Lyophilized recombinant human brain natriuretic peptide for chronic heart failure:Effects on cardiac function and inflammation

BACKGROUND Chronic heart failure(CHF)is a serious and prevalent condition characterized by impaired cardiac function and inflammation.Standard therapy for CHF has limitations,prompting the exploration of alternative treatments.Recombinant human brain natriuretic peptide(BNP)has emerged as a potential therapy,with evidence suggesting that it can improve cardiac function and reduce inflammation in patients with CHF.However,further research is required to determine the efficacy and safety of lyophilized recombinant human BNP in CHF patients and its impact on microinflammatory status.This study aimed to investigate the effects of lyophilized recombinant human BNP therapy on CHF patients’cardiac function and microinflammatory status.AIM To investigate the effects of freeze-dried recombinant human BNP therapy on cardiac function and microinflammatory status in patients with CHF.METHODS In total,102 CHF patients admitted to our hospital from January 2021 to January 2022 were randomly assigned to control and observation groups(n=51 patients/group).The control patients were treated with standard HF therapy for 3 d,whereas the observational patients were injected with the recombinant human BNP for 3 d.Clinical efficacy,inflammatory factor levels,myocardial damage,cardiac function before and after the treatment,and adverse reactions during treatment were compared between the two groups.RESULTS The overall clinical efficacy was higher in the observation group than in the control group.Compared with baseline,serum hypersensitive C-reactive protein,N-terminal proBNP,and troponin I level,and physical,emotional,social,and economic scores were lower in both groups after treatment,with greater reductions in levels and scores noted in the observation group than in the control group.The overall incidence of adverse reactions in the observation group was not significantly different compared with that in the control group(P>0.05).CONCLUSION Freeze-dried recombinant human BNP therapy can improve heart function and enhance microinflammatory status,thereby improving overall quality of life without any obvious side effects.This therapy is safe and reliable.

Lyophilization Process of Hydroxypropyl Tetrahydropyrantriol Liposomes

[Objectives]To enhance the skin permeability of hydroxypropyl tetrahydropyrantriol and provide a reference for the subsequent prevention or treatment of skin aging.[Methods]The lyophilization process of hydroxypropyl tetrahydropyrantriol liposomes was investigated using a single factor method,and a quality evaluation system was established based on the appearance,particle size,PDI,and re-dispersibility of the lyophilized samples.[Results]The lyophilization process of hydroxypropyl tetrahydropyrantriol liposomes was determined by single factor experiments.The pre-freezing period was 16 h at-80℃,the total drying time was 36 h,and the addition of 10%mannitol-sucrose was used as the lyoprotectant.[Conclusions]The product prepared by the lyophilization method exhibits a fluffy and full appearance,with minimal shrinkage and collapse.The volume remains consistent before and after lyophilization,and the re-dispersibility is satisfactory.The re-dissolution process is rapid,and the particle size and polydispersity index(PDI)remain largely unchanged before and after lyophilization.

Clinical Non-inferiority Trial on Treatment of Coronary Heart Disease Angina Pectoris of Xin-blood Stasis Syndrome Type with Lyophilized Salvia Salt of Lithospermic Acid Powder for Injection

Objective： To evaluate the effectiveness and safety of lyophilized Salvia salt of lithospermic acid powder for injection （SSLA） in treating coronary heart diseases angina pectoris （CHD-AP） of Xin-blood stasis syndrome type, and to conduct the non-inferiority trial with Danshen injection （丹参注射液, DSI） as positive control. Methods： An non-inferiority clinical layered, segmented, randomized, and blinded trial on three parallel and multiple centered groups was conducted in 480 patients with stable effort angina grade Ⅰ , Ⅱand Ⅲ, who had two or more times of attack every week. The 240 patients in test group A were treated with SSLA 200 mg added in 250 ml of 5% glucose solution for intravenous dripping every day; the 120 patients in test group B were treated with SSLA but the dosage doubled; and the 120 patients in the control group were treated with DSI 20 ml daily in the same method as SSLA was given. The clinical effectiveness and safety were evaluated after the patients were treated for 14 days. Results： The results showed that the markedly effective rate in test groups A, B and control group was 37.45 %, 36.75 % and 30.09 % respectively, while the total effective rate in them was 88.09%, 89.74% and 67.26% respectively. Statistical significance was shown in comparisons of the therapeutic effect between control group with test group A and test group B, with that in the two test groups superior to that in the control group, and non-inferiority trial showed eligibility （P〈0.01）. Adverse reaction appeared in 8 patients in the test groups and 2 in the control group. Conclusion： SSLA has definite therapeutic effect in treating patients with CHD-AP, with its effect not inferior to that of DSI, and no evident toxic-adverse reaction.

