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1.
Ann Transl Med ; 8(18): 1173, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33241022

RESUMO

Background: Primary bilateral macronodular adrenocortical hyperplasia (PBMAH) is a rare condition with untypical subclinical symptoms of Cushing's syndrome (CS). This study aimed to compare the clinical and pathological features of PBMAH with unilateral cortisol-secreting adrenal adenoma (UAA). Methods: We prospectively included 46 PBMAH patients and 205 UAA patients from January 2000 to February 2014. Cortisol levels and 24 hours urine free cortisol (UFC) were determined at baseline and during dexamethasone suppression test (DST) using the chemiluminescence method. Computed tomography (CT) examination of the adrenal glands was performed in all patients. For patients treated with adrenalectomy, hematoxylin, and eosin, staining was performed for pathological examination. Results: The proportion of patients with autonomous cortisol secretion was significantly higher in PBMAH patients (39.1%) than UAA patients (6.8%). The PBMAH patients showed significantly lower levels of basal cortisol, low dose dexamethasone suppressed cortisol, and high dose dexamethasone suppressed cortisol than the UAA patients (452.6±183.3 vs. 578.7±166.4 nmol/L, P=0.003; 394.5±298.9 vs. 549.2±217.7 nmol/L, P=0.002; 397.3±282.3 vs. 544.3±187.6 nmol/L, P=0.003). Similarly, the PBMAH patients had significantly lower levels of basal 24 hours UFC, low dose dexamethasone suppressed 24 hours UFC, and high dose dexamethasone suppressed 24 hours UFC than the UAA patients (1,144.4±1,048.1 vs. 1,674.9±1,520.4 nmol/24 h, P=0.032; 1,157.3±1,483.5 vs. 1,940.1±1,360.9 nmol/24 h, P=0.003; 1,256.4±1,767.0 vs. 1,969.9±1,361.7 nmol/24 h, P=0.011). Conclusions: PBMAH is often associated with atypical CS symptoms. The clinical and imaging features of PBMAH are useful for the differential diagnosis of this disease.

2.
Drug Des Devel Ther ; 14: 3177-3188, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32821085

RESUMO

Background: Melatonin (MT) has potential protective effect on cerebral ischemia-reperfusion injury (CIRI), but its underlying regulatory mechanism has not been identified. Purpose: This study aimed to explore the role of miR-26a-5p-neuron-restrictive silencing factor (NRSF/REST), Janus kinase-2 (JAK2)-signal transducer and activator of transcription-3 (STAT3) pathway in the protection mechanism of MT against CIRI in vivo and in vitro. Methods: Sprague Dawley rats were induced with ischemia-reperfusion (IR) in vivo model; PC12 cells were induced with oxygen-glucose deprivation/reperfusion (OGD/R) in vitro model; and MT intervention was conducted before the model was established. The effect of MT on autophagy factors (LC3II/LC3I, P62), inflammatory factors (TNF-α, IL-6, IL-10) and oxidative stress indexes (MDA, GSHPx, SOD) was explored, and then the above three indexes were determined by real-time quantitative PCR, ELISA, and detection kit corresponding to oxidative stress indexes. The neuroprotective effect of MT pretreatment on brain IR injury was evaluated by neurological deficit scores and TUNEL method. The levels of miR-26a-5p and NRSF were detected by real-time quantitative PCR and Western blot, and the interaction between them was evaluated by dual luciferase report. The role of JAK2-STAT3 pathway in MT protection mechanism was verified by pathway blocker (AG490) and Western blot. Results: MT pretreatment can significantly reduce neurological deficit score and neuronal apoptosis, inhibit CIRI autophagy, inflammation and oxidative stress in vivo and in vitro, reduce LC3II/LC3I, TNF-α, IL-6, MDA and increase P62, IL-10, GSHPx, SOD. Further analysis identifies that downregulating miR-26a-5p or upregulating NRSF can eliminate the protective effect of MT, and NRSF is the direct target of miR-26a-5p. The protective effect of MT can also be eliminated under AG490 intervention. Conclusion: MT plays a protective role by regulating miR-26a-5p-NRSF and JAK2-STAT3 pathway to improve CIRI autophagy, inflammation and oxidative stress.

