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Objective:To Constructing a nomogram based on clinical, ultrasound and BRAF V600E gene for predicting cervical lymph node metastasis in patients with papillary thyroid carcinoma (PTC).Methods:The clinical data of 287 patients with PTC (374 malignant nodules) from December 2019 to December 2021 in the First Affiliated Hospital of Hunan Normal University were analyzed retrospectively. Among them, there were 205 nodes with cervical lymph node metastasis and 169 nodes without cervical lymph node metastasis. The echo type, capsule, boundary, shape, number, diameter, location, cystic and solid properties, aspect ratio, blood flow signal, echo distribution, ultrasonic classification, microcalcification and enlarged lymph nodes were observed by ultrasound. The mutation of BRAF V600E gene was detected by fluorescence polymerase chain reaction. The nomograph model for predicting neck lymph node metastasis in patients with PTC was constructed and validated by R3.6.3 software.Results:Univariate analysis result showed that gender, age, microcalcifications, aspect ratio, morphology, blood flow signal, diameter, echo distribution, enlarged lymph nodes, ultrasound classification and BRAF V600E gene were the risk factors for cervical lymph node metastasis in patients with PTC ( P<0.05 or <0.01). Multivariate Logistic regression analysis result showed that age (<40 years old), ultrasonic classification (≥4a) and diameter (>1 cm) were independent risk factors for cervical lymph node metastasis in patients with PTC ( OR = 2.847, 1.436 and 2.475; 95% CI 1.827 to 4.436, 1.075 to 1.918 and 1.505 to 4.069; P<0.01 or <0.05). The age, ultrasonic classification and diameter were included as predictors for constructing the nomogram model. The receiver operating characteristic curve analysis result shows that the area under the curve predicted by the nomogram model for neck lymph node metastasis in patients with PTC was 0.692 (95% CI 0.631 to 0.753). Conclusions:Nomogram based on age, ultrasonic classification and diameter is of high value in predicting neck lymph node metastasis in patients with PTC.
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One of the important causes of developmental epileptic encephalopathy (DEE) is the mutation of ion channel genes, including the mutation of the CACNA1E gene. CACNA1E-related DEE cases were first reported in 2018.The mutation types include new missense mutations, nonsense mutations and frameshift mutations, but the correlation between mutation sites and types with the phenotype of DEE is not clear.This review aims to summarize the reported CACNA1E-related DEE cases, and explore the correlation between the clinical phenotype of CACNA1E-related DEE and gene mutation sites and mutation types.Meanwhile, possible pathogenesis of CACNA1E-related DEE and the progress of drug intervention were reviewed to provide references for the diagnosis and precise treatment of DEE.
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ObjectiveTo observe the effect of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis on high-fat diet-induced apolipoprotein E gene knockout （ApoE-/-） mice， and explore its mechanism of treating atherosclerosis by regulating intestinal flora. MethodThirty-two 8-week-old male ApoE-/- mice were randomly divided into model group， rosuvastatin group （10 mg·kg-1）， high-， low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis （75， 25 mg·kg-1）， with 8 mice in each group. Eight C57BL/6 mice were used as blank group. After 8 weeks of continuous administration， blood was taken to determine the blood lipid level. Enzyme-linked immunosorbent assay （ELISA） was used to detect the contents of related indexes in serum of mice. Hematoxylin-eosin （HE） staining was used to observe the formation of aortic plaque in mice. Cecal contents were collected and 16S rRNA amplicon sequencing was used to detect intestinal flora. ResultCompared with the blank group， the plaque area of the model group was significantly increased with inflammatory infiltration， the contents of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein cholesterol （LDL-C）， inflammatory factors and inducible nitric oxide synthase （iNOS） were increased， while the content of high-density lipoprotein cholesterol （HDL-C） was decreased. Compared with the model group， rosuvastatin group and high- and low-dose groups of ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could improve the deposition of aortic plaque， reduce the contents of TG， TC， LDL-C， inflammatory factors and iNOS， and increase the content of HDL-C. Compared with the blank group， the relative abundances of Firmicutes and Proteobacteria in the model group increased， while the relative abundance of Bacteroidetes decreased. Alpha and Beta diversity analysis showed that samples of each group could be significantly isolated， and the total number and abundance of intestinal flora species in the model group were low. Compared with the model group， ethyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis could increase the relative abundance of beneficial bacteria and decrease the relative abundance of pathogenic bacteria. ConclusionEthyl acetate extract of Acanthopanacis Senticosi Radix et Rhizoma seu Caulis was mainly composed of flavonoids， which can treat atherosclerosis by regulating the intestinal flora and improve the pathological changes in the aorta of ApoE-/- mice induced by high-fat diet. The mechanism may be related to its ability to reduce the level of inflammatory factors， improve antioxidant capacity and repair the disorder of intestinal flora structure.
