Clinical and functional analyses of the novel STAR c.558C>A in a patient with classic lipoid congenital adrenal hyperplasia.

Objective: Congenital lipid adrenal hyperplasia (LCAH) is the most serious type of congenital adrenal hyperplasia and is caused by steroid-based acute regulatory (STAR) protein mutations. Herein, we report compound heterozygous mutations c.558C>A (p.S186 R) and c.772C>T (p.Q258*) in a newborn 46 XY patient diagnosed with classic LCAH and explore their clinical and functional characteristics. Methods: Peripheral blood samples were collected from LCAH patient and their families. The pathogenic variant identified by whole-exome sequencing was further confirmed by Sanger sequencing and pedigree verification. The functional consequence and ability to convert cholesterol into progesterone of the identified STAR Q258* and S186 R mutations were analyzed by cell transfection and in vitro assays. Results: The proband was presented with severe glucocorticoid and mineralocorticoid deficiency, high adrenocorticotropic hormone, and enlarged adrenals. Heterozygous mutations p. S186 R and p. Q258* in the STAR gene were identified in the patient, and her parents were carriers, which is consistent with an autosomal recessive disorder. The STAR p. Q258* mutation has been reported and generates a truncated protein. The p. S186 R mutation is a novel variant that disrupts STAR. The residual STAR activities of p. S186R, p. Q258*, and p. S186R/p.Q258* were 13.9%, 7.3%, and 11.2%, respectively, of the wild-type, proving the main negative effects of the mutant proteins. Conclusion: Our findings reveal the molecular mechanisms underlying LCAH pathogenesis, further expanding the genotype and clinical spectrum of LCAH.

Dysbiosis of gut microbiota and decreased propionic acid associated with metabolic abnormality in Cushing's syndrome.

Objective: Chronic hypercortisolism leads to a phenotype resembling metabolic syndrome. We aimed to investigate the association between gut microbiota and metabolic abnormalities in endogenous hypercortisolism (Cushing's syndrome). Methods: A total of 23 patients with Cushing's syndrome (18 female and 5 men, aged 47.24 ± 12.99 years) and 30 age-, sex-and BMI-matched healthy controls (18 female and 12 men, aged 45.03 ± 6.69 years) were consecutively recruited. Differences in gut microbiota and plasma short-chain fatty acid (SCFAs) concentrations between the Cushing's syndrome patients and controls were analyzed by 16S rRNA sequencing and gas chromatography-mass spectrometry (GC-MS). Results: Compared to the controls, the Simpson and Pielou indices of α diversity were dramatically decreased in Cushing's syndrome (P < 0.05). The gut microbiota community structure differed significantly between Cushing's syndrome patients and controls. Compared to controls, the bacterial communities of the Cushing's syndrome patients were enriched in Proteobacteria and Escherichia-Shigella, and depleted in Firmicutes, including Agathobacter, Blautia, Anaerostipes, Eubacterium_eligens_group, and Lachnospira. Spearman analysis demonstrated that HbA1c, SBP, DBP, and cortisol levels were significantly positively correlated with Proteobacteria and Escherichia-Shigella, whereas negatively correlated with Agathobacter, Blautia, Anaerostipes, Eubacterium_hallii_group, and Lachnospira, etc. Cushing's syndrome patients also had a lower propionic acid concentration (0.151±0.054 vs. 0.205±0.032 µg/mL, P=0.039) than controls. Furthermore, the level of propionic acid was negatively correlated with systolic pressure and cortisol levels (P<0.05). Conclusion: Gut microbiota dysbiosis and decreased propionic acid levels were observed in patients with Cushing's, suggesting that the gut microbiota may be a potential therapeutic intervention target to improve hypercortisolism-related metabolic abnormalities.

Highly active Fe-N-doped porous hollow carbon nanospheres as oxygen reduction electrocatalysts in both acidic and alkaline media.

