Association between glucose-lowering drugs and circulating insulin antibodies induced by insulin therapy in patients with type 2 diabetes

BACKGROUND Insulin antibodies(IAs)affect blood glucose control in patients receiving insulin therapy.AIM To investigate the relationship between different hypoglycemic treatments and IAs in patients with type 2 diabetes mellitus(T2DM).METHODS This cross-sectional,retrospective study included 1863 patients with T2DM who were receiving exogenous insulin therapy.All patients received stable antidiabetic therapy in the last 3 months and IA levels were measured using an iodine-125 array.RESULTS A total of 1863 patients were enrolled.There were 902(48.4%)patients who had positive IAs(IA level>5%),with a mean IA level of 11.06%(10.39%-11.72%).IA levels were positively correlated with high fasting blood glucose(odds ratio=1.069,P<0.001).The proportion of positive IAs was lowest in patients using glargine only(31.9%)and highest in patients using human insulin only(70.3%),P<0.001.The IA levels in patients using sulfonylureas/glinides(8.3%),metformin(9.6%),and dipeptidyl peptidase-4 inhibitors(8.2%)were all lower than in patients without these drugs(all P<0.05).CONCLUSION Nearly half of patients on insulin therapy have positive IA antibodies,and IA antibody levels are associated with blood glucose control.Insulin glargine and a combination of oral glucose-lowering drugs were correlated with lower IA levels.

Effects of axylitol-casein complex on insulin resistance and gut microbiota composition in high-fat-diet+streptozotocin-induced type 2 diabetes mellitus mice

This study investigated the effects of a xylitol-casein non-covalent complex(XC)on parameters related to type 2 diabetes mellitus(T2DM),in addition to related changes in gut microbiome composition and functions.High-fat-diet(HFD)+streptozotocin(STZ)-induced T2DM mice were treated with xylitol(XY),casein(CN),and XC,after which fecal samples were collected for gut microbiota composition and diversity analyses based on 16S rRNA high-throughput sequencing and multivariate statistics.XC decreased body weight and improved glucose tolerance,insulin sensitivity,pancreas impairment,blood lipid levels,and liver function in T2DM mice compared to XY-and CN-treated mice.Furthermore,XC modulated theα-diversity,β-diversity and gut microbiota composition.Based on Spearman’s correlation analysis,the relative abundances of Alistipes,Bacteroides,and Faecalibaculum were positively correlated and those of Akkermansia,Lactobacillus,Bifidobacterium,and Turicibacter were negatively correlated with the phenotypes related to the improvement of T2DM.In conclusion,we found that XC alleviated insulin resistance by restoring the gut microbiota of T2DM mice.Our results provide strong evidence for the beneficial effects of XC on T2DM and motivation for further investigation in animal models and,eventually,human trials.

Asiaticoside ameliorates type 2 diabetes mellitus in rats by modulating carbohydrate metabolism and regulating insulin signaling

Objective:To evaluate the effect of asiaticoside on streptozotocin(STZ)and nicotinamide(NAD)-induced carbohydrate metabolism abnormalities and deregulated insulin signaling pathways in rats.Methods:Asiaticoside(50 and 100 mg/kg body weight)was administered to STZ-NAD-induced diabetic rats for 45 days,and its effects on hyperglycaemic,carbohydrate metabolic,and insulin signaling pathway markers were examined.Results:Asiaticoside increased insulin production,lowered blood glucose levels,and enhanced glycolysis by improving hexokinase activity and suppressing glucose-6-phosphatase and fructose-1,6-bisphosphatase activities.Abnormalities in glycogen metabolism were mitigated by increasing glycogen synthase activity and gluconeogenesis was decreased by decreasing glycogen phosphorylase activity.Furthermore,asiaticoside upregulated the mRNA expressions of IRS-1,IRS-2,and GLUT4 in STZ-NAD-induced diabetic rats and restored the beta cell morphology to normal.Conclusions:Asiaticoside has the potential to ameliorate type 2 diabetes by improving glycolysis,gluconeogenesis,and insulin signaling pathways.

