武汉地区葡萄糖-6-磷酸脱氢酶缺乏症患者基因突变分析

目的:探讨武汉地区葡萄糖-6-磷酸脱氢酶(G6PD)缺乏症患者的基因突变特征。方法:对湖北省妇幼保健院1 321例筛查阳性的新生儿及门诊患者进行G6PD基因检测,首选多色探针熔解曲线法(MMCA)检测中国人群常见的12种G6PD基因突变,对于MMCA法未检出者,分析其酶活性和临床信息,必要时进行测序检测。结果:在1 321例受检者中共检出768例突变,检出率58.1%。共检出18种G6PD基因变异类型,包括c.1388G>A、c.1376G>T、c.95G>A、c.1024C>T、c.871G>A、c.392G>T、c.487 G>A、c.1360C>T、c.1004C>A、c.517T>C、c.592C>T、c.94C>G、c.152C>T、c.320A>G、c.1028A>G、c.1316G>A、c.1327G>C和c.1376G>C,其中男性半合子683例,女性纯合子3例,女性杂合子80例,女性复杂杂合2例。结论:本研究共检出18种G6PD基因变异类型,并首次在中国人群中报...     

Complete Deficiency of Leukocyte Glucose-6-Phosphate Dehydrogenase with Defective Bactericidal Activity

A 52 yr old Caucasian female (F. E.) had hemolytic anemia, a leukemoid reaction, and fatal sepsis due to Escherichia coli. Her leukocytes ingested bacteria normally but did not kill catalase positive Staphylococcus aureus, Escherichia coli, and Serratia marcescens. An H(2)O(2)-producing bacterium, Streptococcus faecalis, was killed normally. Granule myeloperoxidase, acid and alkaline phosphatase, and beta glucuronidase activities were normal, and these enzymes shifted normally to the phagocyte vacuole (light and electron microscopy). Intravacuolar reduction of nitroblue tetrazolium did not occur. Moreover, only minimal quantities of H(2)O(2) were generated, and the hexose monophosphate shunt (HMPS) was not stimulated during phagocytosis.These observations suggested the diagnosis of chronic granulomatous disease. However, in contrast to control and chronic granulomatous disease leukocytes, glucose-6-phosphate dehydrogenase activity was completely absent in F. E. leukocytes whereas NADH oxidase and NADPH oxidase activities were both normal. Unlike chronic granulomatous disease, methylene blue did not stimulate the hexose monophosphate shunt in F. E. cells. Thus, F. E. and chronic granulomatous disease leukocytes appear to share certain metabolic and bactericidal defects, but the metabolic basis of the abnormality differs. Chronic granulomatous disease cells lack oxidase activity which produces H(2)O(2); F. E. cells had normal levels of oxidase activity but failed to produce NADPH due to complete glucose-6-phosphate dehydrogenase deficiency. These data indicate that a complete absence of leukocyte glucose-6-phosphate dehydrogenase with defective hexose monophosphate shunt activity is associated with low H(2)O(2) production and inadequate bactericidal activity, and further suggest an important role for NADPH in the production of H(2)O(2) in human granulocytes.     

Should Blood Donors Be Routinely Screened for Glucose-6-Phosphate Dehydrogenase Deficiency? A Systematic Review of Clinical Studies Focusing on Patients Transfused With Glucose-6-Phosphate Dehydrogenase–Deficient Red Cells