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Objective:To investigate the clinical features of patients with Langerhans cell histiocytosis (LCH), and analyze the association between BRAF V600E mutation status and clinical features. Methods:A retrospective analysis was carried out for the clinical data of 60 patients with LCH at the Department of Pediatric Oncology, Sun Yat-sen Memorial Hospital between April 2013 and December 2019.Among them, 39 patients undertook BRAF V600E mutation testing, which in paraffin-embedded tissue samples were detected by quantitative real-time PCR (qRT-PCR), and in peripheral blood and/or bone marrow were tested by high-throughput sequencing, for analyzing the correlation between BRAF V600E mutation and clinical characteristics of LCH. Results:(1)Clinical characteristics: the age of 60 LCH patients was (4.08±0.45) years, with 43 male cases and 17 female cases.Patients at young age (≤2 years) and with risk organ (RO+ ) and central nervous system (CNS) risk lesions involvement were concentrated in the multisystem involvement (MS) group ( P<0.05). (2)Therapeutic response after induction therapy: the response to induction therapy was achieved in 28 of 60 treated patients (41.7%) and 32 (53.3%) did not.After excluding stratification factors of treatment regimen, MS ( OR=6.855, 95% CI: 2.077-22.622, P=0.002) and the age≤2 years ( OR=4.944; 95% CI: 1.601-15.275; P=0.005) were risk factors in poor chemotherapy response.RO+ ( OR=8.250, 95% CI: 1.617-42.090, P=0.005) was a significant risk factor for a poor chemotherapy response in JLSG-02 treatment group.Differently, RO+ had no dramatic effect on chemotherapy response in CCHG-LCH-2019 treatment group.(3) BRAF V600E mutation: 39 patients were determined BRAF V600E status, with the positive rate of BRAF V600E mutation in paraffin-embedded tissue samples reaching 70.3%(26 cases). BRAF V600E mutation was not associated with early treatment response, age, sex, MS and RO+ ( P>0.05). However, the positive rate of BRAF V600E in children with MS and CNS risk lesions was higher than the controls, with 76.0% (19 cases) vs.57.1% (8 cases) and 74.1% (20 cases) vs.58.3% (7 cases), respectively.Totally, 3 of 8 cases were positive in bone marrow, with 2 cases of MS, and 1 case of multiple bone invasions, and 1 of 5 cases was positive in peripheral blood, with liver and spleen being involved. Conclusions:LCH patients with age≤2 years, MS and RO+ exhibited a poor response to initial treatment, required for more aggressive treatment strategy.Lesion with activating BRAF V600E mutations suggests that LCH is a clonal disorder.There may be great variability between BRAF V600E mutations and MS as well as CNS risk lesions.In the mutation dataset, part of patients had positive BRAF V600E mutations in bone marrow/peripheral blood.This might suggest a different pathogenesis in such patients, has a certain clinical sense in some aspect.
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Objective:To analyze the first suspected dengue fever case in Dapeng New District by characterizing the virus and exploring its origin. Methods:Dengue virus nucleic acid in blood sample was identified by real-time fluorescent RT-PCR. The E gene of dengue virus was amplified and sequenced. Homology and phylogenetic tree of this dengue virus were compared with the isolates from other regions. Results:Nucleic acid of dengue virus type 1 was detected in the serum sample from the suspected dengue fever patient. Phylogenetic tree showed that the homology based on nucleotide sequence of E gene of DP-DGR19001 was very similar with the isolates of MG894965/TW/CHN/2016 （99.87%）， LC428079/VNM/2017（99.80%）， MG894867/TW/CHN/2015 （99.60%） and MN512242/GX/CHN/2014 （99.46%）. The deduced amino acid sequence of 5 isolates was 100% identical. The patient had traveled to high-incidence country Cambodia in 14 days before infection. Conclusions:The first suspected dengue fever case in Dapeng New District is caused by dengue virus type 1. The case is an imported dengue fever .