3.
Artigo em Inglês | MEDLINE | ID: mdl-32760349

RESUMO

The anti-thyroglobulin antibody (TgAb) has been suggested to be more common in patients with papillary thyroid cancer (PTC). Here, we performed a retrospective study investigated the correlation between TgAb level and PTC in Chinese patients between 2011 and 2015. Patients with goiter who underwent thyroidectomy and received a confirmed pathological diagnosis were enrolled into the study. Clinical characteristics and preoperative thyroglobulin antibody (TgAb) level data were collected from all enrolled patients. Based on the preoperative TgAb test results, patients were divided into a TgAb negative (TgAb-) group (<60 IU/mL) and a TgAb positive (TgAb+) group (≧60 IU/mL). Of the 4,046 patients, 671 patients were TgAb+ while 3,375 patients were TgAb-. There were 535 (79.7%) patients with PTC in the TgAb+ group, and 2,154 (63.8%) patients with PTC in the TgAb- group. The prevalance of PTC was significantly higher in TgAb+ patients than in TgAb- patients. TgAb+ patients were stratified into four groups based on the TgAb titer. The prevalence of PTC did not increase with TgAb titer. No significant difference in TgAb level was noted in patients with different clinicopathologies, including TNM stage, lymph node metastasis, and multifocal carcinoma. Regression analysis suggested a higher risk of PTC malignancy among TgAb+ patients. Preoperative TgAb level ≥60 IU/mL might be associated with a higher risk of PTC. However, there was no titer-dependent association between elevated TgAb titer and PTC malignancy.

4.
Clin Rheumatol ; 39(12): 3757-3767, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32458241

RESUMO

INTRODUCTION: Our previous cross-sectional study revealed the association between neck circumference (NC) and hyperuricemia. This study aimed to further investigate the longitudinal association between NC and hyperuricemia and to compare the strengths of longitudinal association of NC and waist circumference (WC) with hyperuricemia. METHODS: A total of 4383 subjects without hyperuricemia at baseline were included. Multivariable linear regression was used to assess the association between baseline anthropometric indices and uric acid level at follow-up. Cox regression was used to assess the relationship between baseline anthropometric indices and the risk of future hyperuricemia. A receiver-operating characteristic curve was used to compare the predictive ability of baseline anthropometric indices for future hyperuricemia. RESULTS: Among women, only baseline NC was positively, linearly associated with uric acid level at follow-up (ß = 1.75) and risk of future hyperuricemia (ptrend < 0.05); risk of hyperuricemia in the third and fourth quartile groups of baseline NC significantly increased, compared with the first quartile group of baseline NC (HR = 1.48 and 1.81, respectively). Among men, neither NC nor WC was associated with hyperuricemia after adjusting for confounder factors. The area under the curve of baseline NC and WC for hyperuricemia was comparable in both genders (0.606 for NC and 0.599 for WC in women, 0.578 for NC and 0.602 for WC in men). CONCLUSIONS: For women, only baseline NC is an independent risk factor of hyperuricemia within 3 years. NC and WC have a similar 3-year predictive ability for hyperuricemia in both genders. Key Points • This is the first study to assess the longitudinal association between neck circumference and hyperuricemia. • According to the results of this study, we propose for the first time that neck circumference is an independent risk factor for hyperuricemia in women in the future, rather than waist circumference. • We found for the first time that neck circumference and waist circumference have similar predictive ability for future hyperuricemia in both genders.

5.
Biotechnol J ; 15(8): e1900565, 2020 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-32170810

RESUMO

Chinese hamster ovary (CHO) cells are currently the primary host cell lines used in biotherapeutic manufacturing of monoclonal antibodies (mAbs) and other biopharmaceuticals. Cellular energy metabolism and endoplasmic reticulum (ER) stress are known to greatly impact cell growth, viability, and specific productivity of a biotherapeutic; but the molecular mechanisms are not fully understood. The authors previously employed multi-omics profiling to investigate the impact of a reduction in cysteine (Cys) feed concentration in a fed-batch process and found that disruption of the redox balance led to a substantial decline in cell viability and titer. Here, the multi-omics findings are expanded, and the impact redox imbalance has on ER stress, mitochondrial homeostasis, and lipid metabolism is explored. The reduced Cys feed activates the amino acid response (AAR), increases mitochondrial stress, and initiates gluconeogenesis. Multi-omics analysis reveals that together, ER stress and AAR signaling shift the cellular energy metabolism to rely primarily on anaplerotic reactions, consuming amino acids and producing lactate, to maintain energy generation. Furthermore, the pathways are demonstrated in which this shift in metabolism leads to a substantial decline in specific productivity and altered mAb glycosylation. Through this work, meaningful bioprocess markers and targets for genetic engineering are identified.