The risk factors associated with the use of glucose-6-phosphate dehydrogenase (G6PD)–deficient blood in transfusion have not yet been well established. Therefore, the aim of this review was to evaluate whether whole blood from healthy G6PD-deficient donors is safe to use for transfusion. The study undertook a systematic review of English articles indexed in COCHRANE, MEDLINE, EMBASE, and CINHAL, with no date restriction up to March 2013, as well as those included in articles' reference lists and those included in Google Scholar. Inclusion criteria required that studies be randomized controlled trials, case controls, case reports, or prospective clinical series. Data were extracted following the Preferred Reporting Items for Systematic Reviews using a previously piloted form, which included fields for study design, population under study, sample size, study results, limitations, conclusions, and recommendations. The initial search identified 663 potentially relevant articles, of which only 13 studies met the inclusion criteria. The reported effects of G6PD-deficient transfused blood on neonates and children appear to be more deleterious than effects reported on adult patients. In most cases, the rise of total serum bilirubin was abnormal in infants transfused with G6PD-deficient blood from 6 hours up to 60 hours after transfusion. All studies on neonates and children, except one, recommended a routine screening for G6PD deficiency for this at-risk subpopulation because their immature hepatic function potentially makes them less able to handle any excess bilirubin load. It is difficult to make firm clinical conclusions and recommendations given the equivocal results, the lack of standardized evaluation methods to categorize red blood cell units as G6PD deficient (some of which are questionable), and the limited methodological quality and low quality of evidence. Notwithstanding these limitations, based on our review of the available literature, there is little to suggest that G6PD-deficient individuals should be excluded from donating red blood cells, although transfusions of such blood may potentially have negative impacts on premature neonates or patients who need repeated transfusions, and thus, for this group, screening for G6PD deficiency may be appropriate.     

N-Acetylcysteine Reduces Methemoglobin in an In-vitro Model of Glucose-6-phosphate Dehydrogenase Deficiency

Objective: To determine whether N-acetylcysteine (NAC) reduces methemoglobin (MHB) in an in-vitro model of glucose-6-phosphate dehydrogenase (G6PD) deficiency, given that methylene blue is an ineffective MHB antidote in G6PD deficiency.
Methods: Five volunteers donated blood, which was divided equally into 2 test tubes, centrifuged, and washed with Tris-Mopps buffer (pH 7.4, 15 mmol/L glucose). Both tubes were incubated with epiandrosterone (EA) (400 μmol), a specific inhibitor of G6PD. After 75 μL of 0.18 mol hydroxylamine (HA) was added to induce MHB formation, 150 μL of NAC (20 mg/mL) was added to tube 1 and 150 μL of phosphate-buffered saline (PBS) was added to tube 2 as a volume control. Serial MHB levels are reported as a percentage of total hemoglobin (Hb). G6PD activity was measured at baseline, 15 minutes after EA, and at 5 hours.
Results: Mean G6PD activity at baseline was 9.2 ± 2.9 U/g Hb (normal >4.6 U/g Hb); 15 minutes after EA was 3.0 ± 1.0 U/g Hb; and at experiment's end was 2.3 ± 0.7 U/g Hb. The mean (±SD) areas under the concentration-time curves (AUCs) of NAC-EA-HA and PBS-EA-HA samples were compared using an unpaired t-test and were significantly different: PBS-EA-HA, 20,400 ± 1,100 % min, vs NAC-EA-HA, 10,400 ± 1,000 % min, respectively (p < 0.05).
Conclusion: In this in-vitro model of G6PD deficiency, NAC efficiently reduced MHB.     

Glucose-6-Phosphate Dehydrogenase Status and Risk of Hemolysis in Plasmodium falciparum-Infected African Children Receiving Single-Dose Primaquine

Glucose-6-phosphate dehydrogenase (G6PD) enzyme function and genotype were determined in Ugandan children with uncomplicated falciparum malaria enrolled in a primaquine trial after exclusion of severe G6PD deficiency by fluorescent spot test. G6PD A− heterozygotes and hemizygotes/homozygotes experienced dose-dependent lower hemoglobin concentrations after treatment. No severe anemia was observed.     

葡萄糖-6磷酸异构酶(GPI)在类风湿关节炎中的诊断价值

目的 探讨检测血清葡萄糖-6磷酸异构酶(GPI)在类风湿关节炎中的诊断价值.方法 用双抗体夹心酶联免疫吸附试验ELISA法检测130例RA患者、79例其他自身免疫性疾病患者和54例健康对照者血清中的GPI浓度,并同时检测抗环瓜氨酸肽(CCP)抗体、类风湿因子(RF),RA组还同时检测了C-反应蛋白(CRP)、红细胞沉降率(ESR)以及RA患者的手足X线片检查.结果 RA患者血清中GPI浓度显著高于其他风湿病组和健康对照组(P＜0.05);GPI在类风湿关节炎患者中的阳性率为67.1%,特异度为94.7%;抗CCP抗体阳性率为71.5%,RF阳性率为75.4%.130例RA患者GPI阳性率与抗CCP抗体阳性率比较差异无统计学意义(P>0.05),GPI阳性率与RF阳性率比较差异有统计学意义(P＜0.05),GPI与疼痛关节数和关节肿胀有相关性.结论 GPI有可能成为检测RA的又一个独立而敏感的新指标,从而提高RA诊断的敏感性和可靠性,具有较好的临床研究及应用价值.     

