Accurate Diagnosis of SARS-CoV-2 JN.1 by Sanger Sequencing of Receptor-Binding Domain Is Needed for Clinical Evaluation of Its Immune Evasion

Background: Omicron JN.1 has become the dominant SARS-CoV-2 variant in recent months. JN.1 has the highest number of amino acid mutations in its receptor binding domain (RBD) and has acquired a hallmark L455S mutation. The immune evasion capability of JN.1 is a subject of scientific investigation. The US CDC used SGTF of TaqPath COVID-19 Combo Kit RT-qPCR as proxy indicator of JN.1 infections for evaluation of the effectiveness of updated monovalent XBB.1.5 COVID-19 vaccines against JN.1 and recommended that all persons aged ≥ 6 months should receive an updated COVID-19 vaccine dose. Objective: Recommend Sanger sequencing instead of proxy indicator to diagnose JN.1 infections to generate the data based on which guidelines are made to direct vaccination policies. Methods: The RNA in nasopharyngeal swab specimens from patients with clinical respiratory infection was subjected to nested RT-PCR, targeting a 398-base segment of the N-gene and a 445-base segment of the RBD of SARS-CoV-2 for amplification. The nested PCR amplicons were sequenced. The DNA sequences were analyzed for amino acid mutations. Results: The N-gene sequence showed R203K, G204R and Q229K, the 3 mutations associated with Omicron BA.2.86 (+JN.1). The RBD sequence showed 24 of the 26 known amino acid mutations, including the hallmark L455S mutation for JN.1 and the V483del for BA.2.86 lineage. Conclusions: Sanger sequencing of a 445-base segment of the SARS-CoV-2 RBD is useful for accurate determination of emerging variants. The CDC may consider using Sanger sequencing of the RBD to diagnose JN.1 infections for statistical analysis in making vaccination policies.

Sanger Sequencing for Molecular Diagnosis of SARS-CoV-2 Omicron Subvariants and Its Challenges

Large population passages of the SARS-CoV-2 in the past two and a half years have allowed the circulating virus to accumulate an increasing number of mutations in its genome. The most recently emerging Omicron subvariants have the highest number of mutations in the Spike (S) protein gene and these mutations mainly occur in the receptor-binding domain (RBD) and the N-terminal domain (NTD) of the S gene. The European Centre for Disease Prevention and Control (eCDC) and the World Health Organization (WHO) recommend partial Sanger sequencing of the SARS-CoV-2 S gene RBD and NTD on the polymerase chain reaction (PCR)-positive samples in diagnostic laboratories as a practical means of determining the variants of concern to monitor possible increased transmissibility, increased virulence, or reduced effectiveness of vaccines against them. The author’s diagnostic laboratory has implemented the eCDC/WHO recommendation by sequencing a 398-base segment of the N gene for the definitive detection of SARS-CoV-2 in clinical samples, and sequencing a 445-base segment of the RBD and a 490 - 509-base segment of the NTD for variant determination. This paper presents 5 selective cases to illustrate the challenges of using Sanger sequencing to diagnose Omicron subvariants when the samples harbor a high level of co-existing minor subvariant sequences with multi-allelic single nucleotide polymorphisms (SNPs) or possible recombinant Omicron subvariants containing a BA.2 RBD and an atypical BA.1 NTD, which can only be detected by using specially designed PCR primers. In addition, Sanger sequencing may reveal unclassified subvariants, such as BA.4/BA.5 with L84I mutation in the S gene NTD. The current large-scale surveillance programs using next-generation sequencing (NGS) do not face similar problems because NGS focuses on deriving consensus sequence.