Naringin ameliorates H_(2)O_(2)-induced oxidative damage in cells and prolongs the lifespan of female Drosophila melanogaster via the insulin signaling pathway

Naringin exists in a wide range of Chinese herbal medicine and has proven to possess several pharmacological properties.In this study,PC12,HepG2 cells,and female Drosophila melanogaster were used to investigate the antioxidative and anti-aging effects of naringin and explore the underlying mechanisms.The results showed that naringin inhibited H_(2)O_(2)-induced decline in cell viability and decreased,the content of reactive oxygen species in cells.Meanwhile,naringin prolonged the lifespan of flies,enhanced the abilities of climbing and the resistance to stress,improved the activities of antioxidant enzymes,and decreased malondialdehyde content.Naringin also improved intestinal barrier dysfunction and reduced abnormal proliferation of intestinal stem cells.Moreover,naringin down-regulated the mRNA expressions of inr,chico,pi 3k,and akt-1,and up-regulated the mRNA expressions of dilp2,dilp3,dilp5,and foxo,thereby activating autophagy-related genes and increasing the number of lysosomes.Furthermore,the mutant stocks assays and computer molecular simulation results further indicated that naringin delayed aging by inhibiting the insulin signaling(IIS)pathway and activating the autophagy pathway,which was consistent with the result of network pharmacological predictions.

Outpatient insulin use in type 2 diabetes mellitus and acute respiratory distress syndrome outcomes:A retrospective cohort study

BACKGROUND The impact of type 2 diabetes mellitus(T2DM)on acute respiratory distress syndrome(ARDS)is debatable.T2DM was suspected to reduce the risk and complications of ARDS.However,during coronavirus disease 2019(COVID-19),T2DM predisposed patients to ARDS,especially those who were on insulin at home.AIMTo evaluate the impact of outpatient insulin use in T2DM patients on non-COVID-19 ARDS outcomes.METHODS We conducted a retrospective cohort analysis using the Nationwide Inpatient Sample database.Adult patients diagnosed with ARDS were stratified into insulin-dependent diabetes mellitus(DM)(IDDM)and non-insulindependent DM(NIDDM)groups.After applying exclusion criteria and matching over 20 variables,we compared cohorts for mortality,duration of mechanical ventilation,incidence of acute kidney injury(AKI),length of stay(LOS),hospitalization costs,and other clinical outcomes.RESULTS Following 1:1 propensity score matching,the analysis included 274 patients in each group.Notably,no statistically significant differences emerged between the IDDM and NIDDM groups in terms of mortality rates(32.8%vs 31.0%,P=0.520),median hospital LOS(10 d,P=0.537),requirement for mechanical ventilation,incidence rates of sepsis,pneumonia or AKI,median total hospitalization costs,or patient disposition upon discharge.CONCLUSION Compared to alternative anti-diabetic medications,outpatient insulin treatment does not appear to exert an independent influence on in-hospital morbidity or mortality in diabetic patients with non-COVID-19 ARDS.

Research Progress on the Efficacy and Safety of Different Basal Insulins in the Treatment of Type 2 Diabetes Mellitus

Objective:To evaluate the efficacy and safety of different basal insulins in the treatment of type 2 diabetes mellitus(T2DM).Methods:The current research progress on different basal insulins was evaluated,with efficacy indicators including fasting plasma glucose(FPG)and glycated hemoglobin(HbAic),and safety indicators focusing mainly on weight change and the incidence of hypoglycemia.Results:Several different basal insulins showed similar metabolic control effects in terms of fasting plasma glucose and glycated hemoglobin.However,the risk of hypoglycemia was lower with insulin glargine 300(Glar-300),insulin degludec 100(Deg-100),and insulin degludec 200(Deg-200)compared to insulin glargine 100(Glar-100).Additionally,Glar-300 had the least impact on weight.Conclusion:For the treatment of T2DM,different basal insulins have similar therapeutic effects,but there are differences in the incidence of hypoglycemic events and their impact on weight.Rational insulin selection and dosage adjustments should be made based on the different patient groups.

