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Objective:To study the multi-locus sequence typing (MLST) gene characteristics of Brucella isolates in Guizhou Province. Methods:Brucella strains, which were isolated from 2017 to 2021 in Guizhou Province (preserved in the Bacterial and Viral Seed Bank of Guizhou Center for Disease Control and Prevention) were identified Brucella and species/types by BCSP31-PCR and AMOS-PCR methods, respectively. MLST method was used for genotyping, and Biometrics 8.0 software was used for cluster analysis of the typing results. Results:A total of 32 strains of Brucella were isolated in Guizhou Province and identified as Brucella melitensis ( B.melitensis) by BCSP31-PCR and AMOS-PCR methods. These strains were classified into 2 ST types (ST8 and ST39) by MLST method, with 28 strains of ST8 type(87.5%) and 4 strains of ST39 type (12.5%). The 28 strains of ST8 type were distributed in 7 cities (prefectures) of Guizhou Province, while the 4 strains of ST39 type were only found in Qianxinan Buyi and Miao Autonomous Prefecture. The cluster analysis results showed that ST8 and ST39 types strains were clustered in a group with the reference strain of B.melitensis, and there was only one nucleotide site difference between ST39 and ST8 types in the glk gene, indicating a close genetic relationship. Conclusions:B.melitensis is the main pathogen of the brucellosis epidemic in Guizhou Province in recent years. ST8 is the dominant MLST genotype in Guizhou Province.
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Objective:To analyze the results of the joint screening of newborn hearing and deafness genes in Dalian to provide a reference for the prevention and control of hereditary deafness.Methods:Eight hundred and forty-two neonates born in Dalian Women and Children′s Medical Group from January 1, 2022 to May 30, 2022 were screened retrospectively, using AABR (automatic brainstem evoked potential). And 20 mutation sites of common genetic deafness 4 genes , including GJB2, GJB3, SLC26A4 (PDS) and mitochondrial genes associated with drug-induced deafness (MT-RNRI)(12SrRNA), were detected by high-throughput sequencing.Results:Among the 842 newborns, 840 passed hearing screening (99.8%); 36 cases (4.3%) passed the hearing screening but not the hearing loss gene screening; 804 cases passed through the both screening (95.5%); 2 cases (0.24%) failed in the both screening. 38 cases of deafness gene mutations were detected, with a total carrying rate of 4.51% (38/842). Among them, the carrying rates of heterozygous mutations in GJB2, GJB3, SLC26A4 (PDS), MT-RNRI (12SrRNA) were 1.90%, 0.24%, 1.30%, and 0.95%, respectively. The carrying rates of GJB2/GJB3 composite heterozygous mutations were 0.12%.Conclusions:The combined screening of neonatal hearing and deafness genes can reduce the missed rate of hearing screening. The carrier rate of neonatal deafness gene in Dalian is 4.51%, with the highest GJB 2 carrier rate, followed by SLC26A4 (PDS) carrier rate.
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Uncommon epidermal growth factor receptor (EGFR) mutations account for 10%-20% of all EGFR mutations in non-small-cell lung cancer (NSCLC). The uncommon EGFR-mutated NSCLC is associated with poor clinical outcomes and generally achieved unsatisfactory effects to the current therapies using standard EGFR-tyrosine kinase inhibitors (TKIs), including afatinib and osimertinib. Therefore, it is necessary to develop more novel EGFR-TKIs to treat uncommon EGFR-mutated NSCLC. Aumolertinib is a third-generation EGFR-TKI approved in China for treating advanced NSCLC with common EGFR mutations. However, it remains unclear whether aumolertinib is effective in uncommon EGFR-mutated NSCLC. In this work, the in vitro anticancer activity of aumolertinib was investigated in engineered Ba/F3 cells and patient-derived cells bearing diverse uncommon EGFR mutations. Aumolertinib was shown to be more potent in inhibiting the viability of various uncommon EGFR-mutated cell lines than those with wild-type EGFR. And in vivo, aumolertinib could also significantly inhibit tumor growth in two mouse allograft models (V769-D770insASV and L861Q mutations) and a patient-derived xenografts model (H773-V774insNPH mutation). Importantly, aumolertinib exerts responses against tumors in advanced NSCLC patients with uncommon EGFR mutations. These results suggest that aumolertinib has the potential as a promising therapeutic candidate for the treatment of uncommon EGFR-mutated NSCLC.
