,按照随机数字表法分为研究组与对照组各43例。对照组采用注射用重组人干扰素α2b治疗,研究组在对照组基础上联合舒肝益脾颗粒治疗。比较两组治疗总有效率、乙肝病毒脱氧核糖核酸(HBV-DNA)转阴率、肝功能指标[血清丙氨酸氨基转移酶(ALT)、总胆红素、天门冬氨酸氨基转移酶(AST)和白蛋白]水平、肝纤维化指标[血清Ⅳ型胶原(C-IV)、层粘连蛋白(LN)和透明质酸]水平、炎性指标[血清C反应蛋白(CRP)、降钙素原]水平、T细胞亚群指标(CD3^(+)、CD4^(+))水平和不良反应发生率。结果:研究组治疗总有效率为95.35%,高于对照组的79.07%,差异有统计学意义(P<0.05);研究组HBV-DNA转阴率为93.02%,明显高于对照组的74.42%,差异有统计学意义(P<0.05);治疗后,研究组血清白蛋白和CD3^(+)、CD4^(+)水平高于对照组,血清ALT、总胆红素、AST、C-Ⅳ、LN、透明质酸、CRP和降钙素原水平低于对照组,差异均有统计学意义(P<0.05);两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论:舒肝益脾颗粒联合注射用重组人干扰素α2b治疗慢性乙型肝炎患者可提高治疗总有效率、HBV-DNA转阴率及CD3^(+)、CD4^(+)水平,改善肝功能指标水平,降低肝纤维化指标和炎性指标水平,效果优于单纯注射用重组人干扰素α2b治疗。

流感患儿经重组人干扰素α2b雾化吸入联合帕拉米韦注射液治疗的效果分析

目的 分析流感患儿经重组人干扰素α2b雾化吸入联合帕拉米韦注射液的治疗效果。方法 回顾性分析2020年8月至2023年8月驻马店广济心血管病医院88例流感患儿,按不同治疗方案分为对照组(接受帕拉米韦注射液治疗)、研究组(接受重组人干扰素α2b雾化吸入联合帕拉米韦注射液),各44例。比较两组临床疗效,症状改善及住院时间,不良反应发生率及治疗前后血清炎性因子[肿瘤坏死因子-α(TNF-α)、单核细胞趋化蛋白-1(MCP-1)、C反应蛋白(CRP)、白细胞介素-6(IL-6)],免疫球蛋白[免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、免疫球蛋白G(IgG)]。结果 研究组临床总有效率95.45%高于对照组81.82%(P<0.05);研究组咳嗽、咽痛、高热等症状改善时间及住院时间短于对照组(P<0.05);研究组TNF-α、CRP、IL-6、MCP-1较对照组低(P<0.05);与治疗前比较,两组IgA、IgM、IgG均升高,且研究组更高(P<0.05);研究组不良反应发生率(9.09%)与对照组(6.82%)比较,差异无统计学意义(P>0.05)。结论 流感患儿采用重组人干扰素α2b雾化吸入联合帕拉米韦注射液治疗可减轻机体炎症反应,改善免疫功能,加速病情恢复,效果显著,且安全性高。

重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴HR-HPV感染患者的临床效果及对HPV转阴率的影响

目的分析重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴高危型人乳头瘤病毒(HR-HPV)感染的临床效果。方法选取宫颈低级别上皮内病变伴HR-HPV感染患者80例,采用随机数表法分为对照组及研究组,每组40例。对照组采用重组干扰素α-2b治疗,研究组采用重组干扰素α-2b联合CO_(2)激光气化治疗。比较两组患者的治疗效果。结果研究组治疗总有效率为90.00%,高于对照组的72.50%(P<0.05)。研究组治疗3、6个月的HPV转阴率均高于对照组(P<0.05)。两组不良反应总发生率比较,差异无统计学意义(P>0.05)。结论采用重组干扰素α-2b联合CO_(2)激光气化治疗宫颈低级别上皮内病变伴HR-HPV感染患者,可提高疗效及HPV转阴率。

