Germline PRKACA amplification-associated primary pigmented nodular adrenocortical disease: a case report and literature review

SUMMARY Primary pigmented nodular adrenocortical disease (PPNAD) is a rare adrenocorticotropin hormone (ACTH)-independent Cushing's syndrome (CS). Pediatric patients with PPNAD typically have unusual skin lesions and slow growth with unknown causes. We present a case of a female Chinese patient with PPNAD caused by the germline PRKACA gene copy number gain of chromosome 19. The patient initially presented with kidney stones, short stature, and obesity. After further testing, it was discovered that the patient had diabetes, mild hypertension, low bone mass, a low ACTH level, and hypercortisolemia, and neither the low-dose or high-dose dexamethasone suppression test was able to inhibit hematuric cortisol, which paradoxically increased. PPNAD was pathologically diagnosed after unilateral adrenalectomy. Chromosome microarrays and whole exon sequencing analyses of the peripheral blood, as well as testing of sectioned adrenal tissue, showed a rise in the copy number of the duplication-containing PRKACA gene on chromosome 19p13.13p13.12, a de novo but not heritable gene defect that causes disease. The clinical signs and symptoms supported the diagnosis of Carney complex (CNC). One significant mechanism of CNC pathogenesis may be the rise in germline PRKACA copy number of chromosome 19. When assessing PPNAD patients for CNC, the possibility of PRKACA gene amplification should be considered. The effect of PRKACA gene amplification on the clinical manifestations of CNC needs to be confirmed by more cases.

Effect of atorvastatin on autophagy and cholesterol level in foam cells by activating transcription factor EB / 医学研究生学报

ObjectiveRecent studies revealed that the transcription factor EB (TFEB) plays an important role in regulating autophagy, reducing intracellular lipids, and inhibiting atherosclerosis. This study aims to explore the effects of atorvastatin (ATV) on autophagy and cholesterol levels of foam cells by activating TFEB.MethodsHuman mononuclear cell line THP-1 was cultured in vitro and induced to differentiate into macrophages using phorbol ester. Oxidized low-density lipoprotein (oxLDL) was added to macrophages, which were induced for 48 hours to establish a foam cell model. The experiment was divided into four groups: blank group, model group (oxLDL group), oxLDL+ Chloroquine (CQ) group, oxLDL+ ATV group, and oxLDL+CQ+ ATV group. Cells in each group were treated with drugs for 48 h. The toxicity of ATV and chloroquine on the cells was detected by the CCK8 method. Oil red O staining was used to test the level of lipid droplets. Oxidase method was used to detect levels of intracellular free cholesterol (FC), total cholesterol (TC) and others related to. Cholesterol efflux fluorescence analysis was used to determine the cholesterol efflux rate of the cells. Expression of I, P62, TFEB, LAMP 1 protein was determined by Western blot.ResultsThe results of the CCK8 method showed that the cell survival rate decreased significantly with the increase of ATV and CQ concentrations (P<0.01). Compared to the blank group, the levels of lipid droplets, FC, TC, and CE/TC in the model group significantly increased (P<0.05), and the cholesterol outflow rate significantly decreased (P<0.05). Compared to the model group, the intracellular lipid droplets, FC, TC, and CE/TC levels in the oxLDL+CQ group elevated significantly (P<0.05), while the cholesterol outflow rate decreased significantly (P<0.05). The intracellular lipid droplets, FC, TC, and CE/TC contents in the oxLDL+ATV group decreased significantly (P<0.05), and the cholesterol outflow rate increased significantly (P<0.05). Compared to the oxLDL+CQ group, the intracellular lipid droplets, FC, TC, and CE/TC content in the oxLDL+CQ+ATV group decreased significantly (P<0.05), while the cholesterol outflow rate increased significantly (P<0.05). Western blotting results showed that protein expression levels of LC3II/I, P62 and TFEB were decreased in the model histone in comparison to the blank group (P<0.05). Compared to the expression levels of LC3II/I, P62, TFEB and LAMP1 ((1.006±0.052), (0.183±0.013), (0.333±0.020), and (0.957±0.026)) in the model group, the expression levels of oxLDL+CQ histamine ((1.594±0.017), (0.257±0.006), (0.477±0.024), and (0.957±0.026)) were significantly higher (P<0.05).The protein expression levels of LC3II/I, TFEB and LAMP1 in oxLDL+ATV group ((1.146±0.060), (0.540±0.031), and (1.027±0.054)) were increased, while the protein expression levels of P62 (0.115±0.009) were decreased (P<0.05). Compared to LC3II/I and P62 in oxLDL+CQ group, the expression level of oxLDL+CQ+ATV histone was decreased ((1.419±0.036) and (0.165±0.006)), and the difference was statistically significant (P< 0.05).ConclusionATV can increase the cholesterol outflow rate of macrophages, reduce the level of cholesterol and lipid droplets in macrophages, and reduce the formation of foam cells. The mechanisms behind this are still unknown, which may be related to the activation of TFEB by ATV and influencing the process of autophagy.