Immunogenicity of Lyophilized MVA Vaccine for HIV-1 in Mice Model

Highly attenuated modified vaccinia Ankara（MVA） is sensitive to repeat freeze-thaw cycle and easy to lose activity. In order to make the activity of MVA vaccine remain stable during its manufacturing, storage, and administration, the lyophilization as a good option could be resorted to; through screening, the right stabilizer composition and its production procedure were obtained. The final moisture content of freezing-dried recombinant MVA-HIV vaccine was lower than 3%. It can be reconstituted quickly and shows regular physical appearance and stable potency. In vivo functional experiment, mice were divided randomly into the liquid vaccination group, the lyophilized vaccination group, and the control group. Having been DNA vaccine priming, the mice were boosted with a dose of 10^7 pfu MVA- HIV vaccine, which produced indistinguishable antibody titer and cytotoxic T-lymphocyte（CTL） level compared with those of liquid vaccination group （ P 〉 0.05 ）. These results demonstrate that lyophilized MVA vaccine can induce high immunogenicity in mice.

Reconstruction of Rabbit Corneal Layer Composed of Corneal Fibroblasts and Corneal Epithelium on the Lyophilized Amniotic Membrane

Many researchers have employed the cryopreserved amniotic membrane（CAM） and corneal epithelial cells in the treatment of a severely damaged burned cornea, with corneal epithelial cells cultured on an amniotic membrane （AM）. The lyophilized amniotic membrane（LAM） has a higher graft take and a longer shelf life; it is easier to store and safer because of gamma irradiation. Two Teflon rings（ Ahn＇s supporter） were made for culturing the cells on the LAM, and were then used to support the LAM. To reconstruct a corneal layer composed of corneal fibroblasts and epithelium, the corneal fibroblasts were first cultivated on the stromal side of LAM for five days, foUowed by epithelial cells culture on the epithelial side, by using the air-liquid interface culture. The reconstructed corneal layer composed of corneal fibroblasts and corneal epithelial cells has a much healthier basal layer of corneal epithelium than the reconstructed corneal epithelium, which was got by using only corneal epithelial cells, and resembles the epithelium of normal corneas, without the horny layer. Thus, the reconstruction of the corneal layer by using a LAM is considered to be a good in vitro model, not only for its application in toxicological test kits, but also for transplantation in patients with a severely damaged cornea.

Preparation of ^(188)Re-HEDP lyophilized kit for instant bone metastases therapy

Hydroxyethylidene diphosphonate(HEDP) was prepared and labeled with rhenium-188. Its lyophilized kit was developed for instant preparing of188Re-HEDP.188Re-HEDP showed high focile uptake in normal mice(> 30% ID/g at carrier co-injection) in BABLC/SPF mice. High quality single photon emission computed tomography(SPECT) image of New Zealand rabbit was obtained at 4 h after intravenous injection of 74 MBq radiotracer. The lyophilized HEDP kit affords the new opportunity for routine clinical application in bone metastases therapy.