Analysis of the Correlation Between Visceral Fat Area and Insulin Resistance in Patients with Type 2 Diabetes and Abdominal Obesity

Objective: To analyze the correlation between visceral fat area and insulin resistance index (HOMA-IR) in patients with type 2 diabetes mellitus (T2DM) and abdominal obesity and to provide a reference for screening and related research of such patients. Methods: Two hundred patients with T2DM admitted to Guandu People’s Hospital of Kunming were included. The study was carried out from October 2022 to December 2023. The patients were divided into three groups according to different abdominal visceral fat areas (VFA): Group A (n = 65) was less than 75cm2, Group B (n = 75) was 75-100 cm2, and Group C (n = 60) was greater than 100 cm2. The subjects in the three groups were all tested for glycated hemoglobin (HbA1c), fasting insulin (FINS), and fasting blood glucose (FPG). Height and weight were measured to calculate body mass index (BMI). The HOMA-IR and TYG (fasting triglyceride and glycemic index) were also calculated. Changes in the BMI, VFA, HOMA-IR, and TYG levels were observed in the three groups. Results: The VFA, BMI, HbA1c, FPG, FINS, HOMA-IR, and TYG of the patients all increased, with a more significant increase in the BMI, FINS, HOMA-IR, and TYG levels (P < 0.01). Multiple linear stepwise regression analyses used visceral fat area (VFA) as the dependent variable. The results showed that VFA was closely related to BMI, FINS, HOMA-IR, and TYG. Conclusion: Early reduction of VFA to reduce insulin resistance may be a better treatment and effective method for T2DM, providing powerful measures and new strategies for effective blood sugar control and early prevention in the treatment of metabolic diseases.

Exogenous insulin autoimmune syndrome:A case report and review of literature

BACKGROUND Insulin autoimmune syndrome(IAS)is a severe manifestation of spontaneous hypoglycemia.It is characterized by elevated levels of immune-reactive insulin and highly potent insulin autoantibodies(IAAs),which are induced by endogenous insulin circulating in the bloodstream.It is distinguished by recurring instances of spontaneous hypoglycemia,the presence of IAA within the body,a substantial elevation in serum insulin levels,and an absence of prior exogenous insulin administration.Nevertheless,recent studies show that both conventional insulin and its analogs can induce IAS episodes,giving rise to the notion of nonclassical IAS.Therefore,more attention should be paid to these diseases.CASE SUMMARY In this case report,we present a rare case of non-classical IAS in an 83-year-old male patient who present with symptoms of a psychiatric disorder.Upon symptom onset,the patient exhibited Whipple's triad(including hypoglycemia,blood glucose level less than 2.8 mmol/L during onset,and rapid relief of hypoglycemic symptoms after glucose administration).Concurrently,his serum insulin level was significantly elevated,which contradicted his C-peptide levels.After a comprehensive examination,the patient was diagnosed with exogenous insulin autoimmune syndrome.Considering that the patient had type 2 diabetes mellitus and a history of exogenous insulin use before disease onset,it was presumed that non classical IAS was induced by this condition.The PubMed database was used to search for previous cases of IAS and non-classical IAS to analyze their characteristics and treatment approaches.CONCLUSION The occurrence of non-classical IAS is associated with exogenous insulin or its analogs,as well as with sulfhydryl drugs.Symptoms can be effectively alleviated through the discontinuation of relevant medications,administration of hormones or immunosuppressants,plasma exchange,and lifestyle adjustments.

The Principles of Insulin Management of Type 2 Diabetes Mellitus

Insulin therapy is an integral part of the pharmacological management of Type 2 diabetes mellitus. Guidelines recommended insulin therapy for those patients with suboptimal glycaemic control despite optimal medical treatments. Studies show that insulin therapy with the human and regular insulins improve glycaemic control, reduce the chronic complications, and inevitably improve patient’s quality of life. The new analogue insulin has a better safety profile and efficacies, and has been shown to achieve better outcomes and patient’s acceptability compared with the human and regular insulins. The diabetic guidelines also recommend the intensity of insulin therapy in a personalised glycaemic control strategy based on the patient’s profiles and their preferences. However, the guidelines do not recommend any standardised approach to the principles of insulin initiation, titration, and monitoring. This review summarises the essential principles of insulin initiation, titration, and monitoring in Type 2 diabetes mellitus.