Application of next-generation sequencing in clinical oncology to advance personalized treatment of cancer

With the development and improvement of new sequencing technology,next-generation sequencing(NGS) has been applied increasingly in cancer genomics research over the past decade.More recently,NGS has been adopted in clinical oncology to advance personalized treatment of cancer.NGS is used to identify novel and rare cancer mutations,detect familial cancer mutation carriers,and provide molecular rationale for appropriate targeted therapy.Compared to traditional sequencing,NGS holds many advantages,such as the ability to fully sequence all types of mutations for a large number of genes(hundreds to thousands) in a single test at a relatively low cost.However,significant challenges,particularly with respect to the requirement for simpler assays,more flexible throughput,shorter turnaround time,and most importantly,easier data analysis and interpretation,will have to be overcome to translate NGS to the bedside of cancer patients.Overall,continuous dedication to apply NGS in clinical oncology practice will enable us to be one step closer to personalized medicine.

Association of NFKB1 gene polymorphism (rs28362491) with levels of inflammatory biomarkers and susceptibility to diabetic nephropathy in Asian Indians

AIM To investigate the association of NFKB1 gene-94 ATTG insertion/deletion(rs28362491) polymorphism with inflammatory markers and risk of diabetic nephropathy in Asian Indians.METHODS A total of 300 subjects were recruited(100 each), normoglycemic,(NG); type 2 diabetes mellitus(T2DM) without any complications(DM) and T2 DM with diabetic nephropathy [DM-chronic renal disease(CRD)]. Analysis was carried out by polymerase chain reaction-restriction fragment length polymorphism and ELISA. Pearson's correlation, analysis of variance and logistic regression wereused for statistical analysis.RESULTS The allelic frequencies of-94 ATTG insertion/deletion were 0.655/0.345(NG), 0.62/0.38(DM) and 0.775/0.225(DM-CRD). The-94 ATTG ins allele was associated with significantly increased levels of urinary monocyte chemoattractant protein-1(u MCP-1); u MCP-1(P = 0.026) and plasma tumor necrosis factor-alpha(TNF-α); TNF-α(P = 0.030) and almost doubled the risk of diabetic nephropathy(OR = 1.91, 95%CI: 1.080-3.386, P = 0.025).CONCLUSION-94 ATTG ins/ins polymorphism might be associated with increased risk of developing nephropathy in Asian Indian subjects with diabetes mellitus.

Synergistic anticancer effect of exogenous wild-type p53 gene combined with 5-FU in human colon cancer resistant to 5-FU in vivo

AIM To investigate the anticancer effect of a recombinant adenovirus-mediated p53(r Ad-p53) combined with 5-fluorouracil(5-FU) in human colon cancer resistant to 5-FU in vivo and the mechanism of r Ad-p53 in reversal of 5-FU resistance.METHODS nude mice bearing human colon cancer SW480/5-FU(5-FU resistant) were randomly assigned to four groups(n = 25 each): control group, 5-FU group, r Ad-p53 group, and r Ad-p53 + 5-FU group. At 24 h, 48 h, 72 h, 120 h and 168 h after treatment, 5 mice were randomly selected from each group and sacrificed using an overdose of anesthetics. The tumors were removed and the protein expressions of p53, protein kinase C(PKC), permeability-glycoprotein(P-gp) and multidrug resistance-associated protein 1(MRP1)(Western blot) and apoptosis(TUNEL) were determined.RESULTS The area ratios of tumor cell apoptosis were larger in the r Ad/p53 + 5-FU group than that in the control, 5-FU and r Ad/p53 groups(P < 0.05), and were larger in the r Ad/p53 group than that of the control group(P < 0.05) and the 5-FU group at more than 48 h(P < 0.05). The p53 expression was higher in the r Ad/p53 and the r Ad/p53 + 5-FU groups than that of the control and 5-FU groups(P < 0.05), and were higher in the r Ad/p53 + 5-FU group than that of the r Ad/p53 group(P < 0.05). Overexpression of PKC, P-gp and MRP1 was observed in the 5-FU and control groups. In the r Ad/p53 + 5-FU group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups(P < 0.05), and the expression of PKC was lower than that of the control, 5-FU and r Ad/p53 groups at more than 48 h(P < 0.05). In the r Ad/p53 group, the expression of P-gp and MRP1 was lower that of the control and 5-FU groups at more than 48 h(P < 0.05), and the expression of PKC was lower than that of the control and 5-FU groups at more than 120 h(P < 0.05).CONCLUSION5-FU combined with r Ad-p53 has a synergistic anticancer effect in SW480/5-FU(5-FU resistance), which contributes to reversal of 5-FU resistance.

