Ganciclovir预防异基因造血干细胞移植后巨细胞病毒感染

目的 :评价Ganciclovir在异基因造血干细胞移植 (allo HSCT)后预防巨细胞病毒 (CMV)感染的效果。方法 :观察allo HSCT患者 46例 ,全部病例均系移植前受者和 (或 )供者的CMV IgG阳性 ,分为预防组 2 4例 ,对照组 2 2例。allo HSCT后当患者血中性粒细胞 >1.0× 10 9/L时 ,预防组开始用GCV 10mg·kg-1·d-1,分两次静滴 ,连续 5d ;然后改为 5mg·kg-1·d-1,每周用 5d ,直至 +10 0d。对照组未预防性使用GCV。结果 :在 +10 0d内 ,预防组和对照组的CMV感染率分别为 8% (2 / 2 4)、32 % (7/ 2 2 ) ,P <0 .0 5 ;CMV病发病率分别为 0 %、18% (4 / 2 2 ) ,P <0 .0 5。两组患者在 +10 0d和 +180d内的死亡率分别为 4% (1/ 2 4)和 5 % (1/ 2 2 ) ,P >0 .0 5 ;12 .5 % (3/ 2 4)和 9% (2 / 2 2 ) ,P >0 .0 5。预防组的死因分别为并发细菌和真菌感染、CMV间质性肺炎或原发病复发 ;对照组的死因均是CMV间质性肺炎。结论 :allo HSCT后预防性使用GCV能明显抑制CMV感染 ,减少CMV病发病率。GCV的主要毒副作用是导致中性粒细胞减少 ,使患者继发感染甚至死亡的机率增加。GCV预防性使用的最佳剂量。

单纯疱疹病毒胸苷激酶基因/Ganciclovir系统对脑胶质瘤动物模型的治疗

目的 探讨单纯疱疹病毒Ⅰ型胸苷激酶基因 (HSV1 tk)结合Ganciclovir(GCV)治疗方案 ,在脑胶质瘤动物模型中基因治疗的疗效以及杀伤肿瘤细胞的机制。 方法 采用Southern杂交法 ,证实逆转录病毒载体生产细胞系pN2 A tk VPC(VPC)带有tk基因并稳定整合到基因组DNA上 ;再将VPC与人脑多形性胶质母细胞瘤细胞系DBTRG 0 5MG(0 5MG)分别以 1∶1、1∶4比例混合接种于BALB c裸鼠皮下 (以只接种 0 5MG细胞作为对照 ) ,建立裸鼠皮下胶质瘤动物模型 ,然后腹腔注射GCV治疗 [30mg (kg·d) ],连续 14d ,治疗后 1周 ,取出瘤块作病理组织学分析。 结果 1∶1组瘤块较 1∶4组和对照组明显减小 (P <0 0 1) ,且有 30 %的瘤块完全消失 ,提示HSV1 tk GCV对肿瘤细胞有显著杀伤效应。光镜观察可见 ,1∶1组细胞核固缩、溶解甚至破裂等坏死特征 ,说明HSV1 tk GCV在invivo水平杀伤肿瘤细胞是细胞毒杀伤效应。 结论 HSV1 tk GCV系统能有效杀伤肿瘤细胞 。

Comparison of intravitreal ganciclovir monotherapy and combination with foscarnet as initial therapy for cytomegalovirus retinitis

AIM：To compare the effectiveness between multiple intravitreal injections of ganciclovir alone and combined with foscarnet as initial treatment for patients with newlyonset cytomegalovirus retinitis （CMVR）.METHODS：The retrospective study observed 37 patients（58 eyes） who suffered from CMVR onset between 2013 and 2015. Among them, 35 eyes underwent 4 weekly intravitreal injections of 3.0 mg ganciclovir, and 23 eyes underwent 4 weekly injections of 3.0 mg ganciclovir combined with 2.4 mg foscarnet. Visual acuity, intraocular pressure and viral load of cytomegalovirus （CMV） in aqueous humor measured by real-time polymerase chain reaction were compared before and after each injection.RESULTS：CMV-DNA copies in aqueous humor decreased remarkably in both groups. The average of CMV-DNA copies in patients’ aqueous decreased from 38.3×10~4 copies/mL at baseline to 2.2×10~4 copies/mL after the 4^（th） injection in patients who were treated with ganciclovir monotherapy,and decreased from 76.9×10~4 copies/mL to 11.3×10~4 copies/mL after 4 continuous injections of ganciclovir combined with foscarnet. No significant difference was found in reduction of viral load, change of visual acuities or intraocular pressures between monotherapy or combined therapy.CONCLUSION：Results of this study show that the initial effectiveness of treating CMVR after 4 weekly intravitreal injections is not significantly different from ganciclovir alone or combined with foscarnet. Continuous injection of ganciclovir alone is sufficient in treating immunosuppressive patients with newly-onset CMVR.