Effects of vitamin D supplementation on glucose and lipid metabolism in patients with type 2 diabetes mellitus and risk factors for insulin resistance

BACKGROUND Type 2 diabetes mellitus(T2DM)is a chronic metabolic disease featured by insulin resistance(IR)and decreased insulin secretion.Currently,vitamin D deficiency is found in most patients with T2DM,but the relationship between vitamin D and IR in T2DM patients requires further investigation.AIM To explore the risk factors of IR and the effects of vitamin D supplementation on glucose and lipid metabolism in patients with T2DM.METHODS Clinical data of 162 T2DM patients treated in First Affiliated Hospital of Harbin Medical University between January 2019 and February 2022 were retrospectively analyzed.Based on the diagnostic criteria of IR,the patients were divided into a resistance group(n=100)and a non-resistance group(n=62).Subsequently,patients in the resistance group were subdivided to a conventional group(n=44)or a joint group(n=56)according to the treatment regimens.Logistic regression was carried out to analyze the risk factors of IR in T2DM patients.The changes in glucose and lipid metabolism indexes in T2DM patients with vitamin D deficiency were evaluated after the treatment.RESULTS Notable differences were observed in age and body mass index(BMI)between the resistance group and the non-resistance group(both P<0.05).The resistance group exhibited a lower 25-hydroxyvitamin D_(3)(25(OH)D_(3))level,as well as notably higher levels of 2-h postprandial blood glucose(2hPG),fasting blood glucose(FBG),and glycosylated hemoglobin(HbA1c)than the non-resistance group(all P<0.0001).Additionally,the resistance group demonstrated a higher triglyceride(TG)level but a lower high-density lipoprotein-cholesterol(HDL-C)level than the non-resistance group(all P<0.0001).The BMI,TG,HDL-C,25(OH)D_(3),2hPG,and HbA1c were found to be risk factors of IR.Moreover,the posttreatment changes in levels of 25(OH)D_(3),2hPG,FBG and HbA1c,as well as TG,total cholesterol,and HDL-C in the joint group were more significant than those in the conventional group(all P<0.05).CONCLUSION Patients with IR exhibit significant abnormalities in glucose and lipid metabolism parameters compared to the noninsulin resistant group.Logistic regression analysis revealed that 25(OH)D_(3)is an independent risk factor influencing IR.Supplementation of vitamin D has been shown to improve glucose and lipid metabolism in patients with IR and T2DM.

Persistently High Glycated Hemoglobin in a Subgroup of Type 2 Diabetic Patients Who Failed Usual Oral Antihyperglycemics and Insulin in Côte d’Ivoire

Background: Type II diabetes mellitus is associated with multiple metabolic derangements which can cause secondary pathophysiological changes in multiple organ systems. This in turn can impose a heavy burden of morbidity and mortality from micro‑ and macro‑vascular complications. This study aimed to describe the metabolic and therapeutic profile of a subgroup of type 2 diabetic patients who have treatment failure with oral anti-hyperglycemic agents with persistent hyperglycemia despite insulin treatment. Methods: 60 type 2 diabetic patients in treatment failure with oral antidiabetics and under insulin treatment, aged 35 to 70 years, were recruited at the Diabetes Clinic of the University Teaching Hospital of Treichville in Abidjan, Côte d’Ivoire. Blood samples were collected in tubes containing Ethylenediaminetetraacetic Acid (EDTA) to determine glycated hemoglobin (HbA1c). Results: The average age of the population was 54 ± 9.38 years with a sex ratio (M/F) of 0.3, an average BMI of 30.25 ± 5 kg/m2, and an average HbA1c of 10.1% ± 1.6% for an average diabetes duration of 11.8 ± 5.8 years. The average insulin dose was 74.556 ± 16.21 UI/day, and the average duration of insulin treatment was 5.4 ± 3.1 years. The average HbA1c value was 10.1% ± 1.87% in men against 10.03% ± 1.53% in women with no significant difference (p = 0.1). The mean HbA1c values according to patient weight were 10.08% ± 2.05% for normal weight, 9.55% ± 2.26% for overweight, and 10.57% for obese, with no significant difference between the three groups of patients (p = 0.1). Conclusion: This study showed a persistence increase in glycated hemoglobin regardless of the treatment regimen, duration, and dose of insulin treatment in the subpopulation of type 2 diabetic patients.

