A multicenter prospective study of next-generation sequencing-based newborn screening for monogenic genetic diseases in China

Background Newborn screening(NBS)is an important and successful public health program that helps improve the long-term clinical outcomes of newborns by providing early diagnosis and treatment of certain inborn diseases.The develop-ment of next-generation sequencing(NGS)technology provides new opportunities to expand current newborn screening methodologies.Methods We designed a a newborn genetic screening(NBGS)panel targeting 135 genes associated with 75 inborn disorders by multiplex PCR combined with NGS.With this panel,a large-scale,multicenter,prospective multidisease analysis was conducted on dried blood spot(DBS)profiles from 21,442 neonates nationwide.Results We presented the positive detection rate and carrier frequency of diseases and related variants in different regions;and 168(0.78%)positive cases were detected.Glucose-6-Phosphate Dehydrogenase deficiency(G6PDD)and phenylketonuria(PKU)had higher prevalence rates,which were significantly different in different regions.The positive detection of G6PD variants was quite common in south China,whereas PAH variants were most commonly identified in north China.In addi-tion,NBGS identified 3 cases with DUOX2 variants and one with SLC25A13 variants,which were normal in conventional NBS,but were confirmed later as abnormal in repeated biochemical testing after recall.Eighty percent of high-frequency gene carriers and 60%of high-frequency variant carriers had obvious regional differences.On the premise that there was no significant difference in birth weight and gestational age,the biochemical indicators of SLC22A5 c.1400C>G and ACADSB c.1165A>G carriers were significantly different from those of non-carriers.Conclusions We demonstrated that NBGS is an effective strategy to identify neonates affected with treatable diseases as a supplement to current NBS methods.Our data also showed that the prevalence of diseases has significant regional charac-teristics,which provides a theoretical basis for screening diseases in different regions.

Neuroprotection mediated by the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage

The Wnt/Frizzled signaling pathway participates in many inflammation-linked diseases. However, the inflammatory response mediated by the Wnt/Frizzled signaling pathway in experimental subarachnoid hemorrhage has not been thoroughly investigated. Consequently, in this study, we examined the potential role of the Wnt/Frizzled signaling pathway in early brain injury in rat models of subarachnoid hemorrhage.Simultaneously, possible neuroprotective mechanisms were also investigated. Experimental subarachnoid hemorrhage rat models were induced by injecting autologous blood into the prechiasmatic cistern. Experiment 1 was designed to examine expression of the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage. In total, 42 adult rats were divided into sham(injection of equivalent volume of saline), 6-, 12-, 24-, 48-, 72-hour, and 1-week subarachnoid hemorrhage groups. Experiment 2 was designed to examine neuroprotective mechanisms of the Wnt/Frizzled signaling pathway in early brain injury induced by subarachnoid hemorrhage. Rats were treated with recombinant human Wnt1(rhwnt1), small interfering Wnt1(siwnt1) RNA, and monoclonal antibody of Frizzled1(anti-Frizzled1) at 48 hours after subarachnoid hemorrhage. Expression levels of Wnt1, Frizzled1, β-catenin, peroxisome proliferator-activated receptor-γ, CD36, and active nuclear factor-κB were examined by western blot assay and immunofluorescence staining. Microglia type conversion and inflammatory cytokine levels in brain tissue were examined by immunofluorescence staining and enzyme-linked immunosorbent assay. Our results show that compared with the sham group, expression levels of Wnt1, Frizzled1, and β-catenin were low and reduced to a minimum at 48 hours, gradually returning to baseline at 1 week after subarachnoid hemorrhage. rhwnt1 treatment markedly increased Wnt1 expression and alleviated subarachnoid hemorrhage-induced early brain injury(within 72 hours), including cortical cell apoptosis, brain edema, and neurobehavioral deficits, accompanied by increasing protein levels of β-catenin, CD36, and peroxisome proliferator-activated receptor-γ and decreasing protein levels of nuclear factor-κB. Of note, rhwnt1 promoted M2-type microglia conversion and inhibited release of inflammatory cytokines(interleukin-1β, interleukin-6, and tumor necrosis factor-α). In contrast, siwnt1 RNA and anti-Frizzled1 treatment both resulted in an opposite effect. In conclusion, the Wnt/Frizzled1 signaling pathway may participate in subarachnoid hemorrhage-induced early brain injury via inhibiting the inflammatory response, including regulating microglia type conversion and decreasing inflammatory cytokine release. The study was approved by the Animal Ethics Committee of Anhui Medical University and First Affiliated Hospital of USTC,Division of Life Sciences and Medicine, University of Science and Technology of China(approval No. LLSC-20180202) in May 2017.

