重组人促红细胞生成素对血液透析患者营养状况和免疫功能的影响

目的:研究重组人促红细胞生成素对血液透析患者营养状态和免疫功能的影响。方法:选取2015年8月至2016年4月在我院进行维持性血液透析的患者共139名,随机分为对照组和观察组。对照组患者接受常规铁剂、叶酸等辅助治疗;观察组患者在常规治疗的基础上加用重组人促红细胞生成素。比较两组患者治疗前后血红蛋白、红细胞压积、营养状况(前白蛋白和转铁蛋白)以及免疫功能(CD4~+细胞、CD8~+细胞、CD4~+/CD8~+细胞和免疫球蛋白)等指标。结果:两组患者治疗前血红蛋白、红细胞压积、营养状况指标和免疫功能指标比较均没有统计学差异(P0.05)。治疗12周后,两组患者治疗后的血红蛋白、红细胞压积、前白蛋白、转铁蛋白均较治疗前显著升高(P0.05),且观察组患者以上指标较对照组升高更为明显(P0.05)。对照组患者治疗后的CD4~+细胞比例较治疗前明显升高(P0.05),但CD8~+细胞和CD4~+/CD8~+细胞比例没有显著变化(P0.05),而观察组治疗后这三类细胞的比例均较治疗前明显升高(P0.05)。观察组治疗后IgA、IgM和IgG浓度均较治疗前显著提升(P0.05),而对照组只有IgA和IgM浓度在治疗后有明显提高(P0.05),但仍显著低于观察组(P0.05)。结论:重组人促红细胞生成素能有效纠正血液透析患者的贫血情况,改善其营养状况并提高其免疫功能。     

重组人红细胞生成素在妇科围手术期贫血治疗中的应用研究现状

抗对于妇科围手术期患者,由于术前贫血、术中失血或术后炎症,可能导致贫血状况愈加严重,而旨在改善患者预后和减少异体输血的妇科围手术期血液管理措施越来越被重视。重组人红细胞生成素能显著升高贫血患者血红蛋白水平并减少输血需求,其应用于围手术期贫血的纠正,已成为临床研究的新热点。综述重组人红细胞生成素治疗妇科围手术期贫血的作用机制、国内外临床研究及安全性研究。     

Plasma Levels of Resistin and Ghrelin Before and After Treatment in Patients With Hyperthyroidism

ObjectiveTo investigate ghrelin and resistin concentrations in patients with hyperthyroidism before and after restoration to a euthyroid state and to correlate the 2 peptides with anthropometric and insulin resistance parameters.MethodsThe study included hyperthyroid patients and euthyroid healthy participants as a control group. Hyperthyroid patients were evaluated at the start of the study and after normalization of thyroid function with appropriate antithyroid drugs. Anthropometric parameters, insulin resistance parameters (fasting blood glucose, fasting insulin, and homeostasis model assessment-insulin resistance), thyroid function tests, and measurement of ghrelin and resistin were assessed in patients and control participants.ResultsThe study included 40 hyperthyroid patients (32 women and 8 men, aged 26-42 years) and 30 euthyroid healthy participants (20 women and 10 men, aged 25-43 years) as a control group. In hyperthyroid patients, serum resistin levels and insulin resistance parameters werehigher and plasma ghrelin levels were lower than in control participants (P <.001), and all normalized after treatment. Ghrelin levels were correlated only with insulin resistance parameters, but no correlations with any anthropometric or laboratory data were found. Resistin levels did not correlate with any clinical or laboratory data of hyperthyroid patients.ConclusionIn hyperthyroid patients, resistin was increased and ghrelin was decreased, they were not related to anthropometric parameters, and they normalized after treatment of hyperthyroidism. (Endocr Pract. 2012;18:376-381)     

BMI and Metabolic Profile in Patients With Prolactinoma Before and After Treatment With Dopamine Agonists