Exploration of the molecular mechanism of Qishen decoction in regulating miR-495/FTO pathway mediated macrophage polarization to improve insulin resistance therapy of type 2 diabetes

Objective:To observe the effect of Qishen decoction on macrophage polarization mediated by miR-495/FTO signaling pathway,and to clarify the molecular mechanism of Qishen decoction in improving insulin resistance in the treatment of type 2 diabetes.Methods:THP-1 was induced to differentiate macrophages with phorbol ester.It was divided into the control group,the model group,the Qishen decoction group,the miR-495 inhibitor group,and the Qishen decoction+miR-495 inhibitor group.Except for the control group,the remaining groups were stimulated with 30 mmol/L glucose to construct a macrophage polarization model,and corresponding drugs were given for intervention.Cells were collected from each group for 24 hours and the content of inflammatory factors(IL-6,IL-1β,IL-4,and IL-10)were detected using enzyme-linked immunosorbent assay.The expression of macrophage polarization marker molecules,miR-495,and FTO were detected by flow cytometry,qPCR,and Western blot to detect.Results:Compared with the control group,there was no significant change in the activity of macrophages in the control serum,Qishen decoction containing serum,and miR-495 inhibitor transfected serum,and the difference was not statistically significant(P>0.05).In addition,compared to the control group,the content of IL-6 and IL-1β,the expression levels of CD68,iNOS,COX-2,miR-495,and the ratio of CD68/CD206,were significantly increased(P<0.01).While the content of IL-4 and IL-10,as well as the expression of CD206,Arg-1,YM-1,and FTO were significantly reduced(P<0.01).Compared with the model group,the QiShen decoction significantly reduced the contents of IL-6 and IL-1β,and the expression levels of CD68,iNOS,COX-2,and miR-495,as well as the ratio of CD68/CD206,while the content of IL-4 and IL-10,as well as the expression of CD206,Arg-1,YM-1,and FTO were significantly increased(P<0.01).Conclusion:Qishen decoction upregulate the expression of FTO to promote M2 type polarization of macrophages,thereby inhibiting inflammation and improving insulin resistance by inhibiting the expression of miR-495.

Ghrelin regulates insulin resistance by targeting insulin-like growth factor-1 receptor via miR-455-5p in hepatic cells

Objective: To explore the mechanism by which ghrelin regulates insulin sensitivity through modulation of miR-455-5p in hepatic cells. Methods: HepG2 cells were treated with or without DAG (1 μM). Glucose consumption, intracellular glycogen content, phosphorylation of PI3K and Akt stimulated by insulin, expression of miR-455-5p, as well as IGF-1R protein level were analyzed. In addition, bioinformatic analysis, dual luciferase reporter assay, miR- 455-5p mimic or inhibitor treatment was conducted to investigate the molecular mechanisms. Results: High glucose treatment upregulated miR-455-5p expression but reduced glucose consumption and glycogen content. DAG reversed the effect of high glucose on glucose metabolism, increased protein level of IGF-1R and phosphorylation of PI3K/Akt stimulated by insulin, as well as downregulated miR-455-5p expression. Bioinformatic analysis indicated IGF-1R was the target of miR-455-5p. Dual luciferase reporter assay, as well as transfection with miR-455-5p mimic/inhibitor confirmed that DAG activated IGF-1R/PI3K/Akt signaling via inhibiting miR-455-5p. Conclusion: DAG improves insulin resistance via miR-455-5p- mediated activation of IGF-1R/PI3K/Akt system, suggesting that suppression of miR-455-5p or activation of DAG may be potential targets for T2DM therapy.