Comparative observation of Methylprednisolone and gamma globulin in the treatment of acute myelitis / 中国生化药物杂志

Objective To study and analyze the efficacy of Methylprednisolone and gamma globulin in the treatment of acute myelitis. Methods 100 cases of acute myelitis in the second affiliated hospital of Xi'an Jiaotong University were selected as the research objects, and the control group was treated with Dexamethasone. The experimental group was treated with Methylprednisolone combined with gamma globulin. The clinical efficacy, adverse reactions and related clinical indicators of the experimental group and the control group were compared and analyzed. Results The effective rate of the experimental group (92.0 %) was significantly better than (82.0 %) of the control group with statistical difference (P<0.05). There were no obvious adverse reactions in the two groups, the incidence rate was 8.0% and 10.0 % respectively, and there was no significant difference. Time of muscle strength improved two level in the control group (18.90±5.23) d was significantly longer than that (10.87±4.30) d of the experimental group, the experimental group hospitalization time, sensory recovery time and recovery of urinary function and other indicators are better than the control group, with statistical difference (P<0.05). Conclusion Methylprednisolone combined with gamma globulin in the treatment of acute myelitis has ideal efficacy, could significantly improve the patient's muscle strength with high safety.

Comparative observation of Methylprednisolone and gamma globulin in the treatment of acute myelitis / 中国生化药物杂志

Objective To study and analyze the efficacy of Methylprednisolone and gamma globulin in the treatment of acute myelitis. Methods 100 cases of acute myelitis in the second affiliated hospital of Xi'an Jiaotong University were selected as the research objects, and the control group was treated with Dexamethasone. The experimental group was treated with Methylprednisolone combined with gamma globulin. The clinical efficacy, adverse reactions and related clinical indicators of the experimental group and the control group were compared and analyzed. Results The effective rate of the experimental group (92.0 %) was significantly better than (82.0 %) of the control group with statistical difference (P<0.05). There were no obvious adverse reactions in the two groups, the incidence rate was 8.0% and 10.0 % respectively, and there was no significant difference. Time of muscle strength improved two level in the control group (18.90±5.23) d was significantly longer than that (10.87±4.30) d of the experimental group, the experimental group hospitalization time, sensory recovery time and recovery of urinary function and other indicators are better than the control group, with statistical difference (P<0.05). Conclusion Methylprednisolone combined with gamma globulin in the treatment of acute myelitis has ideal efficacy, could significantly improve the patient's muscle strength with high safety.

Association between Carotid Intima-media Thickness and Aldehyde Dehydrogenase 2 Glu504Lys Polymorphism in Chinese Han with Essential Hypertension / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Aldehyde dehydrogenase 2 (ALDH2) is involved in the pathophysiological processes of cardiovascular diseases. Recent studies showed that mutant ALDH2 could increase oxidative stress and is a susceptible factor for hypertension. In addition, wild-type ALDH2 could improve the endothelial functions, therefore reducing the risk of developing atherosclerosis. The aim of the present study was to explore the frequency of the Glu504Lys polymorphism of the ALDH2 gene and its relation to carotid intima-media thickness (CIMT) in a group of patients with essential hypertension (EH) and to investigate the association between the Glu504Lys polymorphism and CIMT in Chinese Han patients with EH.</p><p><b>METHODS</b>In this study, 410 Chinese Han patients with EH who received physical examinations at the People's Hospital of Sichuan Province (China) were selected. DNA microarray chip was used for the genotyping of the Glu504Lys polymorphism of the ALDH2 gene. The differences in CIMT among patients with different Glu504Lys ALDH2 genotypes were analyzed.</p><p><b>RESULTS</b>The mean CIMT of the patients carrying AA/AG and GG genotypes was 1.02 ± 0.31 mm and 0.78 ± 0.28 mm, respectively. One-way ANOVA showed that the CIMT of the patients carrying the AA/AG genotype was significantly higher than in the ones carrying the GG genotype (P < 0.001). Multivariate logistic regression showed that the Glu504Lys AA/AG genotype of the ALDH2 gene was one of the major factors influencing the CIMT in patients with EH (odds ratio = 3.731, 95% confidence interval = 1.589-8.124, P = 0.001).</p><p><b>CONCLUSIONS</b>The Glu504Lys polymorphism of the ALDH2 gene is associated with the CIMT of Chinese Han patients with EH in Sichuan, China.</p>