SOCS1/JAK2/STAT3 axis regulates early brain injury induced by subarachnoid hemorrhage via inflammatory responses

The SOCS1/JAK2/STAT3 axis is strongly associated with tumor growth and progression,and participates in cytokine secretion in many diseases.However,the effects of the SOCS1/JAK2/STAT3 axis in experimental subarachnoid hemorrhage remain to be studied.A subarachnoid hemorrhage model was established in rats by infusing autologous blood into the optic chiasm pool.Some rats were first treated with JAK2/STAT3 small interfering RNA(Si-JAK2/Si-STAT3)or overexpression plasmids of JAK2/STAT3.In the brains of subarachnoid hemorrhage model rats,the expression levels of both JAK2 and STAT3 were upregulated and the expression of SOCS1 was downregulated,reaching a peak at 48 hours after injury.Simultaneously,the interactions between JAK2 and SOCS1 were reduced.In contrast,the interactions between JAK2 and STAT3 were markedly enhanced.Si-JAK2 and Si-STAT3 treatment alleviated cortical neuronal cell apoptosis and necrosis,destruction of the blood-brain barrier,brain edema,and cognitive functional impairment after subarachnoid hemorrhage.This was accompanied by decreased phosphorylation of JAK2 and STAT3 protein,decreased total levels of JAK2 and STAT3 protein,and increased SOCS1 protein expression.However,overexpression of JAK2 and STAT3 exerted opposite effects,aggravating subarachnoid hemorrhage-induced early brain injury.Si-JAK2 and Si-STAT3 inhibited M1-type microglial conversion and the release of pro-inflammatory factors(inducible nitric oxide synthase,interleukin-1β,and tumor necrosis factor-α)and increased the release of anti-inflammatory factors(arginase-1,interleukin-10,and interleukin-4).Furthermore,primary neurons stimulated with oxyhemoglobin were used to simulate subarachnoid hemorrhage in vitro,and the JAK2 inhibitor AG490 was used as an intervention.The in vitro results also suggested that neuronal protection is mediated by the inhibition of JAK2 and STAT3 expression.Together,our findings indicate that the SOCS1/JAK2/STAT3 axis contributes to early brain injury after subarachnoid hemorrhage both in vitro and in vivo by inducing inflammatory responses.This study was approved by the Animal Ethics Committee of Anhui Medical University and the First Affiliated Hospital of University of Science and Technology of China(approval No.LLSC-20180202)on March 1,2018.

Molecular determinants of response to 5-fluorouracil-based chemotherapy in colorectal cancer: The undisputable role of microribonucleic acids

5-flurouracil(5-FU)-based chemotherapy is the main pharmacological therapy for advanced colorectal cancer(CRC).Despite significant progress in the treatment of CRC during the last decades,5-FU drug resistance remains the most important cause of failure in CRC therapy.Resistance to 5-FU is a complex and multistep process.Different mechanisms including microsatellite instability,increased expression level of key enzyme thymidylate synthase and its polymorphism,increased level of 5-FU-activating enzymes and mutation of TP53 are proposed as the main determinants of resistance to 5-FU in CRC cells.Recently,microribonucleic acids(miRNA)and their alterations were found to have a crucial role in 5-FU resistance.In this regard,the miRNA-mediated mechanisms of 5-FU drug resistance reside among the new fields of pharmacogenetics of CRC drug response that has not been completely discovered.Identification of the biological markers that are related to response to 5-FU-based chemotherapy is an emerging field of precision medicine.This approach will have an important role in defining those patients who are most likely to benefit from 5-FU-based chemotherapy in the future.Thereby,the identification of 5-FU drug resistance mechanisms is an essential step to predict and eventually overcome resistance.In the present comprehensive review,we will summarize the latest knowledge regarding the molecular determinants of response to 5-FU-based chemotherapy in CRC by emphasizing the role of miRNAs.