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@#Aortic valve disease is one of the major diseases threatening human health. Transcatheter aortic valve replacement (TAVR) is a new treatment for aortic disease. Preoperative evaluation is of great significance to the successful operation and the long-term quality of life of patients. The 3D printing technology can fully simulate the cardiac anatomy of patients, create personalized molds for patients, improve surgical efficiency, reduce surgical time and surgical trauma, and thus achieve better surgical results. In this review, the relevant literatures were searched, and the evaluation effect of 3D printing technology on the operation of TAVR was reviewed, so as to provide clinical reference.
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Objective:To analyze the molecular evolution characteristics of HA and NA genes of influenza B/Yamagata (BY) and influenza B/Victoria (BV) lineage viruses in Guizhou Province, aiming to provide reference for scientific prevention and control of influenza. Methods:The prevalence of various types of influenza viruses in Guizhou Province from 2017 to 2021 was analyzed. The nucleic acid of influenza B viruses was extracted, and then the HA and NA genes were amplified by RT-PCR. Fourteen strains were sequenced and the sequences of 83 strains were obtained from GISAID. Homologies between the 97 influenza B viruses as well as the phylogenetic characteristics and amino acid site variations were analyzed. Results:Influenza A, BY and BV lineage viruses co-circulated in Guizhou Province and BV lineage was the predominant type. The homologies of HA and NA genes were 98.7%-99.4% and 98.4%-99.6% between BY lineage viruses and the reference vaccine strain B/PHUKET/3073/2013. BV lineage viruses shared 98.3%-99.3% and 98.9%-99.6% homologies with the reference vaccine strain B/Colorado/06/2017. The BY lineage strains in Guizhou Province mainly belonged to Y3 genetic group with HA gene in two branches of Y3-H1-2 and NA gene in three branches of Y3-N1-3. Three reassortant strains were found in Y3 clade. The isolated BV lineage strains mainly belonged to V1A-2 genetic group with HA gene in four branches of V1A-2 H1-4 and NA gene in five branches of V1A-2 N1-5. Twenty reassortant strains were found in V1A-2 clade and no inter-lineage reassortants were found. Analysis of variations at key amino acid sites showed that there was no mutation at epitopes in Y3 genetic group. However, there were point mutations at four main epitopes and a shift mutation in 190 helix in V1A-2 genetic group. There was no mutation in drug resistance sites. Conclusions:Various types of influenza viruses circulated in Guizhou Province. The homology between influenza B viruses and vaccine strains was decreasing. Different branches of HA and NA genes had been evolved and various forms of mutations were detected in the sequences. Intra-lineage reassortant strains and new varieties emerged. Surveillance of influenza B viruses should be strengthened.
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Objective:To understand the genetic variation and the prevalence of H9N2 subtype avian influenza virus in Guizhou province, and to provide the scientific evidence for the prevention and control of avian influenza virus.Methods:The results of AIV detection in live poultry market(LPM) environment in Guizhou province from October 2018 to March 2019 were statistically analyzed, RNAs were extracted and sequenced from the HA genes of 13 samples of H9N2 positive screened by real-time PCR. Then the homology, the genetic evolution and the mutations of important amino acid were analyzed by bioinformation softwares. Results:The positive rate of AIV was 52.2% and the positive rate of H9N2 was 83.7% in LPM environment. The homology between nucleotides of the HA gene of 21 strains ranged from 91.6% to 100.0%, and the homology between amino acids of the HA gene ranged from 91.0% to 100.0%. All strains belonged to Y280 sublineage and G57 genotype. Key sites analysis showed that they had a common motif PSRSSRGLF and LSRSSRGLF at the cleavage site, which indicated that they were lentogenic and low pathogenic strains. Mutations H191N, E198T/A and Q234L at the receptor binding sites in the HA was found in 21 strains, while indicated the viruses had the potential to bind human-like receptor. The analysis results of glycosylation motifs showed that all 21 strains had 7 glycosylation sites, but had a site deletion at amino acid site 218 and an addition at 313.There was no significant mutation in the key site compared with the human infected strains. Conclusions:The detection rate of AIV in LPM environment in Guizhou province was high, and the pollution was very serious, and H9N2 subtype is the main subtype, All H9N2 subtype AIVs belonged to Y280 sublineage and G57 genotype, and they were low pathogenic avian influenza viruses in Guizhou province, but the genetic gap were widening and mutations of key amino acid site might enhance susceptibility and pathogenicity to human beings. Hence, It is necessary to strengthen the surveillance of molecular characteristic variation of H9N2 subtype AIV.
