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1.
Food Chem Toxicol ; : 112248, 2021 Apr 30.
Artigo em Inglês | MEDLINE | ID: mdl-33940105

RESUMO

Bisphenol B (BPB) has been used as a substitute for bisphenol A (BPA) in plastic materials. Whether BPB disrupts the male reproductive system remains unknown. Here, we report the effect of BPB on Leydig cell maturation in late puberty. Male Sprague-Dawley (35 days old) rats were gavaged with BPB at 0, 10, 100, and 200 mg/kg/day for 21 days. BPB significantly reduced body and epididymis weight at 200 mg/kg. BPB markedly decreased serum testosterone levels at 100 and 200 mg/kg and serum luteinizing hormone and follicle-stimulating hormone levels at 200 mg/kg. BPB significantly increased Leydig cell number at 100 and 200 mg/kg, while down-regulating the expression of Leydig cell genes (Cyp11a1 and Hsd3b1) at ≥100 mg/kg and up-regulating the expression of Sertoli cell genes (Pdgfra, Fshr, Sox9) and cell cycle regulators (Pcna, Ccnb1, Cdk2, and Cdk4) at 10-200 mg/kg. BPB markedly increased the phosphorylation of AKT1, AKT2, and ERK1/2 at 200 mg/kg. BPB increased the proliferation of rat immature Leydig cells via promoting the S/M2 phase shift at 100 and 1000 nM after 24-h culture in vitro. In conclusion, BPB disrupts Leydig cell maturation in late puberty by increasing Leydig cell number while inhibiting its maturation.

2.
Arch Virol ; 2021 May 05.
Artigo em Inglês | MEDLINE | ID: mdl-33950289

RESUMO

A novel positive-stranded RNA virus provisionally named "citrus virus C" (CVC) was discovered in citrus trees displaying mottling symptoms. Its genome comprises 7,215 nucleotides (nt), excluding the 3' poly(A) tail, and contains two open reading frames (ORFs) that encode a replication-associated polyprotein (RP) and a putative coat protein (CP). The CVC genome contains a 16-nt 'marafibox', which is highly conserved in most viruses belonging to the genus Marafivirus of the same family. Sequence analysis suggested that the virus is most closely related to grapevine Red Globe virus (GRGV), which is yet to be officially classified in the family Tymoviridae. The sequence identities between CVC and GRGV in the whole genome (50.7%, nt) and CP (49.4% for amino acid, and 53.9% for nt) are lower than the thresholds (80% in the genome and 90% in the CP) for species demarcation in the family. Therefore, it is legitimate to propose that CVC is a member of new species in the family Tymoviridae.

