Identification of genetic causes in children with unexplained epilepsy based on trio-whole exome sequencing.

Genotype and clinical phenotype analyses of 128 children were performed based on whole exome sequencing (WES), providing a reference for the provision of genetic counseling and the precise diagnosis and treatment of epilepsy. A total of 128 children with unexplained epilepsy were included in this study, and all their clinical data were analyzed. The children's treatments, epilepsy control, and neurodevelopmental levels were regularly followed up every 3 months. The genetic diagnostic yield of the 128 children with epilepsy is 50.8%, with an SNV diagnostic yield of 39.8% and a CNV diagnostic yield of 12.5%. Among the 128 children with epilepsy, 57.0% had onset of epilepsy in infancy, 25.8% have more than two clinical seizure forms, 62.5% require two or more anti-epileptic drug treatments, and 72.7% of the children have varying degrees of psychomotor development retardation. There are significant differences between ages of onset, neurodevelopmental levels and the presence of drug resistance in the genetic diagnostic yield (all p < 0.05). The 52 pathogenic/likely pathogenic SNVs involve 31 genes, with genes encoding ion channels having the largest number of mutations (30.8%). There were 16 cases of pathogenic/possibly pathogenic CNVs, among which the main proportions of CNVs were located in chromosome 15 and chromosome 16. Trio-WES is an essential tool for the genetic diagnosis of unexplained epilepsy, with a genetic diagnostic yield of up to 50.8%. Early genetic testing can provide an initiate appropriate therapies and accurate molecular diagnosis.

Carotid endarterectomy combined with vertebral artery transposition treating vertebral artery V1 segment stenosis combined with ipsilateral carotid artery stenosis / 中国胸心血管外科临床杂志

@#Objective    To explore the treatment outcome of carotid endarterectomy combined with vertebral artery transposition in patients with severe stenosis to occlusion of the vertebral artery V1 segment and the ipsilateral carotid artery. Methods    From June 2017 to September 2020, patients with severe stenosis to occlusion of the vertebral artery V1 segment and the ipsilateral carotid artery treated with carotid endarterectomy combined with vertebral artery transposition in Fuwai Hospital were retrospectively analyzed. Results    Finally 12 patients were enrolled, including 10 males and 2 females with an average age of 67.8±6.0 years. Twelve patients were successfully operated and the follow-up time was 1-3 years. The stenosis degree of the V1 segment of the vertebral artery decreased from 83.5%±11.8% to 24.9%±14.3% (P<0.001). The stenosis degree of carotid artery decreased from 85.6%±11.0% to 0% (P<0.001). Postoperative follow-up showed that the symptoms of symptomatic patients before surgery improved. The 1-year and 3-year patency rates were 100.0%, and there were no peripheral nerve injury complications, perioperative deaths or strokes. Conclusion    Carotid endarterectomy combined with vertebral artery transposition can treat ipsilateral carotid artery  stenosis and vertebral artery stenosis at the same time, improve blood supply to the brain, improve patients' symptoms and has high promotion value.

Clinical Characteristics and Risk Factors in Children with Acute Leukemia Complicated with Multiple Drug Resistant Bacterial Septicemia / 中国实验血液学杂志