Expression of Human Interferon α 2b Gene in Ginseng Cells

The human interferon α 2b（hIFN-α2b） gene was cloned into binary vector pBI121 to obtain plant expression vector pBIFN. The recombinant plasmid pBIFN was transferred into Agrobacterium tumefaciens strain LBA4404. Then the hIFN-α2b gene was introduced into ginseng callus cells via Agrobacterium-mediated transformation and the ginseng cell line carrying hIFN-α2b gene was selected on G418-containing medium. The presence of target gene in transformed cells was confirmed by PCR and RT-PCR. The results indicate that hIFN-α2b gene has been integrated into the ginseng cells＇ genome, with transcription products, hIFN-α2b expressed by the transgenic ginseng cells was detected by Western blot. It was shown that a specific protein band at 19000 could be observed. Cytopathic effect（CPE） inhibition assay using the W1SH-VSV system shows that the mean antiviral activity of expressed hlFN-a2α was 6.0× 10^4 IU/mL.

重组人干扰素α-2b注射液联合CO_(2)激光治疗尖锐湿疣的临床效果研究

目的 探讨重组人干扰素α-2b注射液联合二氧化碳(CO_(2))激光治疗尖锐湿疣(CA)的临床效果。方法 选择82例首次发病的CA患者,根据随机数字表法分为CO_(2)激光组和联合组,每组41例。CO_(2)激光组采用CO_(2)激光治疗,联合组在CO_(2)激光组基础上给予重组人干扰素α-2b注射液治疗。比较两组治疗前后免疫功能指标(CD3^(+)、CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+))及炎性因子血清[肿瘤坏死因子-α(TNF-α)、白细胞介素-1(IL-1)、白细胞介素-6(IL-6)]水平、治疗效果、治疗安全性及复发情况。结果 治疗后,联合组CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)水平分别为(71.5±9.7)%、(39.8±6.5)%、(2.0±0.8),均明显高于CO_(2)激光组的(59.6±8.3)%、(30.6±5.2)%、(1.7±0.4),CD8^(+)水平(21.1±2.7)%明显低于CO_(2)激光组的(23.6±2.5)%,差异有统计学意义(P<0.05)。治疗后,联合组TNF-α、IL-1、IL-6水平分别为(0.7±0.1)、(80.5±13.4)、(63.9±11.7)pg/ml,均明显低于CO_(2)激光组的(0.9±0.1)、(122.7±13.6)、(93.2±11.5)pg/ml,差异有统计学意义(P<0.05)。联合组总有效率95.1%高于CO_(2)激光组的68.3%,差异有统计学意义(P<0.05)。联合组复发率4.8%明显低于CO_(2)激光组的35.7%,差异有统计学意义(P<0.05)。两组不良反应发生率比较,差异无统计学意义(P>0.05)。结论 重组人干扰素α-2b注射液联合CO_(2)激光治疗能明显改善CA患者的免疫功能,减轻机体的炎症反应,提高临床疗效,降低复发风险,值得临床推广借鉴。

