Clinical features and genetic variations of severe neonatal hyperbilirubinemia:Five case reports

BACKGROUND Neonatal hyperbilirubinemia is a common problem faced by pediatricians.The role of genetic factors in neonatal jaundice has been gradually recognized.This study aims to identify genetic variants that influence the bilirubin level in five patients using next-generation sequencing(NGS).CASE SUMMARY Five neonates with severe hyperbilirubinemia were retrospectively studied.They exhibited bilirubin encephalopathy,hypothyroidism,ABO blood type incompatibility hemolysis,glucose-6-phosphate dehydrogenase(G6PD)deficiency and premature birth,respectively.A customized 22-gene panel was designed,and NGS was carried out for these neonates.Eight variations(G6PD c.G1388A,HBA2 c.C369G,ABCC2 c.C3825G,UGT1A1 c.G211A,SPTB c.A1729G,EPB41 c.G520A,c.1213-4T>G and c.A1474G)were identified in these five neonates.Genetic mutations of these genes are associated with G6PD deficiency,thalassemia,Dubin-Johnson syndrome,Gilbert syndrome,hereditary spherocytosis,and hereditary elliptocytosis.One of the neonates was found to have compound variants of the EPB41 splice site c.1213-4T>G and c.G520A(p.E174K),but no elliptocyte was seen on his blood smear of 4 years old.CONCLUSION Pathological factors of severe neonatal hyperbilirubinemia are complicated.Genetic variants may play an important role in an increased risk of neonatal hyperbilirubinemia,and severe jaundice in neonates may be related to a cumulative effect of genetic variants.

Genetic Transformation in Triticeae Crops

Wheat, triticale, tritordeum, barley, oat and rye are the most important crops in human consumptions and industry in the world. Transformation technology supplies a new source of improving Triticeae crops. In the past decade, transformation of wheat crops has considerably progressed. Many transgenic plants of Triticeae crops with various genes were produced via nricroprojectile bombardment, Agrobacterium-mediated transformation, PEG-uptake DNA technique, electroporation, microinjection, injection inflorescence and silicone carbide. Integration and expression of transgenes, inheritance and variation of transgenic plants have been studied. Technical improvements of genetic transformation for wheat crops will be extensively useful in commerce and benefit significantly to human being in the world.

Transcriptomic comparison to identify rapidly evolving genes in Braya humilis

The Brassicaceae species Braya humilis shows broad adaptation to different climatic zones and latitudes. However, the molecular adaptation mechanism of B. humilis is poorly understood. In China, B. humilis is mainly distributed on the QinghaiTibetan Plateau(QTP) and in the adjacent arid region. Previous transcriptome analysis of B. humilis has revealed that 39 salt and osmotic stress response genes are subjected to purifying selection during its speciation. To further explore the adaptation mechanism of B. humilis to an arid environment, OrthoMCL program was employed in this study and 6,268 pairs of orthologous gene pairs with high confidence were obtained between B. humilis and Arabidopsis thaliana. A comparative evolutionary analysis based on nonsynonymous to synonymous substitution ratio(Ka/Ks) was then conducted. There were 64 pairs exhibiting a Ka/Ks ratio more than 0.5 and among which, three instrumental candidate genes, T20487,T22576, and T23757, were identified with strong selection signatures(Ka/Ks >1). The corresponding A. thaliana orthologs are double-stranded RNA-binding domain protein, MADS-box family protein, and NADH-dehydrogenase subunit6, which is encoded by mitochondria genome. This report not only demonstrates the adaptation contribution of fast evolving nuclear genes, but also highlights the potential adaptive value of mitochondria gene to the speciation and adaptation of B. humilis toward the extreme environment in an arid region.