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Objective:To investigate the value of BRAF V600E mutation combined with 2015 American Thyroid Association (ATA) Guidelines ultrasound (US) pattern in fine-needle aspiration (FNA) cytology of thyroid nodules with atypia of undetermined significance or follicular lesion of undetermined significance (AUS/FLUS).Methods:This study retrospectively enrolled 96 consecutive patients with 101 AUS/FLUS thyroid nodules who underwent preoperative US, FNA, and BRAF V600E mutation analysis. All AUS/FLUS nodules were classified based on US pattern-based risk stratification of 2015 ATA Guidelines. With postoperative pathology as the gold standard, the diagnostic value of BRAF V600E mutation, US pattern and the combination of two methods were compared.Results:Postoperative pathology confirmed 33 benign nodules and 68 malignant nodules. The mutation rates of BRAF V600E in AUS/FLUS nodules was 51.5%. The sensitivity, specificity, and accuracy of BRAF V600E in the diagnosis AUS/FLUS nodules were 72.1%, 90.9% and 78.2%, respectively. The ROC curve demonstrated that the best cut-off of US pattern was high suspicion. The sensitivity, specificity, and accuracy of US pattern in the diagnosis of AUS/FLUS nodules were 63.2%, 81.8% and 69.3%, respectively. The accuracy of US pattern in determining AUS/FLUS nodules without BRAF V600E mutation was 70.6%. The sensitivity, specificity, and accuracy of the combination of two methods in the differential diagnosis of AUS/FLUS nodules were 89.7%, 75.8%, and 85.1%, respectively. The combination had the highest sensitivity ( P<0.05). Conclusions:BRAF V600E mutation has a good diagnostic value for differentiating benign and malignant AUS/FLUS nodules. Combined with US pattern, the differential diagnostic value for AUS/FLUS nodules without BRAF V600E mutation can be improved, and the sensitivity can be raised.
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Abstract Objective Permanent hypoparathyroidism can be presented as part of genetic disorders such as Sanjad-Sakati syndrome (also known as hypoparathyroidism—intellectual disability-dysmorphism), which is a rare autosomal recessive disorder. Our aim was to confirm the diagnosis of a group of patients with dysmorphism, poor growth, and hypoparathyroidism clinically labeled as Sanjad-Sakati syndrome and to identify for the first time the genetic variations on Iranian patients with the same ethnic origin. Methods In this study, 29 cases from 23 unrelated Arab kindreds with permanent hypoparathyroidism and dysmorphism indicating Sanjad-Sakati syndrome were enrolled for 10 years in the southwest of Iran. The mutational analysis by direct sequencing of the tubulin folding cofactor E gene was performed for the patients and their families, as well as their fetuses using genomic DNA. Results Twenty-eight out of 29 cases had parental consanguinity. Twenty-seven cases presented with hypocalcemia seizure and two were referred because of poor weight gain and were found to have asymptomatic hypocalcemia. The dysmorphic features, hypocalcemia in the setting of low to normal parathyroid hormone levels and high phosphorus led to the diagnosis of these cases. Sequencing analysis of the tubulin folding cofactor E gene revealed a homozygous 12-bp deletion (c.155-166del) for all patients. Following that, prenatal diagnosis was performed for eight families, and two fetuses with a homozygous 12-bp deletion were identified. Conclusion These results make it much easier and faster to diagnose this syndrome from other similar dysmorphisms and also help to detect carriers, as well as prenatal diagnosis of Sanjad-Sakati syndrome in high-risk families in this population.