Effect of individualized therapy for AIDS patients with cytomegalovirus retinitis in intravitreal ganciclovir injections

The effect of intravitreal ganciclovir injection combined with intravenous infusion on acquired immune deficiency syndrome(AIDS) patients with cytomegalovirus retinitis(CMVR) was investigated. A total of 32 eyes in 23 AIDS patients diagnosed as CMVR from 2017 to 2018 were included in the retrospective study. All patients underwent induction therapy by using intravenous drip of the anticytomegalovirus(CMV) agent ganciclovir(5 mg/kg q12 h) combined with intravitreal ganciclovir injection(3 mg/time, 2 times/wk). The visual acuity, fundus photographs, lesion location, and number of intravitreal injections were observed preoperatively and postoperatively. Totally 14 eyes were cured during induction therapy. The number of injections [4.13(2 to 6)] in CMVR patients with peripherally fundus lesions were significantly lower than those with central lesions (4.89(2 to 6))The individualized therapy of intravitreal ganciclovir injections for AIDS patients with CMVR can effectively reduce the numbers of intravitreal injections.

Severe cytomegalovirus infections in immunocompetent patients at admission as dengue mimic:Successful treatment with intravenous ganciclovir

Cytomegalovirus(CMV) infection is associated with adverse clinical outcomes in immunosuppressed persons.The incidence and association of CMV reactivation with adverse clinical outcomes in critically ill persons lacking evidence of immunosuppression at ICU admission has received great attention in the practice of critical care medicine.Critically ill patients in ICU who had associated risk factors such as mechanical ventilation,severe sepsis, or blood transfusion are more prone to CMV activation,which in turn led to increased mortality and morbidity in tenns of increased ICU stay,longer duration of mechanical ventilation,and higher rates of nosocomial infections.However,severe CMV as initial presentation mimicking dengue infection is rare.We recently came across seven cases with positive CMV serology at ICU admission,which we discuss in the light of current literature.

Analysis of ganciclovir and its related substances using high performance liquid chromatography and liquid chromatography-mass spectrometry methods

Objective High performance liquid chromatography(HPLC)and liquid chromatography-mass spectrometry(LC/MS)methods were developed for the determination of ganciclovir and its related substances.Methods A Hypersil ODS2 column(4.6 mm×250 mm,5 μm)was used with a mobile phase of 0.02 M potassium dihydrogen phosphate buffer(pH 6.0)-methanol(92∶8)at a flow rate of 1.0 mL/min,and UV detector set at 254 nm was used for monitoring the eluents.Results The method was simple,rapid,selective and capable of separating all related substances at trace level with a detection limit of 0.04 μg/mL.It has been validated with respect to accuracy,precision,linearity,and limits of detection and quantification.The linearity range was 10.2-153.0 μg/mL with r=0.9998.The percentage recoveries ranged from 96.7% to 101.6%,and RSD was 1.24%-1.96%(n=5).Conclusion The method was found to be suitable not only for monitoring the reactions during the process development but also for quality control of ganciclovir.For identification of related substances,LC/MS was used.The mainly related substances of ganciclovir active pharmaceutical ingredients(API)were determined as guanine,(1,3-dioxolan-4-yl)methyl acetate,and diacetyl guanine.