Next-generation sequencing and its clinical application

Next-generation sequencing (NGS) is a new technology used for DNA and RNA sequencing and variant/mutation detection. NGS can sequence hundreds and thousands of genes or whole genome in a short period of time. The sequence variants/mutations detected by NGS have been widely used for disease diagnosis, prognosis, therapeutic decision, and follow up of patients. The capacity of its massive parallel sequencing offers new opportunities for personalized precision medicine.

Predictive value of K-ras and PIK3CA in non-small cell lung cancer patients treated with EGFR-TKIs: a systemic review and meta-analysis

Objective: A meta-analysis was performed to augment the insufficient data on the impact of mutative EGFR downstream phosphatidylinositol-3-kinase(PI3K) and mitogen-activated protein kinase(MAPK) pathways on the clinical efficiency of epidermal growth factor receptor tyrosine kinase inhibitor(EGFR-TKI) treatment of non-small cell lung cancer(NSCLC) patients.Methods: Network databases were explored in April, 2015. Papers that investigated the clinical outcomes of NSCLC patients treated with EGFR-TKIs according to the status of K-ras and/or PIK3 CA gene mutation were included. A quantitative meta-analysis was conducted using standard statistical methods. Odds ratios(ORs) for objective response rate(ORR) and hazard ratios(HRs) for progression-free survival(PFS) and overall survival(OS) were calculated.Results: Mutation in K-ras significantly predicted poor ORR [OR =0.22; 95% confidence interval(CI), 0.13-0.35], shorter PFS(HR =1.56; 95% CI, 1.27-1.92), and shorter OS(HR =1.59; 95% CI, 1.33-1.91) in NSCLC patients treated with EGFR-TKIs. Mutant PIK3 CA significantly predicted shorter OS(HR =1.83; 95% CI, 1.05-3.20), showed poor ORR(OR =0.70; 95% CI, 0.22-2.18), and shorter PFS(HR =1.79; 95% CI, 0.91-3.53) in NSCLC patients treated with EGFR-TKIs.Conclusion: K-ras mutation adversely affected the clinical response and survival of NSCLC patients treated with EGFRTKIs. PIK3 CA mutation showed similar trends. In addition to EGFR, adding K-ras and PIK3 CA as routine gene biomarkers in clinical genetic analysis is valuable to optimize the effectiveness of EGFR-TKI regimens and identify optimal patients who will benefit from EGFR-TKI treatment.

The next generation of cancer management

As readers of Cancer Biology and Medicine well know,there has been a seismic shift in human molecular biology over the past few years,as momentous in its own way as the discovery of the double-helical structure of DNA by Watson and Crick 60 years ago,the elucidation of the genetic code shortly thereafter,the advent of recombinant DNA and gene

Whole Exome Sequencing Identifies A Novel Pathogenic Bmpr2 Variant in Pulmonary Atresia

Objective:Pulmonary atresia(PA)is a rare type of complex cyanotic congenital heart defect characterized primarily by an undeveloped pulmonary valve or pulmonary artery.Therefore,defining a disease-causing gene mutation in a pulmonary atresia family is a possible method of genetic counseling,future prenatal diagnosis,and therapeutic approaches for pulmonary atresia.Methods:Blood samples were collected from six PA family members,and genomic DNA was extracted using the QIAamp DNA Blood Mini Kit.Gene detection was performed using a second-generation sequencing gene panel.Results:Genetic testing results indicated that a heterozygous mutation originating from maternal inheritance was detected in the BMPR2 gene of the proband’s genomic DNA.The pathogenic gene was c.2804C>T(p.A935V).The mutation was also detected in the genomic DNA of the proband’s elder brother(III-1),but not in other family members.Conclusion:To the best of our knowledge,this is the first study to report the BMPR2 variant responsible for pulmonary atresia.The frequency of the c.2804C>T(p.A935V)mutation detected in this family is extremely low in the normal population(1/246048).The mutation was highly conserved among different species.Sorting intolerant from tolerant(SIFT)predicts it to be a harmful mutation.