EDTA-K2诱导血小板假性减少症患者的不同检测方式及结果分析

目的分析乙二胺四乙酸二钾(EDTA-K2)诱导血小板假性减少症(EDTA-PTCP)患者的血小板的不同检测方式及结果。方法回顾性选取2016年5月至2017年5月首都医科大学附属北京友谊医院收治的EDTA-K2诱导的血小板假性减少患者35例,采用Sysmex-XE 5000全自动血球分析仪分别对EDTA-K2抗凝电阻抗法、核酸荧光染色法、枸橼酸钠抗凝电阻抗法进行血小板计数和血小板手工计数,并比较标本静置0~5 min、30 min、60 min、120 min不同时间段血小板体积(MPV)、血小板压积(PCT)、血小板分布宽度(PDW)的变化差异。结果 35例患者采集后0~5min,EDTA-K2抗凝电阻抗法、枸橼酸钠抗凝电阻抗法仪器检测和血小板手工计数结果分别为(237.5±89.2)×10~9/L、(268.5±87.8)×10~9/L、(257.2±91.4)×10~9/L,差异无统计学意义(P>0.05);EDTA-K2抗凝电阻抗法静置30 min、60 min、120 min后血小板计数均值分别为(55.2±23.4)×10~9/L、(39.5±25.0)×10~9/L、(37.5±22.0)×10~9/L,结果均显著低于0~5 min内血小板计数结果(P<0.01),亦显著低于同时段枸橼酸钠抗凝电阻抗法和血小板手工计数结果(P<0.01)。电阻抗法和核酸荧光染色法显示,EDTA-K2抗凝标本静置30 min、60 min、120 min后MPV、PDW均显著高于0~5 min的检测结果(P<0.01),PCT则显著低于0~5 min的检测结果(P<0.01);两种方法同一时间段MPV、PDW、PCT差异均无统计学意义(P>0.05)。结论 EDTA-PTCP患者EDTA-K2抗凝在一定程度上引发血小板聚集,MPV、PDW、PCT的数值变化可以提示血小板聚集的发生。应采用末梢血血小板手工计数,从而将可靠数据提供给临床。

Adiponectin Gene Variation -4522C/T Is Associated with Type 2 Diabetic Obesity and Insulin Resistance in Chinese

The authors investigated the possible association of -4522C/T variation of adiponectin gene with coronary heart disease （CHD） and type 2 diabetes mellitus （T2DM）. Genotyping of SNP --4522C/T in 304 patients with CHD, 389 patients with T2DM, and 405 age and sex-matched healthy control subjects was carried out by means of PCR-RFLP approach. No significant difference in the genotype or allele frequencies was found, either between patients with CHD and control subjects, or between patients with T2DM and control subjects. However, in the subgroup analysis, an association of the TAr genotype and T allele with type 2 diabetes combined with obesity （BMI ≥ 25 kg/m2） was found （P = 0.014 and P = 0.034, respectively）. Also the homeostasis model assessment of insulin resistance （HOMA-IR） in T2DM patients with T/T genotype was significantly higher than that in T2DM patients carrying C allele （P = 0.0069）. The authors＇ findings for the first time demonstrated that SNP --4522 in the adiponectin gene was associated with T2DM that combined with obesity and higher insulin resistance index in patients with T2DM. This indicated that the variation might associate with an increased susceptibility to type 2 diabetic obesity and insulin resistance. But -4522C/T polymorphism did not contribute to the susceptibility of CHD.