OBJECTIVE@#To investigate the clinical characteristics and risk factors of acute leukemia complicated with multi-drug resistant bacterial septicemia in children.@*METHODS@#The clinical data of children with acute leukemia complicated with septicemia admitted to the Affiliated Hospital of Guangdong Medical University from January 2013 to May 2021 were retrospectively analyzed. Their flora composition and drug resistance were also analyzed. The children were divided into multi-drug resistant bacteria (MDRB) group and non-multi-drug resistant bacteria (non-MDRB) group according to the drug sensitivity results, and the differences in clinical data between the two group were compared.@*RESULTS@#A total of 108 children had drug sensitivity results, 47 cases in the MDRB group, including 26 strians of Gram-positive bacteria (G+), the most common multi-drug resistant G+ bacteria were coagulase-negative staphylococci (CoNS) and Staphylococcus aureus, and the most common multi-drug resistant Gram-negative bacteria G- bacteria were Escherichia coli and Klebsiella pneumoniae subspecies pneumoniae. Compared with non-MDRB group, children in MDRB group had higher C-reactive protein (CRP) level and mortality rate (P <0.001, P =0.009), lower initial empirical anti-infection efficiency (P <0.001), and were more likely to have septic shock (P =0.003). Logistic analysis showed that the risk factors of acute leukemia complicated with MDRB septicemia in children were previous MDRB infection (OR =6.763, 95% CI: 1.141-40.092, P =0.035), duration of agranulocytosis before infection≥7 days (OR =3.071, 95% CI: 1.139-8.282, P =0.027), and previous use of antimicrobial drugs within 90 days before infection (OR =7.675, 95% CI: 1.581-37.261, P =0.011).@*CONCLUSIONS@#The clinical features of acute leukemia complicated with MDRB septicemia in children include a heavy inflammatory response, significantly elevated CRP, susceptibility to secondary septic shock, low efficiency of initial empirical anti-infective therapy, and high mortality rate. Previous MDRB infection, duration of agranulocytosis before infection≥7 days, and previous use of antimicrobial drugs within 90 days before infection are risk factors of acute leukemia complicated with MDRB septicemia in children.

Modified vertebral-carotid transposition treating stenosis at V1 segment of vertebral artery / 中国胸心血管外科临床杂志

@#Objective    To investigate the treatment of modified vertebral-carotid transposition (VCT) in patients with severe stenosis or occlusion at V1 segment of vertebral artery. Methods    A retrospective study of 13 patients with severe stenosis or occlusion at V1 segment of vertebral artery treated by modified VCT in our hospital from October 2016 to December 2018 was done. There were 10 males and 3 females with an average age of 70.5±7.1 years. Results    The operation was successful in this series of patients. The follow-up duration was 1-3 years. The stenosis degree of the V1 segment of the vertebral artery decreased from 86.8%±7.5% to 17.4%±14.5%. All patients achieved remission of symptoms after the surgery. Temporary peripheral nerve injury occurred in 6 patients. Four patients with neurological complications relieved during follow-up. The patency rate was 100.0% at postoperative 1 and 3 years. There was no perioperative death, stroke or re-intervention. Conclusion    Modified VCT can precisely restore the distal blood flow of patients with severe stenosis or occlusion at V1 segment of vertebral artery, and relieve their symptoms.

Old fusidane-type antibiotics for new challenges: Chemistry and biology / 中国天然药物

The spread of antibiotic-resistant bacteria and exhausted drug leads render some infections untreatable now and in the future. To deal with these "new challenges", scientists tend to re-pick up "old antibiotics". Fusidane-type antibiotics have been known for nearly 80 years as potent antibacterial agents against gram-positive bacteria, especially Staphylococci, and represent the only triterpene-derived antibiotic class in clinical setting. These attractive characteristics have drawn renewed attention on fusidane-type antibiotics in recent decades. Isolation, characterization, biological evaluation, as well as chemical modifications of fusidane-type antibiotics are increasingly being reported. Combinatorial biosynthesis of this type of antibiotics has been successfully utilized not only for elucidating the biosynthetic pathways, but also for expanding their structural diversity. Some isolated and synthetic compounds exhibit comparable or even more potent biological activity than fusidic acid. This review provides an overview of progress on the studies of structure and biology of fusidane-type antibiotics from 1943 to April 2021. The informative structure-activity relationship is also highlighted.

Toxic Effects of Long-term Exposure to PM / 中国实验血液学杂志

OBJECTIVE@#To investigate the toxic damage and possible mechanism of chronic exposure of ambient particulate matter (PM@*METHODS@#Mice were treated with different doses (150, 300, 600 mg/kg) of chitosan after exposure to PM@*RESULTS@#Compared with the mice in control group, IL-2 secretion and CXCL12 expression were decreased in the bone marrow of PM@*CONCLUSION@#Chronic exposure of PM

Combination of physician modified stent-graft fenestration and in-situ needle fenestration during thoracic endovascular aortic repair / 中华普通外科杂志