Some of the experimental and clinical aspects of the effects of the maternal diabetes on developing hippocampus

Diabetes mellitus during pregnancy is associated with an increased risk of multiple congenital anomalies in progeny.There are sufficient evidence suggesting that the children of diabetic women exhibit intellectual and behavioral abnormalities accompanied by modification of hippocampus structure and function.Although,the exact mechanism by which maternal diabetes affects the developing hippocampus remains to be defined.Multiple biological alterations,including hyperglycemia,hyperinsulinemia,oxidative stress,hypoxia,and iron deficiency occur in pregnancies with diabetes and affect the development of central nervous system(CNS) of the fetus.The conclusion from several studies is that disturbance in glucose and insulin homeostasis in mothers and infants are major teratogenic factor in the development of CNS.Insulin and Insulin-like growth factor-1(IGF-1) are two key regulators of CNS function and development.Insulin and IGF-1 receptors(IR and IGF1 R,respectively) are distributed in a highly specific pattern with the high density in some brain regions such as hippocampus.Recent researches have clearly established that maternal diabetes disrupts the regulation of both IR and IGF1 R in the hippocampus of rat newborn.Dissecting out the mechanisms responsible for maternal diabetes-related changes in the development of hippocampus is helping to prevent from impaired cognitive and memory functions in offspring.

Identification of a Novel COL17A1 Compound Heterozygous Mutation in a Chinese Girl with Non-Herlitz Junctional Epidermolysis Bullosa

Summary:Non-Herlitz junctional epidermolysis bullosa(JEB-nH),an autosomal recessive bullous genodermatosis,is characterized by generalized skin blistering from birth onward,dental anomalies,universal alopecia and nail dystrophy.The underlying defect is mutation of the COLI7AI gene encoding the type XVⅡcollagen,resulting in losing structure for attachment of basal epithelial cells to the matrix.In present study,we described one case of congenitally affected female child aged 10 years,with skin blistering.Dermatologic examination revealed sparse,mild blisters on the face and hand,with profound enamel pitting of the teeth.Skin biopsy from proband's bullous skin displayed subepidermal bulla formation without acantholysis.The immunofluorescence of anti-type XVⅡcollagen antibody staining showed loss of type XVⅡcollagen staining at the basement membrane zone.A combination of whole exome sequencing(WES)and Sanger sequencing revealed the novel heterozygous mutations(C.4324C>T;p.Q1442^*and C.I 834G>C;p.G612R)in COLI7AI gene,which could be associated with the observed JEB-nH.One allele had a novel nonsense mutation(c.4324C>T;p.Q1442^*),resulting in nonsense-mediated mRNA decay and truncated collagen XVⅡ;the other allelc had a novel misscnse mutation of c.1834G>C;p.G612R in exon 22,causing a glycine-to-arginine substitution in the Gly-X-Y triple helical repeating motifs and decreasing the thermal stability of collagen XVⅡ.Our findings indicate that the genetic test based on WES can be useful in diagnosing JEB-nH patients.The novel pathogenic mutations identified would further expand our understanding of the mutation spectrum of COLI7AI gene in association with the inherited blistering diseases.

PRKCDBP Methylation is a Potential and Promising Candidate Biomarker for Non-small Cell Lung Cancer

Background and objectives:The occurrence and development of lung cancer are closely linked to epigenetic modification.Abnormal DNA methylation in the CpG island region of genes has been found in many cancers.Protein kinase C delta binding protein(PRKCDBP) is a potential tumor suppressor and its epigenetic changes are found in many human malignancies.This study investigated the possibility of PRKCDBP methylation as a potential biomarker for non-small cell lung cancer(NSCLC).Methods:We measured the methylation levels of PRKCDBP in the three groups of NSCLC tissues.Promoter activity was measured by the dual luciferase assay,with S’-aza-deoxycytidine to examine the effect of demethylation on the expression level of PRKCDBP.Results:The methylation levels of PRKCDBP in tumor tissues and 3 cm para-tumor were higher than those of distant(>10 cm)non-tumor tissues.Receiver operating characteristic(ROC) curve analysis between tumor tissues and distant non-tumor tissues showed that the area under the line(AUC) was 0.717.Dual luciferase experiment confirmed that the promoter region was able to promote gene expression.Meanwhile,in vitro methylation of the fragment(PRKCDBP;e) could significantly reduce the promoter activity of the fragment.Demethylation of 5’-aza-deoxycytidine in lung cancer cell lines A549 and H1299 showed a significant up-regulation of PRKCDBP mRNA levels.Conclusion:PRKCDBP methylation is a potential and promising candidate biomarker for non-small cell lung cancer.