Cytopathic effects and local immune responses in repeated neoadjuvant HSV-tk+ ganciclovir gene therapy for prostate cancer

Objective:Cytopathic effects and local immune response were analyzed histologically in prostatic cancer(PCa)with in situ herpes simplex virus-thymidine kinase(HSV-tk)/ganciclovir(GCV)gene therapy(GT).Methods:Four high-risk PCa patients who received HSV-tk/GCV GT were investigated.After two cycles of intraprostatic injection of HSV-tk and administration of GCV,radical prostatectomy was performed.Formalin-fixed,paraffin-embedded sections were evaluated using immunohistochemistry.PCa with hormone therapy(HT,n=3)or without neoadjuvant therapy(NT,n=4)that were equivalent in terms of risk were also examined as reference.Immunoreactively-positive cells were counted in at least three areas in cancer tissue.Labeling indices(LI)were calculated as percentage values.Results:ssDNA LI in GT increased,indicating apoptosis,as well as tumor-infiltrating lymphocytes and CD68-positive macrophages,compared with their biopsies.GT cases showed significantly higher numbers of single-stranded DNA(ssDNA)LI,CD4/CD8-positive T cells and CD68-positive macrophages including M1/M2 macrophages than HT or NT cases.However,there was no significant difference in CD20-positive B cells among the types of case.There were strong correlations between CD8+T cells and CD68+macrophages(ρ=0.656,p<0.0001)as well as CD4+T cells and CD20+B cells(ρ=0.644,p<0.0001)in PCa with GT.Conclusions:Enhanced cytopathic effect and local immune response might be indicated in PCa patients with HSV-tk/GCV gene therapy.

Cooperative Therapeutic Effects of Herpes Simplex Virus Thymidine Kinase Gene/Ganciclovir System and Chemotherapeutic Agents on Prostate Cancer in vitro

The killing effects of herpes simplex virus thymidine kinase gene/ganciclovir （HSV-tk/GCV） approach by the addition of several commonly clinical chemotherapeutic agents on hormone refractory prostate cancer （HRPC） cells PC-3m were investigated. After transferring of the HSV-tk gene into PC-3m cells, mRNA and protein expression of HSV-tk was detected by reverse-transcript polymerase chain reaction （RT-PCR） and strept avidin-biotin complex （SABC） im- munohistochemical method. The killing effect of GCV, cisplatin （CDDP）, etoposide （VP-16）, vincristine （VCR）, methotrexate （MTX）, 5-fluorouracil （5-Fu）, and suramin on PC-3m cells was evaluated by morphological assessment analysis, trypan blue exclusion assay and MTT assay respectively. Additionally, the cooperative effect of HSV-tk/GCV system combined with the above agents on the target cancer cells was determined by MTT. Furthermore, apoptosis and necrosis induced by GCV plus 5-Fu or suramin was analyzed by flow cytometry （FCM）. The results showed that that there was HSV-tk mRNA and protein expression in pDR2-tk plasmid transduced PC-3m cell. Combination of GCV with VP-16, VCR, 5-Fu or suramin led to an enhanced cellular killing effect, but with CDDP resulted in a reduced one and with MTX in an approximate one. FCM revealed that synergistic use of GCV and 5-fu or suramin resulted in a rather large proportion of apoptosis and necrosis with the apoptosis index being 36.38 % and 35.51%, and the proportion of necrosis being 33.05 % and 28.87 %, respectively. In conclusion, HSV-tk/CGV approach by addition of certain clinical available chemotherapeutic drugs brings on statistically significant enhanced cell killing over single-agent treatment. Our results highlight the potential for such new combination therapies for future treatments of HRPC.

在免疫流行性的宿主中急性巨细胞病毒感染用肠道外Ganciclovir治疗

急性巨细咆病毒(CMV)感染在免疫流行性宿主中是一种常见的急性自限性疾病。因此应用ganciclovir(Z羟基-丙氧基甲基-胍)能很大程度上起预防作用,并对免疫受损的病人感染CMV作为治疗用药。本文报告了两例非免疫受损的急性CMV感染的病人应用ganciciovir短期肠外治疗的情况。

抗巨细胞病毒药Ganciclovir

抗巨细胞病毒药Ganciclovir(GCV)为无环鸟苷(ACV)的衍生物。GCV在美国开发于1983年,1984年开始临床试验。1988年美、法及英国开始出售于15个国家。日本于1988年实施临床试验,1990年作为稀用药而进行制造和销售,其商品名叫做Denosine。由于GCV有骨髓抑制。