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Objective:To study the effect of left atrioventricular interphase (LAVI) via esophageal electrocardiogram on cardiac function after dual-chamber pacemaker implantation in patients with high-degree atrioventricular block.Methods:Using a prospective approach, 40 patients with high-degree atrioventricular block who would undergo dual-chamber pacemaker implantation from January 2017 to March 2018 in Department of Cardiovascular Medicine, Dalian Municipal Central Hospital Affiliated of Dalian Medical University were enrolled. All patients accepted esophageal electrocardiogram tests at 3 months after the implantation, to exam the interatrial conduction time (IACT) of sinus rhythm and pacing rhythm, and interventricular conduction time (IVCT). Then based on the outcome of the echocardiography test, the optimal atrioventricular delay (AVD) of the pacemaker of each patient was determined while the LAVI differed from 100 ms to 150 ms. The left ventricular ejection fraction (LVEF), peak speed of blood flow velocity in early mitral orifice diastole (E), E peak deceleration time (EDT), peak speed of early mitral annular diastolic movement (e′), isovolumic relaxation time (IVRT) and left atrial volume (LAV) were tested by echocardiogram before implantation, before AVD adjustment at 3 months after implantation, after AVD adjustment at 3 months after implantation, and 6, 12, and 18 months after implantation. Then, the left atrial volume index (LAV/body surface area) and E/e′ were calculated.Results:Among the 40 patients, the IACT of sinus rhythm was (55.55 ± 10.33) ms, the IACT of pacing rhythm was (93.95 ± 12.77) ms, and the mean IVCT was (63.20 ± 17.84) ms; the optimal LAVI was 110 to 150 (132.00 ± 10.43) ms, and notably, the optimal LAVI between 120 and 140 ms was 82.5% (33/40). The LVEF, EDT, IVRT, left atrial volume index and E/e′ from before AVD adjustment of 3 months after implantation to follow-up endpoint (18 months after implantation) were significantly improved compared with those before implantation, and there were statistical differences ( P<0.01); the EDT and IVRT after AVD adjustment at 3 months after implantation were significantly improved than those before AVD adjustment at 3 months after implantation: (142.15 ± 35.58) ms vs. (125.94 ± 31.13) ms and (119.52 ± 22.15) ms vs. (133.92 ± 23.87) ms, and there were statistical differences ( P<0.05); the IVRT and left atrial volume index 18 months after implantation were significantly improved compared with those before AVD adjustment at 3 months after implantation: (122.07 ± 16.99) ms vs. (133.92 ± 23.87) and 32.94 ± 3.22 vs. 35.43 ± 5.76, and there were statistical differences ( P<0.05). Conclusions:Optimizing the LAVI after dual-chamber pacemaker implantation via esophageal electrocardiogram can improve the long-term prognosis of patients with high-degree atrioventricular block.
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Objective:To investigate the behavioral problems of children with congenital adrenal hyperplasia (CAH), and to explore the influencing factors, thus providing evidence for their prevention and interventions.Methods:A case-control study was carried out.A total of 25 children with CAH who were aged 4-16 and regularly followed up in the Outpatient Department of the Second Xiangya Hospital, Central South University from June 1, 2017 to March 31, 2019 were enrolled in the study group, and 50 age-and gender-matched healthy children in Hunan Province were selected as the healthy control group.The parents of the selected subjects were investigated with the Achenbach Child Behavior Checklist (CBCL) to evaluate children′s behavior problems.SPSS 22.0 software was applied to statistical analysis.Results:(1) The scores of externalizing behaviors, aggressive factors and behavior problems in 4-to 5-year-old male children in the CAH group were significantly higher than those in the healthy control group [(12.440±8.353) scores vs.(5.060±5.230) scores, (9.670±6.481) scores vs.(4.110±4.157) scores, (22.110±13.062) scores vs.(12.890±9.405) scores] ( t=2.829, 2.711, 2.109, respectively, all P<0.05). There was no significant difference in the scores of other behavior problems and influencing factors between the CAH group and the healthy control group (all P>0.05). (2) The influencing factor of behavioral problems was progesterone ( β=0.567). Testoste-rone not only was the influencing factor of externalizing and internalizing behaviors ( β=0.582, 0.497, respectively), but also affected the behavior of physical complaints, violation of discipline and social withdrawal ( β=0.735, 0.531 and 0.492, respectively). The factor influencing the schizoid behavior was the initial treatment age ( β=0.402). Conclusions:Four- to 5-year-old male children with CAH have behavioral problems, among which aggression and externalizing behaviors are more common.The increase of testosterone may cause the problems of internalizing and externalizing behaviors in children with CAH, and has a great impact on physical complaints, social withdrawal, and discipline violation.The increase of progesterone may lead to the behavior problems of the children.The older the initial treatment age, the more serious the schizoid behavior problem may be.