Hierarchical iron-nitrogen-codoped porous hollow carbon spheres have been synthesized by using melamine-formaldehyde (MF) resin spheres as templates, nitrogen sources and pore-forming agents. FeCl3, 1,10-phenanthroline and carbon black were used as iron, nitrogen and carbon sources. The as-obtained porous hollow carbon spheres possess a high specific surface area of 807 m2 g-1, as well as exhibited excellent electrocatalytic activity for the oxygen reduction reaction (ORR) in both acidic and alkaline media. In 0.1 M HClO4 solution, the onset potential was 0.857 V (vs. RHE) and the half-wave potential was 0.715 V, which are only 78 and 80 mV less than those of the 20% Pt/C catalyst, respectively. In addition, in 0.1 M KOH solution, the onset potential was 1.017 V and the half-wave potential was 0.871 V for the ORR, which are 22 and 28 mV more positive than those of the Pt/C catalyst, respectively. Meanwhile, the catalyst also exhibited excellent methanol tolerance and long-term durability with a more effective four-electron pathway compared to the 20% Pt/C catalyst in both acidic and alkaline media. When used as an air-cathode catalyst for a Zn-air battery, the maximum power density of a Zn-air battery with the MF-C-Fe-Phen-800 cathode was 235 mW cm-2 at a high current density of 371 mA cm-2, and a high open-circuit potential of 1.654 V, superior to that of Pt/C (199 mW cm-2, 300 mA cm-2, 1.457 V). A series of designed experiments suggested that the remarkable performance was attributed to the high specific area, hollow carbon spheres, unique hierarchical micro-mesoporous structures, high contents of pyridinic-N and graphitic-N. The superiority of the as-prepared catalyst makes it promising for use in practical applications.

Hyperglycemia is a strong predictor of poor prognosis in COVID-19.

AIMS: The objective of this study is to explore the association between documented diabetes, fasting plasma glucose (FPG), and the clinical outcomes of Coronavirus disease 2019 (COVID-19). METHODS: This retrospective study included 255 patients with COVID-19. Of these, 214 were admitted to isolation wards and 41were admitted to intensive care units (ICUs). Demographic, clinical, treatment, and laboratory data were collected and compared between ICU and non-ICU patients. Multivariable logistic regression models were used to explore the risk factors associated with poor clinical outcomes (ICU admission or death). RESULTS: There were significant changes in several clinical parameters in ICU patients (leukopenia, lymphopenia, elevated D-dimer, as well as higher levels of FPG, cardiac troponin, serum ferritin, IL-6, and high-sensitivity C-reactive protein)compared with non-ICU patients. The prevalence of known diabetes was substantially higher in ICU than non-ICU patients (31.7% vs. 17.8%, P = 0.0408). Multivariable regression analysis showed that a history of diabetes [odds ratio (OR), 0.099; 95% confidence interval (CI), 0.016-0.627; P = 0.014], high FPG at admission (OR, 1.587; 95% CI, 1.299-1.939, P < 0.001), high IL-6 (OR, 1.01; 95% CI, 1.002-1.018, P = 0.013), and D-dimer higher than 1 mg/L at admission (OR, 4.341; 95% CI, 1.139-16.547, P = 0.032) were independent predictors of poor outcomes. Cox proportional hazards analysis showed that compared with FPG < 7 mmol/L, FPG levels of 7.0-11.1 mmol/L and ≥ 11.1 mmol/L were associated with an increased hazard ratio (HR) for poor outcome (HR, 5.538 [95% CI, 2.269-13.51] and HR, 11.55 [95% CI, 4.45-29.99], respectively). CONCLUSION: Hyperglycemia and a history of diabetes on admission predicted poor clinical outcomes in COVID-19.

Identification of Novel Variants in MEN1: A Study Conducted with Four Multiple Endocrine Neoplasia Type 1 Patients.