A Molecular Epidemiology Analysis of HIV in Shenzhen and HIV Env Gene Variation Replication Analysis

Objective: To analyze molecular trends of the HIV epidemic in Shenzhen. Methods: Serum collected from Shenzhen AIDS patientsbetween 1992-1999 was analyzed using molecular techniques.DNA fragments of the HIV-1 Env gene were amplified bynested PCR from uncultured peripheral blood mononuclearcells (PBMCs) from these serum samples. The C2-C3 region ofthe Env gene was sequenced and analyzed. Specific high-riskbehaviors were also analyzed. Results: We found that the transmission of HIV in the citywas mainly through sexual behaviors (46.0%). There werefour HIV-1 subtypes: B', B, C and E with 6.31%, 7.95%,3.09% and 8.92% gene divergence inside each subtype inShenzhen. These results suggested that epidemic times were 6,8, 3 and 9 respectively. The main cpidemic subtypes were Eand B strains. AIDS patient's antigenic variation was slightlyhigher than that of HIV infected individuals. Conclusion: Surveillance data reflect trends and theepidemic time of HIV which will be useful for policy makersto formulate effeive strategies of HIV/AIDS prevention and control in Shenzhen.

Relationship between HBV genotypes and anti-viral therapeutic efficacy of interferon-alpha

BACKGROUND: Much evidence demonstrates that the genotypes of hepatitis B virus (HBV) present differences in pathogenicity and outcomes owing to differences in genetic structure. This study aimed to investigate the influences of HBV genotypes on the anti-viral therapeutic efficacy of interferon-alpha (IFN-alpha) in chronic hepatitis B patients, and to determine the relationship between HBV genotypes and levels of viral replication or gene variations. METHODS: The chronic hepatitis B patients who were treated with IFN-alpha were selected randomly. Anti-viral therapeutic efficacy was monitored in these patients. The HBV genotypes were detected by PCR microplate hybridization ELISA. The levels of serum HBV-DNA were determined by fluorescence quantitative PCR. HBV gene variation at pre-C and basic core promoter (BCP) regions were assayed by gene chip technology. RESULTS: Genotypes B and C were predominant in 94 chronic hepatitis B patients. A, E and F genotypes were not found in these patients. The HBV-DNA levels of genotype C and mixed genotypes were significantly higher than those of genotype B. The response to IFN-alpha in patients with genotype B was markedly better than in those with genotypes C and D, and the complete response to IFN-alpha was only observed in genotype B. The response to IFN-alpha in patients with mixed genotypes was the least sensitive. The negative transition of HBeAg was correlated with variations in the HBV pre-C and BCP regions in patients with partial or no response to IFN-alpha. The variation rates of HBV pre-C and BCP regions were clearly higher in genotype C than in genotype B. CONCLUSIONS: The results suggest that HBV genotype is correlated with the serum levels of HBV-DNA, HBV gene variations and therapeutic efficacy of IFN-alpha. The regular detection of HBV genotypes in the clinic will be of benefit for disease prognosis and planning of anti-viral therapeutic strategies.

Disease Progression Patterns of SHIV-KB9 in Rhesus Macaques of Chinese Origin in Comparison with Indian Macaques

Objective To develop a model of SHIV-KB9/Chinese origin rhesus （Ch Rh） macaques for vaccine research and to compare the pathogenesis of SHIV-KB9 in Ch Rh macaques with that reported in Indian rhesus （Ind Rh） macaques. Methods Seven mamu-A＊01 negative Ch Rh macaques were inoculated intravenously with 1-10000 MID50 of SHIV-KB9. The monkeys were monitored for viral load, CD4, CDS, SHIV-specific antibody and virus genetic variation. The results were compared with those previously observed in Ind Rh macaques. Results As compared to that observed in Ind Rh macaques, SHIV-KB9 in Ch Rh macaques displayed three identical disease progression patterns. However, the primary pattern was not identical between the two subspecies. The level of plasma viremia differed in SHIV-KB9-infected Ch Rh macaques which exhibited different outcomes from those in Ind Rh macaques. Generally, the values of viral load and the maintenance of CD4^＋ T cells were associated with humoral responses. Otherwise, the viral genetic distances （divergence, diversity） were larger in animals （M419, M425） with their CD4^＋ T cells profoundly depleted. Conclusion The model of SHIV-KB9/Ch Rh macaques displays a relatively slow progression to AIDS compared with Ind Rh macaques, which may more accurately reflect the potential of candidate vaccines in humans.