Ampligen及其与Ganciclovir和Coumermycin Al联合应用对血清中鸭乙型肝炎病毒抑制效果

Ａｍｐｌｉｇｅｎ是一种免疫调节剂或干扰素诱导剂。单独和联合Ｇａｎｃｉｃｌｏｖｉｒ和／或ＣｏｕｍｅｒｍｙｃｉｎＡｌ治疗了鸭乙型肝炎病毒（ＤＨＢＶ）阳性鸭模型。结果表明，上述三种治疗方。案对鸭血清中ＤＨＢＶ均有抑制作用，但治疗结束ＤＨＢＶ回升到治疗前水平。任何一种方案对血清循环ＤＨＢＶ表面抗原（ＤＨＢｓＡｇ）均未产生明显抑制效果。本研究结果为人乙型肝炎治疗时合理使用抗病毒药物和免疫调节、诱导剂提供了重要参考。

Management of cytomegalovirus infection after liver transplantation

Cytomegalovirus(CMV)infection is one of the primary causes of morbidity and mortality following liver transplantation(LT).Based on current worldwide guidelines,the most effective strategies for avoiding post-transplant CMV infection are antiviral prophylaxis and pre-emptive treatment.CMV-IgG serology is the established technique for pretransplant screening of both donors and recipients.The clinical presentation of CMV infection and disease exhibits variability,prompting clinicians to consistently consider this possibility,partic-ularly within the first year post-transplantation or subsequent to heightened immunosuppression.At annual symposia to discuss CMV prevention and how treatment outcomes can be improved,evidence on the incorporation of immune functional tests into clinical practice is presented,and the results of studies with new antiviral treatments are evaluated.Although there are ongoing studies on the use of letermovir and maribavir in solid organ transplantation,a consensus reflected in the guidelines has not been formed.Determining the most appro-priate strategy at the individual level appears to be the key to enhancing out-comes.Although prevention strategies reduce the risk of CMV disease,the disease can still occur in up to 50%of high-risk patients.A balance between the risk of infection and disease development and the use of immunosuppressants must be considered when talking about the proper management of CMV in solid organ transplant recipients.The objective of this study was to establish a compre-hensive framework for the management of CMV in patients who have had LT.

更昔洛韦联合雾化重组人干扰素α1b治疗对呼吸道合胞病毒感染肺炎患儿炎症因子及免疫功能的影响

目的:分析更昔洛韦联合雾化重组人干扰素α1b治疗对呼吸道合胞病毒感染肺炎患儿炎症因子及免疫功能的影响。方法:选取本院收治的呼吸道合胞病毒感染肺炎患儿100例作为研究对象。根据随机数表法分为对照和观察两组,各50例;比较两组治疗前、后的炎性因子降钙素原(PCT)、白介素-6(IL-6)、C反应蛋白(CRP)与免疫球蛋白IgA、IgG、IgM水平,详细记录症状体征消失时间并分析治疗期间发生的不良症状。结果:治疗后两组比较,观察组持续发热、咳嗽、憋喘以及肺啰音消失时间短(P<0.05),血清PCT、IL-6与CRP水平低(P<0.05),血清IgG、IgA、IgM水平明显升高(P<0.05);治疗期间,两组恶心呕吐、白细胞降低、皮疹的总发生率比较无统计学意义(P>0.05)。结论:对呼吸道合胞病毒感染肺炎患儿实施更昔洛韦联合雾化重组人干扰素α1b联合用药可缩短症状体征消失时间,减轻机体炎症反应,提高机体免疫力,安全性相对较好。

1例白细胞及血小板降低的巨细胞病毒肺炎患者抗感染治疗分析

临床药师在工作中需评估用药风险,协助临床医生合理用药。该文介绍了临床药师参与1例白细胞及血小板(PLT)降低的巨细胞病毒肺炎患者使用更昔洛韦治疗的过程。临床药师评估患者PLT及白细胞降低的原因、使用更昔洛韦导致白细胞及PLT降低的风险以及肾功能减退情况,制定个体化方案,使患者得到有效治疗。通过此案例分析,以期为临床救治巨细胞病毒肺炎积累经验,为制定用药监护方案提供借鉴和参考。