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Objective:To investigate the clinical characteristic, gene mutations and genotype-phenotype correlation of 21-hydroxylase deficiency (21-OHD) in Hunan.Methods:A total of 48 patients with 21-OHD who were admitted to the Department of Pediatrics, the Second Xiangya Hospital, Central South University from March 2016 to March 2017 were collected.According to the clinical manifestations and biochemical characteristics of the patients, they were divided into salt wasting (SW) and simple virilizing (SV). Sanger sequencing combined with multiplex ligation-dependent probe amplification(MLPA) were used to detect the mutations of CYP21A2 gene.The patients were divided into 3 groups according to their mutations severity: severe mutation group, moderate mutation group and unknown mutation group.Then, the correlation between genotype and phenotype was analyzed. Results:(1) Forty-eight 21-OHD patients included 28 SW cases and 20 SV cases, and the first visiting age of SW was younger than that of SV, and the difference was statistically significant ( U=44.5, P<0.05). The SW cases had high incidence rate of adrenal crisis and the SV patients were liable to advanced bone age and precocious puberty.(2) Forty-four patients were detected abnormal gene mutation and the positive rate of genetic diagnosis was 91.7%.Fourteen mutation types including I2G, Del, I173N, R357W, R484fs(c.1451_1452delGGinsC, c.1450dupC), R483fs, G111Vfs*21, Q319X, c.292+ 1G>A, c.377C>G, E6Cluster, p.H393Q and m. 1647C>T, were found in 88 alleles.The most frequent mutations were I2G(36.4%), I173N(20.4%), and Del(22.7%). p.H393Q and m. 1647C>T were 2 novel mutations.I2G (47.3%) and Del (27.3%) were the most frequent mutations in SW cases, and I173N (48.5%) was the most frequent mutation in SV cases.(3) Severe mutation was in 29 patients, including 26 SW, and moderate mutation was in 13 patients, including 12 SW.The percentage of SW in severe mutation group was 89.7% and SV in moderate mutation was 92.3%. Conclusions:I2G, I173N and Del were the frequent mutations of 21-OHD in Hunan, and the total percen-tage was 79.5%.Genotype of 21-OHD has strong correlation with clinical phenotype, which can effectively predict SW by severe mutation and predict SV by moderate mutation.
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Objective:To investigate the effects of caspase-11 non-canonical inflammasome on the Leptospira interrogans ( L. interrogans)-induced secretion of inflammatory cytokines in J774A.1 cells. Methods:Murine mononuclear macrophage cells (J774A.1) were infected with L. interrogans strain 56601. Expression of caspase-11, IL-1β, IL-1α and IL-18 at mRNA level in J774A.1 cells were detected by real-time RT-PCR. The levels of caspase-11, IL-1β, IL-1α and IL-18 in the culture supernatants of J774A.1 cells were detected by ELISA. Results:Real-time RT-PCR showed that caspase-11 expression at mRNA level was 5.12, 14.21, 8.94, 14.06, 18.58 and 0.93 times of that in uninfected cells after 1, 2, 4, 8, 12 and 24 h of L. interrogans infection, and respectively decreased to 0.10, 0.07, 0.10, 0.09, 0.07 and 0.45 times after caspase-11 inhibitor intervention ( P<0.05). Expression of IL-1β, IL-1α and IL-18 at mRNA level was significantly increased after infection ( P<0.05). After the intervention with caspase-11 inhibitor, IL-1β mRNA decreased to 0.05, 0.03, 0.02, 0.05, 0.06 and 0.02 times ( P<0.05); IL-1α mRNA decreased to 0.14, 0.07, 0.15, 0.10, 0.03 and 0.06 times ( P<0.05); IL-18 mRNA decreased to 0.08, 0.10, 0.16, 0.18, 0.10 and 0.07 times ( P<0.05). ELISA results showed that the expression of caspase-11, IL-1β, IL-1α and IL-18 at protein level was significantly increased. After the intervention with caspase-11 inhibitor, caspase-11 level decreased to 43.07, 41.64, 51.96, 86.56, 105.36, and 129.95 pg/ml ( P<0.05); IL-1β level decreased to 15.01, 14.19, 68.02, 31.20, 173.13 and 104.98 pg/ml ( P<0.05); IL-1α level decreased to 12.14, 15.40, 38.01, 21.97, 24.48 and 27.09 pg/ml ( P<0.05); IL-18 level decreased to 96.27, 102.21, 85.34, 116.28, 155.36 and 114.03 pg/ml ( P<0.05). Conclusions:Caspase-11 non-canonical inflammasome was involved in the mediation of IL-1β, IL-1α and IL-18 secretion in mouse mononuclear macrophages after L. interrogans infection.