A three-year clinical investigation of a Chinese child with craniometaphyseal dysplasia caused by a mutated ANKH gene

BACKGROUND Craniometaphyseal dysplasia(CMD)is a rare genetic disorder.Autosomal dominant CMD(AD-CMD)is caused by mutations in the ANKH gene.Affected individuals typically have distinctive facial features including progressive thickening of the craniofacial bones.Treatment for AD-CMD primarily consists of surgical intervention to release compression of the cranial nerves and the brain stem/spinal cord.To alleviate progression of the clinical course and improve the quality of life in children waiting to undergo the necessary surgery,we investigated clinical changes in a diagnosed patient with AD-CMD over three years.CASE SUMMARY A 17-mo-old boy presented with progressive nasal obstruction,snoring and hearing loss symptoms.Physical examination showed enlargement of the head circumference and clinical features such as wide nasal bridge,paranasal bossing,widely spaced eyes with an increased bizygomatic width,and a prominent mandible.The patient underwent otolaryngological examination,endoscopy,hearing test,laboratory examination of phosphorus and bone metabolism,cranial and femoral computed tomography,X-ray and next-generation sequencing.The patient was diagnosed with AD-CMD due to p.Phe377 deletion(c.1129_1131del)on exon 9 of the ANKH gene.After adherence to a prescribed low-calcium diet,the boy’s alkaline phosphatase(ALP)levels continuously decreased to within the normal range.However,after 14 mo of dietary intervention,his parents altered his diet to an intermittent low-calcium diet to include milk and eggs.The patient’s ALP was slightly higher than normal after the dietary change but remained close to the normal range.His serum osteocalcin changed to within normal levels after dietary regulation for 33 mo.His serum combined beta C-terminal telopeptide of type I collagen also continuously decreased after the nutritional intervention,although still slightly higher than normal levels.Despite fluctuating blood test results,the boy’s nasal symptoms were markedly relieved and steadily improved after dietary intervention.No significant changes were found in the craniofacial bones by cranial radiography.Close monitoring of clinical features is still ongoing.Calcitriol treatment is currently under consideration and a surgical procedure is planned as necessary in the future.CONCLUSION We herein report the first Chinese case of AD-CMD with heterozygous mutation of p.Phe377 deletion(c.1129_1131del)on the ANKH gene.Biochemical alterations were significantly improved after dietary intervention indicating that a lowcalcium diet may be applied in pediatric AD-CMD patients with ANKH mutations to help alleviate phenotypic manifestations and improve the quality of life before surgical intervention.Further large scale studies are needed to replicate these findings and to establish the appropriate timing for nutritional and surgical interventions。

Towards a Pan-Anti-Allergy Vaccine

Allergic manifestations affect 20% - 30% of the population in industrialized countries. The global market for asthma and allergy medications has been estimated to exceed USD 6 billion, since 40% of the human population has some form of IgE sensitization to diverse proteins. Most therapeutic intervention strategies cope with the symptoms of allergy without eliminating the underlying cause and many are associated with undesirable and often long-term debilitating side effects. We designed a peptide immunogen encompassing sequences of the human Cε2-3 linker region to prime rat (Rattus norvegicus) immune systems, we then designed a chimeric human-dog-human IgE antibody and used it to boost the immune system and produce high-affinity antibodies that targets native IgE. The investigation showed that this peptide immunogen elicit the formation of antibodies recognizing the native IgE of human, canine and equine origin. The current investigation describes novel approaches aimed at the development of safe anti-allergy vaccines based on active immunization with IgE-derived peptides that are involved in the complementary interaction with the high affinity receptor. The immunization strategy was successful but did not fully work as predicted, thus we propose that peptides described in the current study may lead to the development of a pananti-allergy vaccine with applications for the treatment of all IgE-mediated allergic response independent of the nature of the offending allergen.