Germline mutations in Thai patients with nonmucinous epithelial ovarian cancer

BACKGROUND Genetic testing is widely recommended for all epithelial ovarian cancer(EOC)patients.However,an increased probability of identifying germline mutations has been reported in selected patients with risk factors such as a family history or personal history of cancer and high-grade serous carcinoma(HGSC)subtype.HGSC has been reported to be the most common subtype of EOC worldwide(approximately 70%).However,this subtype is less prevalent in Thai patients(reported as only 20%).The difference in the distribution of various subtypes of EOC may reflect the incidence of germline mutations in Thai EOC patients.AIM To evaluate the frequencies of germline mutations in EOC patients and to compare the frequencies in those with and without clinical risk factors for hereditary ovarian cancer.METHODS This cross-sectional study included 112 nonmucinous EOC patients who underwent primary surgery at our tertiary care hospital.Clinical risk factors for hereditary ovarian cancer were defined as follows:Age below 40 years,a significant family history of cancer,synchronous ovarian and endometrial cancer,and HGSC.Comprehensive germline mutations were detected by nextgeneration sequencing.RESULTS Of a total of 112 patients,82(73.2%)patients had≥1 risk factor and 30(26.8%)patients had no risk factors.Germline mutations were detected in 26 patients:20(17.8%)patients had BRCA1/2 mutations,but 6(5.4%)patients had mutations in other genes,including 1 in MLH1,1 in MSH2,1 in RAD51C,2 in ATM and 1 in CDH1.Germline mutations were only detected in patients with risk factors(26 of 82,31.7%),not in patients without risk factors(P<0.001).A significant family history of cancer and HGSC were the only two significant risk factors associated with a higher proportion of germline mutations(56.3%vs 10%for those with and without a history of cancer,respectively,40.8%vs 9.5%for those with and without HGSC).Germline BRCA mutations were detected in 38.8%of patients with HGSC but in only 1.6%of those with non-HGSC.An age below 40 years,personal history of breast cancer,and synchronous ovarian and endometrial cancer were not significant factors(14.3%vs 23.5%,33.3%vs 21%,22.2%vs 22.3%).CONCLUSION Approximately one-third of EOC patients with risk factors had germline mutations.Almost all germline BRCA mutations were found in patients with the HGSC subtype.Selected patients with HGSC and a family history of cancer should be initially considered for genetic analysis in Thailand.

PI3K/AKT signaling pathway as a critical regulator of Cisplatin response in tumor cells

Chemotherapy is one of the main therapeutic modalities for cancer patients.Cisplatin(CDDP),as one of thefirst-line drugs,is of great importance in the chemotherapy of various tumors.However,a significant percentage of cancer patients are resistant to CDDP treatment.Due to the CDDP side effects on normal tissues,the diagnosis of CDDP resistance is required to suggest the most efficient therapeutic strategies for cancer patients.Several molecular mechanisms and signaling pathways are associated with CDDP response.The PI3K/AKT signaling pathway has a pivotal role in the transmission of extracellular signals into the cells to regulate various pathophysiological processes such as cell proliferation,migration,and drug resistance.In the present review,we summarized all of the studies which have been reported on the role of PI3K/AKT pathway in regulation of CDDP response.It was shown that the PI3K/AKT pathway is mainly involved in CDDP response in lung,ovarian,and gastrointestinal cancers.It was also observed that the non-coding RNAs have a key role in CDDP response by regulation of PI3K/AKT pathway.This review paves the way for suggesting a PI3K/AKT-related panel marker for the prediction of CDDP response in different cancer patients.