Resumo Objetivo O hipoparatireoidismo permanente pode estar presente como parte das doenças genéticas como na síndrome de Sanjad-Sakati (também chamada de síndrome de hipoparatireoidismo, retardo e dismorfismo), que é um distúrbio autossômico recessivo raro. Nosso objetivo foi confirmar o diagnóstico de um grupo de pacientes com dismorfismo, crescimento deficiente e hipoparatireoidismo clinicamente identificado como síndrome de Sanjad-Sakati e identificar as variações genéticas, pela primeira vez, em pacientes iranianos com a mesma origem étnica. Métodos Neste estudo, foram inscritos 29 casos de 23 famílias árabes sem parentesco com hipoparatireoidismo e dismorfismo indicando síndrome de Sanjad-Sakati, durante 10 anos no sudoeste do Irã. Foi feita a análise mutacional por sequenciamento direto do gene do cofator E de dobramento da tubulina dos pacientes e de suas famílias e também de seus fetos com o DNA genômico. Resultados Apresentaram consanguinidade parental 28 dos 29 casos. Desses, 27 casos apresentaram convulsão por hipocalcemia e dois foram encaminhados devido ao baixo ganho de peso, considerando diagnóstico de hipocalcemia assintomática. As características dismórficas, hipocalcemia na configuração de níveis de hormônio da paratireoide baixos a normais e alto nível de fósforo levaram ao diagnóstico dos casos. A análise de sequenciamento do gene do cofator E de dobramento da tubulina revelou deleção homozigótica de 12 pares de base (pb) (c.155-166del) em todos os pacientes. Após isso, foi feito o diagnóstico pré-natal em oito famílias e dois fetos foram identificados com deleção homozigótica de 12 pb. Conclusão Esses resultados tornam o diagnóstico dessa síndrome muito mais fácil e rápido do que outros dismorfismos semelhantes e também ajudam a detectar portadores, bem como o diagnóstico pré-natal da síndrome de Sanjad-Sakati em famílias de alto risco nessa população.
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Humans , Osteochondrodysplasias , Seizures , Abnormalities, Multiple , Growth Disorders , Hypoparathyroidism , Intellectual Disability , Tubulin , Molecular Chaperones , IranABSTRACT
Objective To evaluate the diagnostic efficacies of contrast-enhanced ultrasound(CEUS) combined with BRAF V600E mutation detection in ultrasound-guided fine-needle aspiration cytology of thyroid nodules with atypia of undetermined significance . Methods A total of 129 thyroid nodules underwent the examinations of CEUS and BRAF V600E mutation were analyzed retrospectively . With surgical pathology as the gold standard ,ROC curve was used to investigate the diagnostic values of CEUS , BRAF V600E and the combination of the two methods . Results The sensitivity ,specificity and accuracy of CEUS and BRAF V600E gene detection for thyroid nodules with atypia of undetermined significance diagnosed by ultrasound-guided fine-needle aspiration biopsy were 86 .7％ ,83 .3％ ,85 .3％ and 72 .0％ , 100％ ,83 .7％ ,respectively . The sensitivity and accuracy of CEUS were higher than those of BRAF V 600E gene detection ( all P < 0 .001 ) ,however its specificity was lower than BRAF V 600E with statistically significance( P < 0 .001) ,those of the combined test of CEUS and BRAF V600E mutation analysis were 94 .7％ ,83 .3％ ,89 .9％ ,respectively . The combination of two methods had the highest diagnostic efficacy , with statistically difference ( P <0 .001) ,and the area under the ROC curve ( AUC) was higher than that for each test(0 .951 vs 0 .860 vs 0 .901) . Conclusions The combined test of CEUS and BRAF V600E mutation has a higher diagnostic efficacy for cytologically indeterminate thyroid nodules compared with CEUS or BRAF V600E mutation alone .
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OBJECTIVE: To investigate the correlation between clinicopathological features and BRAF~(V600E) gene mutation in papillary thyroid carcinoma(PTC). METHODS: The data of 290 cases of PTC admitted from August 2016 to May 2018 in Beijing Tongren Hospital,Capital Medical University were analyzed retrospectively. All the cases of PTC were examined by BRAF~(V600E) immunohistochemistry after operation. Mutation positive group included 192 cases. Mutation negative group included 59 cases and partial mutation group included 39 cases. The cases of partial mutantion were excluded. The clinicopathological differences between the two groups were compared. RESULTS: By statistical analysis,there was a statistical difference between two groups in the invasion of the capsule(P<0.05). In the two groups(microcarcinoma and non-microcarcinoma),there was the same result.There was no statistical difference in other indexes. CONCLUSION: In patients with PTC,BRAF~(V600E) gene mutation is not associated with the clinicopathological features of the PTC. Patients with non-BRAF~(V600E) gene mutations are more likely to invade the membranes of microcarcinomas and non-microcarcinomas.