3.
BMC Geriatr ; 21(1): 238, 2021 04 09.
Artigo em Inglês | MEDLINE | ID: mdl-33836651

RESUMO

BACKGROUND: Quadratus lumborum block (QLB) is a novel and effective postoperative analgesia method for abdominal surgeries. However, whether QLB can affect early postoperative cognitive function by inhibiting surgical traumatic stress and the inflammatory response remains unclear. This study aimed to explore the effect of QLB on postoperative cognitive function in elderly patients undergoing laparoscopic radical gastrectomy. METHODS: Sixty-four elderly patients who underwent laparoscopic radical gastrectomy were randomly divided into the QLB group (Q group, n = 32) and control group (C group, n = 32). The Mini-Mental State Examination (MMSE) and Montreal Cognitive Assessment (MoCA) were used to measure cognitive function 1 day before and 7 days after surgery. Postoperative cognitive dysfunction (POCD) was defined as a decline of ≥ 1 SD in both tests. The visual analog scale (VAS) scores 6 h (T1), 24 h (T2), and 48 h (T3) after surgery were measured. The serum levels of high mobility group box protein 1 (HMGB1), interleukin-6 (IL-6), and tumor necrosis factor-α (TNF-α) were evaluated 1 day before surgery (baseline), and 1 day (day 1) and 3 days after surgery (day 3). The intraoperative remifentanil dosage, sufentanil consumption 24 h after surgery, recovery time from anesthesia, and adverse effects were also compared. RESULTS: POCD was present in two patients in the QLB group and eight patients in the C group 7 days after surgery (6.7 % vs. 27.6 %, P = 0.032). The MMSE and MoCA scores were similar in both groups preoperatively, and the two scores were higher in the QLB group than in the C group 7 days after surgery (P < 0.05). The VAS scores were significantly lower in the Q group at all times after surgery (P < 0.05). Compared with the C group, the levels of HMGB1, TNF-α, and IL-6 were significantly decreased 1 and 3 days after surgery in the QLB group (P < 0.05). The remifentanil consumption intraoperatively and sufentanil 24 h postoperatively were significantly lower in the QLB group (P < 0.05). The recovery time from anesthesia was shorter in the QLB group (P < 0.05). No severe adverse effects occurred in either group. CONCLUSIONS: QLB could improve postoperative cognitive function in elderly patients undergoing laparoscopic radical gastrectomy. This may be related to the suppression of the inflammatory response after surgery. TRIAL REGISTRATION: Chictr.org.cn identifier ChiCTR1900027574 (Date of registry: 19/11/2019, prospectively registered).


Assuntos
Laparoscopia , Dor Pós-Operatória , Idoso , Anestésicos Locais , Cognição , Gastrectomia/efeitos adversos , Humanos , Laparoscopia/efeitos adversos
4.
Zhongguo Zhen Jiu ; 41(3): 295-302, 2021 Mar 12.
Artigo em Chinês | MEDLINE | ID: mdl-33798313

RESUMO

OBJECTIVE: To screen protein target in prevention and treatment with electroacupuncture (EA) for Alzheimer's disease (AD) and explore the potential mechanism of EA in prevention of AD. METHODS: A total of 40 APP/PS1 transgenic young male mice, 1.5-month old, were randomized into an EA group and a model group, 20 mice in each one, and 20 C57BL/6J mice were chosen as the normal control group. After adaptive housing for 1 week, the mice in the EA group were stimulated with EA at "Baihui" (GV 20), "Fengfu" (GV 16) and "Shenshu" (BL 23), with intermittent wave, 10 Hz in frequency and 2 mA in electric intensity. EA was given once daily, 20 min each time. There was 1 day at interval after EA for 6 days each week. Totally, the intervention lasted for 16 weeks. On day 3 after the end of EA intervention, Morris water maze test was adopted to detect learning and memory abilities of mice in each group. After water maze test, the label-free method was used to measure the difference expressions in cerebral cortex and hippocampus. Using Western blot method, the expressions of guanylate binding protein beta 5 (GNB 5) and histone-H 3 in cerebral cortex and hippocampus were verified. Using immunohistochemical method, the expressions of amyloid beta protein (Aß) in cerebral cortex and hippocampus were detected. RESULTS: Compared with the normal control group, the escape latency (on day 2, 3 and 4) was prolonged, the frequency of crossing platform and the duration of platform stay were decreased in the mice of the model group (P<0.05). Compared with the model group, the escape latency (on day 3 and 4) was shortened, the frequency of crossing platform and the duration of platform stay were increased in the mice of the EA group (P<0.05). By the comparison among the three groups, the high mobility group nucleosome-binding domain-containing protein 5, band 3 anion transport protein, histone-H 3, epoxide hydrolase 4 (fragment), neurolysin (mitochondria), phosphoglycerate mutase 2, GNB5 and Aß were the differential proteins with the larger fold-change difference in expression. Compared with the normal control group, the expression of histone-H 3 in cerebral cortex and hippocampus was reduced (P<0.001) and the expressions of GNB 5 and Aß were increased (P<0.001, P<0.01) in the mice of the model group. Compared with the model group, the expression of histone-H 3 in cerebral cortex and hippocampus was increased (P<0.001) and the expressions of GNB 5 and Aßwere reduced (P<0.001, P<0.05) in the mice of the EA group. CONCLUSION: The intervention with EA effectively prevents from the decline of learning and memory ability and the formation of Aß senile plaques in cerebral cortex and hippocampus in young mouse models of AD after growing up. Besides, EA plays a regulatory function for protein expression differences induced by AD model.