6.
J Int Med Res ; 48(3): 300060519887832, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31779500

RESUMO

OBJECTIVE: Autoimmune hypothalamitis (AHT) is a rare inflammatory disorder that involves the hypothalamus. It remains unclear whether autoimmune hypophysitis (AH) and AHT represent different diseases or different aspects of the same disease. Thus, further investigation of AHT is required. METHODS: A retrospective review of medical and pathological records of AHT patients from the Chinese PLA General Hospital were examined from January 1, 2005 to May 1, 2017. Clinical data, treatments, and outcomes were investigated. RESULTS: Five female patients were identified (median age, 42.6 years). Symptoms included central diabetes insipidus, hypopituitarism, hyperprolactinemia, headache, and hypothalamic syndrome. The following hormonal deficits were noted: follicle-stimulating hormone, luteinizing hormone, adrenocorticotropic hormone, thyroid stimulating hormone, and growth hormone. One patient underwent high-dose methylprednisolone pulse treatment (HDMPT) and azathioprine plus intensity modulated radiation therapy (IMRT), and two patients underwent HDMPT and two rounds of replacement therapy. During follow-up, one patient died because of non-compliance with therapy and the others were in remission or they recovered. CONCLUSIONS: AHT had similar MRI results, pathology, and treatment compared with AH. Thus, it may be a subtype of AH, and AHT patients may also show hypothalamic syndrome.

7.
Stem Cells Int ; 2019: 8628027, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31781248

RESUMO

Nonalcoholic fatty liver disease (NAFLD) is increasingly common among patients with type 2 diabetes mellitus (T2DM). The two conditions can act synergistically to produce adverse outcomes. However, the therapeutic options for patients with NAFLD and T2DM are currently limited. Human umbilical cord-derived mesenchymal stem cells (UC-MSCs) have shown therapeutic potential for diabetes and hepatic disorders such as liver cirrhosis and fulminant hepatic failure. The present study is aimed at investigating the effect of human UC-MSCs on a mouse model of NAFLD and T2DM, characterized by obesity-induced hyperglycaemia, dyslipidaemia, hepatic steatosis, and liver dysfunction. Thirty-week-old male C57BL/6 db/db mice were infused with human UC-MSCs or phosphate-buffered saline (PBS) via the tail vein once a week for six weeks. Age-matched male C57BL/6 wild-type db/+ mice were used as controls. Body weight and random blood glucose were measured every week. One week after the sixth infusion, intraperitoneal glucose tolerance tests and insulin tolerance tests were performed and the blood and liver were harvested for biochemical and histopathological examinations. Quantitative real-time reverse transcriptase polymerase chain reaction (qRT-PCR), immunofluorescence staining, and western blot were performed to monitor the expression of the lipid metabolism- and regulatory pathway-related genes. UC-MSC infusions significantly ameliorated hyperglycaemia, attenuated the elevation of hepatic transaminases, and decreased lipid contents, including triglyceride, total cholesterol, and low-density lipoprotein cholesterol. Moreover, histological lesions in the liver diminished markedly, as evidenced by reduced lipid accumulation and attenuated hepatic steatosis. Mechanistically, UC-MSCs were found to regulate lipid metabolism by increasing the expression of fatty acid oxidation-related genes and inhibiting the expression of lipogenesis-related genes, which were associated with the upregulation of the HNF4α-CES2 pathway. Our results demonstrate that human UC-MSCs can ameliorate NAFLD and reverse metabolic syndrome in db/db mice. Thus, UC-MSCs may serve as a novel therapeutic agent for T2DM patients with NAFLD.

8.
Adv Exp Med Biol ; 1140: 225-236, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31347050

RESUMO

Selection of high-producing lead and backup cell lines with high-fidelity primary structure is a major goal of cell line development of protein therapeutics. Conventional techniques for sequence variant analysis, such as mass spectrometry (MS) and next-generation sequencing (NGS) have limitations on the sample number and turnaround time, thus often are only applied at the final stages of development, where an undesired lead or backup clone could cause a significant delay in project timeline. Here we presented a high-throughput (HT) peptide mapping workflow which can be applied at early stages of cell line selection for testing of a batch of 30-40 clones within 2-week turnaround while reporting valuable information on sequence variants and posttranslational modifications (PTMs). The successful application of this workflow was demonstrated for two mAb programs. Multiple clones were removed from a total of 33 mAb-1 clones using various criteria: nine clones contained at least one >1% upregulated unknown peptide ions, 11 clones contained at least eight >0.1% upregulated unknowns, and six clones contained upregulated critical PTMs. For mAb-2, light chain (LC) sequence extension of approximately 30 amino acids were detected in 6 out of 36 clones at levels up to 11%. Besides, a Q to H mutation at ~30% was detected in the heavy chain (HC) of a single clone. Q to H mutation has mass change of 9.00 Da and failed to be detected by intact mass analysis. Rapid PTM quantitation also facilitated the selection of clones with desirable quality attributes, such as N-glycan profile. Hence, we demonstrated a risk-reducing strategy where abnormal clones could be detected at earlier stages of cell line selection, which should result in reduced and more predictable timeline of cell line development.