Objective:To evaluate the safety and feasibility of the in-situ needle fenestration combined with the in vitro physician modified fenestration technique to reconstruct supra-aortic branches during thoracic endovascular aortic repair (TEVAR) for aortic arch lesions requiring landing at Z0 and Z1.Methods:From Nov 2017 to Dec 2019, eighteen patients who underwent both the in-situ needle fenestration and the in vitro physician modified fenestration techniques to extend the proximal landing zone to Z0 and Z1 during TEVAR were included in our study.Results:Sixteen patients underwent in vitro physician modified fenestration ,two patients underwent in vitro physician modified fenestration to reconstruct both the left common carotid artery and the innominate artery. All eighteen patients received in-situ needle fenestration to preserve the left subclavian artery. Supra aortic branches were preserved in all patients (38/38, 100%). There was no Type Ⅰ endoleak. Type Ⅱ endoleak was found in four paitnets (4/18). Type Ⅲ endoleak occurred in one patient (1/18). Type Ⅳ endoleak in four patients (4/18). Type Ⅲ endoleak needed open aortic arch repair 6 months later. The median follow-up time was 12 months. One (1/18) died in 12 months and the other patients were doing well.Conclusions:The joint application of the in-situ needle fenestration and the in vitro physician modified fenestration to reconstruct supra-aortic branches during TEVAR for aortic arch pathologies requiring landing at Z0 and Z1 was satisfactory.

The Comparison of Iron Deficiency Anemia Rat Treated with Hydroxypropyl Chitosan Ferrous Ion Complex and Ferrous Sulfate / 中国实验血液学杂志

OBJECTIVE@#To observe and compare the therapeutic effects of hydroxypropyl chitosan ferrous ion complex solution and ferrous sulfate solution in iron deficiency anemia rats and their effects on gastric mucosa.@*METHODS@#Seven rats were randomly selected from thirty five SPF grade SD rats as control group, and were fed with normal diet, distilled water (E). The rest of SD rats were fed with low iron feed and distilled water plus continuous tail vein bloodletting to establish the iron deficiency anemia model. After the model was established successfully, the rats were randomly divided into four groups: blank control group (A), iron deficiency anemia control group (B), ferrous sulfate group (C), hydroxypropyl chitosan ferrous ion complex (HPCTS-Fe@*RESULTS@#After modeling, except the normal control group, the hair color of the rats in the four groups showed dark yellow and the belly of the toes became white gradually. HGB, HCT, Ret%, MCV, MCH, MCHC and SF decreased significantly (P < 0.05). After treatment, the rats with dark yellow hair in group C and D were improved, and the toe abdomen turned pink gradually. RBC, HGB, HCT, Ret%, MCV, MCH, MCHC and SF in rats in group C and D increased, which were higher than those in group B (P < 0.05). The HGB of the rats in group D was higher than that of group C in day 28th during treatment and the Ret% was higher than that in group C at day 10th (P<0.05).After treatment, the liver and spleen of the rats in group C and D were lighter than those in group B (P<0.05).The gastric mucosa in group A, B, D and E was not damaged obviously, while it was slightly irritated and damaged in group C.@*CONCLUSION@#Hydroxypropyl chitosan ferrous complex solution can improve the hemoglobin level of SD rats with iron deficiency anemia, which is stronger than ferrous sulfate solution and shows no damage to gastric mucosa.

Identification of CXCR4 and CXCL10 as Potential Predictive Biomarkers in Triple Negative Breast Cancer (TNBC).

BACKGROUND Efficacious therapy for triple negative breast cancer (TNBC) continues to be a profound clinical challenge, but the key driven genes and convoluted signaling pathways are still unknown. MATERIAL AND METHODS A total of 223 samples (163 TNBC and 60 healthy breast tissues) were taken and deeply integrated analyzed by R software from 4 expression profiles in the study, including GSE53752, GSE45827, GSE65194, and GSE38959. We examined differentially expressed genes (DEGs) and screen for critical genes and pathways enrichment. The protein­protein interaction (PPI) network of DEGs-associated was built through the STRING Version: 11.0 database and Cytoscape software to filter the hub gene. Then, we verified hug gene expression levels through the Oncomine database. Also, we analyzed the prognostic value of TNBC patient's hub genes using the Kaplan-Meier plotter database. RESULTS In our study, we filter out 365 DEGs, including 212 upregulated genes and 153 downregulated genes. Then, 10 hub genes were picked out by the intersection of 12 algorithms. At the same time, we discovered that CXCR4 and CXCL10 overexpression are favorable prognostic factors for recurrence-free survival of TNBC through the Kaplan-Meier plotter database. CONCLUSIONS Our research found that CXCR4 and CXCL10 overexpressed, and they were a favorable prognostic factor in patients with TNBC. CXCR4 and CXCL10 might be effective targets for TNBC therapy.