Hyperprolactinemia might be related to weight gain, metabolic syndrome (MS), and insulin resistance (IR). Treatment with dopamine agonist (DA) has been shown to reduce body weight and improve metabolic parameters. The objectives of this study were to determine the prevalence of obesity, overweight, MS, and IR in patients with prolactinoma before and after therapy with DA and to evaluate the relation between prolactin (PRL), body weight, fat distribution, leptin levels, IR, and lipid profile before treatment. In addition, we investigated the correlation of the reduction in PRL levels with weight loss and metabolic profile improvement. Twenty‐two patients with prolactinoma completed 6 months of treatment with DA. These patients were submitted to clinical (BMI, waist circumference, blood pressure (BP)), laboratory evaluation (leptin, glucose, low‐density lipoprotein (LDL)‐cholesterol, and triglyceride (TG) levels) and abdominal computed tomography (CT) before and after treatment. The statistical analyses were done by nonparametric tests. At the beginning of the study, the prevalence of obesity, overweight, MS, and IR was 45, 27, 27, and 18%, respectively. After 6 months of treatment with DA, PRL levels normalized, but no significant difference in BMI was observed. However, there was a significant decrease on homeostasis model assessment of insulin resistance (HOMA_IR) index, glucose, LDL‐cholesterol, and TG levels. This study suggests a possible involvement of prolactinoma on the prevalence of obesity. We should consider that DA may be effective on improving metabolic parameters, and we speculate that a period longer than 6 months of treatment is necessary to conclude whether this drug can interfere in the body weight of patients with prolactinoma.     

Molecular Design,Expression and Evaluation of PASylated Human Recombinant Erythropoietin with Enhanced Functional Properties

Erythropoietin (EPO) is the principal hormone which, has somewhat short half-life involved in the differentiation and regulation of circulating red blood cells. The present study was carried out to evaluate the capability of a polyethylene glycol mimetic technology as a biological alternative to improve pharmaceutical properties of human recombinant EPO. In silico models of EPO fused to 200 amino acids of proline, alanine, and serine (PAS) were initially generated and assessed by molecular dynamic (MD) simulation. The fluctuations of the modeled structure reached a plateau after 6000 ps of MD simulation. The Phi and psi analysis showed >99.2% of residues were located in the allowed regions. An expression vector consisting of EPO cDNA tagged to PAS coding sequences was synthesized and expressed in CHO-K1 Cells. The produced PASylated molecule was purified and characterized by standard analytical methods. The molecular weight of fusion protein was expanded to 70 kDa using sodium dodecyl sulfate polyacrylamide gel electrophoresis method. Analytical size exclusion chromatography revealed an approximately sevenfold increase in apparent size of produced protein. Although the in vitro potency of the fusion protein was significantly reduced (1.26?±?0.05 vs. 0.24?±?0.03 ng/ml) but, the in vivo activity was considerably increased up to 1.58?×?10⁵ IU/ml in normocythemic mice assay. Pharmacokinetic animal studies revealed strongly 15.6-fold plasma half-life extension for the PASylated EPO (83.16?±?13.28 h) in comparison to epoetin α (8.5?±?2.4 h) and darbepoetin α (25.3?±?2.2h).     

Dynamic Control of Oligosaccharide Modification in the Mammary Gland: Linking Recombinant Human Erythropoietin

We analyzed two transgenic mouse lines that secrete rhEPO in their milk to assess the dynamic control of N-linked oligosaccharides. Since pharmaceutically available epoetin α and β are produced in CHO cells, we compared transgenic mammary gland-derived rhEPO to its CHO cell-derived counterpart. The major glycosyltransferases that determine the N-oligosaccharides patterns of rhEPO include N-acetylglycosaminyltransferase (GnT) and α1,3/4 fucosyltransferase (Fuc-TIV), GnT-III, -V and Fuc-TIV expression in the mouse mammary gland is significantly higher than that in Chinese hamster ovary (CHO)-derived cells, where the protein is not detectable. The data suggest that N-linked sugar chain patterns of recombinant glycoproteins, produced by the mammary gland differ, since GnT-III alters the sugar pattern extensively. In our experiments, rhEPO produced by the transgenic mice contains more tetra-acidic oligosaccharide structures than epoetin α derived from CHO cells, a rhEPO that is widely used therapeutically. Accordingly, we examined milk-derived rhEPO activity, both in vitro and in vivo. The rhEPO protein purified from the milk of mammary glands upregulates the EPO receptor-mediated expression of the STAT5 gene in MCF-7 cells in a dose-dependent manner, similar to the effects of epoetin α. Furthermore, direct injection of rhEPO into the mouse tail vein leads to an increase in the levels of blood components, such as red blood cells and platelets. In light of these findings, we suggest that the mammary glands of transgenic animals provide a sufficient environment to generate rhEPO with post-translational modifications for biopharmaceutical use. These authors are equal contributors to this work.     