In Vitro Study on the Repair Effect of Fibronectin Lyophilized Powder

Fibronectin has a good repair effect on skin,but its practical application is limited by its easy degradation and difficult preservation.Freeze-drying technology can extend the shelf life of biological products,but may cause damage to some of their biological activities.Therefore,it is necessary to conduct comprehensive and scientific testing of fibronectin lyophilized powder prepared by freeze-drying technology to evaluate its actual efficacy.In this study,by means of in vitro cell experiment,zebrafish animal model experiment and the experiment on the human skin,the safety of fibronectin lyophilized powder prepared by the new formula and the actual efficacy of skin repair were preliminarily and multidimensionally evaluated,so as to provide reference and basis for further research,human clinical trial and application.The results showed that fibronectin lyophilized powder prepared by the new formula had low cytotoxicity,significant ability of promoting cell migration and proliferation,and had stronger skin repair effect.The new Fibronectin lyophilized powder has the characteristics of good safety and strong skin repair effect,which shows that it has good development prospects.These results provide a new strategy for the development of skin repair products.

Biological Characteristics of Trehalose and Its Protective Effect on Food Fermentation Starters during Lyophilization

This paper reviewed the unique biological function of trehalose and its mechanism of stabilizing biological macromolecules and the research progress in the protective effect of trehalose on lactic acid bacteria fermentation starters during lyophilization in food production.The application of trehalose in food industry was prospected.

组合浸渍和超低温冻融预处理对真空冷冻干燥蓝莓品质的影响

为探究组合浸渍冻干保护剂(海藻糖-CaCl_(2))和超低温冻融预处理对蓝莓活性物质和品质的影响,该研究以直接冷冻干燥为对照组,以普通冻融(-20℃)、超低温快速冻融(-80℃)、超声波辅助冻干保护剂浸渍冻融(-20℃)、(-80℃)预处理后进行真空冷冻干燥为试验组,比较分析蓝莓活性物质(多酚氧化酶和过氧化物酶活性)、营养物质(维生素C、花青素、总酚、类黄酮)、质构特性(硬度、咀嚼性)等指标的变化情况,并在此基础上进行BoxBehnken三因素三水平试验设计。结果表明:单一的冻干保护剂浸渍处理或者超低温处理均不如两者组合效果理想,两者组合浸渍后-80℃冻融处理可以有效维持蓝莓硬度,提高蓝莓营养物质的保留度并减少真空冷冻干燥时间。冻融次数为2次,浸渍时间为3.7 h,单次冷冻时间为4.2 d时蓝莓综合指标最高,营养物质保留度最高。该研究阐述了冻干保护剂浸渍预处理与超低温冻融两者联合处理对真空冷冻干燥蓝莓品质提升的机理,探究了冻融次数、冷冻时间、浸渍时间对品质的影响,得出了真空冷冻干燥预处理的最佳参数,为蓝莓真空冷冻干燥技术的发展提供了新思路。

真空冻干/快速溶剂萃取-气相色谱-质谱法测定土壤中35种不同种类半挥发性有机物

建立了真空冻干/快速溶剂萃取-气相色谱-质谱法(GC-MS)测定土壤中35种不同种类半挥发性有机物(SVOCs)的方法。样品经真空冷冻脱水、快速溶剂萃取、旋转蒸发浓缩以及固相萃取SPE小柱净化后,氮吹定容,采用GC-MS上机测定。结果表明:土壤中35种SVOCs在10~1000μg/L范围内线性相关系数r^(2)≥0.991,方法检出限为0.034~0.40μg/kg,方法定量限为0.14~1.20μg/kg;以空白样品为基体进行加标回收实验,所得加标回收率为71.2%~113%,相对标准偏差(RSDs,n=7)为0.38%~13.3%,替代物荧蒽-D 10回收率为84.0%~92.5%;经实际样品检测验证,该方法灵敏、快速、准确可靠,适用于土壤中35种不同种类SVOCs的分析检测。