Effect of neoadjuvant chemoradiotherapy on perioperative immune function of patients with locally advanced esophageal cancer.

This study aims to evaluate the effect of neoadjuvant chemoradiotherapy (NCRT) on perioperative immune function during surgery to treat resectable locally advanced esophageal cancer. Records were retrospectively analyzed for 220 patients with locally advanced esophageal cancer, of whom 112 received surgery alone and 98 received neoadjuvant NCRT plus surgery. The two groups were compared in terms of proportions of CD3+, CD4+, CD8+, and natural kill (NK) cells, as well as the ratio of CD4+ to CD8+ cells. These measurements were made using flow cytometry on preoperative day 1 and on postoperative days 1 and 7. Subgroup analysis were performed in terms of degrees of pathological response of NCRT. When the entire NCRT and no-NCRT (surgery alone) cohorts were compared, no significant differences in propocrtions of CD3+, CD4+, CD8+, or NK cells or in the CD4+/CD8+ ratio occurred at any of the three time points. Similar results were obtained using the subgroup of NCRT patients who were NCRT-sensitive, but the subgroup of NCRT-insensitive patients showed significantly lower CD4+ and NK proportions and lower CD4+/CD8+ ratio than the no-NCRT group. Our findings suggest that NCRT does not affect perioperative immune function in patients who are NCRT-sensitive, but it does significantly reduce such function in patients who are NCRT-insensitive.

Mid-term outcomes in patients undergoing tricuspid valve repair with rheumatic tricuspid valve disease / 中华胸心血管外科杂志

Objective The aim of this study was to summarize the characteristics of rheumatic tricuspid valve disease ( RTVD) and to evaluate the mid-term outcomes in patients undergoing tricuspid valve repair with RTVD. Methods Between January 2009 and June 2016, 251 consecutive patients with rheumatic heart disease( RHD) underwent left-sided valvular re-placement by a single surgeon. We analyzed 39 patients with RTVD which was diagnosed during the operation. Among them, 32 patients, with moderate or higher tricuspid regurgitation( TR) , were compared with other 59 patients of functional tricuspid regurgitation( FTR) for a better understanding of the features of the RTVD. A total of 39 patients were categorized into 2 groups:Ring annuloplasty group(n=33) and non-Ring annuloplasty group(n=6) which consisted of modified De Vega annu-loplasty for 4 patients and edge-to-edge repair for 2. Meanwhile, 13 of them underwent concomitant tricuspid commissurotomy and 1 patient had a tricuspid leaflet augmentation procedure. We analyzed the mid-term outcomes of 22 patients( follow-up du-ration>1 year)with a mean follow-up duration of(45.5 ±25.1) months. Results Compared with FTR, patients with RTVD had higher preoperative TR grade(3.1 ±0.8 vs. 2.6 ±0.7, P=0.004) but with lower preoperative PASP[(53.8 ±19.4) mmHgvs.(63.6±21.5)mmHg,P=0.037)](1mmHg=0.133kPa) andtricuspidannulusdiameter(TAD) thatobserved bothinpreoperativeechocardiogramtests[(37.0±5.7)mmvs.(41.9±6.7)mm,P=0.018)]andintraoperativedetection [(35.6±4.1)mmvs.(39.9±6.5)mm,P=0.000)] . TherewasnoearlymortalityandresidualmoderateorhigherTR grades in either group. Compared with patients in non-ring annuloplasty group, patients in ring annuloplasty group showed low-er postoperative TR grade(0. 2 ± 0. 4 vs. 0. 7 ± 0. 5, P=0. 039) and acceptable TR grade(0. 8 ± 0. 5 vs. 1. 3 ± 1. 9, P>0. 050) during the mid-term follow-up. PASP, the peak diastolic velocity and pressure gradient across tricuspid valve were not different between groups in preoperative, postoperative and follow-up. Conclusion Compared with FTR, Patients with RTVD had lower preoperative PASP and TAD, but with a higher preoperative TR grade. In our study, ring annuloplasty showed simi-lar mid-term outcomes compared with other procedures.