Potential Use of Recombinant Human Thyrotropin in the Treatment of Distant Metastases in Patients with Differentiated Thyroid Cancer

ObjectiveIn order to effectively treat differentiated thyroid cancer (DTC) with radioiodine (RAI) it is necessary to raise serum TSH levels either endogenously by thyroid hormone withdrawal (THW) or exogenously by administration of recombinant human TSH (rhTSH). The goal of this review is to present current data on the relative efficacy and side effects profile of rhTSH-aided versus THW-aided RAI therapy for the treatment of patients with distant metastases of DTC.MethodsWe have searched the PubMed database for articles including the keywords "rhTSH", "thyroid cancer", and "distant metastases" published between January 1, 1996 and January 7, 2012. As references, we used clinical case series, case reports, review articles, and practical guidelines.ResultsExogenous stimulation of TSH is associated with better quality of life because it obviates signs and symptoms of hypothyroidism resulting from endogenous TSH stimulation. The rate of neurological complications after rhTSH and THW-aided RAI therapy for brain and spine metastases is similar. The rate of leukopenia, thrombocytopenia, xerostomia, and pulmonary fibrosis is similar after preparation for RAI treatment with rhTSH and THW. There is currently a controversy regarding RAI uptake in metastatic lesions after preparation with rhTSH versus THW, with some studies suggesting equal and some superior uptake after preparation with THW. Analysis of available retrospective studies comparing survival rates, progression free survival, and biochemical and structural response to a dosimetrically-determined dose of RAI shows similar efficacy after preparation for therapy with rhTSH and THW.ConclusionThe rhTSH stimulation is not presently approved by the FDA as a method of preparation for adjunctive therapy with RAI in patients with metastatic DTC. Data on rhTSH compassionate use suggest that rhTSH stimulation is as equally effective as THW as a method of preparation for dosimetry-based RAI treatment in patients with RAI-avid metastatic DTC. (Endocr Pract. 2013;19:139-148)     

重组人红细胞生成素二聚体不同真核表达载体的构建及表达

为构建重组人红细胞生成素(recombinant human erythropoietin,rhEPO)二聚体真核表达载体,应用PCR方法扩增EPO cDNA,PCR产物克隆入T载体后,经酶切、连接、转化等过程分别构建了3个EPO二聚体的真核表达载体pBT-1c、pBT-2s及pBT-3c,经测序序列完全正确。然后将3个真核表达载体分别转染于CCS-7细胞及CHO-dhfr-细胞中,用ELISA方法检测,它们在COS-7的瞬时表达量分别为4IU／ml、11.5IU／ml和7.2IU／ml。其中EPO二聚体真核表达载体pBTsv稳定转染CHO-dhfr-细胞后,用氨甲喋呤(MTX)逐渐加压的方法筛选到阳性克隆,表达量可达到4000IU／106cells／72h。     

Metabolic Changes in Rat Brain After Prolonged Ethanol Consumption Measured by 1H and 31P MRS Experiments

SUMMARY 1. In vivo ¹H and ³¹P magnetic resonance spectroscopy techniques were applied to reveal biochemical changes in the rat brain caused by prolonged ethanol consumption.2. Three models of ethanol intoxication were used.3. ¹H MRS showed a significant decrease in the concentration of myo-inositol in the brain of rats fed with 20% ethanol for 8 weeks. This change is consistent with perturbances in astrocytes. On the other hand, N-acetyl aspartate and choline content did not differ from controls.4. ³¹P MRS did not reveal any significant changes in the high-energy phosphates or intracellular free Mg²⁺ content in the brain of rats after 14 weeks of 20% ethanol drinking. The intracellular pH was diminished.5. By means of a ³¹P saturation transfer technique, a significant decrease was observed for the pseudo first-order rate constant k _for of the creatine kinase reaction in the brain of rats administered 30% ethanol for 3 weeks using a gastric tube.6. The ¹H MRS results may indicate that myo-inositol loss, reflecting a disorder in astrocytes, might be one of the first changes associated with alcoholism, which could be detected in the brain by means of in vivo ¹H MRS.7. The results from ³¹P MRS experiments suggest that alcoholism is associated with decreased brain energy metabolism.8. ³¹P saturation transfer, which provides insight into the turnover of high-energy phosphates, could be a more suitable technique for studying the brain energetics in chronic pathological states than conventional ³¹P MRS.     