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Objective To understand the differences in engulfing ability and phagolysosome for-mation between mononuclear-macrophages and neutrophils during Leptospira interrogans infection. Methods Human THP-1 monocytes and HL-60 cells were pretreated with PMA ( phorbol-12-myristate-13-acetate) and ATRA ( all-trans retinoic acid) to differentiate them into mononuclear-macrophages and neutrophils, respec-tively. The phagocytosis of Leptospira interrogans in THP-1-PMA mononuclear-macrophages and HL-60-AT-RA neutrophils was detected by confocal microscopy. The morphology of intracellular Leptospira was deter-mined by transmission electron microscopy. The viability of phagocytized Leptospira and the percentages of dead Leptospira were analyzed by confocal microscopy and spectrofluorimetry, respectively. Confocal micros-copy was used to measure the formation of phagolysosomes in different phagocytes. Results Both THP-1-PMA mononuclear-macrophages and HL-60-ATRA neutrophils could phagocytize Leptospira interrogans, but the phagocytic ability of the former was notably stronger than that of the latter (P<0. 05). Intracellular Lep-tospira were surrounded by phagocytic vesicles in both types of phagocytes. THP-1-PMA mononuclear-mac-rophages were better than HL-60-ATRA neutrophils in killing intracellular Leptospira (P<0. 05). More phagolysosomes were formed in THP-1-PMA mononuclear-macrophages than in HL-60-ATRA neutrophils ( P<0. 05). Conclusions Human mononuclear-macrophages but not neutrophils act as major phagocytes that play an important role in phagocytizing and killing Leptospira during infection. Less fusion of the phagosomes with lysosomes may be responsible for the lower Leptospira-killing ability of neutrophils.
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Objective To understand the genetic variations of neuraminidase (NA) genes of avian influenza virus H9N2 in Weining,Guizhou Province,and to provide the scientific evidence for the prevention and control of avian influenza virus.Methods Ribonucleic acids (RNA) were extracted and NA genes were amplified and sequenced from 13 randomly selected H9N2 positive samples from the live poultry market (LPM)environments in north of Weining Yi and Hui and Miao autonomous county (Weining),Guizhou Province during 2015 to 2017.Then the homology,genetic evolution,and sites of stalk deletion areas,potential N-glycosylation,receptor binding regions and drug resistance of H9N2 subtype avian influenza viruses were analyzed by a series of bioinformation software.Results Homology analysis revealed that there were 93.0％-100.0％ and 92.1％-100.0％ similarity among 13 strains H9N2 avian influenza viruses in nucleotide and amino acid of the NA gene,respectively.All strains belonged to DK/HK/Y280/97 sub-lineage,but their genetic sources were complex and diverse.Thirteen strains had a stalk deletion of 3 amino acid residues TEI at positions 63-65 and 3 isolates had mutation QN to QK at positions 39-40.The potential N-glycosylation sites at amino acid residues 86,146,200,and 234 of the NA protein of all strains were highly conserved,while other N-glycosylation sites had quantity and site mutations.There were different mutation types at the three sialic acid binding site areas,especially at 399-404 area.All NA protease activity sites and key sites of the 13 strains had no mutations associated with resistance to the neuraminidase inhibitor drugs.Conclusions All 13 strains H9N2 viruses belongs to DK/HK/Y280/97 sub-lineage in Weining,Guizhou Province during 2015-2017,and their genetic sources are complex and diverse.The mutations on sites of stalk areas,potential N-glycosylation and sialic acid binding site areas are presented at different degrees.Hence,enhancing surveillance and controlling H9N2 avian influenza virus is necessary.