Adrenal pheochromocytoma impacts three main pathways: cysteine-methionine, pyrimidine, and tyrosine metabolism

Pheochromocytomas and paragangliomas(PPGLs)cause symptoms by altering the circulation levels of catecholamines and peptide hormones.Currently,the diagnosis of PPGLs relies on diagnostic imaging and the detection of catecholamines.In this study,we used ultra-performance liquid chromatography(UPLC)/quadrupole time-of-flight mass spectrometry(Q-TOF MS)analysis to identify and measure the perioperative differential metabolites in the plasma of adrenal pheochromocytoma patients.We identified differentially expressed genes by comparing the transcriptomic data of pheochromocytoma with the normal adrenal medulla.Through conducting two steps of metabolomics analysis,we identified 111 differential metabolites between the healthy group and the patient group,among which 53 metabolites were validated.By integrating the information of differential metabolites and differentially expressed genes,we inferred that the cysteine-methionine,pyrimidine,and tyrosine metabolism pathways were the three main metabolic pathways altered by the neoplasm.The analysis of transcription levels revealed that the tyrosine and cysteine-methionine metabolism pathways were downregulated in pheochromocytoma,whereas the pyrimidine pathway showed no significant difference.Finally,we developed an optimized diagnostic model of two metabolites,L-dihydroorotic acid and vanylglycol.Our results for these metabolites suggest that they may serve as potential clinical biomarkers and can be used to supplement and improve the diagnosis of pheochromocytoma.

Common genetic heterogeneity of human interleukin-37 leads to functional variance

Interleukin-37(IL-37)is an inhibitory member of the IL-1 family of cytokines.We previously found that balanced selection maintains common variations of the human IL37 gene.However,the functional consequences of this selection have yet to be validated.Here,using cells expressing exogenous IL-37 variants,including IL-37 Ref and IL-37 Var1 and Var2,we found that the three variants of IL-37 exhibited different immunoregulatory potencies in response to immune stimulation.The protein level of IL-37 Var2 was found to be significantly less than that of IL-37 Ref or Var1,despite the comparable mRNA levels of all three variants.Further study showed that IL-37 Var2 was rapidly degraded by a proteasome-dependent mechanism mediated by enhanced polyubiquitination,leading to a transient upregulation of IL-37 Var2 after immune stimulation.Finally,when ectopically expressed in cells,human IL-37 Var2 exerted less inhibition on proinflammatory cytokine production than did other IL-37 variants.Conversely,purified extracellular IL-37 variant proteins demonstrated comparable inhibitory abilities in vitro.In conclusion,our study reveals that common genetic variants of IL37 lead to different immune-inhibitory potencies,primarily as a result of differences in IL-37 protein stability,suggesting the possible involvement of these variants in various human diseases.

Studying gene duplication and genetic variation of budding yeast:a systems and interdisciplinary approach

Budding yeast (Saccharomyces cerevisiae) is a single cell model organism that is amenable to genome wide experimental interrogation using high-throughput genomics, proteomics

Preparation, identification, and clinical application ofanti-HBs monoclonal antibody that binds both wild-typeand immune escape mutant HBsAgs