凝聚专家的力量提高低收入国家的妇幼保健质量--美国出生缺陷基金会资助的全球妇幼保健协作项目

This short report describes a model for international collaboration on perinatal health that is innovative,highly-productive and challenging. The model,funded by the U.S. March of Dimes Foundation and entitled the 'March of Dimes Global Network for Maternal and Infant Health(GNMIH)' ,allows developing country experts to more easily share their knowledge,experience,skills and materials in ways that can improve women's,maternal,newborn and child health in lower-income countries. This report begins with a brief description of the March of Dimes and its Global Programs which oversees the GNMIH. It then discusses the structure of the GNMIH,with an emphasis on the benefits and challenges of working within the network,and concludes with a brief description of the acti-vities of network members.

New-onset diabetes in COVID-19 and clinical outcomes: A systematic review and meta-analysis

BACKGROUND Diabetes mellitus(DM)is associated with adverse clinical outcomes and high mortality in patients with coronavirus disease 2019(COVID-19).The relationship between diabetes and COVID-19 is known to be bidirectional.AIM To analyze the rate of new-onset diabetes in COVID-19 patients and compare the clinical outcomes of new-onset diabetes,pre-existing diabetes,hyperglycemic,and non-diabetes among COVID-19 patients.METHODS We used the Meta-analysis of Observational Studies in Epidemiology statement for the present meta-analysis.Online databases were searched for all peerreviewed articles published until November 6,2020.Articles were screened using Covidence and data extracted.Further analysis was done using comprehensive meta-analysis.Among the 128 studies detected after thorough database searching,seven were included in the quantitative analysis.The proportion was reported with 95%confidence interval(CI)and heterogeneity was assessed using I2.RESULTS Analysis showed that 19.70%(CI:10.93-32.91)of COVID-19 patients had associated DM,and 25.23%(CI:19.07-32.58)had associated hyperglycemia.The overall mortality rate was 15.36%(CI:12.57-18.68)of all COVID-19 cases,irrespective of their DM status.The mortality rate was 9.26%among non-diabetic patients,10.59%among patients with COVID-19 associated hyperglycemia,16.03%among known DM patients,and 24.96%among COVID-19 associated DM patients.The overall occurrence of adverse events was 20.52%(CI:14.21-28.70)among COVID-19 patients in the included studies,15.29%among non-diabetic patients,20.41%among patients with COVID-19 associated hyperglycemia,20.69%among known DM patients,and 45.85%among new-onset DM.Metaregression showed an increasing rate of mortality among new hyperglycemic patients,known diabetics,and new-onset DM patients in comparison to those without diabetes.CONCLUSION A significantly higher rate of new onset DM and hyperglycemia was observed.Higher mortality rates and adverse events were seen in patients with new-onset DM and hyperglycemia than in the non-diabetic population.

Molecular aspects of MERS-CoV

Middle East respiratory syndrome coronavirus （MERS-CoV） is a betacoronavirus which can cause acute respiratory distress in humans and is associated with a relatively high mortality rate. Since it was first identified in a patient who died in a Jeddah hospital in 2012, the World Health Organization has been notified of 1735 laboratory-confirmed cases from 27 countries, including 628 deaths. Most cases have occurred in Saudi Arabia. MERS-CoV ancestors may be found in Old World bats of the Vespertilionidae family. After a proposed bat to camel switching event, transmission of MERS-CoV to humans is likely to have been the result of multiple zoonotic transfers from dromedary camels. Human-to-human transmission appears to require close contact with infected persons, with outbreaks mainly occurring in hospital environments. Outbreaks have been associated with inadequate infection prevention and control implementation, resulting in recommendations on basic and more advanced infection prevention and control measures by the World Health Organization, and issuing of government guidelines based on these recommendations in affected countries including Saudi Arabia. Evolutionary changes in the virus, particularly in the viral spike protein which mediates virus-host cell contact may potentially increase transmission of this virus. Efforts are on-going to identify specific evidence-based therapies or vaccines. The broad-spectrum antiviral nitazoxanide has been shown to have in vitro activity against MERS-CoV. Synthetic peptides and candidate vaccines based on regions of the spike protein have shown promise in rodent and non-human primate models. GLS-5300, a prophylactic DNA-plasmid vaccine encoding S protein, is the first MERS-CoV vaccine to be tested in humans, while monoclonal antibody, m336 has given promising results in animal models and has potential for use in outbreak situations.