Multiple endocrine neoplasia type 1 (MEN1) is an autosomal dominant inherited endocrine tumor syndrome caused by inactivating variants of the MEN1 gene. The aim of this study is to explore the clinical and genetic characteristics of four MEN1 patients. We isolated genomic deoxyribonucleic acid from lymphocytes, parathyroid, and thymic tumoral tissue specimens from the MEN1 patients. All exons of the MEN1 and CDNK1B genes and adjacent exon-intron sequences were amplified by polymerase chain reaction and subsequently sequenced. Further, the splice alterations were studied by sequencing the amplified RT-PCR products for MEN1 cDNA. We identified four heterozygous MEN1 germline variants: c.564delC, c.1268G>A, IVS5+5delG, and c.1546_1547insC. Both c.564delC and IVS5+5delG were novel variants. The impact of the MEN1 splice variant, IVS5+5delG, was evaluated using bioinformatics and in vitro analyses. The analyses indicated that this variant resulted in skipping of the neighboring exon and was disease-causing. Two novel somatic variants, c.249_252delGTCT and c.313_314insC, were found. Additionally, loss of heterozygosity (LOH) for the MEN1 locus (IVS5+5delG and c.564delC) was found in tumor tissue samples from the MEN1 patients, consistent with Knudson's two-hit mechanism. We identified four MEN1 germline variants and two novel somatic variants. Early recognition of the phenotype coupled with variant screening of the MEN1 gene is the key to diagnosing and treating MEN1 effectively at an early stage.

A Novel Role of VEGFC in Cerebral Ischemia With Lung Injury.

Cerebral ischemia (CI) is a severe brain injury resulting in a variety of motor impairments combined with secondary injury in remote organs, especially the lung. This condition occurs due to insufficient blood supply to the brain during infancy. However, it has a molecular linkage that needs to be thoroughly covered. Here, we report on the role of vascular endothelial growth factor C (VEGFC) in lung injury induced by CI. The middle cerebral artery occlusion (MCAO) was depended to establish the animal model of CI. Rats were used and brain ischemia was confirmed through TTC staining. Serum was used for protein chip analysis to study the proteomic interaction. Immunohistochemistry analyses were used to quantify and locate the VEGFC in the lung and brain. The role of VEGFC was detected by siVEGFC technology in SY5Y, HUCEV, and A549 cell lines, under normal and oxygen glucose deprivation (OGD) conditions in vitro. As a result, the TTC staining demonstrated that the model of brain ischemia was successfully established, and MPO experiments reported that lung damage was induced in MCAO rats. VEGFC levels were up-regulated in serum. On the other hand, immunohistochemistry showed that VEGFC increased significantly in the cytoplasm of neurons, the endothelium of small trachea and the lung cells of CI animals. On a functional level, siVEGFC effectively inhibited the proliferation of SY5Y cells and decreased the viability of HUVEC cells in normal cell lines. But under OGD conditions, siVEGFC decreased the growth of HUVEC and increased the viability of A549 cells, while no effect was noticed on SYSY cells. Therefore, we confirmed the different role of VEGFC played in neurons and lung cells in cerebral ischemia-reperfusion injury. These findings may contribute to the understanding the molecular linkage of brain ischemia and lung injury, which therefore provides a new idea for the therapeutic approach to cerebral ischemia-reperfusion.

Transcriptomic analysis of Pak Choi under acute ozone exposure revealed regulatory mechanism against ozone stress.

BACKGROUND: Ground-level ozone (O3) is one of the major air pollutants, which cause oxidative injury to plants. The physiological and biochemical mechanisms underlying the responses of plants to O3 stress have been well investigated. However, there are limited reports about the molecular basis of plant responses to O3. In this study, a comparative transcriptomic analysis of Pak Choi (Brassica campestris ssp. chinensis) exposed to different O3 concentrations was conducted for the first time. RESULTS: Seedlings of Pak Choi with five leaves were exposed to non-filtered air (NF, 31 ppb) or elevated O3 (E-O3, 252 ppb) for 2 days (8 h per day, from 9:00-17:00). Compared with plants in the NF, a total of 675 differentially expressed genes (DEGs) were identified in plants under E-O3, including 219 DEGs with decreased expressions and 456 DEGs with increased expressions. Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses revealed that O3 stress invoked multiple cellular defense pathways to mitigate the impaired cellular integrity and metabolism, including 'glutathione metabolism', 'phenylpropanoid biosynthesis', 'sulfur metabolism', 'glucosinolate biosynthesis', 'cutin, suberine and wax biosynthesis' and others. Transcription factors potentially involved in this cellular regulation were also found, such as AP2-ERF, WRKY, JAZ, MYB etc. Based on the RNA-Seq data and previous studies, a working model was proposed integrating O3 caused reactive oxygen burst, oxidation-reduction regulation, jasmonic acid and downstream functional genes for the regulation of cellular homeostasis after acute O3 stress. CONCLUSION: The present results provide a valuable insight into the molecular responses of Pak Choi to acute O3 stress and the specific DEGs revealed in this study could be used for further functional identification of key allelic genes determining the O3 sensitivity of Pak Choi.