Exploration of the BF2*15 major histocompatibility complex class I binding motif and identification of cytotoxic T lymphocyte epitopes from the H5N1 influenza virus nucleoprotein in chickens.

The binding motif of BF2*15 major histocompatibility complex (MHC) class I was explored by analyzing the interaction between an infectious bronchitis virus octapeptide and BF2*15, and the cytotoxic T lymphocyte (CTL) epitope from the nucleoprotein (NP) of H5N1 virus was identified using experimental methods. Computational methods, including homology modeling, molecular dynamics simulation, and molecular docking analysis, were used. The recombinant plasmid pCAGGS-NP was constructed, and NP expression was confirmed by indirect immunofluorescence and Western blot in transfected 293T cells. Antibodies against NP in pCAGGS-NP-inoculated specific-pathogen-free chickens were detected by enzyme-linked immunosorbent assay (ELISA). Interferon γ (IFN-γ) mRNA was quantified, and IFN-γ production was evaluated using quantitative reverse transcription PCR and capture ELISA, respectively. CD8(+) T-lymphocyte proliferation was detected using flow cytometric analysis. The BF2*15 MHC class I binding motif "x-Arg/Lys-x-x-x-Arg/Lys" was explored. Quantification of chicken IFN-γ mRNA, evaluation of IFN-γ production, and measurement of CD8(+) T-lymphocyte proliferation confirmed that the peptide NP67-74 of H5N1 was the BF2*15 MHC-class-I-restricted CTL epitope.

Comparison of postoperative immune function in patients with thoracic esophageal cancer after video-assisted thoracoscopic surgery or conventional open esophagectomy.

AIM: The aim of the study was to compare postoperative immune function in patients with thoracic esophageal cancer (EC) after video-assisted thoracoscopic surgery (VATS) or conventional open esophagectomy. PATIENTS AND METHODS: Medical records were retrospectively analyzed for 228 patients with thoracic EC treated at a single hospital using VATS (n = 52) or conventional open esophagectomy (n = 176). Proportions of CD3(+), CD4(+), CD8(+), and natural kill (NK) cells, as well as the ratio of CD4(+) to CD8(+) cells, were measured in the two groups using flow cytometry on preoperative day (PrD) 1 and postoperative days (PoD) 1 and 7. RESULTS: Proportions of CD3(+), CD4(+), and NK cells as well as the CD4+/CD8+ ratio decreased significantly from PrD1 to PoD1 in both the VATS and open esophagectomy groups. In the VATS group, these parameters had returned to preoperative levels (PrD1) by PoD7. These parameters in open esophagectomy group increased from PoD1 to PoD7 but also lowered significantly to PrD1 by PoD7. The proportion of CD8(+) cells was similar between the two groups at all time points tested. CONCLUSION: Patients may experience less postoperative immune suppression after VATS than after conventional open esophagectomy, and they may recover preoperative immune function more quickly.

Interactive mechanism between avian infectious bronchitis S1 protein T cell peptide and avian MHC I molecule.

This study aims to construct a 3D structure of the avian major histocompatibility complex (MHC)-ß2M complex through homology modelling technology, perform molecular docking of the predicted infectious bronchitis virus (IBV) S1 protein potential epitope peptide Sp6 (NQFYIKLT) and the avian MHC-ß2M complex, and demonstrate the interactive mechanism between Sp6 and MHC using molecular dynamical simulations. The peptide Sp6 and the non-related peptide NP89-97 (PKKTGGPIY) were used to stimulate in vitro recombinant plasmid (pCAGGS-S1) avian splenic lymphocytes. Flow cytometric results show that CD8(+) T lymphocytes reproduce stimulated by the Sp6 and the nonrelated peptide proliferate by 34.8% and 2.6%, respectively. Meanwhile, fluorescent quantitative PCR results show that the secretion of IFN-γ in avian splenic lymphocytes increases after Sp6 stimulation. These data suggest that Sp6 can induce the activated avian lymphocytes in vitro to produce CTL, which is the CTL epitope in IBV S1.