淡紫拟青霉及哈茨木霉混配冻干粉剂的制备及药效研究

以淡紫拟青霉和哈茨木霉为研究对象,选择合适的冻干保护剂及浓度制备淡紫拟青霉和哈茨木霉冻干粉剂,同时以苹果轮纹病为防治对象,进行混配菌剂的室内离体试验。结果表明:淡紫拟青霉冻干保护剂最佳配方组合为蔗糖10%、乳糖10%、甘油5%,当-80℃预冻18 h,菌体∶保护剂=2∶3时,淡紫拟青霉冻干存活率达85.23%;哈茨木霉冻干保护剂组成为蔗糖15%、蛋白胨2%、甘油5%,当-80℃预冻18 h,菌体∶保护剂=1∶1时,哈茨木霉冻干存活率可达95.25%。哈茨木霉与淡紫拟青霉冻干粉以6∶4的比例混配时,对苹果轮纹病防效最佳,防治效率可达70.24%。该结果有助于解决冻干菌剂在加工过程中容易失活、环境中存活时间短、药效不稳定等问题,可为开发良好的生防菌新剂型提供依据,对推动生物农药行业的发展有重要意义。

阿法替尼脂质体冻干粉的制备与性质

设计一种新型脂质体冻干粉,作为吸入剂有望解决阿法替尼因口服给药产生的胃肠道不良反应等问题,发挥局部定位治疗效果。根据外观形态、溶解性、粒径和包封率确定最佳冻干工艺,并对制备的冻干粉性质进行表征。实验结果显示,采用冷冻干燥法制备的以甘露醇+乳糖为联合冻干保护剂的阿法替尼脂质体冻干粉为白色蓬松粉末,溶解性好,略有引湿性,流动性好,复溶后的平均粒径为(167.0±3.7)nm,包封率为(91.3±1.2)%,稳定性较好。体外释放实验表明,该制剂在PBS和含20%FBS的PBS中初始阶段释放较为缓慢,48 h释放率均为60%左右。体外细胞毒性实验表明,阿法替尼脂质体冻干粉与阿法替尼脂质体的肿瘤细胞毒性作用相当,IC_(50)值为(4.9±0.6)μg/mL。

牛多杀性巴氏杆菌病疫苗效力检验用攻毒菌的研究

为提高我国牛多杀性巴氏杆菌病疫苗效力评价方法的稳定性、可靠性及检验效率,研究制备了活菌数定量的攻毒用牛多杀性巴氏杆菌C45-2株的冻干菌。对其性状、纯粹、真空度、剩余水分、均一性、活菌数稳定性及毒力稳定性进行了检验和评价。冻干攻毒菌在疫苗效力检验中进行应用,并与-70℃冻存的攻毒菌液进行了比较研究。结果表明,定量冻干菌的性状、纯粹、真空度和剩余水分均符合冻干制品的要求;均一性检查变异系数与低温冻存菌液变异系数均小于6%,均一性良好;保存15个月活菌数稳定性良好,较冻存菌液有明显的优势;保存12个月内毒力略有下降,但不影响疫苗的效力检验,可有效评价疫苗质量。试验表明冻干菌株能满足牛多杀性巴氏杆菌效力检验使用,且简化了细菌培养步骤,提高了检验结果的稳定性和工作效率。

注射用益气复脉(冻干)对心肌梗死后慢性心力衰竭的改善作用

目的观察注射用益气复脉(冻干)对心梗后心衰小鼠以及OGD诱导的H9C2细胞损伤的影响。方法30只C57BL/6J小鼠随机分为假手术组、模型组、益气复脉组,采用冠脉左前降支结扎法制备心梗后慢性心衰模型,连续给药4周。将H9C2细胞分为正常组、OGD组、益气复脉低、中、高剂量组和JQ1(BRD4抑制剂)组。心脏超声检测小鼠心功能;HE染色观察心肌组织病理变化;分光光度计检测ATP水平;JC-1法检测线粒体膜电位;RT-qPCR法检测ANP、BNP、BRD4 mRNA表达;TUNEL法观察心肌细胞的凋亡水平;Western blot法检测BRD4、AMPK、p-AMPK蛋白表达。结果与假手术组比较,模型组小鼠心功能降低(P<0.01),心肌细胞增大,排列不规则,心肌组织ANP、BNP、BRD4 mRNA表达升高(P<0.01),心肌细胞凋亡率升高(P<0.01),心肌组织p-AMPK/AMPK蛋白表达降低(P<0.01),BRD4蛋白表达升高(P<0.01);与模型组比较,益气复脉组以上指标均有改善(P<0.05,P<0.01)。在H9C2细胞中,与正常组比较,OGD组ANP、BNP、BRD4 mRNA表达升高(P<0.01),H9C2细胞凋亡率升高(P<0.01),p-AMPK/AMPK蛋白表达降低(P<0.01),BRD4蛋白表达升高(P<0.01),线粒体膜电位和ATP水平降低(P<0.01);与OGD组比较,益气复脉各剂量组和JQ1组以上指标均有改善(P<0.05,P<0.01)。结论注射用益气复脉(冻干)具有改善心梗后慢性心力衰竭小鼠心功能,改善心肌细胞线粒体功能和能量代谢的作用,其机制可能与BRD4/AMPK介导的信号通路相关。