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Objective To investigate the imaging features of the cerebral complications of infective endocarditis (IE)and the evolution process of infective cerebral infarction.Methods The clinical and imaging data from 5 patients diagnosed as IE with neurological complications were retrospectively and comprehensively reviewed,so that the imaging features of cerebral complications and infective cerebral infarctions were summarized.Results Among the 5 cases,3 showed multiple acute infarctions,including 2 massive ones.All the 5 cases showed multiple hemorrhagic lesions at different stages.The infarction and hemorrhage were mostly located at the corticalGmedullary junction.1 case of subarachnoid hemorrhage and 1 case of meningitis were also observed.Two massive infarctions mentioned above showed irregular patchy shape,which evolved into cerebral abscesses after 1 2 and 1 5 days of neurological symptoms showing up,then abscesses started shrinking after 33 and 3 1 days,respectively.Conclusion MRI can accurately reflect the features of cerebral complications of IE and the evolution process of infective cerebral infarctions,which provides evidences for physicians to make correct diagnoses and the treatment plans.
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The number of H7N9 bird flu cases was high and the situation was grim in guizhou province in 2017. To understand the molecular characteristics of the hemagglutinin gene (HA) and the risk of human infection with avian influenza virus A(H7N9) in Guizhou Province, 2017. Homology, genetic evolution and pivotal sites related to receptor binding regions, pathogenicity and potential glycosylation of 14 avian influenza viruses A(H7N9) were analyzed by a series of bioinformation softwares. It was cleared that there was 95.9%-100% similarity among 14 strains in nucleotide of the HA gene, and there were 96.8%-97.8% and 96.8%-97.9% similarities with vaccine strains A/Shanghai/2/2013 and A/Anhui/1/2013 recommended by WHO, respectively. Phylogenetic analysis showed that 14 HA genes were directly evolved in the Yangtze River Delta evolution branch, but they could be derived from five diffenrent strains. Then 13 of 14 strains cleavage site sequences of HA protein revealed they were low pathogenic avian influenza viruses, while A/Guizhou-Weining/CSY01/2017 was high pathogenic avian influenza virus. Mutation G186V at the receptor binding sites in the HA was found in all 14 strains, and mutation Q226L in 13 strains besides A/Guizhou-Weining/CSY01/2017. All five potential glycosylation motifs in the HA were conservative.
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Objective To understand the molecular characteristics of hemagglutinin (HA) and neuraminidase (NA) as well as the disease risk of influenza virus A H7N9 in Guizhou province.Methods RNAs were extracted and sequenced from HA and NA genes of H7N9 virus strains obtained from 18 cases of human infection with H7N9 virus and 6 environmental swabs in Guizhou province during 2014-2017.Then the variation and the genetic evolution of the virus were analyzed by using a series of bioinformatics software package.Results Homology analysis of HA and NA genes revealed that 2 strains detected during 2014-2015 shared 98.8％-99.2％ and 99.2％ similarities with vaccine strains A/Shanghai/2/2013 and A/Anhui/1/2013 recommended by WHO,respectively.Two strains detected in 2016 and 14 strains detected in 2017 shared 98.2％-99.3％ and 97.6％-98.8％ similarities with vaccine strain A/Hunan/02650/2016,respectively.Other 6 stains detected in 2017 shared 99.1％-99.4％ and 98.9％-99.3％ similarities with strain A/Guangdong/17SF003/2016,respectively.Phylogenetic analysis showed that all the strains were directly evolved in the Yangtze River Delta evolution branch,but they were derived from different small branch.PEVPKRKRTAR ↓ GLF was found in 6 of 24 strains cleavage site sequences of HA protein,indicating the characteristic of highly pathogenic avian influenza virus.Mutations A134V,G186V and Q226L at the receptor binding sites were found in the HA.All the strains had a stalk deletion of 5 amino acid residue "QISNT" in NA protein,and drug resistance mutation R294K occurred in strain A/Guizhou-Danzhai/ 18980/2017.In addition,potential glycosylation motifs mutations NCS42NCT were found in the NA of 9 of 24 strains.Condusions HA and NA genes of avian influenza A (H7N9) virus showed genetic divergence in Guizhou province during 2014-2017.The mutations of key sites might enhance the virulence of the virus,human beings are more susceptible to it.Hence,the risk of infection is increasing.