深圳宝安区孕期夫妇红细胞葡萄糖-6-磷酸脱氢酶缺乏现状情况调查

目的 了解宝安区孕期夫妇红细胞葡萄糖-6-磷酸脱氢酶(G6PD)缺乏的现状,以便引起社会的重视。方法 收集2013年2月～11月来深圳市宝安区人民医院产前门诊进行孕检的宝安区育龄夫妇共6 574例,其中男性3 192例,女性3 382例,分别采集静脉血用EDTA抗凝混匀,采用改良红细胞葡萄糖-6-磷酸脱氢酶定量比值法测定抗凝全血中G6PD/6PGD的比值,以比值＜1.0判为G6PD缺乏。结果 3 192例男性受检者中,G6PD缺乏的176例,3 382例孕妇受检者中检出135例,检出率分别是5.51%和3.99%,总检出率为4.73%(311/6 574),G6PD缺乏率男性高于孕妇,G6PD缺乏男:女为1.30:1(176:135),两者之间差异有统计学意义(χ²=7.16,P＜0.05)。结论 宝安区孕期夫妇G6PD缺乏有较高的发生率,男性明显高于女性,应引起社会关注。孕期夫妇孕检时应提倡双方G6PD活性筛查,这对优生优育和保健母婴身体健康有着重要的意义。     

湖南长沙地区G6PD基因突变与新生儿黄疸关系的研究

【目的】了解湖南长沙地区葡萄糖 6 磷酸脱氢酶(G6PD)缺乏症的基因突变类型。探讨 G6PD缺乏症基因突变与新生儿黃疸的关系。【方法】采用突变特异性扩增系统对65例湖南长沙地区G6PD缺乏症的新生儿进行基因突变检测,并对其临床表现进行分析。【结果】65 例 G6PD缺乏症的新生儿中检出 G1388A16例(24.6%), G1376T 33 例(50.8%), A95G 13 例(20%),未定型 3 例(4.6%)。G1388A的血红蛋白与G1376T的血红蛋白比较差异有显著性(P<0.05);各基因型患儿黃疸出现时间、血总胆红素值及间接胆红素值比较差异均无显著性(P>0.05)。【结论】G1388A、G1376T、A95G基因突变型是湖南长沙地区最常见的G6PD基因突变型,且均可引起新生儿黃疸,引起新生儿黃疸临床表现的严重程度从重至轻的次序为G1376T、A95G、G1388A。     

A Survey of Schoolchildren from a Server Endemic Goitre Area in Spain

All the boys and girls between six and fourteen years old regularlyattending school in three villages from an area in Spain knownas Las Hurdes were studied with respect to thyroid size, bodyweight and height. Casual urine samples were obtained for thedetermination of iodine and creatinine concentrations. Capillaryblood was spotted on filter paper and used for the determinationof thyroxine and thyrotrophin by radioimmunoassay. The samesurvey was carried out in 354 school children from Madrid, whosedata were used as a reference of the adequacy of the methodsemployed in Las Hurdes. Goitre prevalence was very high, the overall frequency; being86 per cent The concentration of iodine in the urine was <20µg/lin 71 per cent of the school children. The concentration ofcreatinine in the urine was also half that found in childrenfrom Madrid, and the daily creatinine excretion in childrenfrom Las Hurdes was lower than expected from their body weightTheir somatic development, as measured by height and weight,was retarded markedly, both as compared to the Madrid, referencegroup and to international charts. Serum T4 was less than 78nmol/1 (6 µg/d1) in 46 per cent of the children from LasHurdes, and serum TSH greater than 7·5 mU/l in 40 percent A relatively low stature (below the 10th percentile) wasassociated with either a low serum T4, an elevated serum TSH,or both, In 30 per cent of the children. These results indicate that the persistence of endemic goitrein Las Hurdes is of great significance and that there is a continuingrisk of the birth of cretins. Observations made a decade earlierin the same area show that the goitre endemic is not diminishing,and that an iodization programme is urgently needed. It alsoappears that a high proportion of the ‘normal’ schoolchildrenhave features found in hypothyroidism.     