水蛭冻干粉与微粉的研究进展

水蛭是目前常用的活血化瘀药,其体内含有较高的抗凝血酶活性物质以及多种活性成分,从《神农本草经》开始记载使用,至今已有2000余年的历史,在医学领域有着广阔的应用实践和发展前景。近年来随着对水蛭研究的不断深入,对其药理药效的研究和实践应用的探索越来越多,而水蛭一些有效成分的不耐热性问题和对药材饮片质量提升等方面也越来越被重视,为解决这些矛盾,在炮制方式和药材饮片呈现形式也不断改进创新,水蛭冻干粉和微粉等新型饮片形式应运而生,目前已经在市场上广泛流通并受到认可。本文对水蛭冻干粉及微粉的研究成果进行综述,为水蛭冻干粉及微粉的进一步研究和使用提供参考。

复方中药提取物冻干精华液对头皮组织衰老的影响

目的:探究复方中药提取物冻干精华液对头皮组织衰老的影响。方法:以体外培养的正常人体头皮组织作为实验对象,并分为对照组和精华液处理组。采用William完全培养基进行组织培养,然后通过酶联免疫吸附试验、免疫荧光检测等技术,评估精华液对头皮组织衰老相关关键蛋白的影响。结果:在体外培养的头皮组织中,精华液在x/5、x/3、x/2浓度下均能将白介素-6表达水平降至基线的1%以下。此外,精华液在x/10浓度下可分别降低25%的8-羟基-2'-脱氧鸟苷(8-hydroxy-2'-deoxyguanosine,8-OHdG)表达、48%的硫酸软骨素蛋白多糖versican表达和19%的髓鞘蛋白零样蛋白3(myelinproteinzero-like3,MPZL3)表达(均P<0.01),在x/5浓度下可分别降低24%的8-OHdG表达、24%的versican表达和33%的MPZL3表达(均P<0.01)。结论:复方中药提取物冻干精华液对体外培养的头皮组织具有抗炎、抗氧化和抗衰老的作用,具有开展进一步临床研究的价值。

双水平气道正压通气呼吸机辅助冻干重组人脑钠肽对肺心病患者的影响

目的探讨双水平气道正压通气(BPAP)呼吸机辅助冻干重组人脑钠肽对肺心病患者的影响。方法纳入中国人民解放军联勤保障部队第九六〇医院2020年10月至2022年10月收治的肺心病患者236例,采用随机数字表法将其分为对照组(118例)与试验组(118例)。对照组给予冻干重组人脑钠肽治疗,试验组给予BPAP呼吸机辅助冻干重组人脑钠肽治疗,两组均治疗10 d。比较两组临床疗效,检测两组治疗前后心功能、肺功能、血气指标,记录两组不良反应。结果试验组疗效优于对照组,差异有统计学意义(P<0.05)。治疗后,两组每搏输出量(SV)、左室射血分数(LVEF)高于治疗前,N-末端脑钠肽前体(NT-proBNP)低于治疗前,且试验组SV、LVEF高于对照组,NT-proBNP低于对照组,差异有统计学意义(P<0.05)。治疗后,两组第1秒用力呼气容积占预计值百分比(FEV1%)、FEV1/用力肺活量(FVC)、动脉血氧分压(PaO_(2))高于治疗前,动脉血二氧化碳分压(PaCO_(2))低于治疗前,且试验组FEV1%、FEV1/FVC、PaO_(2)高于对照组,PaCO_(2)低于对照组,差异有统计学意义(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BPAP呼吸机辅助冻干重组人脑钠肽对肺心病有较好的疗效,可改善患者心肺功能及动脉血气指标。