Novel variants of SYNGAP1 associated epileptic encephalopathy: two cases report and literature review

Background SYNGAP1 is a signifcant genetic risk factor for global developmental delay,autism spectrum disorder,and epileptic encephalopathy.De novo loss-of-function variants in this gene cause a neurodevelopmental disorder,for example,early-onset and drug-refractory seizures.We report two children with global developmental delay and epileptic encephalopathy,which are caused by SYNGAP1 gene novel mutations,and drug treatment is efective.Case presentation We report a boy and a girl presented with global developmental delay when they were young babies;as they grew up,cognitive impairment and social-communication disorder became more and more prominent;unfortunately,the patients developed into various seizure types,including eyelid myoclonia,myoclonic and absences when the boy was 1 year 8 mouths old and the girl was 3 years old.The two patients were found two previously unknown mutations by high throughput sequencing[c.3271_c.3272insT;(p.L1091L fs^(*)62),c.2515A>T(p.K839^(*))]in exon 15 of the SYNGAP in the proband.Sanger sequencing confrmed the heterozygous nature,and neither of their parents carried the same mutation.The girl treated with valproic acid and prednisone became seizure-free,and valproic acid and levetiracetam combined with clonazepam were infuential in the other.Conclusions The global developmental delay and epileptic encephalopathy of the children were probably due to the pathogenic mutation of the SYNGAP1 gene,and prednisone and clonazepam may be efective in achieving seizure-free.

Prevalence of Depression in Elderly People Living With HIV:A Systematic Review and Meta-analysis

We conducted a systematic review andmeta-analysis to determine the prevalence of depression among elderly people living with HIV.We searched electronic databases and included 12 observational studies that reported on depression among people who were 50 years of age or older and had HIV.The studies were mostly hospital-based and fromWestern and African countries.They included a total of 5868 older peoplewith HIV,ofwhom 1667 had depression.Pooled prevalence of depression was 28%with heterogeneity of 95.2%.Prevalence ranged from 14.5%in the Netherlands to 42.4%in South Africa.Although therewas asymmetry in the funnel plot,the Egger’s test showed no significant publication bias.Our meta-analysis found a high burden of depression among older people with HIV.Comprehensive health services addressing psychosocial needs and interventions to prevent and treat depression are needed,especially in community settings.Healthcare providers and policymakers should address the prevalence of depression in such communities.

Cholera:an overview with reference to the Yemen epidemic

Cholera is a secretory diarrhoeal disease caused by infection with Vibrio cholerae, primarily the V. cholerae O1 El Tor biotype. There are approximately 2.9 million cases in 69 endemic countries annually, resulting in 95 000 deaths. Cholera is associated with poor infrastructure and lack of access to sanitation and clean drinking water. The current cholera epidemic in Yemen, linked to spread of V. cholerae O1 (Ogawa serotype), is associated with the ongoing war. This has devastated infrastructure and health services. The World Health Organization had estimated that 172 286 suspected cases arose between 27th April and 19th June 2017, including 1170 deaths. While there are three oral cholera vaccines prequalified by the World Health Organization, there are issues surrounding vaccination campaigns in conflict situations, exacerbated by external factors such as a global vaccine shortage. Major movements of people complicates surveillance and administration of double doses of vaccines. Cholera therapy mainly depends on rehydration, with use of antibiotics in more severe infections. Concerns have arisen about the rise of antibiotic resistance in cholera, due to mobile genetic elements. In this review, we give an overview of cholera epidemiology, virulence, antibiotic resistance, therapy and vaccines, in the light of the ongoing epidemic in Yemen.