不同剂量更昔洛韦治疗新生儿先天性巨细胞病毒感染的疗效

目的:比较不同剂量更昔洛韦治疗新生儿先天性巨细胞病毒感染的疗效及安全性。方法:选取郑州大学第三附属医院2019年2月至2022年2月期间收治的50例新生儿先天性巨细胞病毒感染患儿,依据不同剂量更昔洛韦用药分为小剂量组(6.0 mg·kg^(-1))、大剂量组(7.5 mg·kg^(-1)),各25例,比较两组患儿临床症状评分、肝功能、病毒学检查结果、临床疗效、副作用发生情况。结果:两组患儿治疗后发热、呕吐腹泻、黄疸、肝大、原始反射异常评分均低于治疗前,血清谷草转氨酶(AST)、谷丙转氨酶(ALT)、总胆红素(TBIL)水平及人类巨细胞病毒的脱氧核糖核酸(CMV-DNA)、基因拷贝数变异的免疫球蛋白G(CNV-IgG)、基因拷贝数变异的免疫球蛋白M(CNV-IgM)阳性率均低于治疗前,差异具有统计学意义(P<0.05),但两组组间比较,差异均无统计学意义(P>0.05);小剂量组患儿的副作用发生率低于大剂量组,差异具有统计学意义(P<0.05)。两组患儿总有效率比较,差异均无统计学意义(P>0.05)。结论:大剂量和小剂量更昔洛韦治疗新生儿先天性巨细胞病毒感染的疗效相当,但小剂量安全性更高。

新型冠状病毒引起的病毒性角膜炎病例观察

目的:报告3例严重急性呼吸综合征冠状病毒2型(SARS-CoV-2)引起的病毒性角膜炎。方法:对3例新冠感染确诊患者行裂隙灯、眼压、角膜荧光染色、前段照相、活体共聚焦显微镜(IVCM)和常规眼底筛查。治疗包括更昔洛韦眼用凝胶、人工泪液和糖皮质激素滴眼液。结果:3例SARS-CoV-2角膜炎(SCK)患者经标准治疗后恢复良好。结论:SARS-CoV-2角膜炎通常表现为角膜上皮下浸润,可导致角膜上皮下神经纤维密度降低和树突状细胞(DC)增加。抗病毒治疗联合糖皮质激素已被证明是有效的。

Clinical Efficacy of Oral Ganciclovir for Prophylaxis and Treatment of Recurrent Herpes Simplex Keratitis

Background：Herpes simplex keratitis （HSK） caused by herpes simplex virus 1 （HSV-1）,which has high recurrent rate and incidence of severe vision loss,is the leading cause of infectious blindness in the world.The aim was to explore the clinical efficacy of oral ganciclovir （GCV） in the prevention of recurrent HSK.Methods：A multicenter,prospective,randomized,single-blind,and controlled clinical trial was conducted from April 2010 to June 2013.One hundred seventy-three patients （173 eyes involved） who were diagnosed as recurrent HSK definitely,including stromal keratitis and corneal endotheliitis,were divided into three groups randomly：negative control （placebo） group was topically administered with 0.15% GCV ophthalmic gel,4 times per day and 0.1% fluorometholone eye drops,3 times per day until resolution of HSK; positive control acyclovir （ACV） group was topically adopted the same ophthalmic gel and eye drops and additionally received oral ACV 400 mg 5 times a day for 10 weeks and followed by 400 mg 2 times per day for 6 months; test GCV group was topically adopted the same treatment as negative control group and additionally received oral GCV 1000 mg 3 times per day for 8 weeks.The symptoms and signs were evaluated before and after the therapy 1^st week,2^nd week and then followed up every 2 weeks until recovery.Furthermore,we followed up recurrence of HSK for every 3 months after recovery and then assessed the cure time,recurrent rate and adverse reactions.Results：One hundred and seventy-three patients were followed up 7-48 months （mean 32.1 ± 12.3 months）,but 34 patients were failed to follow-up.The cure time was 12.1± 4.3,11.9 ± 4.0 weeks in negative control （placebo） group and positive control ACV group respectively （P =0.991）,which was longer than that in test GCV group （8.6 ± 2.8 weeks） and there was a significant difference between test GCV group and negative control （placebo） group or positive control ACV group （P =0.000）.Furthermore,the recurrent rate was higher in negative control （placebo） group （47.3%） than that in positive control group ACV （26.7%） and test GCV group （17.2%）,and there was a great significant difference among the three groups （P =0.007）,but there was no significant difference between positive control ACV group and test GCV group （P =0.358）.In addition,there was no obvious adverse reaction expect neutropenia （only one patient in test GCV group）.Conclusion：Short-term oral GCV could cure recurrent HSK and endotheliitis,shorten the course,reduce recurrent rate of HSK and have confirmed safety.