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Diffuse axonal injury (DAI) is a common primary brain injury.The main mechanism of DAI is the relative displacement of brain tissue junction caused by shearing force during cranial rotation.The pathological changes are the deformation and rupture of white matter nerve fiber bundles.The mechanism of injury and recovery is still unclear,and unified diagnostic criteria are also lacking.Apolipoprotein E (APOE) gene polymorphisms are associated with nervous system development,injury,and repair process.The APOE gene contains six genotypes by the combination among three genes and synthesizes three proteins including apoE2,apoE3 and apoE4.The abnormal spatial structure of apoE4 decreases its ability of transporting lipids,resulting in reduced repair ability after nerve injury.The molecular mechanism of ApoE in the development of axonal injury is complicated.apoE protein can bind to receptor proteins such as low density lipoprotein receptor related protein 1 (LRP1) and heparan sulfate proteoglycan (HSPG) to mediate axonal repair,and it can also damage nerves by influencing calcium influx and producing toxic fragments through hydrolysis.Correcting the abnormal structure of apoE4 is helpful to nerve repair,and apoE analogues also have certain neuroprotective effects.This article reviews the injury characteristics,pathological characteristics,APOE gene polymorphisms,and the role of apoE synthesis in DAI,to provide new insights for elucidation of mechanism of DAI and related clinical application.
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Objective To study the screening of trace amount mutation of BRAF V 600E gene for avoiding the appearance of ineffective treatment in cancer patients .Methods The internal competitive amplification fragments were used to improve the inhibition of wild-type blocking (WTB) probe on wile-type BRAF V600E gene to increase the detection efficiency of BRAF V600E genotype of trace amount mutation occurrence .Re-sults When the template DNA concentration was 50 -200 ng/μL ,the constructed trace amount gene muta-tion real time fluorescence quantitative detection method could completely block the amplification of the wild-type BRAF V600E gene .The sensitivity of this assay reached as high as 0 .1％ ,which was in line with the sen-sitivity requirement for the gene trace amount mutation detection technique .In the colorectal biopsy tissues from 50 cases of suspected colorectal cancer ,8 cases (16 .0％ ) of BRAF V600E gene trace amount mutation were detected by using this constructed method ,which had higher detection rate .Conclusion The constructed gene trace amount mutation detection method can make the rapid ,simple and low cost quantitative analysis for BRAF V600E gene trace amount mutation in clinical samples .
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Objective@#To determine the first dengue fever case in Taizhou and trace probable transmission sources.@*Methods@#Collected serum of three patients for antigen, antibody and nucleic acid detection. Dengue viruses were isolated and cultured in C6/36 cell. The whole length of E gene was amplified by reverse transcriptase-polymerase chain reaction(RT-PCR) and then sequenced. The phylogenetic tree was drawn.@*Results@#Three cases were positive in nucleic acid detection. Two cases were IgM positive. One case was NSI antigen postive. Three strains of type I dengue virus were isolated from samples. The phylogenetic trees shown that the three strains were on the same branch. The identities of nucleotide were 99.87%. The identities of amino acid were 99.6%-99.8%.@*Conclusions@#The dengue virus strains isolated in Taizhou was imported from Guangdong or Southeast Asia and caused location infection.
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Objective To evaluate the relationship between clinicopathological features and central lymph nodes metastasis in patients with papillary thyroid microcarcinoma (PTMC),and aim to provide an appropriate operation in clinic.Methods The clinical data of 536 patients undergoing surgical treatment in Department of Thyroid Surgery,the First Affiliated Hospital of Zhengzhou University from January 2015 to December 2017,who had pathologically confirmed popillary thyroid microcarcinoma (PTMC) were retrospectively analyzed.Thyroidectomy and isthmectomy with central compartment neck dissection were performed in all patients.The patients were divided into two groups according to whether there existed central lymph node metastasis,and compared the differences of gender,age,number of cancer lesions,tumor breakthrough envelope,BRAFV600E gene mutation,and tumor maximum diameter in the central lymph node metastasis,respectively,by t-test x2-test.Univariate logistic regression analysis and multivariate binary logistic regression analysis were conducted to find risky factors.Results There existed difference between two groups by completing the gender,age,the BRAFV600E gene mutation and maximal tumor diameter(P ＜ 0.05).Univariate logistic regression analysis indicated that gender (P =0.046),age (P ＜ 0.01),maximal tumor diameter(P ＜0.01) and the BRAFV600E gene mutation(P =0.016) were significant predictors for central lymph nodes metastasis.And multivariate binary logistic regression analysis revealed that the rate of lymph node metastasis significantly increased in cases of larger tumor diameter(P ＜0.01),BRAFV600E gene mutation(P =0.035) and ageing below 45 years old (P ＜ 0.01).Conclusions The treatment for central lymph node metastasis of PTMC should be different considering elements including BRAF600E gene mutation prophylactic,ageing below 45 years old and larger tumor diameter.Therefore central lymphadenectomy should be performed when the primary lesion was resected.