重组人促红细胞生成素在国内外药典中质量标准不同点的对比分析

目的:对比重组人促红细胞生成素在《欧洲药典》(第5版)和《中国药典》(2005年版)中标准的不同点,为国内研发机构对基因重组蛋白产品的研发及国内企业对该产品的进出口提供参考。方法:按2部药典要求,对进口产品或国产产品实样进行部分关键指标的对比分析。结果:2部药典对该产品的标准描述上存在具体内容的区别。结论:在现代药物的研发和生产中,对药品的国际及国内标准要进行综合分析而拟定。     

Serum Monocyte Chemokine Protein-1 Levels Before and After Parathyroidectomy in Patients with Primary Hyperparathyroidism

ObjectiveTo investigate the effects of parathyroidectomy on serum monocyte chemokine protein-1 (MCP- 1) levels in patients with primary hyperparathyroidism (PHPT).MethodsForty-three PHPT patients, age 56 ± 12 years, underwent minimally invasive parathyroidectomy. Serum samples were collected at 0 and 15 to 20 minutes after parathyroid adenoma removal. Serum samples were stored at -70°C until time of assay.ResultsIn 40 PHPT patients with a single adenoma, MCP-1 levels decreased from 342 ± 103 to 250 ± 77 pg/ mL (P < .001) 15 to 20 minutes after parathyroid adenoma removal. MCP-1 levels were positively correlated with intact parathyroid hormone (PTH) levels (R = 0.47; P < .01). In 3 PHPT patients with double parathyroid adenoma, MCP-1 levels did not decrease after removal of the first adenoma but decreased 15 to 20 minutes after second adenoma removal.ConclusionOur results provide evidence that the decrease in serum intact PTH due to minimally invasive parathyroidectomy results in an immediate decrease in serum MCP-1 levels. (Endocr Pract. 2014;20:1165-1169)     

Generation and Phenotypic Analysis of a Transgenic Line of Rabbits Secreting Active Recombinant Human Erythropoietin in the Milk

Production of recombinant human erythropoietin (rhEPO) for therapeutic purposes relies on its expression in selected clones of transfected mammalian cells. Alternatively, this glycoprotein can be produced by targeted secretion into the body fluid of transgenic mammals. Here, we report on the generation of a transgenic rabbits producing rhEPO in the lactating mammary gland. Transgenic individuals are viable, fertile and transmit the rhEPO gene to the offspring. Northern blot data indicated that the expression of the transgene in the mammary gland is controlled by whey acidic protien (WAP) regulatory sequences during the period of lactation. While the hybridization with total RNA revealed the expression only in the lactating mammary gland, the highly sensitive combinatory approach using RT-PCR/hybridization technique detected a minor ectopic expression. The level of rhEPO secretion in the founder female, measured in the period of lactation, varied in the range of 60–178 and 60–162 mIU/ml in the milk and blood plasma, respectively. Biological activity of the milk rhEPO was confirmed by a standard [³H]-thymidine incorporation test. Thus, we describe the model of a rhEPO-transgenic rabbit, valuable for studies of rhEPO glycosylation and function, which can be useful for the development of transgenic approaches designed for the preparation of recombinant proteins by alternative biopharmaceutical production.     