Assuntos
Anticorpos Monoclonais/química , Sequenciamento de Nucleotídeos em Larga Escala , Mapeamento de Peptídeos , Processamento de Proteína Pós-Traducional , Animais , Células CHO , Cricetinae , Cricetulus , Espectrometria de Massas
9.
Diabetes Ther ; 10(4): 1391-1405, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31222594

RESUMO

INTRODUCTION: The efficacy and safety of vildagliptin alone or with metformin is well established by randomized trials, but it is unknown whether it can be extrapolated to the real-world setting in Chinese patients with type 2 diabetes mellitus (T2DM). This study aimed to assess the effectiveness and safety of vildagliptin add-on to metformin versus dual oral antidiabetes drug (OAD), non-vildagliptin combination therapies in real-world Chinese patients with T2DM. METHODS: The China Prospective Diabetes Study was a post-marketing, prospective, multicenter, observational, real-world study conducted in 52 centers. Patients inadequately controlled with OAD monotherapy and who initiated vildagliptin add-on to metformin (VM cohort) or two OADs other than vildagliptin (comparator cohort) were included for the present analysis. The composite primary endpoint was glycated hemoglobin (HbA1c) < 7% and without tolerability events (hypoglycemia, weight gain ≥ 3%, or discontinuation due to gastrointestinal events) at 12 months. Secondary endpoints included change in HbA1c from baseline, subgroup analysis, and tolerability. Propensity score matching analysis was performed to adjust for baseline covariates imbalance (body mass index (BMI) and HbA1c). RESULTS: A total of 604 patients received VM and 670 received comparator therapy. Patients who received VM were younger, more obese, and had a higher baseline HbA1c and a shorter duration of T2DM. After propensity score matching, there were 530 patients per cohort. After 12-month treatment, the success rates of the composite primary endpoint were 50.9% and 33.0% in the VM and comparator cohorts, respectively (P < 0.001; odds ratio = 2.10, 95% confidence interval (CI) 1.64-2.70). Furthermore, the success rates of the composite endpoint were higher with VM across geographic area, BMI, and baseline HbA1c subgroups. Fewer tolerability events occurred in the VM cohort versus the comparator cohort (8.3% vs. 16.2%, P < 0.001; relative risk = 0.51, 95% CI 0.36-0.72). CONCLUSION: Compared with dual OAD non-vildagliptin combination therapies, vildagliptin add-on to metformin is effective and safe to achieve glycemic control in Chinese patients with T2DM. FUNDING: Novartis.

10.
Dis Aquat Organ ; 133(3): 195-205, 2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-31187734

RESUMO

The lined seahorse Hippocampus erectus is an economically important aquaculture species; however, the low survival rate of juvenile seahorses severely restricts their large-scale cultivation. According to previous research, dead juvenile seahorses (4-6 cm) showed symptoms of suspected enteritis, including abdominal depression, raised cloaca, partial hepatic congestion, and yellow sticky liquid filling the intestine. Here, we isolated a Gram-negative bacterium from diseased juvenile seahorses and tentatively named the strain HEL-5. Healthy juvenile seahorses were then challenged with the strain through intraperitoneal injection, with results confirming that HEL-5 was pathogenic for seahorses at a median lethal dose of 5.81 × 105 CFU g-1 fish weight. Based on morphological observations, biochemical characteristics, and sequence analysis of 16S rRNA and housekeeping genes (gyrB, ftsZ, and gapA), we identified HEL-5 as Vibrio tubiashii. Histopathological observations revealed that V. tubiashii was capable of causing lytic necrosis of hepatocytes and forming obvious necrotic foci, and renal pathology was characterized by tubular collapse and tubular epithelial-cell shedding into the lumen accompanied by a large number of inflammatory cells infiltrating the tissues of the intestines and kidneys. Antimicrobial-susceptibility testing showed that the strain was highly sensitive to macrolides, chloramphenicol, sulfonamides, aminoglycosides, and cephalosporins. These findings represent the first report of isolation of V. tubiashii from diseased juvenile seahorses and provide a foundation for the prevention and treatment of vibrio disease in seahorse aquaculture.