精蛋白重组人胰岛素混合注射液40/60治疗老年2型糖尿病的疗效及对患者预后的影响

目的:探讨精蛋白重组人胰岛素混合注射液40/60(简称甘舒霖40/60)治疗老年2型糖尿病(T2DM)的疗效及对患者预后的影响。方法:采用简单随机分组法将西部战区总医院2022年12月至2023年12月收治的106例T2DM患者分为对照组(n=53)和治疗组(n=53)。对照组予常规二联降糖方案治疗(二甲双胍+达格列净),治疗组予甘舒霖40/60皮下注射治疗。12周后评价疗效及安全性,测定两组治疗前、后血糖指标、体重指数(BMI)、胰岛β细胞功能指标及血清单核细胞趋化因子(MCP-1)、超敏C反应蛋白(hs-CRP)、白细胞介素6(IL-6)水平差异,并统计随访6个月预后。结果:治疗组治疗总有效率为90.57%,较对照组的71.70%更高(P<0.05)。治疗后,与对照组相比,治疗组空腹血糖(FPG)、餐后2h血糖(2hPG)、糖化血红蛋白(HbAlc)、胰岛素抵抗指数(HOMA-IR)及血清MCP-1、hs-CRP、IL-6水平更低,胰岛β细胞功能指数(HOMA-β)更高(P<0.05)。治疗组(3.77%)、对照组(7.55%)总不良反应发生率比较差异无统计学意义(P>0.05);两组随访6个月微血管并发症发生率差异无统计学意义(P>0.05)。结论:采用甘舒霖40/60皮下注射治疗老年T2DM安全有效,不仅能降低患者血糖水平,促进胰岛β细胞功能恢复,减轻炎症反应,但并未改善患者短期预后。

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.

不同剂量重组人干扰素α2b雾化吸入治疗儿童病毒性肺炎的疗效及其对血清炎性因子和免疫功能的影响

目的:探讨雾化吸入不同剂量重组人干扰素α2b注射液对病毒性肺炎患儿血清炎性因子水平及免疫功能的影响。方法:选择2015年1月至2017年6月我院收治的病毒性肺炎患儿140例,按照给药剂量分为高剂量组(n=46)、低剂量组(n=48)和对照组(n=46)。三组患儿均接受病毒性肺炎常规治疗,高剂量组给予重组人干扰素α2b注射液每次20万U/kg雾化吸入治疗,低剂量组给予重组人干扰素α2b注射液每次10万U/kg雾化吸入治疗,对照组给予0.9%生理盐水2 m L雾化吸入治疗。观察三组患儿临床疗效、不良反应以及治疗前、治疗3周后血清炎症因子水平和免疫功能的变化。结果:高剂量组和低剂量组总有效率高于对照组(P<0.05),高剂量组和低剂量组总有效率比较差异无统计学意义(P>0.05)。三组患儿不良反应发生率比较差异无统计学意义(P>0.05)。治疗3周后三组患儿血清白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、C反应蛋白(CRP)水平均较治疗前降低(P<0.05),高剂量组IL-6、TNF-α、CRP水平低于低剂量组和对照组(P<0.05),低剂量组IL-6、TNF-α、CRP水平低于对照组(P<0.05)。治疗3周后高剂量组和低剂量组血清CD3^+、CD4^+、CD4^+/CD8^+较治疗前升高,CD8^+较治疗前降低(P<0.05);对照组CD3^+、CD4^+/CD8^+较治疗前显著升高,CD8^+较治疗前显著降低(P<0.05),CD4^+比较差异无统计学意义(P>0.05)。高剂量组CD3^+、CD4^+、CD4^+/CD8^+高于低剂量组和对照组,CD8^+低于低剂量组和对照组(P<0.05),低剂量组CD3^+、CD4^+、CD4^+/CD8^+高于对照组,CD8^+低于对照组(P<0.05)。结论:雾化吸入重组人干扰素α2b注射液可以降低病毒性肺炎患儿血清炎性因子水平,增强患儿细胞免疫功能,每次20万U/kg雾化吸入的效果优于10万U/kg,值得临床推广应用。