The Variation of Cross-Sectional Area of the Sciatic Nerve in Flexion-Distraction Technique: A Cross-Sectional Study

ObjectiveThe purpose of this study was to compare the cross-sectional area of the sciatic nerve in different positions of spinal manipulation using flexion-distraction technique.MethodsThirty healthy participants were assessed in 6 different flexion-distraction technique positions of varying lumbar, knee, and ankle positions. Participants stood in the following 3 positions with the lumbar in the neutral position: (A) with knee extended, (B) with knee flexed, and (C) with the knee extended and ankle dorsiflexion. Participants then stood in the following 3 positions with the lumbar flexed: (D) with the knee extended, (E) with the knee flexed, and (F) with knee extended and ankle dorsiflexion. The cross-sectional area (CSA) of the sciatic nerve was measured with ultrasound imaging in transverse sections in the posterior medial region of the left thigh. The CSA values measured at each position were compared.ResultsWe analyzed 180 ultrasound images. The cross-sectional area of the sciatic nerve (in mm²) in position B (mean; standard deviation) (59.71-17.41) presented a higher mean cross-sectional area value compared with position D (51.18-13.81; P =.005), position F (48.71-15.16; P = .004), and position C (48.37-16.35; P = .009).ConclusionThe combination of knee extension and ankle dorsiflexion reduced the CSA of the sciatic nerve, and flexing the knee and keeping the ankle in the neutral position increased it.     

The Chiropractic Scope of Practice in the United States: A Cross-Sectional Survey

Objective

The purpose of this study was to assess the current status of chiropractic practice laws in the United States. This survey is an update and expansion of 3 original surveys conducted in 1987, 1992, and 1998.

Methods

A cross-sectional survey of licensure officials from the Federation of Chiropractic Licensing Boards e-mail list was conducted in 2011 requesting information about chiropractic practice laws and 97 diagnostic, evaluation, and management procedures. To evaluate content validity, the survey was distributed in draft form at the fall 2010 Federation of Chiropractic Licensing Boards regional meeting to regulatory board members and feedback was requested. Comments were reviewed and incorporated into the final survey. A duplicate question was imbedded in the survey to test reliability.

Results

Partial or complete responses were received from 96% (n = 51) of the jurisdictions in the United States. The states with the highest number of services that could be performed were Missouri (n = 92), New Mexico (n = 91), Kansas (n = 89), Utah (n = 89), Oklahoma (n = 88), Illinois (n = 87), and Alabama (n = 86). The states with the highest number of services that cannot be performed are New Hampshire (n = 49), Hawaii (n = 47), Michigan (n = 42), New Jersey (n = 39), Mississippi (n = 39), and Texas (n = 30).

Conclusion

The scope of chiropractic practice in the United States has a high degree of variability. Scope of practice is dynamic, and gray areas are subject to interpretation by ever-changing board members. Although statutes may not address specific procedures, upon challenge, there may be a possibility of sanctions depending on interpretation.     

Russian Physicians Burnout during the COVID-19 Pandemic: A Cross-Sectional Survey Study