Assuntos
Doença de Alzheimer , Eletroacupuntura , Doença de Alzheimer/genética , Doença de Alzheimer/terapia , Peptídeos beta-Amiloides/genética , Animais , Modelos Animais de Doenças , Hipocampo , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Proteômica
6.
Neurosci Bull ; 37(5): 597-610, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33900570

RESUMO

Mesocorticolimbic dopaminergic (DA) neurons have been implicated in regulating nociception in chronic pain, yet the mechanisms are barely understood. Here, we found that chronic constructive injury (CCI) in mice increased the firing activity and decreased the KCNQ channel-mediated M-currents in ventral tegmental area (VTA) DA neurons projecting to the nucleus accumbens (NAc). Chemogenetic inhibition of the VTA-to-NAc DA neurons alleviated CCI-induced thermal nociception. Opposite changes in the firing activity and M-currents were recorded in VTA DA neurons projecting to the medial prefrontal cortex (mPFC) but did not affect nociception. In addition, intra-VTA injection of retigabine, a KCNQ opener, while reversing the changes of the VTA-to-NAc DA neurons, alleviated CCI-induced nociception, and this was abolished by injecting exogenous BDNF into the NAc. Taken together, these findings highlight a vital role of KCNQ channel-mediated modulation of mesolimbic DA activity in regulating thermal nociception in the chronic pain state.

7.
Front Immunol ; 12: 624230, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33868241

RESUMO

Cervical cancer is the fourth most common cancer and fourth leading cause of cancer death among women worldwide. In low Human Development Index settings, it ranks second. Screening and surveillance involve the cytology-based Papanicolaou (Pap) test and testing for high-risk human papillomavirus (hrHPV). The Pap test has low sensitivity to detect precursor lesions, while a single hrHPV test cannot distinguish a persistent infection from one that the immune system will naturally clear. Furthermore, among women who are hrHPV-positive and progress to high-grade cervical lesions, testing cannot identify the ~20% who would progress to cancer if not treated. Thus, reliable detection and treatment of cancers and precancers requires routine screening followed by frequent surveillance among those with past abnormal or positive results. The consequence is overtreatment, with its associated risks and complications, in screened populations and an increased risk of cancer in under-screened populations. Methods to improve cervical cancer risk assessment, particularly assays to predict regression of precursor lesions or clearance of hrHPV infection, would benefit both populations. Here we show that women who have lower risk results on follow-up testing relative to index testing have evidence of enhanced T cell clonal expansion in the index cervical cytology sample compared to women who persist with higher risk results from index to follow-up. We further show that a machine learning classifier based on the index sample T cells predicts this transition to lower risk with 95% accuracy (19/20) by leave-one-out cross-validation. Using T cell receptor deep sequencing and machine learning, we identified a biophysicochemical motif in the complementarity-determining region 3 of T cell receptor ß chains whose presence predicts this transition. While these results must still be tested on an independent cohort in a prospective study, they suggest that this approach could improve cervical cancer screening by helping distinguish women likely to spontaneously regress from those at elevated risk of progression to cancer. The advancement of such a strategy could reduce surveillance frequency and overtreatment in screened populations and improve the delivery of screening to under-screened populations.