雾化吸入干扰素α2b注射液治疗小儿呼吸道合胞病毒肺炎的疗效

目的观察雾化吸入重组人干扰素α2b注射液治疗小儿呼吸道合胞病毒肺炎的疗效。方法选择2013年9月至2014年8月保定市第一中心医院收治的60例呼吸道合胞病毒肺炎患儿,按照随机数字表法随机分为对照组和观察组,每组各30例。对照组患儿给予常规治疗,观察组患儿在常规治疗的基础上,雾化吸入干扰素α2b注射液10万IU/kg,1次/天。观察治疗1周后两组患儿咳嗽、喘憋、肺部啰音等症状消失的时间,血清肿瘤坏死因子-α(TNF-α)、白介素-10(IL-10)、白介素-18(IL-18)水平的变化以及不良反应发生例数。结果治疗1周,观察组患儿咳嗽、喘憋及肺部啰音消失时间均短于对照组[(4.11±0.89)d vs(5.19±1.27)、(4.01±1.17)d vs(5.22±1.35)d、(4.35±1.09)d vs(5.67±1.32)];观察组血清TNF-α、IL-18水平低于对照组[(43.33±5.65)ng/L vs(58.27±6.21)ng/L、(176.39±88.31)ng/L vs(248.25±91.78)ng/L];IL-10水平高于对照组[(19.74±6.88)ng/L vs(15.26±6.60)ng/L],差异均有统计学意义(P<0.05)。观察组患儿治疗总有效率(96.42%)高于对照组(80.00%),差异有统计学意义(P<0.05),观察组不良反应发生率为6.67%,与对照组比较差异无统计学意义(P>0.05)。结论雾化吸入干扰素α2b注射液治疗小儿呼吸道合胞病毒肺炎能降低患者炎症因子水平,不良反应较轻,临床疗效显著。

NLRP3、AIM2、IFI16炎症小体在慢性乙型病毒性肝炎患者PBMC中的活化水平和与HBV感染的相关性分析

目的:探讨乙肝病毒(HBV)是否激活了慢性乙型病毒性肝炎(CHB)患者外周血单个核细胞(PBMCs)内核苷酸结合寡聚化结构域样受体蛋白3(NLRP3)、黑色素瘤缺乏因子2(AIM2)和干扰素诱导蛋白16(IFI16)炎症小体,分析HBV影响炎症小体活化的可能机制.方法:收集感染内科临床确诊CHB患者35例.同时选取健康住院医师28例为对照.以常规淋巴细胞分层液密度梯度离心法分离健康对照组和CHB患者组静脉血得到PBMCs,采用逆转录、实时荧光定量PCR检测CHB患者组和健康对照组PBMCs NLRP3、AIM2、IFI16、凋亡相关的斑点样蛋白(ASC)、半胱天冬酶1(CASP1)、IL-1β、IL-18 mRNA表达水平,ELISA法检测两组血清中IL-1β蛋白分泌水平.结果:CHB患者组和健康对照组PBMCs ASC、NLRP3、AIM2、IL-1β、IL-18 mRNA表达水平及两组血清IL-1β蛋白分泌水平无显著性差异.CHB患者组PBMCs IFI16、CASP1 mRNA表达水平显著上调,且IFI16 mRNA表达水平与患者血清HBV DNA载量显著正相关(r=0.699 8,P<0.01).结论:慢性HBV感染未导致CHB患者PBMCs NLRP3、AIM2炎症小体的活化;尽管HBV DNA可能诱导了CHB患者IFI16炎症小体的高表达,但通过抑制pro-caspase-1的活化、IL-1β的表达,HBV阻断了IFI16炎症小体的活化效应.