Changes in Muscle Volume and Composition After Treatment of Hip Dysplasia with Periacetabular Osteotomy

BackgroundPeriacetabular osteotomy (PAO) is a common treatment for pre-arthritic hip dysplasia in young adults. The purpose of this study was to better understand changes in muscle volume and composition after PAO visualized using magnetic resonance imaging (MRI).MethodsA prospectively collected series of individuals that underwent PAO for hip dysplasia were reviewed to identify subjects with pre- and postoperative MRI. In our practice, MRI was obtained preoperatively and greater than 6 months after PAO for persistent hip pain. MRI sequences were selected to optimize visualization of the muscle volume, fatty infiltration, and hip joint cartilage. MRI images were selected at predetermined bony landmarks and analyzed using 3D Slicer (©2021, www.slicer.org) software to measure muscle diameter and calculate muscle cross-sectional area (CSA) in 17 individual muscles surrounding the hip. Muscle atrophy was graded using the Goutallier classification for fatty infiltration and acetabular cartilage condition was graded using the Outerbridge classification. We compared pre- and postoperative muscle area and composition as well as cartilage for each case.ResultsA series of six female patients met our inclusion criteria. Mean age was 26 years at time of surgery. All cases had MRI sequences adequate for muscle volume measurements. Fatty infiltration and cartilage changes were recorded in four subjects with appropriate MRI sequences. Separating muscle groups, external rotators underwent the largest volume increase. Hip flexors demonstrated mild volume decrease. CSA change among external rotators averaged +12%, hip flexors -9.3%, and hip abductors -9.2% after PAO. All muscles had either the same or increased fatty infiltration after surgery, with gluteus medius and iliacus undergoing the most average increase. Similarly, cartilage condition worsened by a small margin in this series.ConclusionOur results provide preliminary indication that PAO may have noticeable effects on muscle characteristics and cartilage in the early postoperative period. This was a limited case series of subjects with adequate pre- and post-operative MRI imaging.Level of Evidence: IV     

Recombinant Human Erythropoietin Protects Against Hippocampal Damage in Developing Rats with Seizures by Modulating Autophagy via the S6 Protein in a Time-Dependent Manner

Neurochemical Research - Epilepsy is among the most common neurological disorders. Recurrent seizures result in neuronal death, cognitive deficits and intellectual disabilities in children....     

Changes in the Serum Levels of Trace Elements Before and After the Operation in Thyroid Cancer Patients

The present study aims to examine the changes in the serum levels of trace elements before and after the operation in thyroid cancer patients. The study registered 50 individuals, of whom 25 were female and 25 were male. The patients were allocated to four groups: group 1: male thyroid cancer patients group (n = 15), group 2: female thyroid cancer patients group (n = 15), group 3: male control group (n = 10), group 4: female control group (n = 10). The subjects in groups 1 and 2 were the patients who were post-operatively diagnosed with a pathological malignancy in the thyroid tissue samples. Blood samples were collected from all subjects before the operation, immediately after the operation, and on the post-operative day 15. Additionally, thyroid tissue samples were taken from all subjects post-operatively. Some elements in the blood and tissue samples were determined using the atomic emission method. Zinc and selenium levels of groups 1 and 2 in the pre- and post-operative measurements were significantly lower than those in the control groups (p < 0.05), but were higher in the thyroid tissue (p < 0.05). Serum zinc and selenium levels measured in the subjects on the post-operative day 15 were similar to those measured in the controls. Our study show that changes in the serum and thyroid tissue levels of trace elements like zinc and selenium, which play a critical role in thyroid function, might be associated with the pathogenesis of thyroid cancer.     

丁酸钠对CHO-EPO工程细胞株rhEPO表达量的影响

以稳定整合有ｐＥＤＥＰＯ的ＣＨＯＥＰＯ工程细胞株为研究对象,在无血清条件下,系统观察了０５、１０、２５和５０ｍｍｏｌ／Ｌ４个浓度的丁酸钠作用于该细胞株的情况,结果表明：丁酸钠对ＣＨＯＥＰＯ工程细胞的生长有明显的抑制作用;影响ＣＨＯＥＰＯ工程细胞ＥＰＯ表达,浓度１０ｍｍｏｌ／Ｌ可提高ＥＰＯ表达量２５倍左右,并可持续较长的一段时间;延缓ＣＨＯＥＰＯ工程细胞在无血清培养时的细胞脱落;提高ＣＨＯＥＰＯ工程细胞ＥＰＯｍＲＮＡ水平     