Efficacy of Chinese Medicine Acupoint Application Combined with Montelukast on Children with Perennial Allergic Rhinitis:A Randomized Controlled Trial

Objective To evaluate the efficacy of Chinese medicine acupoint application(CMAA)combined with Western medicine for perennial allergic rhinitis(PAR)in children.Methods In this prospective,parallel,randomized,placebo-controlled and single-blind trial from August to September,2017,180 children with PAR were randomly assigned to an integrative group(CMAA and Montelukast),CMAA group(CMAA and placebo tablet),or Montelukast group(placebo CMAA and Montelukast).Participants were applied with CMAA for 6 sessions over 2 weeks,and/or Montelukast Chewable Tablet orally once daily for 12 weeks.The changes in severity of symptoms were measured by Visual Analog Scale(VAS)and rhinitis control assessment test(RCAT)at 0,2,4 and 12 weeks of treatment.Blood samples were collected for serum interleukin-4,interferon gammaγand T helper type 1(Th1)/Th2 flow cytometric analysis at the time points of 0,4 and 12 weeks.Results Eight cases dropped out from the trial,3 in the integrative group,2 in the CMAA group and 3 in the Montelukast group.The VAS scores decreased significantly while the RCAT scores increased significantly in all three groups at 4 and 12 weeks compared with baseline(P<0.01 or P<0.05).The VAS scores were significantly lower while the RCAT scores were significantly higher in the integrative and CMAA groups than the Montelukast group at 2 and 4 weeks(P<0.01 or P<0.05).At 2,4 and 12 weeks,the scores of nasal congestion,sneezing,sleep problem,and rhinitis symptom control in the integrative and CMAA groups increased significantly compared with baseline(P<0.01 or P<0.05).The least percentages of Th2 and the most alleviated Th2 shift(highest Th1/Th2)were observed in the integrative group at 12 weeks compared with the other two groups(P<0.05).Conclusion The combination of CMAA with Montelukast might be more effective and appropriate than either option alone for children with PAR.(Registered at Chinese Clinical Trial Register,registration No.ChiCTR-IOR-17012434).

Cancer, circulating tumor cells, and metastasis: could protein-derived peptide fragments impede brain metastases?

The majority of cancer deaths can be attributed to cancer cell metastases that migrate to distant target organs.Brain metastases constitute one of the leading causes of morbidity and mortality among cancer patients,occurring in about 40%of patients with metastatic disease.Thus,there exists an unmet need for early detection,diagnosis,and treatment directed against early stage cancer cell metastasis.Previous studies have reported the development of methods to detect and identify early circulating tumor cells(CTCs)in the bloodstream prior to their seeding into distant organs.Using a comprehensive analysis of total CTCs mRNA content,investigators have developed a mRNA“transcriptome signature”of 126 genes involved in CTC metastatic events.The genes were parsed into various metastatic-related activities indicating that CTCs sustained a semi-dormancy state bent on:(1)stress survival;(2)metabolic maintenance;(3)DNA and translational stability;and(4)chemotactic pro-inflammatory capabilities.These activities suggested that CTCs might be susceptible to interactions with protein-derived peptide segments whose actions are involved with metastatic activities such as cell invasiveness,contact,adhesion,motility,spreading,and migration.The use of protein-derived(encrypted)peptides to impede CTC metabolic activities and disrupt signaling pathways could have therapeutic potential in patients with early metastatic disease.

Developing a one-step triplet-repeat primed PCR assay for diagnosingmyotonic dystrophy

Myotonic dystrophy type 1 (DM1),or Steiner’s disease,is an autosomal dominant disorder caused by the expansion of unstable trinucleotide repeats (CTG) in the 3’ untranslated region of the myotonic dystrophy protein kinase gene (DMPK) (Brook et al.,1992;Mahadevan et al.,1992).The number of CTG repeats observed in normal individuals is in a range of 5-34,while the individuals with 35-49 CTG repeats are usually asymptomatic but at risk of