Assuntos
Smegmamorpha , Vibrio , Animais , Filogenia , RNA Ribossômico 16S
11.
Sichuan Da Xue Xue Bao Yi Xue Ban ; 50(2): 252-255, 2019 Mar.
Artigo em Chinês | MEDLINE | ID: mdl-31106548

RESUMO

OBJECTIVE: To ivestigate the effect of daily walking number on clinical, inflammatory and nutritional indexes for patients with chronic kidney disease. METHODS: 90 non-dialysis patients with chronic kidney disease were selected and randomly divided equally into three groups, for groups A, B and C. 30 patients for group A were asked for the number of daily walk should less than 5 000 steps, while group B patients were asked for 5 000-9 999 steps of walk and group C for more than 10 000 steps. Basic treatments were given for each group of patients and basic information, clinical, inflammatory and nutritional datas of patients were collected. RESULTS: 87 patients with chronic kidney disease completed the study, with baseline data between group A, B, C (n=30, 29, 28) consistently. After 3 months of exercise, there were no significant changes on blood lipids, serum uric acid and parathyroid hormone (PTH) for three groups, with serum creatinine of three groups stably. However, in group C, hemoglobin, ferritin, transferrin saturation were found increased significantly (P<0.05). For nutritional indexes, increasing of albumin and prealbumin level were found in three groups, while significant differences were only found in group B and C (P<0.05) and group C increased most. There were no significant change on body mass index (BMI), upper arm skinfold thickness and SGA score in three groups. For inflammatory data, significant decrease of C-reactive protein (CRP) and interleukin-6 (IL-6) were only seen in group C (P<0.05). CONCLUSION: Walking does not increase the burden of the kidney, but can improve the nutrition and clinical indicators of patients, reduce inflammation.


Assuntos
Inflamação/terapia , Estado Nutricional , Insuficiência Renal Crônica/sangue , Caminhada , Proteína C-Reativa/análise , Humanos , Inflamação/sangue , Interleucina-6/sangue , Lipídeos/sangue , Hormônio Paratireóideo/sangue , Diálise Renal , Ácido Úrico/sangue
12.
Artigo em Inglês | MEDLINE | ID: mdl-31040822

RESUMO

Objective: To investigate the role of PSMA in the differential diagnosis of adrenocortical carcinoma samples (ACCs) and adrenocortical adenoma samples (ACAs), to validate the prognostic role of PSMA in patients with ACCs, and to explore the possibility that this marker can differentiate localized ACCs from adrenal metastases from other sites. Methods: PSMA protein expression in tissue samples from 50 ACCs, 90 ACAs (including 20 from patients who presented with Cushing's syndrome, 20 aldosterone-producing adenomas and 50 non-functional tumors) and 10 tissues that were metastases from other primary sites was assessed by immunohistochemistry. The clinical and pathological characteristics were compared, the intensity and density were analyzed, and the prognostic role was evaluated. Results: The analysis of clinical and pathological features revealed that the size of ACCs was greater than that of benign tissues and the ACC patients were older than the ACA patients (p < 0.01). The percentage of PSMA-positive vessels, the mean intensity and the degree of staining density were found to be significantly lower in ACAs than in ACCs (p < 0.01). In these 140 samples, 60% of the ACCs were grouped in the positive category. The samples were negative for metastases that were from other primary sites. The ENSAT stage and Ki-67 were correlated with PSMA expression. The survival distribution revealed that high PSMA expression did not show any prognostic relevance in the current ACCs series. Those samples with a score of > 3.5 were 75 times more likely to be malignant (OR = 75). We established a cut-off score of 3.5 (p < 0.05), which had 46% sensitivity and 99% specificity. Paralleling PSMA and Ki-67 maximized the area under the curve, with 72% sensitivity and 100% specificity. Conclusions: Our results strongly confirm that PSMA is helpful for distinguishing benign from malignant tumors and that its high expression levels correlate with a high ENSAT stage and high proliferation. The combination of PSMA and Ki-67 can be particularly useful. Furthermore, PSMA might be a useful tool for the identification of localized adrenal carcinoma and metastatic carcinoma.