Objective: To study burnout of Russian physicians in the conditions of COVID-19 pandemic and how their work with coronavirus-infected patients influenced it. According to a three-factor model of burnout developed by Maslach and Jackson, this syndrome includes emotional exhaustion, depersonalization, and reduction of personal accomplishment. Design: A cross-sectional survey study. Setting: Large medical practice. Participants: Physicians of different specialties. Methods: Data collection was conducted from June 23 to July 12, 2020. We developed a Google form including a questionnaire and psychological inventories and placed it in a medical portal. Maslach Burnout Inventory — Human Services Survey for Medical Personnel was used to study burnout; the Hospital Anxiety and Depression Scale was used to determine anxiety and depression. Results: Of all the physicians who took part in the study (N = 599), 31.2 % worked with coronavirus-infected patients. Of the medical personnel who treated COVID-19 patients, 63.6% noted increased workload during the pandemic. Compared to other physicians, they more often had a high degree of emotional exhaustion (43.3 % vs 33.0 %, φ* = 2.404, P ≤ 0.01) and depersonalization (41.7 % vs 34, 0%, φ* = 1.803, P ≤ 0.05). An overwhelming majority of physicians, without any dependence on work with infected patients, had an absence of anxiety and depression. The identified interrelations between the symptoms of burnout, anxiety, depression; age and career stage in medical personnel were identical, except for weak correlations between age and emotional exhaustion (rs = -0.097, P ≤ 0.05), as well as career stage and personal accomplishment (rs = 0.102, P ≤ 0.05) in those physicians who worked with COVID-19 patients. The structure of burnout was identical in all physicians and did not depend on interaction with the infected patients. Conclusion: Public health authorities should reduce the workload on physicians involved in treating infected patients against the backdrop of the pandemic. Psychotherapeutic measures focused on preventing burnout should reduce its number among physicians interacting with patients infected with the coronavirus.     

Dynamics of Malaria Drug Resistance Patterns in the Amazon Basin Region following Changes in Peruvian National Treatment Policy for Uncomplicated Malaria

Monitoring changes in the frequencies of drug-resistant and -sensitive genotypes can facilitate in vivo clinical trials to assess the efficacy of drugs before complete failure occurs. Peru changed its national treatment policy for uncomplicated malaria to artesunate (ART)-plus-mefloquine (MQ) combination therapy in the Amazon basin in 2001. We genotyped isolates collected in 1999 and isolates collected in 2006 to 2007 for mutations in the Plasmodium falciparum dihydrofolate reductase (Pfdhfr) and dihydropteroate synthase (Pfdhps) genes, multidrug resistance gene 1 (Pfmdr-1), the chloroquine (CQ) resistance transporter gene (Pfcrt), and the Ca²⁺ ATPase gene (PfATP6); these have been shown to be involved in resistance to sulfadoxine-pyrimethamine (SP), MQ, CQ, and possibly ART, respectively. Microsatellite haplotypes around the Pfdhfr, Pfdhps, Pfcrt, and Pfmdr-1 loci were also determined. There was a significant decline in the highly SP resistant Pfdhfr and Pfdhps genotypes from 1999 to 2006. In contrast, a CQ-resistant Pfcrt genotype increased in frequency during the same period. Among five different Pfmdr-1 allelic forms noted in 1999, two genotypes increased in frequency while one genotype decreased by 2006. We also noted previously undescribed polymorphisms in the PfATP6 gene as well as an increase in the frequency of a deletion mutant during this period. In addition, microsatellite analysis revealed that the resistant Pfdhfr, Pfdhps, and Pfcrt genotypes have each evolved from a single founder haplotype, while Pfmdr-1 genotypes have evolved from at least two independent haplotypes. Importantly, this study demonstrates that the Peruvian triple mutant Pfdhps genotypes are very similar to those found in other parts of South America.Several countries in South America have been obliged to change their malaria treatment policies (14, 15, 27), because drug resistance has spread rapidly across much of the continent. Chloroquine (CQ) resistance was first documented in 1960 in Venezuela (4), with widespread resistance across the Amazon by the mid-1980s (4). Sulfadoxine-pyrimethamine (SP) was introduced in the 1970s to counter CQ resistance, but in the early 1980s low levels of SP resistance were reported in in vivo efficacy trials, and failure rates had increased to >25% by the mid-1990s. As a result, CQ and SP were removed as first-line therapies for Plasmodium falciparum in many countries and were replaced by the more expensive artesunate (ART)-plus-mefloquine (MQ) combination therapy (ACT) (14). Until the early 1990s, CQ was the first-line therapy for uncomplicated P. falciparum malaria in Peru, and it remains the first-line therapy for Plasmodium vivax (15). Peru''s national policy for the treatment of P. falciparum malaria changed from CQ to SP therapy for the Amazon basin region of Loreto in 1995 (13, 14) and for the North Coast region in June 1999 (13), but the usefulness of SP in the Peruvian Amazon was short-lived. ACT was implemented in 2001 and remains the primary treatment for uncomplicated P. falciparum infections in the Peruvian Amazon.As malaria treatment policies have shifted to drug therapies with as yet unknown targets (i.e., artemisinin derivatives), it has become vital to monitor efficacy using passive surveillance, in vitro assays, and, when possible, molecular markers. Yet it is equally important that active surveillance be used to monitor how changes in public and open-market antimalarial usage influence circulating malaria parasite populations (12). In some instances, changes in drug policy have led to an increase in sensitive phenotypes, suggesting that mutant parasites have lower biological fitness than sensitive parasites in the absence of drug pressure (12, 34). Therefore, it is possible that antimalarials retired due to the emergence of drug resistance may eventually regain some usefulness. To this end, molecular surveillance data can provide highly useful information for policy makers in understanding the population dynamics of drug-resistant genotypes.Genetic markers are being utilized to monitor the origins and spread of antimalarial drug resistance (1). Several loci have been identified for the monitoring of resistance to SP, CQ, atovaquone, MQ, and other important drugs that are still being utilized to treat uncomplicated malaria worldwide. Specific point mutations in P. falciparum crt (Pfcrt) and in Pfdhfr and Pfdhps have been well characterized for CQ and SP resistance, respectively (7). Genotyping of specific polymorphisms before and during drug use is an excellent means of tracking the onset of resistance. Additionally, the characterization of microsatellite loci around these genes gives further traction to the molecular epidemiological approach in determining the origin and spread of the resistant phenotypes. Microsatellite loci have shown that both CQ-resistant and predominant SP-resistant genotypes originated independently in South America and Southeast Asia and that the Asian resistant genotypes spread to other parts of the world, including Africa (25, 33). Therefore, microsatellite markers are a useful tool for documenting the origins and tracking the spread of drug-resistant genotypes.In this study, we have compared the frequencies of mutations in the Pfdhfr and Pfdhps, Pfmdr-1, Pfcrt, and PfATP6/SERCA (sarco/endoplasmic reticulum Ca²⁺ ATPase) genes from parasite isolates collected in the Amazon basin of Peru in 1999 (before ACT implementation) and in 2006 to 2007 (5 to 6 years after ACT implementation) in concert with an examination of the limited in vitro drug sensitivity data. Our findings demonstrate important shifts in the drug-resistant genotypes associated with resistance to SP, MQ, CQ, and possibly ART after the introduction of ACT. Additionally, we characterized microsatellite markers surrounding the genes in order to address the origins of drug-resistant genotypes in the Amazon basin.     