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Background and purpose:The BRAF V600E mutation is a highly attractive drug target. Therefore, determining the BRAF gene mutation status of patients is essential in order to assess patients' eligibility for targeted BRAF geneinhibitor therapy. The aim of this study was to validate the utility of immunohistochemistry to rapidly obtain the BRAF gene mutation status. This study aimed to analyze the correlation of the BRAF V600E gene mutation and VE1 protein ex-pression with the clinical pathological characteristics in papillary thyroid carcinoma (PTC).Methods:The mutation status of BRAF V600E was detected by DNA sequencing. Immunohistochemistry was used to detect the expression of BRAF V600E protein in 108 cases of PTC, 54 cases of thyroid adenoma and 54 cases of normal thyroid tissue.Results:The gene mutation rate of BRAF V600E is 67.6%, and VE1 protein expression rate is 64.8% in 108 cases of PTC. The differences were statistically significant compared with thyroid adenoma and goiter (P<0.05), but have no correlation with the clinical pathological characteristics.Conclusion:BRAF V600E gene mutation and VE1 protein expression are useful biomarkers for the pathological diagnosis of PTC. High consistency was observed between the immunohistochemical staining results and the DNA sequencingresults of BRAF V600E gene mutations. Immunohistochemical technique detecting the BRAF V600E protein expression can effectively reflect indirectly BRAF V600E gene mutation status in PTC.BRAF V600E gene mutation has no contribution to the development of papillary thyroid carcinoma.
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[Objects]To isolate and identify the pathogen of the large outbreak of dengue in Guangdong province in 2014. To understand the origin and the phylogenetic characteristics of the isolates ,and provide scientific foundation for the surveillance and prevention of dengue fever.[Methods]Collected the patient serum samples over all the Guangdong province during the 2014 outbreakperiod,isolated and identified the virus from these samples. Amplified complete E gene and complete genome with certain primers and sequenced all the products. Then the Phylogenetic ,Bayesian phylogeography and mutations analysis were carried.[Results]40 DENV-1 strains were isolated and identified. 40 complete E gene sequences and 6 complete genome sequences of DENV-1 were obtained. Phylogenetic analysis with E gene sequences revealed that the 40 isolates were classified into two genotypes including 16 genotypeⅠ(Asia)and 24 genotypeⅤ(America/Africa). 14 genotypeⅠisolates were clustered closest with isolates from Guangdong province(2013)and Sigapore(2013)which share the nucletide identities of 99.6% ~ 99.9%,other two genotypeⅠisolates were clustered with strains from Malaysia (2013) and both share the nucletide identities of 99.7%;24 genotypeⅤisolates were all classified in one clade with striains from Bangladesh(2009),China(2009)and Bhutan(2013)which share nucletide identities of 99.0%-99.9%. Further analysis with six complete genome sequences showed that five isolates were clustered closest with strains isolated from Guangdong province(2013)share the nucletide identities of 99.6%-99.8% while the sixth stains closest with strains isolated from Myanmar(2002)share the nucletide identities of 98.8%. The isolates have five amino acid mutations compared with strains epidemic in Guangdong province in 2013,three mutations(S88V,E203G,T275R)are in the EⅡdomain and one mutation (S305P)is in the EⅢdomain which associated with virulence.[Conclusions]During the outbreak in Guangdong province in 2014, DENV-1 is the predominant causative serotype,and there are at least two different kinds of genotypes of DENV-1 largely epidemiced in the whole province. Evolution analysis reveals the multiple origins of the isolates which may origin from Guangdong province , Sigapore,Malaysia,Myanmar so that we should enhance the study and surveillance of autochthonous and vectors in order to understand the epidemic way of dengue in Guangdong province. The isolates have had four mutations in the domain associated with virulence which remain further study to know their biological effects.