32例EDTA-K2抗凝血引起血小板假性减少的结果分析

目的分析由于EDTA-K2抗凝导致血小板假性减少的原因及纠正的方法。方法应用Sysmex公司生产的XE-2100全自动血球计数仪进行全血测定,对1 800例血小板减少患者进行人工血涂片染色,显微镜下观察,其中32例血小板低于70×109/L的血涂片中,发现血小板有聚集现象,少则几个,多则达数百聚集在一起,改用枸橼酸钠抗凝剂采血重新测定,血小板值均在正常范围。结果 32例血小板的测定结果中,最低者为21×109/L,最高者为69×109/L,所有患者均为无血倾向。结论 32例血小板减少患者均系由于EDTA-K2抗凝剂的影响,改用不同的抗凝剂重新测定后,血小板均在正常范围,仅1例为89×109/L。

20例EDTA-K2致假性血小板减少动态分析

EDTA-K2抗凝血时导致血小板计数假性减少现象及动态观察。EDTAK2做为抗凝剂又确有促使或诱导导致血小板发生凝集，从而导致血小板计数假性低下的问题也偶有出现。在《诊断学》教科书上及国外出版的血液学检验书籍中都有简要说明，国外的有关文献报道也很多，而在国内检验书籍中和文献中却很少提到，值得引起检验界同行的重视。我科采用全自动血球计数仪对血小板计数值异常偏低的5000多例患者中的20例标本采血，进行0、30、60、90、120min检测，不抗凝末梢血手工计数，血涂片观察血小板分布情况等方法进行复检。现将结果分析报道如下。

EDTA-K2使网织红细胞溶血造成手工计数失败原因

目前,全国绝大多数医院进行网织红细胞计数仍然采用的是煌焦油蓝染色手工计数法,正常情况下,采用干粉EDTA-K2子弹头离心管采集新鲜离体末梢血进行网织红细胞染色计数对结果是没影响的,但我院在更换了一新批号的干粉EDTA-K2子弹头离心管后,却发现每次进行煌焦油蓝染色网织红细胞计数时全都会失败,镜下均无网织红细胞,而换以前剩下的子弹头离心管同样染色,结果却很好.这一现象以前无人报道过,因此我们就围绕离心管、染液和干粉EDTA-K2这几个因素进行了一系列的试验,以探讨染色失败的原因.

EDTA-K2对溶血样本胰岛素测定的作用研究

目的探讨EDTA—K2是否减弱溶血对胰岛素测定的影响。方法制备10例溶血样本（溶血程度2g／L），每例分为3份（500μl／份），分别加入0，3．8，7．6mg的EDTA—K2，测定在室温放置0，1，2h的胰岛素浓度，每例以其相应未溶血样本为对照。结果溶血与未溶血样本比较，各时间点所测胰岛素浓度均显著降低（P〈0．01），而溶血样本加入7．6mgEDTA—K2则未发生以上改变，加入3．8mgEDTA-K2，只有放置2h的胰岛素浓度仍然显著低于对NN（P〈0．05）。结论EDTA—K2可有效防止溶血对胰岛素测定的影响，其效果与样本中EDTA—K2含量有关。

抗凝剂EDTA-K2对胶体硒免疫层析试验的影响

目的了解不同浓度的EDTA-K2抗凝剂对胶体硒免疫层析试验（胶体硒法）的影响，指导人类免疫缺陷病毒（HIV）抗体的筛查工作。方法分别在不同的一次性使用的真空抽血管（抗凝剂为EDTA-K2）中，加入不等量的新鲜血液，使用胶体硒法检测标本中的HIV抗体，同时用与标本中EDTA-K2抗凝剂的浓度相应的试剂作对照。结果当EDTA-K2抗凝剂的最终浓度为60mg／ml时，标本和试剂对照的试验结果无效；在HIV抗体阴性标本中，浓度在40～20mg／ml时结果为阳性，小于17．14mg／ml后结果为阴性；在HIV抗体阳性标本中，浓度小于40mg／ml后的结果为阳性；在试剂对照中，40～30mg／ml时的试验结果为阳性，小于24mg／ml后结果为阴性。结论较高浓度的EDTA-K／抗凝剂可导致胶体硒法试验的结果无效，或阴性标本的试验结果呈现阳性反应。提示采集足量的血液标本可避免高浓度的EDTA-K2抗凝剂对胶体硒法的影响，是HIV抗体检测能够顺利进行的前提和保证。