Attitudes Toward Influenza Vaccination Administration in the Emergency Department Among Patients: A Cross-Sectional Survey

IntroductionInfluenza is a serious, vaccine-preventable illness. The current vaccination rates in Canada are below target rates, highlighting the potential need for more convenient ways to receive vaccinations. Wait times to be seen in Canadian emergency departments are escalating, and using the time spent waiting to offer and administer an influenza vaccine could potentially improve ease of access to immunization for some Canadians.MethodsThe aim of this cross-sectional study was to gauge public interest and identify perceived barriers and facilitators to influenza vaccine availability in a Canadian emergency and trauma center. Anonymous questionnaires were completed by a convenience sample of adult patients classified as low acuity (n = 151) as 1 arm of a 2-arm study.ResultsOf the unvaccinated patients, 34.6% expressed willingness to be vaccinated in the emergency department. The patients who had received a vaccine in the previous year were significantly more willing to accept the vaccine in the emergency department (χ² [1] = 23.78, P < 0.001). The 3 top factors associated with having received vaccination in the previous year include trust in vaccine information (χ² [2] = 27.34, P < 0.001), immunity preferences (χ² [2] = 32.25, P < 0.001), and beliefs about efficacy (χ² [2] = 44.90, P < 0.001).DiscussionPatients classified as low acuity were supportive of ED influenza vaccination. In addition, some of the unvaccinated participants had unmet education needs (ie, regarding trustworthy sources of vaccine information, immunity, and vaccine efficacy) that would require addressing before they would likely consider receiving influenza vaccination in future during their ED visit.