8.
Drug Des Devel Ther ; 15: 721-733, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33654378

RESUMO

RNAi therapeutics have been growing. Patisiran and givosiran, two siRNA-based drugs, were approved by the Food and Drug Administration in 2018 and 2019, respectively. However, there is rare news on the advance of miRNA drugs (another therapeutic similar to siRNA drug). Here we report the existing obstacles of miRNA therapeutics by analyses for resources available in a drug target perspective, despite being appreciated when it began. Only 10 obtainable miRNA drugs have been in clinical trials with none undergoing phase III, while over 60 siRNA drugs are in complete clinical trial progression including two approvals. We mechanically compared the two types of drug and found that their major distinction lay in the huge discrepancy of the target number of two RNA molecules, which was caused by different complementary ratios. One miRNA generally targets tens and even hundreds of genes. We named it "too many targets for miRNA effect" (TMTME). Further, two adverse events from the discontinuation of two miRNA therapeutics were exactly answered by TMTME. In summary, TMTME is inevitable because of the special complementary approach between miRNA and its target. It means that miRNA therapeutics would trigger a series of unknown and unpreventable consequences, which makes it a considerable alternative for application.

9.
Brief Bioinform ; 22(2): 1137-1149, 2021 03 22.
Artigo em Inglês | MEDLINE | ID: mdl-33675361

RESUMO

Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a severe and rapidly evolving epidemic. Now, although a few drugs and vaccines have been proved for its treatment and prevention, little systematic comments are made to explain its susceptibility to humans. A few scattered studies used bioinformatics methods to explore the role of microRNA (miRNA) in COVID-19 infection. Combining these timely reports and previous studies about virus and miRNA, we comb through the available clues and seemingly make the perspective reasonable that the COVID-19 cleverly exploits the interplay between the small miRNA and other biomolecules to avoid being effectively recognized and attacked from host immune protection as well to deactivate functional genes that are crucial for immune system. In detail, SARS-CoV-2 can be regarded as a sponge to adsorb host immune-related miRNA, which forces host fall into dysfunction status of immune system. Besides, SARS-CoV-2 encodes its own miRNAs, which can enter host cell and are not perceived by the host's immune system, subsequently targeting host function genes to cause illnesses. Therefore, this article presents a reasonable viewpoint that the miRNA-based interplays between the host and SARS-CoV-2 may be the primary cause that SARS-CoV-2 accesses and attacks the host cells.


Assuntos
/metabolismo , MicroRNAs/genética , /genética , Interações Hospedeiro-Patógeno , Humanos , /isolamento & purificação
10.
Artigo em Inglês | MEDLINE | ID: mdl-33739587

RESUMO

The development of efficient and low-cost electrocatalysts toward the oxygen evolution reaction (OER) is critical for improving the efficiency of several electrochemical energy conversion and storage devices. Here, we report an elaborate design and synthesis of porous Co-based trimetallic spinel oxide nanoboxes (NiCo2-x Fex O4 NBs) by a novel metal-organic framework engaged strategy, which involves chemical etching, cation exchange, and subsequent thermal oxidation processes. Owing to the structural and compositional advantages, the optimized trimetallic NiCo2-x Fex O4 NBs (x is about 0.117) deliver superior electrocatalytic performance for OER with an overpotential of 274 mV at 10 mA cm-2 , a small Tafel slope of 42 mV dec-1 , and good stability in alkaline electrolyte, which is much better than that of Co-based bi/monometallic spinel oxides and even commercial RuO2 .

11.
Technol Health Care ; 29(S1): 57-63, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33682745

RESUMO

BACKGROUND: Oral glucose tolerance test (OGTT) is a standard for the diagnosis of gestational diabetes mellitus (GDM). However, clinically, some cases with normal results were diagnosed as GDM in the third trimester. OBJECTIVE: To establish a risk model based on energy metabolism, epidemiology, and biochemistry that could predict the GDM pregnant women with normal OGTT results in the second trimester. METHODS: Qualitative and quantitative data were analyzed to find out the risk factors, and the binary logistic backward LR regression was used to establish the prediction model of each factor and comprehensive factor, respectively. RESULTS: The risk factors including the rest energy expenditure per kilogram of body weight, oxygen consumption per kilogram of body weight, if more than the weight gain criteria of the Institute of Medicine, the increase of body mass index between the second trimester and pre-pregnancy, and fasting blood glucose. By comparison, the comprehensive model had the best prediction performance, indicating that 85% of high-risk individuals were correctly classified. CONCLUSION: Energy metabolism, epidemiology, and biochemistry had better recognition ability for the GDM pregnant women with normal OGTT results in the second trimester. The addition of metabolic factors in the second trimester also improved the overall prediction performance.