Comparison of morphine and codeine concentration in urines of heroin abusers and codeine users / 法医学杂志

OBJECTIVE@#To compare morphine and codeine concentration in urines of heroin abusers and codeine users and to discuss the judgment index to distinguish between heroin abuser and codeine user.@*METHODS@#The urines of heroin abusers and codeine users were collected at different time periods. After protein precipitation, the urine samples were conducted for the qualitative and quantitative analysis of morphine and codeine by UPLC-MS/MS. And the results were all statistically analyzed.@*RESULTS@#Statistical analysis showed that morphine and codeine concentration in urines of heroin abusers and codeine users were both abnormal distributions. The probability of the heroin abuser would be more than 95% and less than 5% for the codeine user when the concentration of morphine in urine sample was more than 67 ng/mL. The probability of the codeine user would be more than 95% and less than 5% for the heroin abuser when the concentration of morphine in urine sample was less than 67 ng/mL.@*CONCLUSION@#The morphine concentration in urine could be used as a criterion to distinguish the heroin abuser from the codeine user, while the codeine concentration could not.

Simultaneous determination of opioid compounds in human urine by UPLC-MS/MS / 法医学杂志

OBJECTIVE@#To propose a method for simultaneous determination of codeine(COD), 6-monoacetyl-morphine (6-MAM), morphine (MOR), morphine-3-glucuronide (M3G) and morphine-6-glucuronide (M6G) in human urine by ultra performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS).@*METHODS@#After precipitation of protein by acetonitrile, the urine samples, with added the morphine-d3 (MOR-d3) and morphine-3-Glucuronide-d3 (M3G-d3) as internal standards, were pre-treated by Sirocco protein precipitation plate, and then analyzed by UPLC-MS/MS.@*RESULTS@#The limit of detection was 0.2 ng/mL for both COD and MAM, the limit of quantitation was 0.5 ng/mL for both COD and MAM. The limit of detection was 0.5 ng/mL for MOR, M3G and M6G, the limit of quantitation was 1 ng/mL for them. The linear correlation coefficients were not less than 0.9997, both the inter-day and intra-day precisions were less than 10%, the recoveries were in the range of 70.0% to 98.3%, the matrix effects were about 50.5% to 99.0%.@*CONCLUSION@#This proposed method is simple, rapid and accurate, it could be applied in forensic toxicological analysis.

Spine disc MR image analysis using improved independent component analysis based active appearance model and Markov random field / 生物医学工程学杂志

As there are not many research reports on segmentation and quantitative analysis of soft tissues in lumbar medical images, this paper presents an algorithm for segmenting and quantitatively analyzing discs in lumbar Magnetic Resonance Imaging (MRI). Vertebrae are first segmented using improved Independent component analysis based active appearance model (ICA-AAM), and lumbar curve is obtained with Minimum Description Length (MDL); based on these results, fast and unsupervised Markov Random Field (MRF) disc segmentation combining disc imaging features and intensity profile is further achieved; finally, disc herniation is quantitatively evaluated. The experiment proves that the proposed algorithm is fast and effective, thus providing doctors with aid in diagnosing and curing lumbar disc herniation.