Construction of plasmid vector pAFP-HSVtk-IRES2-EGFP and its effect on the cytotoxicity of ganciclovir to hepatocellular carcinoma

Background Herpes simplex virus thymidine kinase phosphorylates ganciclovir to ganciclovir monophosphate,which is then converted to ganciclovir triphosphate by endogenous cellular nucleoside kinases.The ganciclovir triphosphate acts as a DNA chain terminator due to the lack of a functional 3＇-OH group and terminates the process of DNA replication,hence leading to cell apoptosis.At present,HSVtk gene usually acts as suicide gene to kill tumor cells.The aim of this study was to investigate the selective cytotoxicity of the herpes simplex virus thymidine kinase/ganciclovir （HSVtK/GCV） suicide gene system controlled by the α-fetoprotein （AFP） promoter on hepatocellular carcinoma （HCC） cells in vitro.Methods pAFP-HSVtk-IRES2-EGFP recombinant plasmid vectors driven by the AFP promoter were constructed.HL-7702 liver cells,HUH-7 HCC,and HepG2 HCC were transfected with the recombinant plasmids.HSVtK gene expression was detected using Western blotting analysis.HepG2 cells line stably expressing HSVtk gene was selected by G418 reagent.The cytotoxicity of HSVtK/GCV suicide gene system on hepatoma cells was measured by CCK-8 reagents when different doses of ganciclovir were added.Results Plasmid pAFP-TK-IRES2-EGFP-expressed HSVtk gene was constructed successfully.HSVtk gene expression level was significantly higher in AFP-positive hepatoma cells than in AFP-negative liver cells.After G418 selection,a HepG2 cells line stably expressing HSVtk gene was acquired.With the increase of the dose of ganciclovir the optical density at 450 nm of HepG2 cells stably expressing HSVtk gene gradually decreased （P ＜0.05）.Conclusion The HSVtK gene-specific expression in hepatoma cells as well as the cytotoxicity of the suicide gene system in HepG2 cells provided the basis for the targeted gene therapy of HCC.

布拉氏酵母菌联合更昔洛韦治疗巨细胞病毒感染相关性溃疡性结肠炎的疗效

目的 分析布拉氏酵母菌联合更昔洛韦治疗巨细胞病毒(CMV)感染相关性溃疡性结肠炎(UC)的临床疗效。方法 60例巨细胞病毒感染相关性溃疡性结肠炎患者为研究对象,将患者的入院序列数作为基础,使用随机数字分组的方式将患者分为对照组与观察组,每组30例。对照组患者采用更昔洛韦治疗,观察组患者采用布拉氏酵母菌联合更昔洛韦治疗。比较两组患者治疗前后炎症因子[肿瘤坏死因子-α(TNF-α)、白介素-6(IL-6)]水平,临床症状好转时间及巨细胞病毒感染转阴率,临床治疗效果。结果 治疗后,观察组TNF-α(15.23±2.19)pg/ml、IL-6(145.93±11.02)pg/ml低于对照组的(25.32±2.63)、(198.72±11.51)pg/ml,差异具有统计学意义(P<0.05)。观察组患者的腹泻消失时间(6.17±1.58)d、血便消失时间(4.32±1.35)d短于对照组的(9.87±1.69)、(7.21±1.86)d,巨细胞病毒感染转阴率93.3%高于对照组的63.3%,差异具有统计学意义(P<0.05)。观察组患者的总有效率96.7%高于对照组的80.0%,差异具有统计学意义(P<0.05)。结论 在对巨细胞病毒感染相关性溃疡性结肠炎患者进行治疗的过程中,采用布拉氏酵母菌联合更昔洛韦进行治疗,能够改善患者的炎症因子水平、临床症状好转时间与临床治疗效果,值得推广与应用。