Single-copy Loss of Rho Guanine Nucleotide Exchange Factor 10(arhgef10)Causes Locomotor Abnormalities in Zebrafish Larvae

Objective To determine if ARHGEF10 has a haploinsufficient effect and provide evidence to evaluate the severity,if any,during prenatal consultation.Methods Zebrafish was used as a model for generating mutant.The pattern of arhgef10 expression in the early stages of zebrafish development was observed using whole-mount in situ hybridization(WISH).CRISPR/Cas9 was applied to generate a zebrafish model with a single-copy or homozygous arhgef10 deletion.Activity and light/dark tests were performed in arhgef10^(−/−),arhgef10^(+/−),and wild-type zebrafish larvae.ARHGEF10 was knocked down using small interferon RNA(siRNA)in the SH-SY5Y cell line,and cell proliferation and apoptosis were determined using the CCK-8 assay and Annexin V/PI staining,respectively.Results WISH showed that during zebrafish embryonic development arhgef10 was expressed in the midbrain and hindbrain at 36-72 h post-fertilization(hpf)and in the hemopoietic system at 36-48 hpf.The zebrafish larvae with single-copy and homozygous arhgef10 deletions had lower exercise capacity and poorer responses to environmental changes compared to wild-type zebrafish larvae.Moreover,arhgef10^(−/−) zebrafish had more severe symptoms than arhgef10^(+/-) zebrafish.Knockdown of ARHGEF10 in human neuroblastoma cells led to decreased cell proliferation and increased cell apoptosis.Conclusion Based on our findings,ARHGEF10 appeared to have a haploinsufficiency effect.

Pulmonary Hypertension Induced by Thalidomide (and Derivatives) in Patients with Multiple Myeloma: A Systematic Review

Thalidomide is widely used in the treatment of multiple myeloma (MM). In recent years, several cases of pulmonary hypertension have been reported following treatment with thalidomide. The aim of this review was to evaluate the published literature on multiple myeloma patients with pulmonary hypertension following thalidomide treatment. A literature search was performed between 2000 and 2016. A total of 7 eligible studies were identified and deemed eligible, including 11 cases—approximately 37% (4 cases) with IgA (k), 27% (3 cases) with IgG (λ) MM, 27% (3 cases) with IgG (k) MM, and one case (9%) with primary plasma cell leukemia (PPCL). The vast majority of cases—82% (9 cases)—are associated with thalidomide, while only 18% (2 cases) are related to thalidomide derivatives (lenalidomide and pomalidomide). In conclusion, pulmonary hypertension induced by thalidomide or derivatives in multiple myeloma (MM) patients is related to a multifactorial etiology including the pathophysiology of the disease, thromboembolic events, preexisted cardiovascular conditions, comorbidities, and combination with other chemo- or bio-therapeutic agents. MM patients should be evaluated for signs and symptoms underlying cardiopulmonary disease before initiating, and during treatment with thalidomide.

Gene therapy for monogenic disorders: challenges, strategies, and perspectives

Monogenic disorders refer to a group of human diseases caused by mutations in single genes. While disease-modifying therapies have offered some relief from symptoms and delayed progression for some monogenic diseases, most of these diseases still lack effective treatments. In recent decades, gene therapy has emerged as a promising therapeutic strategy for genetic disorders. Researchers have developed various gene manipulation tools and gene delivery systems to treat monogenic diseases. Despite this progress, concerns about inefficient delivery, persistent expression, immunogenicity, toxicity, capacity limitation, genomic integration, and limited tissue specificity still need to be addressed. This review gives an overview of commonly used gene therapy and delivery tools, along with the challenges they face and potential strategies to counter them.

CircPTEN-MT from PTEN regulates mitochondrial energy metabolism

Phosphatase and tensin homolog(PTEN)is a multifunctional gene involved in a variety of physiological and pathological processes.Circular RNAs(circRNAs)are generated from back-splicing events during mRNA processing and participate in cell biological processes through binding to RNAs or proteins.However,PTEN-related circRNAs are largely unknown.Here,we report that circPTEN-mitochondria(MT)(hsa_circ_0002934)is a circular RNA encoded by exons 3,4,and 5 of PTEN and is a critical regulator of mitochondrial energy metabolism.CircPTEN-MT is localized to mitochondria and physically associated with leucine-rich pentatricopeptide repeat-containing protein(LRPPRC),which regulates posttranscriptional gene expression in mitochondria.Knocking down circPTEN-MT reduces the interaction of LRPPRC and steroid receptor RNA activator(SRA)stem-loop interacting RNA binding protein(SLIRP)and inhibits the polyadenylation of mitochondrial mRNA,which decreases the mRNA level of the mitochondrial complex I subunit and reduces mitochondrial membrane potential and adenosine triphosphate production.Our data demonstrate that circPTEN-MT is an important regulator of cellular energy metabolism.This study expands our understanding of the role of PTEN,which produces both linear and circular RNAs with different and independent functions.