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Objective@#To understand the serotypes, genotypes and transmission source of dengue viruses(DENV) isolated in Yunnan from 2013 to 2015.@*Methods@#Viral RNA was extracted from serum samples of dengue fever(DF) cases at the acute stage in Yunnan, then the gene fragments of envelope protein(E) region were amplified by RT-PCR. The homology and phylogenetic analysis was made on the nucleotide and deduced amino acid sequences by bioinformatics softwares including Clustal X, DNAStar and MEGA5.@*Results@#Viral nucleic acid detection and sequencing indicated that 40 E genes of DENV were obtained. The serotypes and genotypes of DENV were revealed by homology and phylogenetic analysis based on E genes of DENV. Fifteen virus strains belonged to DENV serotype 1(DENV-1), of these, 14(11 from Ruili, 1 from Lincang and 2 from Kunming) were genotype I(G-I), 1 from Kunming was G-V. Twenty-two virus strains belonged to DENV serotype 2(DENV-2), of these, 10 from Ruili were G-I and 12 from Xishuangbanna were G-IV. Two virus strains belonged to DENV serotype 3(DENV-3) and G-II. One virus strain belonged to DENV serotype 4(DENV-4) and G-I. All detected DENV genotypes were mainly predominant in Southeast Asia. All the 40 Yunnan DENV strains shared high homology with the DENV strains in Southeast Asia countries.@*Conclusions@#Four serotypes and multiple genotypes of DENV had been co-circulating in Yunnan from 2013 to 2015. The DENV transmitted from Southeast Asia countries was the main cause of DF in Yunnan. It is necessary to strengthen the surveillance and management on the imported cases of DF in Yunnan.
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OBJECTIVE: To establish a PC12 cell line with stable expression of human apolipoprotein E( ApoE4) gene by transfection with a lentiviral vector carrying human ApoE4 gene and to investigate the effect of maltol aluminum on the viability of transfected PC12 cells. METHODS: The lentiviral vector carrying human ApoE4 gene was transfected into PC12 cells. PC12 cells with overexpression of ApoE4 gene and negative control vector were obtained after puromycin screening.The mRNA relative expression of R-Apo E and( or) H-Apo E-FLAG of cells in PC12,PC12-NC and PC12-ApoE4 groups were detected by real-time fluorescent quantitative polymerase chain reaction,and the effect of cell construction was identified. PC12-ApoE4 cells and PC12 cells were exposed to maltol aluminum solution at concentrations of 0. 00,100. 00,200. 00 and 400. 00 μmol/L respectively for 24 hours,and cell viability was detected by Cell Counting Kit-8( CCK-8)assay. RESULTS: PC12-ApoE4 and PC12-NC cells under the fluorescence microscope showed fluorescence expression,suggesting that transfection was successful. The expression of PC12 cells showed no fluorescence. The relative expression of H-Apo E-FLAG gene mRNA( the median amount) of PC12-ApoE4 cells was 148. 74,which was higher than the R-Apo E gene in PC12 cells( 1. 00) and PC12-NC cells( 1. 01)( P < 0. 01). After exposure to maltol aluminum,the cell survival rates in terms of the main effect and interaction effect of dose and cell type were statistically significant( P < 0. 01),among them,the cell viabilities were decreased in the concentration range of 0. 00-400. 00 μmol/L with the dose of maltol aluminum exposure increased,showing dose-effect relationship( P < 0. 01). CONCLUSION: The cell line stably expressed human ApoE4 gene was constructed successfully. There was interaction between the effects of maltol aluminum and ApoE4 gene on the survival rate of PC12 cells,and ApoE4 gene could enhance the cytotoxicity of maltol aluminum on PC12 cells.