Network Pharmacology-Based Dissection of the Bioactive Compounds and Pharmacological Mechanisms of Yiqi Fumai Lyophilized Injection for the Treatment of Heart Failure

Objective:Yiqi Fumai Lyophilized Injection(YQFM),a Chinese medicine injection,has been widely used for the treatment of cardiovascular diseases,especially heart failure(HF).However,bioactive compounds and underlying mechanisms of YQFM in treating HF remain poorly understood.Materials and Methods:Network pharmacology was employed to investigate the bioactive compounds and mechanisms of YQFM.A compound-target network was constructed to screen bioactive compounds based on contribution index calculation.Then,an adriamycin-induced HF rat model was established to evaluate the cardio-protective effects of YQFM by hematoxylin and eosin staining and enzyme-linked immunosorbent assays.Results:Network pharmacology indicated that YQFM may alleviate HF through 36 compounds and 109 targets.Particularly,ginsenosides Rb1,Rg1,Re,Rf,Rb2,Rh1,schisandrin,and ginsenoside Rc were indicated as the top contributors of YQFM in treating HF.YQFM was predicted to act on multiple targets such as vascular endothelial growth factor A,interleukin-2(IL-2),IL-6,and IL-1β,as well as to regulate signaling pathways such as hypoxia-inducible factor 1,tumor necrosis factor,VEGF,and PI3K-Akt.The pharmacological study suggested that YQFM could attenuate cardiac injury and up-regulate plasma concentrations of VEGFR-1 and NO in HF rats.Ginsenoside Rb1,as the major contributor from network pharmacology analysis,also showed a cardioprotective effect and up-regulation of VEGFR-1 in plasma.Conclusions:Ginsenosides and schisandrin were predicted as the most important contributors to the cardioprotective effect of YQMF.Ginsenoside Rb1 was proved to alleviate HF and increase the plasma concentration of VEGFR-1.

注射用重组人脑利钠肽与左西孟旦治疗缺血性心肌病致心力衰竭的效果

目的 分析缺血性心肌病致心力衰竭患者联合使用注射用重组人脑利钠肽及左西孟旦对肺毛细血管楔压(PCWP)、肺动脉压(PAP)水平及心肌细胞功能的影响。方法 选择160例于2021年4月至2023年4月就诊在安阳市人民医院收治的缺血性心肌病致心力衰竭患者,对其行随机方法,使用方法为随机数字表法,分组结果为患者分别进入例数为80例的联合组和对照组。对照组接受左西孟旦等基础治疗,联合组在对照组基础上接受重组人脑利钠肽联合治疗。比较两组疗效。比较两组治疗前后血流动力学参数、心肌细胞功能、心功能、药物不良反应发生情况。结果 联合组治疗总有效率达93.75%,同对照组相比较更高(P<0.05)。两组PCWP、PAP及氨基末端脑钠肽前体(NT-proBNP)水平同治疗前相较均下降,且联合组相对更低(P<0.05)。两组左心射血分数(LVEF)同治疗前相较升高,左心收缩末期内径(LVESD)及左心舒张末期内径(LVEDD)通治疗前相较下降,且联合组LVEF水平相对更高,LVESD及LVEDD则相对更低(P<0.05)。联合组治疗后1个月内不良反应发生率为10.00%,较对照组不良反应总发生率差异无统计学意义(P>0.05)。结论 缺血性心肌病致心力衰竭患者联合应用重组人脑利钠肽及左西孟旦,其PCWP、PAP水平及心肌细胞功能的调节效果均得到增强,增强患者心功能改善效果,且不增加药物不良反应发生风险。