Objective:To analyze phylogenetic structure and molecular characteristics of H5N6 avian influenza virus (AIVs) isolated from live poultry market (LPM).Methods:Oropharyngeal and cloacal swabs from poultry, and environmental samples were collected from LPM in Urumqi in December 2018, AIVs were isolated and identified by inoculation of chicken embryo, hemagglutination test and RT-PCR, the viral whole genome was amplified with the universal primers of influenza A virus, and then sequenced, pairwise sequence alignments, phylogenetic and molecular characteristics analysis were performed by BLAST, Clustal W, MEGA-X and DNAStar software.Results:Six strains of H5N6 AIVs were isolated from poultry samples, the identity between the viral genes was high (99.4%-100.0%), so the isolates were the same source. BLAST analysis revealed that the viral NP sequence had the highest identity (99.7%) with H5N6 AIVs isolated from poultry in Suzhou, while the sequence of the remaining 7 viral genes had the highest identity (99.0%-100.0%) with H5N6 AIVs isolated from environment in Guangdong during 2017 to 2018. Phylogenetic analysis showed that the viral HA belonged to Clade 2.3.4.4C, and the viral HA, NA, PB1, PA, NP, and MP were all clustered together with H5N6 AIVs isolated from mink in Eastern China in 2018, while the PB2 and NS were clustered together with H5N6 AIVs isolated from environment in Guangdong from 2017 to 2018. The HA cleavage site contained multiple basic amino acid residues, which was highly pathogenic AIVs (HPAIVs). S137A and T160A mutations of HA could increase binding to human-type receptor SAα2, 6-Gal. Additionally, the viral multiple mutations, including 59-69 deletion in NA, the L89V, G309D, R477G, I495V, I504V, D391E, and A661E in PB2, as well as the P42S, D92E, and 80-84 deletion in NS1, could enhance the viral virulence and pathogenicity to mammals. Conclusions:The 6 strains of H5N6 HPAIVs isolated from LPM have relatively close genetic relationship with H5N6 AIVs isolated from mink in Eastern China and environment in Guangdong during 2017 to 2018, the viral multiple mutations could increase its pathogenicity to mammals, which could pose a potential risk to public health.

BACKGROUND: Heat shock proteins (HSP), especially the HSP 70 family, may play certain roles in the immunophysiology of some skin diseases such as psoriasis, pemphigus, and lichen planus. HSPs generally induce down-regulation of the process of apoptosis that is considered to be one of the acantholysis-producing pathways in pemphigus. OBJECTIVE: We planned to examine possible roles of HSPs 70/105 in the blistering process in pemphigus vulgaris (PV) and pemphigus foliaceus (PF), in connection with the detection results of apoptosis in local tissue specimens. METHODS: Immunohistochemical stainings and Western blot analysis were performed for the detection and semiquantitation of HSPs 70/105 in skin specimens from lesional, nonlesional, and normal control sites. Hoechst 33342 staining was simultaneously carried out to examine features of apoptosis in lesional skin specimens. RESULTS: The findings on expression of HSP were as follows. In PV, the expression of HSP 70 was minimum or negative; however, in PF, the expression was obvious and recognizable in lesional and perilesional normal skin. In contrast, HSP 105 was not detected in all cases of PV and PF. The features of apoptosis were evident at the lesional skin of all cases of pemphigus with acantholytic changes. CONCLUSION: PV and PF had different relative intensities of HSPs in lesional tissue stainings, especially in cases with HSP 70. This suggests that there may be subtle differences in the mechanisms causing acantholysis between PV and PF.
Acantholysis ; Apoptosis ; Benzimidazoles ; Blister ; Blotting, Western ; Down-Regulation ; Heat-Shock Proteins ; Hot Temperature ; HSP70 Heat-Shock Proteins ; Humans ; Lichen Planus ; Pemphigus ; Psoriasis ; Skin ; Skin Diseases