Attenuation mechanism of virulent infectious bronchitis virus strain with QX genotype by continuous passage in chicken embryos.

The virulent isolate SDZB0808 of QX-type infectious bronchitis virus (IBV) was continuously passaged in chicken embryos for 110 generations. The safety and immune efficacy of the 110th generation of IBVs (P110) were evaluated. Damage was not found in the appearance of the 3-day-old specific-pathogen-free (SPF) chicks immunized with 10(4.5) EID50 (median embryo infective dose) of P110 by intranasal and ocular administration. At 14 d after the vaccination with 10(4.5) EID50 of P110, all the 3-day-old SPF chicks were immune from the attack of the homologous virulent strain SDZB0808 and the heterologous virulent strain SDIB821/2012. The whole genome sequencing of SDZB0808 of different generations (P1-P110) indicated that the replicase 1a sequences of P60-P110 all lost a length of 30bp in the same region. Specific primers were designed according to the differences in the genomes of P1-P110. SYBR Green I real-time quantitative PCR was adopted to analyze the proportion of the viruses with 30bp deletion in P60, P100, and P110. Results showed that with the passage in chicken embryos, the proportion of the viruses with 30bp deletion gradually increased. Almost 100% of the viruses in the P110 had 30bp deletion in the replicase 1a sequence. Therefore, the attenuation of IBV's virulence may be the outcome of directional screening in the chicken embryos. This work confirmed the high safety and immune efficacy of P110 in SPF chickens. Thus, P110 can serve as an attenuated IBV vaccine candidate.

Mechanisms of FOXM1 in carcinogenesis and the progression of the anti-FOXM1 target drugs / 国际肿瘤学杂志

Forkhead box M1 (FOXM1),one member of the Forkhead family,plays an important role in tumor development,invasion,metastasis and angiogenesis by regulating the expression of tumor related genes.Moreover,The specific mechanism of FOXM1 is not fully understood in the development of tumors.Currently,the study of anticancer drugs targeting FOXM1 is in the initial stage.Exploring the mechanism of FOXM1 in carcinogenesis and the development of drugs targeting FOXM1 and its downstream signaling pathways have broad clinical application prospects.

Analysis of the key factors in SCID mouse-human leukemia model / 中国实验血液学杂志

<p><b>OBJECTIVE</b>This study was to explore the key factors in leukemia model through the analysis of mouse with bad life state in the modeling process of leukemia so as to provide the theoretical reference for improving the success rate of modeling.</p><p><b>METHODS</b>At 1 week after inoculation of leukemia cells into SCID mice, the life status and peripheral hemogram of SCID mice were tested, the bone marrow smears, splean biopsy and spleen index of mice were examined after dissecting mormal and agoned/died mice during modoling, and the examined results were compared.</p><p><b>RESULTS</b>As compared with control mice, the life status of experimental mice was poor; the blood smear test showed juvenile cells, slightly more white blood cells with irregular shape and partial rupture, the lymphocytes and band cells obviously increased, the neutrophile granulocytes showed nuclear left shift; the bone marrow smears showed larger cell volume, smaller mulcoplasm, abnormal morphology of cells and cell nuclei and serious cell rapture; the spleen examination showed that the spleen diplayed enlargement and hyperemia to varying degree, the spleen index obviously increase, the spleen interstitial expansion, cell disordered arragement and irregular cell shope were observed, however there was no infiltration of leukemia cells in control and experimental mice.</p><p><b>CONCLUSION</b>The mouse age, pathway of inoculating the leukemia cells, sterile condition in breading and avoiding the rejection and inflammatory response in modeling process are the key foctors influencing the modeling success.</p>

[Complete genomic analysis of a novel infectious bronchitis virus isolate].

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.