12.
Technol Health Care ; 29(S1): 165-170, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33682756

RESUMO

BACKGROUND: The prediction of hypertensive disorders in pregnancy (HDP) mainly involves various aspects such as maternal characteristics and biomarkers. OBJECTIVE: We aimed to study the effect of the HDP prediction model with or without placental growth factor (PlGF). METHODS: This study used maternal factors and PlGF, and standardized the data uniformly. At 12-20 weeks, the comprehensive comparison of model quality with or without PlGF was conducted by logistic regression. RESULTS: The area under curve and the model accuracy of the model with PlGF were higher than those of the model without PlGF. The accuracy of the model with PlGF was above 90%. CONCLUSIONS: Adding PlGF to the model for predicting HDP improved the accuracy and effectiveness of the model. This study confirmed the predictive performance of PlGF.

13.
Technol Health Care ; 29(S1): 311-317, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33682767

RESUMO

BACKGROUND: Fetal weight is one of the important indicators for judging whether fetal growth and development are normal. Fetal weight exceeding the normal range may lead to poor delivery outcomes. OBJECTIVE: We aimed to establish a personalized fetal growth curve in order to effectively monitor fetal growth during pregnancy. Fetal weight can be monitored while fetal growth and development are assessed. METHODS: This study retrospectively analyzed the birth weight and ultrasound database of 3,093 newborns delivered at normal term. The personalized fetal growth curve model was generated based on the birth weight formula established by Gardosi combined with the proportional weight equation. RESULTS: (1) The average birth weight of the single fetus at normal term was 3,457g. (2) According to the regression results of the proportion of fetal weight in full-term pregnancy and gestational week, the proportional weight equation is Weight% = 500.9 - 51.60GA + 1.727GA2- 0.01718GA3 (GA is gestational week), R2 is 98%, P< 0.001. CONCLUSIONS: In this study, the normal birth weight of newborns and normal range of fetal weight can be estimated by using the personalized fetal growth curve model.

14.
Technol Health Care ; 29(S1): 345-350, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33682771

RESUMO

BACKGROUND: Monitoring fetal weight during pregnancy has a guiding role in prenatal care. OBJECTIVE: To establish a personalized fetal growth curve for effectively monitoring fetal growth during pregnancy. METHODS: (1) This study retrospectively analyzed the birth weight database of 2,474 singleton newborns delivered normally at term. The personalized fetal growth curve model was formed by combining the estimating birth weight of newborns with the proportional weight formula. (2) Multiple linear stepwise regression method was used to estimate the birth weight of newborns. RESULTS: (1) Delivery gestational age, weight at first visit, maternal height, pre-pregnancy body mass index, fetal sex, parity had significant effects on birth weight. Based on these parameters, the formula for calculating term optimal weight was obtained (R2= 22.8%, P< 0.001). (2) The personalized fetal growth curve was obtained according to the epidemiological factors input model of each pregnant woman. CONCLUSIONS: A model of personalized fetal growth curve can be established, and be used to evaluate fetal growth and development through estimated fetal weight monitoring.

15.
Technol Health Care ; 29(S1): 427-432, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33682779

RESUMO

BACKGROUND: Placental growth factor (PlGF), one of the biomarkers, has a certain predictive effect on hypertensive disorders in pregnancy (HDP). OBJECTIVE: To study the HDP prediction effect of different methods for variable selection and modeling for models containing PlGF. METHODS: For the model containing PlGF, the appropriate range of PlGF parameters needed to be selected. Step-logistic regression and lasso were used to compare the model effect of twice range selection. The PlGF model with good predictive effect and appropriate detecting gestational age was selected for the final prediction. RESULTS: The effect of the model containing PlGF tested at 15-16 weeks was better than the PlGF value without comprehensive screening. The sensitivity of both methods was over 92%. By comprehensive comparison, the final model of lasso method in this study was more effective. CONCLUSIONS: In this study, a variety of methods were used to screen models containing PlGF parameters. According to clinical needs and model effects, the optimal HDP prediction model with PlGF parameters in the second trimester of 15-26 weeks of pregnancy was finally selected.