Using a standard cellular fusion technique and indirect enzyme-linked immunosorbent assay(ELISA),a hybridoma cell line strain secreting anti-HBs monoclonal antibody(mAb)(defined G6 mAb)was obtained.The cells grew and secreted mAb stably.Antibody titers in the culture supernatant and ascites were 2.048�106 and 4.096�106,respectively.By applying the anti-HBs G6 mAb and horseradish peroxidase(HRP)-labeled goat anti-HBs antibody,we developed a sandwich ELISA(defined G6m ELISA)for detecting both wild-type and immune escape mutant HBsAgs(IEM HBsAg).The assay was performed to detect 17 species of genome recombinant expression HBsAg,including two wild-type species and 15 IEM HBsAg species,which varied in the“a”determinant,in a group of patients infected with hepatitis B virus(HBV).The patients previously had a lower ELISA detection signal[(absorbance of patients/absorbance of normal people(P/N):1.0–4.5)].The results demonstrated that the sensitivity of this assay to wild-type HBsAg was no less than 0.125μg/L;12 of 15 IEM HBsAg species(P/N≥2.5)were positive for G6 mAb.Of the positive IEM HBsAg species,two had a low absorbance value at 450 nm(A450),one had an intermediate A450 value and nine had a high A450 value,which was 7.55%(mean),59.4%and 92.1%–109.4%of the wild-type A450 value,respectively.The two species with low A450 value and the three negative species mutated at the bases 120–124 in thefirst loop of the HBV“a”determinant.Using the G6 ELISA and two commercial ELISA kits(A and B),177 patients were tested.The G6 ELISA had a significantly higher detection rate than either commercial ELISAs(19.21%vs 14.89%and 6.21%,respectively;P<0.01,P<0.05,respectively).

One Review on the Latest Etiology Research Progress of Primary Dysmenorrhea

Primary dysmenorrhea(PDM) is the painful menses with spasmodic cramping in the lower abdomen in the absence of any discernable macroscopic pelvic pathology. The prevalence of dysmenorrhea changes between 16% and 91% in women. About 2%-29% of the women studied suffer from severe pain. This review focuses on the current knowledge, particularly with regard to the latest research on the etiology of PDM. Full-text manuscripts on PDM were searched on PubMed and Google Scholar. One or more of the following search terms were used to obtain articles published: PDM, pain, functional magnetic resonance imaging, brain image, etiology, epidemiology, metabolism, hormone,gene variation, and quality of life. In this review, we detailed four potential etiology aspects of PDM: brain abnormality, gene expression,metabolism, and hyperalgesia. We highlighted the latest brain research on PDM patients and investigated genetic aspects. We are dedicated to identifying more metabolic variations and expand the previous knowledge on the sensitive pain threshold.

Successful medical treatment of west syndrome with a KCNA2 variant: a case report

Background:West syndrome is a devastating disorder characterized by a triad of epileptic spasms,abnormal electroencephalography(EEG),and developmental arrest or psychomotor delay.In addition to early diagnosis,knowing the etiology of the condition is also important for its treatment.Among various etiologies,the genetic factors,especially mutations of ion channel genes,are very common and strongly linked to West syndrome.Case presentation:A boy who had epileptic spasms from the age of 4 months was diagnosed with West syndrome based on the clinical manifestation and EEG results in Shenzhen Children's Hospital in June 2019.Trios whole-exome sequencing(WES)test and protein structural model prediction were performed.We also reviewed the clinical and genetic features of this syndrome and the mechanisms of action of topiramate(TPM)by literature search in databases of Online Mendelian Inheritance in Man,Clinical Genome Resource,PubMed,Chinese National Knowledge Infrastructure and Wanfang database using keywords"KCNA2""West syndrome"and"Topiramate"by December 2020.The relationship between the effect of TPM and the pathogenesis of the KCNA2 variant was also assessed.The WES test revealed c.244C>T/p.Arg82Cys varaint of KCNA2(NM_004974.3)in this patient,and Sanger sequencing identified this was a de novo mutation.As far as we know,this is the first report of the C.244C>T/p.Arg82Cys variant in KCNA2,which was likely a pathogenic mutation.The seizures were successfully controlled for 10 months by TPM after failure of sodium valproate,large doses of vitamin B6,and adrenocorticotropic hormone.We speculate that the therapeutic effect of TPM in this patient is partially due to the inhibition of carbonic anhydrase.Conclusions:Mutations in the KCNA2 gene should be considered for patients with West syndrome.The TPM treatment is probably effective for KCNA2-associated disorders.

New Trend of Management Science - Managerial Bioengineering System

This article seeks to depict the management science＇s new trend - managerial bioengineering system which is making decisive influence to enterprise management, generalizes the characteristics and categories of managerial gene variation, and brings forward a managerial genome plan with 5-＂All＂： participation of all members, enjoyment of all aspects, accumulation of all time, devotion of all strength and operation of all speed.