Umbilical cord blood application analysis of Guangdong Cord Blood Bank

To the Editor:With the development of clinical research and the application of cord blood,the clinical value of umbilical cord blood(UCB)has drawn more and more attention.UCB has been increasingly applied to treatments of hematological diseases,autoimmune diseases,and malignant tumors in children and adults[1]and has achieved high overall survival.[1–3]UCB contains substantial amounts of various stem cells,including hematopoietic stem cells(HSC),mesenchymal stem cells,endothelial progenitor cells,and so on[4];among them,HSC are most widely used in study and clinical application.Transplantation,where UCB HSC are implanted into patients,requires ruining the immune system and hematopoietic system before the procedure.Another therapeutic method is UCB infusion,where UCB is regarded as a nutrient,to improve the outcome of peripheral blood HSC transplantation[5]or symptoms via paracrine pathway of UCB stem cells.[6]In addition to the application of allogeneic UCB,autologous UCB has also been used in clinical treatment research.Autologous UCB is not limited to treating hemopathy,but also used to treat non-genetic metabolic disease,immune disease,and nervous system injury.[6]

Uncompromised NK cell activation is essential for virus-specific CTL activity during acute influenza virus infection

Natural killer (NK) cells are indispensable components of both the innate and adaptive immune response. However,their precise roles in the cross-talk between innate and adaptive immunity during influenza virus infection remaincontroversial. By comparing NK cell dynamics and activity under a sub-lethal dose and high dose of influenza virusinfection, we showed that influenza virus PR8 directly infected NK cells during natural infection, which wasconsistent with our previous findings obtained from an in vitro investigation of human NK cells. The impairments incytotoxicity and IFN-γ production by spleen NK cells following high-dose infection were accompanied by decreasedvirus-specific killing mediated by cytotoxic T lymphocytes (CTLs). Importantly, the weakened CTL activity could bereversed by adoptive transfer of spleen NK cells harvested from low-dose-infected mice but not healthy donors.Taken together, our data provide direct evidence supporting the contribution of NK cells to antiviral T-cellresponses. This study also indicates that a novel NK-targeted immune evasion strategy is used by influenza virusto shrink both innate and adaptive immune responses.

Complete genome sequence of high-yield strain S. lincolnensis B48 and identification of crucial mutations contributing to lincomycin overproduction

The lincosamide family antibiotic lincomycin is a widely used antibacterial pharmaceutical generated by Streptomyces lincolnensis,and the high-yield strain B48 produces 2.5 g/L lincomycin,approximately 30-fold as the wild-type strain NRRL 2936.Here,the genome of S.lincolnensis B48 was completely sequenced,revealing a^10.0 Mb single chromosome with 71.03%G+C content.Based on the genomic information,lincomycinrelated primary metabolism network was constructed and the secondary metabolic potential was analyzed.In order to dissect the overproduction mechanism,a comparative genomic analysis with NRRL 2936 was performed.Three large deletions(LDI-III),one large inverted duplication(LID),one long inversion and 80 small variations(including 50 single nucleotide variations,13 insertions and 17 deletions)were found in B48 genome.Then several crucial mutants contributing to higher production phenotype were validated.Deleting of a MarRtype regulator-encoding gene slinc377 from LDI,and the whole 24.7 kb LDII in NRRL 2936 enhanced lincomycin titer by 244%and 284%,respectively.Besides,lincomycin production of NRRL 2936 was increased to 7.7-fold when a 71 kb supercluster BGC33 from LDIII was eliminated.As for the duplication region,overexpression of the cluster situated genes lmbB2 and lmbU,as well as two novel transcriptional regulator-encoding genes(slinc191 and slinc348)elevated lincomycin titer by 77%,75%,114%and 702%,respectively.Furthermore,three negative correlation genes(slinc6156,slinc4481 and slinc6011)on lincomycin biosynthesis,participating in regulation were found out.And surprisingly,inactivation of RNase J-encoding gene slinc6156 and TPR(tetratricopeptide repeat)domain-containing protein-encoding gene slinc4481 achieved lincomycin titer equivalent to 83%and 68%of B48,respectively,to 22.4 and 18.4-fold compared to NRRL 2936.Therefore,the comparative genomics approach combined with confirmatory experiments identified that large fragment deletion,long sequence duplication,along with several mutations of genes,especially regulator genes,are crucial for lincomycin overproduction.