16.
Medicine (Baltimore) ; 100(10): e24670, 2021 Mar 12.
Artigo em Inglês | MEDLINE | ID: mdl-33725826

RESUMO

ABSTRACT: To report the rationale, design, and baseline demographic characteristics of TuYou-County Pediatric Eye study, which mainly aimed to determine the retinal microvascular changes with optical coherence tomography angiography (OCTA) and its association with eye abnormalities in school aged children and adolescents at suburban location in Northern China.TuYou-County Pediatric Eye study was a school-based survey conducted in TuYou-County. Multi-ethnic (Mongol, Han, and Hui) participants will be followed up for 5 years. Standardized ophthalmological examinations include visual acuity, ocular biometry, retinal photography, and OCTA. A questionnaire survey was conducted to collect variables regarding to eye disease such as parental history of eye diseases, near work, outdoor activities, living and eating habits, etc.After sampling, 687 participants were eligible for investigation, and 20 students did not attend the investigation, living 667 (response rate, 97.1%) students completed questionnaires and all ocular examinations. The average age of all participants was 14.9 ±â€Š5.11.TYPE study is the first large-scale school-based multi-ethnic survey in suburban site of Northern China. Continuous identification of retinal microvascular changes with eye diseases will provide new insights into the control related diseases in school-age children and adolescents.


Assuntos
Microvasos/diagnóstico por imagem , Projetos de Pesquisa , Retina/diagnóstico por imagem , Vasos Retinianos/diagnóstico por imagem , Transtornos da Visão/diagnóstico , Adolescente , Angiografia , Criança , China/epidemiologia , Feminino , Humanos , Incidência , Masculino , Prevalência , Instituições Acadêmicas/estatística & dados numéricos , População Suburbana/estatística & dados numéricos , Tomografia de Coerência Óptica , Transtornos da Visão/epidemiologia , Acuidade Visual
17.
J Heart Lung Transplant ; 40(5): 377-386, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33648871

RESUMO

BACKGROUND: Air trapping (AT) is one of the hallmarks of allograft dysfunction after lung transplantation (LT). Inert gas‒based ventilation‒perfusion (VQ) lung scintigraphy has excellent sensitivity in the detection of AT. METHODS: We reviewed the charts of patients who underwent single or double LT between January 2012 and December 2014 (N = 193). Patients without a VQ scintigraphy at the first annual visit (n = 16) and those who did not survive till 1 year (n = 26) were excluded (final n = 151, mean age = 55.8 [SD =14] years, male = 85, female = 66). VQ scintigraphy was independently reviewed and reconciled for the presence and severity of AT by 2 investigators blinded to the clinical data (D.F.P. and D.M.). A 3-year post-transplant survival was the primary end-point. RESULTS: AT was common (n = 73, 48.3%). Patients with obstructive lung diseases as the underlying diagnosis (adjusted odds ratio [OR], 4.36, 95% CI: 1.64‒11.6; p = 0.003) and those with lower body mass index (BMI) (BMI < 25 kg/m2 and 25‒30 kg/m2; p < 0.001) had an increased risk of developing AT in the allograft. The presence of AT (adjusted OR, 2.33, 95% CI: 1.01‒5.36; p = 0.04) and peak forced expiratory volume in 1 sec (FEV1) <60% predicted during the first year after LT were independently associated with 3-year mortality. The association of AT with post-transplant mortality was the strongest among patients with BMI <30 kg/m2 and peak FEV1 <60% predicted. CONCLUSIONS: The finding of AT on VQ scintigraphy at the first annual visit after LT is independently associated with worse post-transplant mortality. The sub-group of patients who fail to achieve a peak FEV1 of 60% predicted during the first year after LT appears to be the key driver of this association.