13.
Aging (Albany NY) ; 11(10): 3080-3093, 2019 05 22.
Artigo em Inglês | MEDLINE | ID: mdl-31118311

RESUMO

Gibberellin, a plant growth regulator, is widely used to increase the shelf life and quality of fruits and vegetables. In this study, human semen samples were exposed to different concentrations of gibberellin, which reduced spermatozoa motility in vitro. Gibberellin exposure also increased levels of reactive oxygen species and the protein levels of apoptosis markers in human sperm. Gibberellin inhibited the activity of Na+/K+-adenosine triphosphatase (ATPase) and Ca2+-ATPase, which maintain the stability of ions inside and outside the membranes of spermatozoa. Moreover, gibberellin exposure suppressed adenosine triphosphate production and reduced the protein levels of adenosine triphosphate synthases, which may have induced the protein expression of adenosine 5'-monophosphate-activated protein kinase (AMPK) and its phosphorylated form. These results suggest that gibberellin reduces human sperm motility in vitro by increasing reactive oxygen species levels and reducing ATPase activity, which may upregulate AMPK and consequently reduce the fertilization potential of spermatozoa.

14.
World J Clin Cases ; 7(8): 961-971, 2019 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-31119141

RESUMO

BACKGROUND: Adrenocorticotropic hormone (ACTH)-independent Cushing's syndrome (CS) is mostly due to unilateral tumors, with bilateral tumors rarely reported. Its common causes include primary pigmented nodular adrenocortical disease, ACTH-independent macronodular adrenal hyperplasia, and bilateral adrenocortical adenomas (BAAs) or carcinomas. BAAs causing ACTH-independent CS are rare; up to now, fewer than 40 BAA cases have been reported. The accurate diagnosis and evaluation of BAAs are critical for determining optimal treatment options. Adrenal vein sampling (AVS) is a good way to diagnose ACTH-independent CS. CASE SUMMARY: A 31-year-old woman had a typical appearance of CS. The oral glucose tolerance test showed impaired glucose tolerance and obviously increased insulin and C-peptide levels. Her baseline serum cortisol and urine free cortisol were elevated and did not show either a circadian rhythm or suppression with dexamethasone administration. The peripheral 1-deamino-8-D-arginine-vasopressin (DDVAP) stimulation test showed a delay of the peak level, which was 1.05 times as high as the baseline level. Bilateral AVS results suggested the possibility of BAAs. Abdominal computed tomography showed bilateral adrenal adenomas with atrophic adrenal glands (right: 3.1 cm × 2.0 cm × 1.9 cm; left: 2.2 cm × 1.9 cm × 2.1 cm). Magnetic resonance imaging of the pituitary gland demonstrated normal findings. A left adenomectomy by retroperitoneoscopy was performed first, followed by resection of the right-side adrenal mass 3 mo later. Biopsy results of both adenomas showed cortical tumors. Evaluations of ACTH and cortisol showed a significant decrease after left adenomectomy but could still not be suppressed, and the circadian rhythm was absent. Following bilateral adenomectomy, this patient has been administered with prednisone until now, all of her symptoms were alleviated, and she had normal blood pressure without edema in either of her lower extremities. CONCLUSION: BAAs causing ACTH-independent CS are rare. AVS is of great significance for obtaining information on the functional state of BAAs before surgery.

15.
J Chromatogr B Analyt Technol Biomed Life Sci ; 1114-1115: 93-99, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30939413

RESUMO

Characterization of free thiol variants in antibody therapeutics is important for biopharmaceutical development, as the presence of free thiols may have an impact on aggregate formation, structural and thermal stability, as well as antigen-binding potency of monoclonal antibodies. Most current methods for free thiol quantification involve labeling of free thiol groups by different tagging molecules followed by UV, fluorescence or mass spectrometry (MS) detection. Here, we optimized a label-free liquid chromatography (LC)-UV/MS method for free thiol quantification at a subunit level and compared this method with two orthogonal and conventional approaches, Ellman's assay and peptide mapping with differential alkylation. This subunit unit approach was demonstrated to be able to provide domain-specific free thiol quantification and comparable results with labeling approaches, using a relatively simple and efficient workflow.


Assuntos
Anticorpos Monoclonais/análise , Anticorpos Monoclonais/química , Cromatografia Líquida/métodos , Compostos de Sulfidrila/análise , Compostos de Sulfidrila/química , Animais , Ácido Ditionitrobenzoico , Imunoglobulina G/análise , Imunoglobulina G/química , Espectrometria de Massas/métodos , Mapeamento de Peptídeos , Reprodutibilidade dos Testes
16.
BMJ Open ; 9(1): e023334, 2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30782703