薄芝糖肽与胸腺五肽分别联合重组人干扰素α2b治疗HBeAg阳性的慢性乙型肝炎的临床观察

目的:观察薄芝糖肽和胸腺五肽分别联合重组人干扰素α2b治疗慢性乙型肝炎的疗效及安全性。方法:选择2014年1月-2015年1月我院90例慢性乙型肝炎患者,随机分为A、B、C组,各30例。A组患者给予注射用重组人干扰素α2b(假单胞菌)500万IU皮下注射,qod;B组患者在A组基础上加用薄芝糖肽注射液4 m L加入5%葡萄糖注射液250 m L中,ivgtt,qd;C组患者在A组基础上加用注射用胸腺五肽2 mg加入5%葡萄糖注射液250 m L中,ivgtt,qd。3组患者均治疗24周。比较3组患者治疗4、8、12、24周丙氨酸转氨酶(ALT)复常率、HBeAg阴转率、HBeAg/抗HBeAg血清转换率(以下简称"HBeAg转换率")、HBV-DNA阴转率、乙肝表面抗原(HBsAg)和HBV-DNA下降量,治疗24周时的HBsAg阴转率,并记录不良反应的发生情况。结果:治疗4、8、12周,3组患者ALT复常率、HBeAg阴转率、HBeAg转换率、HBsAg下降量比较,差异均无统计学意义(P>0.05)。治疗4周,B组和C组患者HBV-DNA转阴率显著高于A组,C组患者HBV-DNA下降量显著大于A组和B组,差异有统计学意义(P<0.05)。治疗8、12周,B组和C组患者HBV-DNA阴转率和HBV-DNA下降量显著高于A组,差异有统计学意义(P<0.05),但B组与C组间比较,差异无统计学意义(P>0.05)。治疗24周,3组患者ALT复常率、HBeAg转换率、HBsAg下降量及HBsAg阴转率比较,差异均无统计学意义(P>0.05);B组和C组患者HBeAg阴转率、HBV-DNA阴转率和HBV-DNA下降量显著高于A组,差异有统计学意义(P<0.05),但B组与C组间比较,差异无统计学意义(P>0.05)。3组患者不良反应发生率比较,差异无统计学意义(P>0.05)。结论:薄芝糖肽和胸腺五肽分别联合重组人干扰素α2b对慢性乙型肝炎具有较好的抑制病毒增殖作用,且在ALT复常率、HBeAg转换率、HBsAg下降量及HBsAg阴转率方面效果相当。

高效液相色谱法测定重组人干扰素α-2b注射液中乙二胺四乙酸二钠含量

目的：利用高效液相色谱（HPLC）法测定重组人干扰素α-2b注射液中EDTA二钠（乙二胺四乙酸二钠）的含量。方法：将EDTA二钠与氯化铁溶液于70℃水浴中反应20 min左右;色谱柱为SunFire C18（250 mm×4.6 mm,5μm,Waters）,流动相为5%甲醇＋95%0.64 g/L四丁基溴化铵和4.1 g/L三水合乙酸钠混合液,用冰乙酸调pH值至4.0,流速1 mL/min,检测波长254 nm。结果：该测定方法线性范围为0.025-0.5 g/L,线性关系良好（r=0.9997）,加样回收率为98.27%（n=9,RSD=2.55%）。结论：本方法准确、快速、可靠,可用于重组人干扰素α-2b注射液中EDTA二钠含量的测定。

重组人干扰素α-2b注射液联合ALA-PDT疗法对尖锐湿疣患者外周血CD4^+/CD8^+水平及预后的影响

目的探讨重组人干扰素α-2b注射液联合5-氨基酮戊酸光动力(ALA-PDT)疗法对尖锐湿疣患者外周血CD4^+/CD8^+水平及预后的影响。方法选取2015年2月至2018年2月陕西省地方病防治研究所诊治的265例尖锐湿疣患者为研究对象,采用随机数字表法分将其分为研究组(n=133)与对照组(n=132)。研究组患者在对照组治疗的基础上采取重组人干扰素α-2b注射液治疗,对照组患者采取ALA-PDT疗法治疗,观察比较两组患者疗效、复发率和不良反应发生率,比较治疗前后两组患者血清肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)水平和T淋巴细胞(CD4^+、CD8^+、CD4^+/CD8^+)水平。结果(1)疗效:研究组患者总有效率(92.5%)高于对照组患者(82.6%),其差异具有统计学意义(P<0.05);(2)免疫功能:研究组患者治疗后血清CD4^+、CD4^+/CD8^+水平高于对照组患者,血清CD8^+水平低于对照组患者,其差异均具有统计学意义(均P<0.05);(3)炎性因子水平:研究组患者治疗后血清TNF-α水平低于对照组患者,血清IL-6水平高于对照组患者,其差异均具有统计学意义(均P<0.05);(4)预后:研究组患者复发率(0.8%)低于对照组患者(9.8%),其差异具有统计学意义(P<0.05);(5)安全性:研究组患者不良反应总发生率(6.0%)高于对照组患者(3.0%),其差异无统计学意义(P>0.05)。结论尖锐湿疣患者采取重组人干扰素α-2b注射液联合ALA-PDT疗法,治疗效果确切,能够改善其免疫功能和炎性因子水平,降低复发率,且具有安全性。