重组人血管内皮抑制素联合顺铂化疗治疗老年晚期非小细胞肺癌的临床疗效

目的:分析重组人血管内皮抑制素联合顺铂化疗方案治疗老年晚期非小细胞肺癌(NSCLC)的疗效和安全性。方法:选取82例老年晚期NSCLC患者作为研究对象,应用随机数字表将患者分为观察组和对照组,每组各41例。对照组患者给予含顺铂的两药化疗方案进行治疗,观察组患者在对照组疗法的基础上加用重组人血管内皮抑制素治疗。对两组患者的临床疗效、临床有效率(CRR)、临床受益率(CBR)进行评价。对两组患者治疗前后的Karnofsky评分、血清癌胚抗原(CEA)水平变化进行观察和比较。对两组患者进行随访,对患者的总生存期(OS)和疾病进展时间(TTP)进行观察和比较。对两组患者治疗期间不良反应发生率进行观察和比较。结果:观察组患者CRR和CBR均显著高于对照组,差异均有统计学意义(P0.05)。两组患者治疗前、后Karnofsky评分的上升幅度和血清CEA水平的下降幅度的差异无统计学意义(P0.05)。观察组患者和对照组患者的OS中位数估计值分别为16.720月和14.590月,TTP中位数估计值分别为6.260月和4.770月,两组患者OS和TTP中位数估计值的差异均有统计学意义(P0.05)。两组患者不良反应发生率差异无统计学意义(P0.05)。结论:在老年晚期NSCLC患者的治疗中,在含顺铂治疗方案基础上加用重组人血管内皮抑制素进行治疗,能够提高患者的临床受益和治疗有效率,延长患者的生存期,改善患者的预后,且未增加不良反应的发生率。     

Altered Plasma Concentrations of Trace Elements in Ulcerative Colitis Patients Before and After Surgery

Ileal pouch-anal anastomosis (IPAA) is a classical surgery for ulcerative colitis patients. However, knowledge on trace element alteration in patients who had undergone this surgery is limited. This study was conducted to assess trace element alteration in patients with ulcerative colitis before and after ileal pouch-anal anastomosis. Preoperative (40) and postoperative (35) ulcerative colitis patients were studied. The dietary assessment of trace element intake was undertaken by a semiquantitative food frequency questionnaire. Patients' trace element status of zinc, copper, manganese, selenium, calcium, iron, and vitamin D₃ was assessed by measuring their blood concentrations. We found that with the similar dietary intake, there was no statistical difference in the concentrations of plasma copper, iron, calcium, and vitamin D₃ in the two groups (P?>?0.05). Compared with preoperative patients, postoperative patients had higher concentrations of plasma zinc (14.51?±?4.75 μmol/l) and manganese (0.21?±?0.11 μmol/l) and lower concentrations of plasma selenium (0.86?±?0.58 μmol/l). Both preoperative and postoperative mean concentrations of plasma calcium and vitamin D₃ were below their reference range, respectively. We conclude that IPAA does not seem to alter patients' abnormal trace elements completely. It is important to monitor and supply some specified trace elements even in postoperative patients.     

重组人血管内皮抑素联合同步放化疗治疗晚期非小细胞肺癌临床研究

目的:探讨重组人血管内皮抑素联合同步化疗治疗晚期非小细胞肺癌(NSCLC)的临床疗效.方法:将我院收治的40例NSCLC患者随机分为治疗组和对照组,对照组患者采用紫杉醇化疗方案联合同步放射治疗,治疗组患者给予重组人血管内皮抑素联合同步放化疗进行治疗,治疗2个周期后对患者的近期疗效、毒副反应以及无肿瘤进展时间(TTP)等进行比较.结果:治疗组CR+PR的患者为15例,明显高于对照组的5例(P＜0.05);治疗组患者TTP为154±11d,中位生存时间为274d,而对照组TTP为108±8d,中位生存时间为179d,两组患者比较存在明显的差异性(P＜0.05);治疗组患者QOL评分明显优于对照组(P＜0.05),而两组患者毒副反应无明显的差异性(P＞0.05).结论:对晚期NSCLC患者采用重组人血管内皮抑素联合同步化疗进行治疗,可提高患者生存时间,且不会增加患者毒副反应的发生.