Complete genomic analysis of a novel infectious bronchitis virus isolate / 病毒学报

The genome of CK/CH/SD09/005, an isolate of infectious bronchitis virus (IBV), was characterized to enable the further understanding of the epidemiology and evolution of IBV in China. Twenty-five pairs of primers were designed to amplify the full-length genome of CK/CH/SD09/005. The nucleotide sequence of CK/CH/SD09/005 was compared with reference IBV strains retrieved from GenBank. The phylogenic relationship between CK/CH/SD09/005 and the reference strains was analyzed based on S1 gene sequences. The complete genome of CK/CH/SD09/005 consisted of 27691 nucleotides (nt), excluding the 5' cap and 3' poly A tail. The whole-genome of CK/CH/SD09/005 shared 97 - 99% nucleotide sequence homology with the GX-NN09032 strain, which was the only complete genome that was closely related to CK/CH/SD09/005. When compared with all reference strains except GX-NN09032, CK/CH/SD09/005 showed the highest similarity to ck/CH/LDL/091022 and SDIB821/2012 (QX-like) in the replicase gene (Gene 1) and 3'UTR, with a sequence identity rate of 97% and 98%, respectively. However, CK/CH/SD09/005 exhibited lower levels of similarity with ck/CH/LDL/091022 and SDIB821/2012 in S-3a-3b-3c/ E-M-5a-5b-N with a sequence identity of 72% - 90%. CK/CH/SD09/005 showed the highest level of nucleotide identity with Korean strain 1011, and Chinese strains CK/CH/LXJ/02I, DK/CH/HN/ZZ2004 and YX10, in ORF 3c/E (97%), 5a (96%), 5b (99%) and N (96%), respectively. ORFs 3a, 3b and M of CK/CH/SD09/005 exhibited no more than 90% homology with the reference strains, excluding GX-NN09032. The phylogenic analysis based on the S1 gene revealed that CK/CH/SD09/005 and 39 published strains were classified into seven clades (genotypes). CK/CH/SD09/005 was distributed in clade IV with several isolates collected between 2007 and 2012. CK/CH/SD09/005 showed 66% - 69% and 72% - 81% nucleotide identities with the IBV strains of other six clades in the S1 and S2 subunits, respectively. More over, multiple substitutions were found throughout the entire S gene of CK/CH/SD09/005, while insertions and deletions were located within the S1 gene. These results indicated that CK/CH/SD09/005 is a novel variant that may be derived from the QX-like strains that are prevalent in China. Multiple genetic mechanisms, including recombinations, mutations, insertions and deletions, are likely to have contributed to the emergence of this IBV strain.

Effect of notch signaling pathway on VEGF promoting rat mesenchymal stem cell proliferation / 中国实验血液学杂志

This study was purposed to investigate the effect of Notch signaling pathway on VEGF promoting the proliferation of rat mesenchymal stem cells (MSC). Rat MSC were cultured in vitro, and the cells in logarithmic growth phase were used for experiments. The inhibitor DAPT was used to block Notch signaling pathway, and the effect of the pathway on VEGF promoting proliferation of MSC was observed. The experiment was divided into 4 groups: control, VEGF, DAPT and VEGF+DAPT. The CCK-8 was used to assay the cells proliferation of each group, while RT-PCR was used to detect the changes of related genes (Notch1, Notch2, Flk-1, Hes-1) at mRNA levels. The results indicated that the cells survival rate MSC in DAPT group and VEGF+DAPT group was low in each time point (24 h, 48 h, 72 h), the cell number decreased, and the cells became rounded. The survival rate of MSC in VEGF group was the highest; the difference of cell survival rate was statistically significant between the groups (P < 0.01); Compared with the control group, the mRNA expression level of Notch1, Notch2 and Flk-1 in VEGF group was raised, while the expression level of Notch1 and Notch2 in DAPT group and VEGF+DAPT group come down, with statistically significant differences (P < 0.05); whereas the mRNA expression level of Hes-1 in VEGF group was down-regulated, but that in DAPT group and VEGF+DAPT group was up-regulated, and the difference was statistically significant (P < 0.05). Flk-1 mRNA level in DAPT group and VEGF+DAPT group was slightly lower, but the difference was not statistically significant (P > 0.05). It is concluded that Notch signaling pathway plays an important role in promoting the proliferation of rat MSC, treated with VEGF, however, the DAPT can weaken this effect.