RESUMO

OBJECTIVES: This study aims to summarise the features and trends of thyroid carcinoma in the past two decades in China. DESIGN, SETTING AND PARTICIPANTS: Clinical data obtained from 10 798 patients treated by thyroidectomy from 1994 to 2015 at the Department of General Surgery of the People's Liberation Army General Hospital, Beijing, China were retrospectively analysed. OUTCOME MEASURES: Incidence and histopathological features of thyroid cancer were compared and the risk factors for local lymph node metastasis analysed. RESULTS: Our data indicated a significant increase in the detection of thyroid cancer (from 16.8% to 69.8%, p<0.01). Among the 5235 thyroid cancer cases, papillary thyroid carcinoma (PTC) was the most common histotype, accounting for 95.1% of all malignancies over the 22-year period. Among the 4979 PTCs, micro-PTCs (mPTC) with the largest diameter ≤10 mm has gradually become the dominant form, and its percentage in PTCs has increased from 13.3% in the biennial period of 1994-1995 to 51.2% in 2010-2011. Furthermore, the size of the tumour has decreased significantly from 2.3±1.1 cm in 1994 to 1.2±0.9 cm in the largest diameter (p<0.01), while the average age at diagnosis and female dominance remained unchanged during the period. Logistic regression showed that tumour nodules>1 cm and male gender were the main risk factors for local lymph node metastasis (LNM), whereas patients over 45 years had lower risk. CONCLUSIONS: During the 22-year period, an increased detection of thyroid cancer, particularly mPTC, was found while the occurrence of LNM decreased. Our results suggest that the current preoperative diagnosis and risk stratification are adequate, supporting the published guidelines for the diagnosis of thyroid cancer.


Assuntos
Câncer Papilífero da Tireoide/epidemiologia , Neoplasias da Glândula Tireoide/epidemiologia , Tireoidectomia/estatística & dados numéricos , Adulto , Fatores Etários , Pequim/epidemiologia , Estudos Transversais , Bases de Dados Factuais , Humanos , Incidência , Metástase Linfática/patologia , Pessoa de Meia-Idade , Estudos Retrospectivos , Fatores Sexuais , Câncer Papilífero da Tireoide/patologia , Câncer Papilífero da Tireoide/cirurgia , Neoplasias da Glândula Tireoide/patologia , Neoplasias da Glândula Tireoide/cirurgia
17.
Anal Biochem ; 571: 25-36, 2019 04 15.
Artigo em Inglês | MEDLINE | ID: mdl-30653944

RESUMO

Monoclonal antibody (mAb), one of the major types of therapeutic proteins in the pharmaceutical industry, is predominantly manufactured using mammalian cell culture [1]. Oxidative stress, potentially present during cell culture process, may increase the protein carbonyl content in the mAb product, which was reported to positively correlate with aggregate burst rate during storage [2]. In order to monitor carbonyl content during mAb process development, we developed a high-throughput screening method for therapeutic mAbs using size-exclusion chromatography followed by ultraviolet and fluorescence detection (SEC-UV/FL), optimized from a fluorescein thiosemicarbazide (FTC) semi-microplate method. The method demonstrated a good correlation with conventional ELISA assay and FTC-based fluorometric semi-microplate method with improved throughput and precision. The method was successfully applied in three case studies to improve our understanding of mAb carbonylation, including the impact of metal-catalyzed oxidation on an IgG4 mAb, comparison of carbonyl content between several mAbs expressed by CHO cell culture with human serum antibody pool, as well as the surface charge property of carbonylated mAb assessed by ion-exchange chromatography.


Assuntos
Anticorpos Monoclonais/sangue , Ensaios de Triagem em Larga Escala , Animais , Células CHO , Células Cultivadas , Cromatografia em Gel , Cricetulus , Fluorescência , Humanos , Carbonilação Proteica , Raios Ultravioleta
18.
Diagn Cytopathol ; 47(3): 187-193, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30499203

RESUMO

BACKGROUND: Thyroid nodules are very common. Ultrasound (US) and fine needle aspiration (FNA) are both integral in cancer screening. This study investigated the concordance between thyroid nodule sizes measured by US and gross pathologic examination and their relationship with malignancy. METHODS: A retrospective design was used to select consecutive patients with proven carcinoma of the thyroid. The number and maximum diameter of nodules, rates and types of malignancy, as well as predictors of malignancy were determined. RESULTS: The 10 944 patients examined had 15 283 thyroid nodules, 44.6% of which were malignant. Of the 4449 nodules sampled by FNA and the 8748 not sampled by FNA, 76.5% and 30.5% were malignant, respectively. The sensitivity, specificity, positive and negative predictive values (NPVs), and overall accuracy of FNA based on final pathology were 97.9%, 96.3%, 98.8%, 93.5%, and 97.5%, respectively. Nodule sizes determined by US were comparable with most nodules having either the same size range (n = 2959, 77.7%) or differing only by one size range (n = 770, 20.5%). CONCLUSIONS: Thyroid nodule size is inversely related to malignancy risk, as larger nodules have lower malignancy rates. Nodule size estimated by US shows relatively good correlation with final pathologic size. However, thyroid nodules should undergo FNA regardless of size. If the FNA is not benign, nodule size should influence therapeutic decision making.