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Objective To understand the molecular characteristics of hemagglutinin (HA) and neuraminidase (NA) as well as the disease risk of influenza virus A H7N9 in Guizhou province.Methods RNAs were extracted and sequenced from HA and NA genes of H7N9 virus strains obtained from 18 cases of human infection with H7N9 virus and 6 environmental swabs in Guizhou province during 2014-2017.Then the variation and the genetic evolution of the virus were analyzed by using a series of bioinformatics software package.Results Homology analysis of HA and NA genes revealed that 2 strains detected during 2014-2015 shared 98.8％-99.2％ and 99.2％ similarities with vaccine strains A/Shanghai/2/2013 and A/Anhui/1/2013 recommended by WHO,respectively.Two strains detected in 2016 and 14 strains detected in 2017 shared 98.2％-99.3％ and 97.6％-98.8％ similarities with vaccine strain A/Hunan/02650/2016,respectively.Other 6 stains detected in 2017 shared 99.1％-99.4％ and 98.9％-99.3％ similarities with strain A/Guangdong/17SF003/2016,respectively.Phylogenetic analysis showed that all the strains were directly evolved in the Yangtze River Delta evolution branch,but they were derived from different small branch.PEVPKRKRTAR ↓ GLF was found in 6 of 24 strains cleavage site sequences of HA protein,indicating the characteristic of highly pathogenic avian influenza virus.Mutations A134V,G186V and Q226L at the receptor binding sites were found in the HA.All the strains had a stalk deletion of 5 amino acid residue "QISNT" in NA protein,and drug resistance mutation R294K occurred in strain A/Guizhou-Danzhai/ 18980/2017.In addition,potential glycosylation motifs mutations NCS42NCT were found in the NA of 9 of 24 strains.Condusions HA and NA genes of avian influenza A (H7N9) virus showed genetic divergence in Guizhou province during 2014-2017.The mutations of key sites might enhance the virulence of the virus,human beings are more susceptible to it.Hence,the risk of infection is increasing.
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Objective To explore the value of serum IgA,serum C3 and IgA/C3 ratio in the diagnosis and clinical monitoring of IgA nephropathy,which provide new insight for the alternatives of renal biopsy. Methods In the first affiliated hospital of Dalian Medical University,60 patients with IgAN,60 patients with non-IgAN and 40 healthy people were detected by blood biochemical indexes and immune indexes,such as the Urea,serum IgA and serum C3,IgA/C3 ratio. According to the healthy condition and Lee grading,we divided all the 160 patients into different groups. By comparing the blood biochemical indexes and immune indexes among these different groups,we investigated the relationship between the serum IgA/C3 ratio and the IgAN.Results Firstly,the level of serum IgA and IgA/C3 were obviously different among the patients with IgAN,patients with non-IgAN and healthy people(P<0.01).Secondly,the serum IgA and IgA/C3 have high specificity and sensitivity when distin-guishing between IgAN and non IgAN. Thirdly,the level of serum IgA and IgA/C3 have significant deference be-tween the patients with Lee Ⅰ-Ⅱ grading and the patients with Lee Ⅲ-Ⅴ grading,while the level of C3 is very similar between different groups in all the three situations mentioned above. Conclusion Our research provides stronge evidence demonstrates that the level of serum IgA/C3 ratio have close relationship with IgAN and they can serve as the reference index when diagnosising and grading IgAN.
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Objective@#To investigate the molecular characteristics and tracing of the hemagglutinin (HA) gene, and to analyze the risk of human infection with influenza virus A (H7N9) in Guizhou Province, so that to provide evidence for the prevention and control of highly pathogenic avian influenza A (H7N9).@*Methods@#Nucleic acids of 5 strains of H7N9 including 1 sample of the patient′s nasopharyngeal swab and 4 samples of the live poultry market (LPM) environment were extracted and HA genes were amplified and sequenced. Then the homology, genetic evolution and the pivotal sites related to receptor binding regions, pathogenicity and potential glycosylation of the avian influenza A (H7N9) viruses were analyzed by a series of bioinformatics softwares.@*Results@#Homology analysis revealed that the homologies of nucleotide and amino-acid of the HA gene of H7N9 strains from the patient and LPM in Weining County, Guizhou Province were 99.8% and 99.6%, respectively, while those of 4 strains from LPM were both 100%. The homologies of nucleotide and amino-acid of the HA gene of H7N9 strains were the highest with the strain of A/Guangxi/5/2017 isolated from a Guangxi infected patient (99.7%-99.9% and 99.4%-99.8%, respectively), while those with the strain isolated from LPMs environment at the end of 2016 (A/Environment/Guangdong/C16283222/2016) were 99.0%-99.2% and 98.9%-99.2%, respectively. However, the homologies of nucleotide and amino-acid of the HA gene of H7N9 strains with A/Shanghai/2/2013 recommended by world health organization and the candidate vaccine strain A/Anhui/1/2013 were 96.8%-97.0% and 95.8%-96.2%, respectively. Phylogenetic analysis showed that the 5 strains had the nearest genetic distance to the strain A/Guangxi/5/2017. All the 5 strains cleavage site sequences of HA protein showed mutation of PEVPKRKRTAR↓GLF, and they were highly pathogenic avian influenza viruses mutant strains, which all had mutation of G186V at the receptor binding sites of HA gene, while no Q226L mutation was found. All 5 strains had new mutation of A363S, and new mutations of R56K and I297V were only found in the strain isolated from the patient. Among the five potential glycosylation motifs in the HA, only 421NWT and 493NNT had variation of the position post shift.@*Conclusions@#All the 5 H7N9 strains isolated in Weining County, Guizhou Province are highly pathogenic avian influenza mutative viruses. The current candidate vaccine may not provide a very good protection. The mutations of cleavage site of HA protein, G186V as well as other new mutation sites of HA may enhance the susceptibility and pathogenicity to human beings.