Interleukin­6 RNA knockdown ameliorates acute lung injury induced by intestinal ischemia reperfusion in rats by upregulating interleukin­10 expression.

Acute lung injury (ALI) is a common complication following intestinal ischemia/reperfusion (II/R) injury and contributes to the associated high mortality rate. However, the underlying mechanism is poorly understood and treatments are limited. RNA interference (RNAi) has been demonstrated to provide a promising disease treatment strategy both in vitro and in vivo. Therefore, the present study aimed to test whether blocking the proinflammatory cytokine IL­6 by RNAi may protect the lungs from remote organ injury following II/R, and to investigate the potential underlying mechanisms. A total of 176 adult healthy male Sprague­Dawley rats were randomly divided into sham, II/R, negative­control and IL­6­short hairpin (sh)RNA groups. The rats underwent II/R injury with occlusion of the superior mesenteric artery and coeliac artery to induce ischemia for 40 min, and were subsequently reperfused for 0­48 h. The negative­control group received a control lentiviral vector containing scrambled or non­specific sequences, and the IL­6­shRNA groups were administered with a vector containing an IL­6 shRNA sequence to affect RNAi­mediated knockdown of IL­6. ALI severity was determined by lung edema (lung wet/dry ratio) and histological analysis (lung injury scores). IL­6 localization, and mRNA and protein expression levels, were detected by immunofluorescence, reverse transcription­quantitative polymerase chain reaction and western blot analysis, respectively. IL­10 expression induced by IL­6 knockdown in lung tissues was additionally detected. IL­6 RNAi was revealed to significantly reduce the expression of IL­6, which was associated with upregulated IL­10 expression in lung tissues. Consequently, the severities of ALI and edema induced by II/R were substantially improved. In conclusion, the present study demonstrated that IL­6 RNAi may protect the lung from ALI induced by II/R, and that this protective role may be associated with upregulation of IL­10. These findings may contribute to the development of an IL­6­RNAi­based therapeutic strategy for the treatment of II/R­induced ALI.

Comparison of biomechanical effect between oblique Ban-pulling manipulation and lumbar erection-rotation manipulation in sitting position for lumbar intervertebral disc herniation / 针灸推拿医学（英文版）

Objective:To compare the biomechanical effects between oblique Ban-pulling manipulation and lumbar erection-rotation manipulation in sitting position in treating lumbar intervertebral disc herniation (LIDH). Methods:A three-dimensional finite element model of L3-S1 was developed to carry out a comparative study between oblique Ban-pulling manipulation and lumbar erection and rotation manipulation in sitting position. The disc protrusion was assumed to be on the rear left of L4 disc, and the manipulations were performed on the right side. The loading process was simulated by two steps. In the first step, only the compression loading was imposed, and in the second step, both the compression loading and axial rotation moment were imposed. The displacement and stress distribution in L4 disc were investigated. Results:The values of stress and displacement in the second step were lower than those in the first step in each manipulation. The stress and displacement differences between the two steps were respectively 1.79 times and 3.03 times larger in oblique Ban-pulling manipulation than those in lumbar erection-rotation manipulation in sitting position. Conclusion: Oblique Ban-pulling manipulation may result in a better biomechanical effect than lumbar erection-rotation manipulation in sitting position for LIDH.