The role of 11C-choline positron emission tomography-computed tomography and videomediastinoscopy in the evaluation of diseases of middle mediastinum / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Middle mediastinal masses comprise a wide variety of tumors but may also reflect lymphadenopathy, and thus remain an interesting diagnostic challenge. We performed positron emission tomography (PET) of mediastinal masses in order to evaluate the ability of PET to predict the malignancy of these tumors. We compared histologic findings, videomediastinoscopy, computed tomography (CT), and PET-CT in patients with mediastinal disease.</p><p><b>METHODS</b>Thirty-two patients were evaluated with CT, PET-CT and videomediastionoscopy, and all studies were performed within four weeks in each patient. (11)C-choline as a PET tracer was used to visualize masses. PET data were evaluated using the standardized uptake value (SUV) and were compared with pathologic data.</p><p><b>RESULTS</b>There were 13 men and 19 women aged from 21 to 74 (mean 45.2) years. Among the patients with mediastinal diseases, sarcoidosis was diagnosed in 12 patients, tuberculosis in 5 patients, lymphoma in 5 patients, and noncaseating granulomata without classical "sarcoid" finding in 3 patients. N2 or N3 nodal metastasis was revealed in 6 of 7 patients who had non-small cell lung cancer or suspected lung cancer, and one was negative (the pathological diagnosis was reactive hyperplasia). The accuracies for correctly diagnosing mediastinal masses for CT, PET-CT and videomediastinoscopy were 38% (12/32), 63% (20/32), and 91% (29/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 11.130, P < 0.001). The SUVs were similar among these diseases. On the other hand, if the diagnostic classification was benign vs malignancy, the accuracies for CT, PET-CT and videomediastinoscopy were 53% (17/32), 75% (24/32), 100% (32/32) respectively. The diagnostic accuracy of videomediastinoscopy was superior to that of PET-CT (chi(2) = 22.042, P < 0.001). The SUV of malignant lesions (6.9, 3.2 - 9.8; n = 11) appeared to be higher than that of benign lesions (4.9, 2.9 - 8.3; n = 21), however, this difference was not statistically significant (P = 0.054).</p><p><b>CONCLUSIONS</b>To diagnose lesions located in the middle mediastinum, videomediastinoscopy possesses the highest diagnostic accuracy, and therefore remains the gold standard. PET-CT is valuable for differential diagnosis of benign vs malignant lesions, CT alone or PET alone (SUV) may provide misdiagnosis in a substantial proportion of patients with mediastinal masses.</p>

Mutational studies of adenomatous polyposis coli gene in carcinomas from patients with hereditary non-polyposis colorectal cancers / 中华医学遗传学杂志

<p><b>OBJECTIVE</b>To analyze the mutational features of adenomatous polyposis coli (APC) gene and to explore the effect of mismatch repair (MMR) deficiency on its mutations in hereditary non-polyposis colorectal cancers (HNPCC).</p><p><b>METHODS</b>PCR-based in vitro synthesized protein test (IVSP) assay and sequencing analysis were used to confirm somatic mutations of whole APC gene in 19 HNPCC patients.</p><p><b>RESULTS</b>Eleven cases with thirteen mutations were determined. The frequency of APC mutation was 58%(11/19). The exhibiting mutations consisted of 9 frameshift mutations and 4 nonsense ones, indicating the existence of more frameshift mutations (69%). All of frameshift mutations were deletion or insertion of 1-2 bp and most of them (7/9) happened at simple nucleotide repeat sequences, particularly within (A) n tracts (5/9). All of four nonsense mutations resulted from C to T transitions at CpG sites.</p><p><b>CONCLUSION</b>Mutational inactivations of APC gene were detected in more than half of HNPCC patients in this study, indicating that APC mutation is a common molecular event in the tumorigenesis of HNPCC. According to the location of frameshift mutations at simple nucleotide repeat sequences and point mutations at CpG sites, it was suggested that endogenous mechanisms like MMR deficiency might exert an effect on the nature of APC mutations in most HNPCC.</p>

Study on antioxidant effect of Astragalus polysaccharide / 中国药理学通报

Xanthine-Xanthine oxidase ( X-XOD) added to the culture medium of cultured cardiac cells in rats may damage cell membrane. Electrophysiological findings indicated that APA, MDP, OS,Vmax, were decreased and the SDF was increased. Astragalus polysaccharide (APS) could protect the cells from being damaged by X-XOD. APS recoverted all the de-creased cardiac functional parameters in free-radical-damaged rats by X-XOD. APS had anti-free-radical damage action on the cultured my-ocardiocytes and the myocardial contractility of the isolated rat working heart.