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Objective To investigate the epidemiological characteristics of dengue fever in Shenz-hen city in 2014 and to analyze the evolutional characteristics of the epidemic dengue virus(DENV)strains in order to provide scientific guidelines for the prevention and control of dengue fever. Methods Descrip-tive epidemiological analysis was used to analyze the prevalence of dengue fever in Shenzhen city in 2014. Immunochromatography and real-time PCR were performed to detect the specific antibodies(IgM and IgG) and DENV nucleic acids in serum samples collected from suspected cases of dengue fever. Serum samples collected from the patients at early stage of dengue fever were used to infect the C6 / 36 cell line for further isolation of DENV strains. The types of isolated DENV strains were determined by using real-time PCR. E genes of the isolated DENV strains were amplified by RT-PCR and then sequenced. DNAStar and Clustslx (1. 83)softwares were used to analyze the homology between DENV strains isolated in Shenzhen and other areas. A phylogenetic tree based on the sequences of E genes of Shenzhen strains and other sequences of DENV reference strains downloaded from GenBank was constructed for further analysis. Results A total of 454 cases of dengue fever were reported in Shenzhen in 2014 with a male to female ratio of 1. 43 ∶ 1. Local patients accounted for 76. 21% and the rest 23. 79% were imported cases mainly from Southeast Asian and surrounding cities. There were 441 cases reported from September to November,accounting for 97. 14% of all reported cases. Most of the infected subjects were aged 20 to 50,accounting for 76. 73% . Of the 270 samples positive for DENV nucleic acids,strains of DENV-1,DENV-2,DENV-3 and DENV-4 accounted for 87. 41% ,8. 89% ,0. 37% and 2. 22% ,respectively. The phylogenetic tree analysis revealed that the DENV-1 strains belonged to two genotypes,which were genotypeⅠ and genotype Ⅴ. The DENV strains of genotypeⅠ were highly similar to the epidemic strain isolated in Shenzhen in 2010 and the genotype Ⅴstrains were first reported in Shenzhen. The homology analysis of the nucleotides of E genes showed that mi-nor differences in the nucleotide sequences were found between DENV-2 strains. All of the DENV-2 strains belonged to the genotype Ⅳ as indicated by the phylogenic tree. Conclusion There were 454 cases of den-gue fever(including both local and imported cases)reported in Shenzhen city in 2014,reaching an all-time high. DENV-1 was the predominant pathogen in combination with an increased infection of DENV-2. This study indicated that the prevalent DENV strains might be imported from Southeast countries and neighboring cities. Further researches should be conducted to analyze whether dengue fever is endemic in Shenzhen City.
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Objectives To analysis the expression difference of eIF4E in bone marrow between the acute myeloid leukemia (AML) and the non-tumor patient,before and after induction therapy,and the different subtypes of AML,and to explore the relation between eIF4E and other molecular biology abnormalities in AML.Methods The bone marrow specimens of newly diagnosed AML and non-tumor control patients were collected.The expression level of eIF4E was detected by RT-PCR.Results The positive rate of eIF4E was 65.2 ％ (101/155) in AML.eIF4E turned to negative in 12 patients (17.6 ％) who got complete remission after induction therapy.eIF4E was negatively expressed in bone marrow of the nontumor control patients.The positive rates of eIF4E were 75.0 ％ (15/20) and 80.8 ％ (21/26) in M4 and M5 type AML,respectively,and was 59.6 ％ (65/109) in other subtype AML (P ＞ 0.05).FLT3/ITD gene mutation was found in 26 cases of newly diagnosed AML.The eIF4E expression and FLT3-ITD gene mutation were independent each other (P ＞ 0.05).Conclusions eIF4E is positive in most of the newly diagnosed AML and turns to negative in part of AML achieving complete remission.The expression of eIF4E is no difference among different subtype of AMLs.eIF4E and FLT3-ITD gene mutation are independent each other.
ABSTRACT
Objective:To study the Xinjiang Kazakh,Han nationality patients with coronary heart disease apolipoprotein E gene promoter region rs405509 (G-T),rs449647 (A-T),rs7259620 (G-A) whether there exist differences in the two national distribution between loci polymorphism.Methods:201 cases were studied.DNA product was extracted by using the PheNol-chloroform method and the PCR outcome was purified.We take advantage of multiple single base extension reaction to make DNA Sequencing on The ABI3130XL.Results: The Kazak and Han patients in Xinjiang area in the apolipoprotein E gene promoter rs449647 ( A-T) was statistically significant differences in genotype and allele in two ethnic groups (P0.05).Conclusion:Apolipoprotein E promoter rs449647 ( A-T ) genotype and allele polymorphism have significant differences between Han nationality and Kazak nationality in Xinjiang,others have no statistic difference between the two ethnic groups.