Comparison of biomechanical effect between oblique Ban-pulling manipulation and lumbar erection-rotation manipulation in sitting position for lumbar intervertebral disc herniation / 针灸推拿医学（英文版）

Objective:To compare the biomechanical effects between oblique Ban-pulling manipulation and lumbar erection-rotation manipulation in sitting position in treating lumbar intervertebral disc herniation (LIDH). Methods:A three-dimensional finite element model of L3-S1 was developed to carry out a comparative study between oblique Ban-pulling manipulation and lumbar erection and rotation manipulation in sitting position. The disc protrusion was assumed to be on the rear left of L4 disc, and the manipulations were performed on the right side. The loading process was simulated by two steps. In the first step, only the compression loading was imposed, and in the second step, both the compression loading and axial rotation moment were imposed. The displacement and stress distribution in L4 disc were investigated. Results:The values of stress and displacement in the second step were lower than those in the first step in each manipulation. The stress and displacement differences between the two steps were respectively 1.79 times and 3.03 times larger in oblique Ban-pulling manipulation than those in lumbar erection-rotation manipulation in sitting position. Conclusion: Oblique Ban-pulling manipulation may result in a better biomechanical effect than lumbar erection-rotation manipulation in sitting position for LIDH.

Analysis on the maternal HBsAg carrying rate and the neonatal immunization in Huangpu District of Shanghai / 上海预防医学

Objective To understand the maternal positive rate of HBsAg and the neonatal immunization in Huangpu District of Shanghai so as to provide bases for developing strategies of preventing hepatitis B and reducing the rate of neonatal hepatitis B virus infection.Methods By adopting cluster sampling research methods, retrospective surveys were conducted for the detection of HBsAg of the pregnant women from all the obstetric institutions(7 institutions) in Huangpu District of Shanghai as well as for the neonatal hepatitis B vaccinations therein from 2013 to 2015, and statistical analyses were made for the results thereof by utilizing the SPSS 20.0 software.Results A total of 29 425 pregnant women were surveyed and the positive rate of HBsAg thereof was 3.95%.The detection rate of HBsAg(98.42% & 96.76%) as well as the positive rate of HBsAg (2.58% & 4.50%) for both the local and external pregnant women in Shanghai, respectively, showed statistical differences (P<0.01).The newborns of mothers with positive rates of HBsAg who used combined immunization(HBIG+hepatitis B vaccine) amounted to 1 130, with the rate of combined immunization reaching 98.52%.Conclusion The positive rate of HBsAg for the pregnant women in Huangpu District of Shanghai is below the level thereof for those in either the whole country or the other cities and provinces, which shows that Shanghai has made effective achievements in hepatitis B vaccination as well as in the health education relevant thereto.However, active and positive combined immunization measures should still be strengthened for the newbornsof mothers with positive rates of HBsAg.In addition, with the growth of pregnant women under new policies, better maternal health care mechanism should be established and various prenatal inspections should be well implemented, thus reducing the rate of the people without taking the detection of HBsAg and preventing newborns from being infected by hepatitis B virus as a result of failing to take timely immunization measures.

Toxic effects of HgCl2 on activities of SOD, AchE and relative expression of SOD, AChE, CYP1A1 of zebrafish.

In order to explore the impact of mercury on fish, zebrafish were exposed in various levels of HgCl2 and the superoxide dismutase (SOD), acetylcholinesterase (AChE) activities were measured after 1 and 7 days exposure. Muscle RNA was also extracted for semi-quantitative RT-PCR determination of the expression of AChE, CYP1A1, Cu/Zn-SOD. It indicated that 96 h-LC50 of HgCl2 to zebrafish is 244.06 µg L(-1). The AChE activity was decreased on 1st day and 7th day. The SOD activity was almost unchanged on 1st day, it showed a dose-response effect on 7th day. The expression of AChE was down-regulated on 1st day, it is consistent with the AChE activity, but the expression was showed a significant difference between exposed group and the control group on 7th day. The expression of CYP1A1 was decreased compared with the control group on 1st day. After 7 day exposure, the expression of CYP1A1 was up-regulated in 0.1LC(50) group, and down-regulated in 0.4LC(50) group, 0.8LC(50) group, showing a stimulation effect at low concentration and inhibition at high concentration. The expression of Cu/Zn-SOD was down-regulated on 1st day, also showing an effect of stimulation at low concentration and inhibition at high concentrations. Thus HgCl2 was highly toxic to zebrafish, both the enzymatic activities and the gene transcription were changed significantly. But the change of gene transcription is earlier than enzymes activities. It indicates that gene transcript can be used as an early warning for environmental risk assessment.