The paradoxical roles of miR-4295 in human cancer:Implications in pathogenesis and personalized medicine

MiR-4295,located on chromosome 10q25.2,is a unique miRNA with a wide range of biological functions.miR-4295 is widely expressed in vivo,participating in the biological processes of multiple cancers.Although miR-4295 is dysregulated in various cancers,it has also been found to have the function of inhibiting cancer.At the same time,the expression of miR-4295 is related to prognosis and can be affected by numerous factors connecting to the therapeutic effects of various drugs.This article is to better summarize the role of miR-4295 in cancer and review the potential diagnostic,prognostic,and therapeutic value of miR-4295,which may provide insight into subsequent research.

Update of the spectrum of mucopolysaccharidoses type Ⅲ in Tunisia: identification of three novel mutations and in silico structural analysis of the missense mutations

Background:Mucopolysaccharidoses type Ⅲ (MPS Ⅲ) are a group of autosomai recessive lysosomal storage diseases,caused by mutations in genes that code for enzymes involved in the lysosomal degradation of heparan sulphate:heparan sulfate sulfamidase (SGSH),a-N-acetylglucosaminidase (NAGLU),heparan sulfate acetyl-CoA:a-glucosaminide N-acetyltransferase (HGSNAT),and N-acetylglucosamine-6-sulfatase (GNS).Methods:In this study,we have performed the molecular analysis of the SGSH,NAGLU and HGSNAT genes in 10 patients from 6 different MPS Ⅲ Tunisian families.Results:In the SGSH gene,two mutations were identified:one novel (p.D477N) and one already described (p.Q365X).In the NAGLU gene,two novel mutations were discovered (p.L550P and p.E153X).For the novel missense mutations found in these two genes we performed an in silico structural analysis and the results were consistent with the clinical course of the patients harboring those mutations.Finally,in HGSNAT gene,we found the splicesite mutation c.234+1G>A that had already been reported as relatively frequent in MPS ⅢC patients from countries surrounding the basin of the Mediterranean sea.Its presence in two Tunisian MPS ⅢC families points to the hypothesis of its peri Mediterranean origin.With the exception of the c.234+1G>A mutation,that was identified in two unrelated MPS ⅢC families,the other identified mutations were family-specific and were always found in homozygosity in the patients studied,thus reflecting the existence of consanguinity in MPS Ⅲ Tunisian families.Conclusions:Three novel mutations are reported here,further contributing to the knowledge of the molecular basis of these diseases.The results of this study will allow carrier detection in affected families and prenatal molecular diagnosis,leading to an improvement in genetic counseling.

Identification of a novel nonsense mutation in kyphoscoliosis peptidase gene in an Iranian patient with myofibrillar myopathy

Myofibrillar myopathies(MFMs)are rare genetic and slowly progressive neuromuscular disorders.Several pathogenic mutations have been reported in MFM-related genes including DES,CRYAB,MYOT,LDB3 or ZASP,FLNC,BAG3,FHL1 and DNAJB6.Although MFMs is commonly inherited in an autosomal dominant manner,the inheritance pattern and novel mutated genes are not thoroughly elucidated in some cases.Here,we report discovery of a novel nonsense mutation in a 29-year-old Iranian male patient with motor disorders and deformity in his lower limbs.His parents are second cousins.Hereditary Motor Sensory Neuropathy as initial genetic diagnosis was ruled out.Whole exome sequencing using NGS on Illumina Hi-Seq4000 platform was performed to identify the disease and possible mutated gene(s).Our data analysis identified a homozygous nonsense unreported c.C415T(p.R139X)variant on kyphoscoliosis peptidase(KY)gene(NM_178554:exon4).Sanger sequencing of this mutation has been performed for his other related family members.Sequencing and segregation analysis was confirmed the NGS results and autosomal recessive inheritance pattern of the disease.