LC-MS/MS法测定注射用重组人干扰素α2b中氨苄西林残留

目的建立LC-MS／MS法测定注射用重组人干扰素ct2b中氨苄西林残留。方法选择阿莫西林为内标，直接沉淀蛋白。采用AgilentEclipse XDB-C18（4．6mm×150mm，5μm）柱，以乙腈-水（甲酸调pH3．1）为流动相进行梯度洗脱。采用电喷雾离子源（ESI），正离子模式，多反应监测（MRM）。氨苄西林、内标监测离子对的m／z分别为350．0／106．1，350．0／192．0和366．0／114．0，366．0／208．0。结果氨苄西林在0．0442-17．6472ng／mL浓度范围内线性关系良好（r=0．9991），最低定量限为0．0442ng／mL，方法回收率在95．26％-102．40％之间，批内、批间RSD分别小于3．71％及6．88％。结论本方法准确可靠，重复性好，灵敏度高，适用于注射用重组人干扰素α2b中氨苄西林残留检测。

自体CIK细胞联合IL-2治疗老年B细胞恶性淋巴瘤的价值探讨

目的:探讨自体细胞因子诱导的杀伤细胞(cytokine-induced killer,CIK)联合重组人白介素-2(recombinant human interleukin-2,rh IL-2)治疗老年B细胞恶性淋巴瘤的价值。方法:对本院血液科收治的85例老年B细胞恶性淋巴瘤患者实施自体CIK细胞联合rh IL-2治疗(CIK^+IL-2组),选取同时期未接受自体CIK细胞联合rh IL-2治疗老年B细胞性恶性淋巴瘤患者85例作为对照组。CIK^+IL-2组和对照组按淋巴瘤类型分为4亚组:A组:弥漫性大B细胞淋巴瘤(DLBCL);B组:黏膜相关淋巴组织淋巴瘤(MALT);C组:淋巴浆细胞淋巴瘤(LPL);D组:霍奇金淋巴瘤(HL)。观察患者临床疗效、治疗前后T淋巴细胞亚群、β2微球蛋白水平和生存质量变化情况以及远期生存情况。结果:CIK^+IL-2组4亚组患者治疗后CD3^+、CD3^+/CD8^+、CD3^+/CD56^+水平均显著高于同组治疗前和对照组,β2微球蛋白水平均显著低于同组治疗前和对照组,差异具有统计学意义(P<0.05);CIK^+IL-2组4亚组患者经8个疗程治疗后除1例死亡外,其余CRu、PR患者疗效均转变为CR,各亚组间CR率差异无统计学意义(P>0.05);4亚组患者治疗后躯体功能、角色功能、认知功能、情绪功能及社会功能评分均显著高于同组治疗前,差异具有统计学意义(P<0.05);所有自体CIK联合IL-2治疗组患者中位生存时间为22.36±5.38(8-76)个月,对照组患者中位生存时间为(16.15±3.62(7-55)个月(P<0.05)。结论:自体CIK细胞联合rh IL-2治疗老年B细胞恶性淋巴瘤疗效显著,可有效改善患者T细胞亚群及β2微球蛋白水平,提高生存质量,延长生存时间。