Assuntos
Nódulo da Glândula Tireoide/diagnóstico por imagem , Nódulo da Glândula Tireoide/patologia , Adulto , Idoso , Biópsia por Agulha Fina , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Sensibilidade e Especificidade , Ultrassonografia
19.
Biotechnol J ; 14(4): e1800352, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30485675

RESUMO

There is continual demand to maximize CHO cell culture productivity of a biotherapeutic while maintaining product quality. In this study, a comprehensive multi-omics analysis is performed to investigate the cellular response to the level of dosing of the amino acid cysteine (Cys) in the production of a monoclonal antibody (mAb). When Cys feed levels are insufficient, there is a significant decrease in protein titer. Multi-omics (metabolomics and proteomics, with support from RNAseq) is performed over the time course of the CHO bioprocess producing an IgG1 mAb in 5 L bioreactors. Pathway analysis reveals that insufficient levels of Cys in the feed lead to Cys depletion in the cell. This depletion negatively impacts antioxidant molecules, such as glutathione (GSH) and taurine, leading to oxidative stress with multiple deleterious cellular effects. In this paper, the resultant ER stress and subsequent apoptosis that affects cell viability and viable cell density has been considered. Key metabolic enzymes and metabolites are identified that can be potentially monitored as the process progresses and/or increased in the cell either by nutrient feeding or genetic engineering. This work reinforces the centrality of redox balance to cellular health and success of the bioprocess as well as the power of multi-omics to provide an in-depth understanding of the CHO cell biology during biopharmaceutical production.


Assuntos
Anticorpos Monoclonais/biossíntese , Técnicas de Cultura de Células , Meios de Cultura/farmacologia , Cisteína/farmacologia , Animais , Anticorpos Monoclonais/efeitos dos fármacos , Reatores Biológicos , Células CHO , Sobrevivência Celular/efeitos dos fármacos , Cricetinae , Cricetulus , Cisteína/química , Estresse do Retículo Endoplasmático/efeitos dos fármacos , Glutationa/química , Imunoglobulina G/biossíntese , Imunoglobulina G/química , Estresse Oxidativo/efeitos dos fármacos , Proteômica , Taurina/química
20.
Cell Physiol Biochem ; 51(6): 2591-2603, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-30562745

RESUMO

BACKGROUND/AIMS: Chronic inflammation contributes to the development of type 2 diabetes mellitus by targeting the insulin receptor substrate protein-1 (IRS-1) signaling pathway. Previous studies showed that Leukemia related protein 16 (LRP16) reduced insulin stimulated glucose uptake in adipocytes by impairing the IRS-1 signaling pathway. We explored the mechanism by which LRP16 promotes the inflammatory response. METHODS: We screened LRP16 induced proteins in the lipopolysaccharide (LPS)-stimulated inflammatory response using liquid chromatography-mass spectrometry (LC-MS) and analyzed the potential biological functions of these proteins using online bioinformatics tools. mRNA expression and protein expression of target genes were measured by real time PCR and Western blot, respectively. RESULTS: A total of 390 differentially expressed proteins were identified. The mitogen-activated protein kinase (MAPK) signaling pathway was the primary activated pathway in LRP16-expressing cells. Overexpression of LRP16 activated ERK1/2 and Rac1, which are two key players related to the MAPK signaling pathway. Furthermore, knock down of endogenous LRP16 by RNA interference (RNAi) reduced Rac1 expression, ERK activation, and inflammatory cytokine expression in human adipocytes stimulated by LPS. The stimulatory effect of LRP16 was diminished by suppressing Rac1 expression and treating the cells with the ERK specific inhibitor, PD98059. CONCLUSION: These findings revealed the functions of LRP16 in promoting the inflammatory response through activating the Rac1-MAPK1/ERK pathway in human adipocytes.


Assuntos
Inflamação/imunologia , Lipopolissacarídeos/imunologia , Sistema de Sinalização das MAP Quinases , Proteínas de Neoplasias/imunologia , Transdução de Sinais , Proteínas rac1 de Ligação ao GTP/imunologia , Adipócitos , Linhagem Celular , Diabetes Mellitus Tipo 2/imunologia , Humanos
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