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Objective To investigate the molecular characteristics of H5 subtype avian influenza viruses (AIV) in Weining, Guizhou Province. Methods Nine representative strains were randomly select-ed from H5 subtype AIV that were identified by real-time PCR in Weining, Guizhou Province from 2015 to 2017. Nucleic acid was extracted from each sample and hemagglutinin (HA) genes were amplified and then sequenced. Homology, genetic evolution and the sites related to pathogenicity, receptor binding regions as well as potential glycosylation of H5 AIV were analyzed by bioinformation software. Results Homology analysis revealed that there was 96. 1%-99. 9% and 95. 7%-100% similarity among the nine strains in nu-cleotide and amino acid of HA gene, respectively. These strains belonged to two branches, H5-1 and H5-2. The cleavage site motifs were PLREKRRKR↓GLF for five strains in H5-1 branch and PQRERRRKR↓GLF for four strains in H5-2 branch, which made them high pathogenic. QSG and QRG at the key receptor bind-ing sites were found in H5-1 and H5-2 branch strains, respectively. They were responsible for receptor bind-ing specificity of AIV. Mutations of 138Q, 139G and 53K were all detected in the nine strains. 129K, 189T, 140K and 282V mutations were discovered in the five strains of H5-1 branch, while 189N, 140M and 282I mutations were found in the four strains of H5-2 branch. Results of the glycosylation motif analysis showed that six sites were conservative, but there was an addition of 124NHT site in two strains of H5-2 branch isolated in 2017. Conclusion Two high pathogenic H5 subtypes of AIV could be epidemic in Wein-ing, Guizhou Province during 2015 to 2017. Although H5 subtype AIV did not possess specific receptor binding regions like human influenza viruses, they were in continuous variation with an increase in glycosyla-tion motifs, which might enhance their virulence and pathogenicity to human beings. Hence, surveillance and study on the molecular properties of H5 subtype AIV should be strengthened.
ABSTRACT
Objective To investigate the influences of Leptospira interrogans (L.interrogans) in-fection on the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion mole-cule-1 (VCAM-1) on endothelial cells. Methods Expression of ICAM-1 and VCAM-1 at mRNA level was detected by reverse transcription-polymerase chain reaction (RT-PCR) after infecting human umbilical vein endothelial cells (HUVEC) with L.interrogans strain Lai. Silver staining was used to detect leptospires in lung,liver and kidney tissues of L.interrogans-infected C3H/HeJ mice. Expression of ICAM-1 and VCAM-1 in lung,liver and kidney tissues of L.interrogans-infected mice was measured with immunohistochemistry. Results L.interrogans infection increased the expression of ICAM-1 and VCAM-1 on HUVEC(P<0.05). Moreover,the expression of VCAM-1 at mRNA level was significantly higher than that of ICAM-1 (P<0.05). Silver-stained leptospires could be found in lung,liver and kidney tissues of L.interrogans-infected C3H/HeJ mice. Results of the immunohistochemical examination showed that increased expression of both ICAM-1 and VCAM-1 could be detected in ling,liver and kidney tissues of L.interrogans-infected mice,and the VCAM-1 level was significantly higher than that of ICAM-1 in every tissue sample(P<0.05). Conclu-sion L.interrogans infection could induce the expression of ICAM-1 and VCAM-1 on endothelial cells and increase the expression of VCAM-1 to a level significantly higher than that of ICAM-1, which mediated the infiltration of specific inflammatory cells to the site of infection.