18.
J Sep Sci ; 44(9): 1852-1865, 2021 May.
Artigo em Inglês | MEDLINE | ID: mdl-33646615

RESUMO

Low-dose methotrexate is the first-line therapy for juvenile idiopathic arthritis. In vivo, methotrexate is converted into a series of methotrexate polyglutamates whose intracellular levels contribute significantly to its efficacy and toxicity. In this study, a novel high-performance liquid chromatography-tandem mass spectrometry method was developed and validated to simultaneously determine erythrocyte methotrexate polyglutamates using stable isotope-labeled internal standards. Erythrocyte samples were precipitated by perchloric acid and then determined on an XBridge BEH C18 column with an XP vanguard precolumn in 12 min. The mobile phase consisted of 10 nM ammonium acetate (pH 10) and methanol under gradient elution. The detection was carried out in multiple reaction monitoring mode via an electrospray ionization source in positive ionization mode. The calibration curve for each metabolite was linear from 2.0 to 500.0 nmol/L (r2  > 0.99). The intraday and interday accuracies were between 93.0 and 107.0%, and the corresponding precisions were between 0.8 and 5.2%. The relative recovery ranged from 82.7 to 105.1%, and the relative matrix effect varied from 96.5 to 104.4%. The erythrocyte metabolites were stable for 30 days at -80°C. This simple and accurate method is applicable to routine monitoring of the concentration of erythrocyte methotrexate polyglutamates in patients to achieve individualized treatment.

19.
Anal Chim Acta ; 1154: 338325, 2021 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-33736808

RESUMO

New technologies permit determining metabolomic profiles of human diseases by fingerprinting metabolites levels. However, to fully understand metabolomic phenotypes, metabolite levels and turnover rates are necessary to know. Krebs cycle is the major hub of energy metabolism and cell signaling. Traditionally, 13C stable isotope labeled substrates were used to track the carbon turnover rates in Krebs cycle metabolites. In this study, for the first time we introduce H2[18O] based stable isotope marker that permit tracking oxygen exchange rates in separate segments of Krebs cycle. The chromatographic and non-chromatographic parameters were systematically tested on the effect of labeling ratio of Krebs cycle mediators to increase selectivity and sensitivity of the method. We have developed a rapid, precise, and robust GC-MS method for determining the percentage of 18O incorporation to Krebs cycle metabolites. The developed method was applied to track the cancer-induced shift in the Krebs cycle dynamics of Caco-2 cells as compared to the control FHC cells revealing Warburg effects in Caco-2 cells. We demonstrate that unique information could be obtained using this newly developed 18O-labeling analytical technology by following the oxygen exchange rates of Krebs cycle metabolites. Thus, 18O-labeling of Krebs cycle metabolites expands the arsenal of techniques for monitoring the dynamics of cellular metabolism. Moreover, the developed method will allow to apply the 18O-labeling technique to numerous other metabolic pathways where oxygen exchange with water takes place.

20.
Cell Rep ; 34(12): 108889, 2021 Mar 23.
Artigo em Inglês | MEDLINE | ID: mdl-33761348

RESUMO

Spinal cord injury (SCI) often results in spasticity. There is currently no effective therapy for spasticity. Here, we describe a method to efficiently differentiate human pluripotent stem cells from spinal GABA neurons. After transplantation into the injured rat spinal cord, the DREADD (designer receptors exclusively activated by designer drug)-expressing spinal progenitors differentiate into GABA neurons, mitigating spasticity-like response of the rat hindlimbs and locomotion deficits in 3 months. Administering clozapine-N-oxide, which activates the grafted GABA neurons, further alleviates spasticity-like response, suggesting an integration of grafted GABA neurons into the local neural circuit. These results highlight the therapeutic potential of the spinal GABA neurons for SCI.

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