Objective To summarize the CT imaging features of peritoneal metastasis of gastric cancer (GC).Methods The retrospective cross-sectional study was conducted.The clinicopathological data of 78 GC patients with peritoneal metastasis who were admitted to the First Affiliated Hospital of Nanjing Medical University from January 2013 to September 2016 were collected.All the patients underwent plain and enhanced scans of CT,and imaging data were converted to the multiplanar reformation (MPR) and maximum intensity projection (MIP).Observation indicators:(1) CT imaging features of primary lesion of GC;(2) CT diagnostic accurary and imaging features of GC with peritoneal metastasis;(3) CT imaging features of lymph nodes and other abdominal organ metastasis;(4) treatment and follow-up.Patients selected treatment plan according to results of laboratory and imaging examinations and patient's will.Follow-up using outpatient examination,telephone interview and correspondence was performed to detect the treatment method and patients' prognosis up to December 2016.The survival time was from post-treatment to death or end of follow-up.Measurement data with skewed distribution were described as M (range).Results (1) CT imaging features of primary lesion of GC:primary tumor location of 78 patients:tumor located in the antrum,leather bottle stomach,gastric cardia,lesser curvature,gastric antrum and body and greater curvature were detected in 40,11,9,9,5 and 4 patients,respectively.All the 78 patients were in T4 stage,including 43 with T4a stage and 35 with T4b stage.(2) CT diagnostic accurary and imaging features of GC with peritoneal metastasis:of 78 patients,57 were found with peritoneal metastasis by CT examinations before chemotherapy,15 with peritoneal metastasis by exploratory laparotomy or open surgery and 6 with peritoneal metastasis by follow-up CT re-examinations after gastrectomy.Seventy-two patients were diagnosed as GC with peritoneal metastasis by CT scans,and final diagnosis,missed diagnosis and overdiagnosis were detected in 78,9 and 3 patients,respectively.The precision,sensitivity,specificity,positive and negative predictive values of CT diagnosis were respectively 98.7％,88.5％,99.6％,95.8％ and 98.9％.Location and manifestation of 78 GC with peritoneal metastasis patients:① Peritoneal effusions:71 patients were accompanied with peritoneal effusions,including 21 with the slight peritoneal effusions and 50 with moderate and massive peritoneal effusions.② Greater omentum thickening:greater omnentum of 59 patients showed sheet,flocculent and nodular thickening,with a mass and cake-like change.③ Peritoneal thickening:54 patients had peritoneal thickening,with the main of nodulelike and thick lines-like thickening;thickening occurred mainly in around the liver,peritoneal cavity and pelvic floor;11 patients were found with obvious left anterior renal fascia thickening.④ Mesenteric thickening:46 patients were found with mesenteric thickening and edema,showing increased fat density with multi-node shadow.⑤ Lesser omentum and hepatogastric ligament:18 patients were found with increased fat density of lesser omental bursa,showing striped and nodular shadow.⑥ Transverse mesocolon:increased local fat density with striped-and nodular-like changes were seen in 15 patients after coronal and sagittal planes reconstruction,including 5 with local wall thickening of transverse colon.⑦ Ovary:8 patients had ovarian metastases,including 6 with bilateral metastases and 2 with unilateral metastasis;diameter of metastatic tumor was 3-12 cm.⑧ Intestinal canal:6 patients had local intestinal wall thickening,including 3 in small intestine and 3 in transverse colon,thickening tissues were mainly located in the mesentery,showing obvious intestinal wall enhancement.⑨ Liver capsule node:2 patients showed multi-node abnormal enhanced lesions under liver capsule.(3) CT imaging features of lymph nodes and other abdominal organ metastasis:78 patients were accompanied with lymph nodes enlargement,including 41 in N2 stage and 37 in N3 stage;liver metastases were detected in 13 patients;5 had adrenal metastases,including 3 with bilateral metastases and 2 with unilateral metastasis;4 had lower GC invading the pancreatic head and body;2 had upper GC invading the liver and spleen;1 had leather bottle stomach invading the pancreatic head and neck,and inducing to obstruction of biliary tract.(4) Treatment and follow-up:of 78 patients,62 underwent systemic chemotherapy,6 underwent systemic chemotherapy and intraperitoneal hyperthermic perfusion chemotherapy,5 underwent systemic chemotherapy and local radiotherapy and 5 underwent palliative operations due to gastrointestinal tract obstruction or bleeding.Of 78 patients,69 were followed up for 15 months (range,3-21 months),and 9 lost to follow-up.The median survival time of 69 follow-up patients was 12 months (range,3-19 months).Conclusions CT imaging features of peritoneal metastasis of GC show specific sites of metastasis and performance.Combining with CT axial images and images of coronal and sagittal planes reconstruction,adjusting appropriate window width and level would be benefit to observe primary lesions of GC and peritoneal metastasis.

Exosomes are vesicles with a double globular membrane of lipids that can be secreted by a variety of cells, including stem cells. Exosomes have unique biological characteristics and irreplaceable powerful functions which play an important role in intercellular communication. The various cytokines, signal proteins, lipids and regulatory nucleic acids contained in stem cell exosomes can play a protective role against the injury of kidney, liver, heart, blood vessels and nerves. Stem cell exosomes delay the process of intervertebral disc degeneration by inhibiting the apoptosis of nucleus pulposus cells and increasing the synthesis of extracellular matrix, etc. The mechanism of its role is mainly through miRNA and related signaling pathways. Exosomes contain complex components. Although the mechanism of action of exosomes in intervertebral discs has been preliminarily explored, the components contained in exosomes are complex and the specific situation has not been fully understood, which still needs further study. In this review, the characteristics and functions of stem cell exosomes, extraction, identification and storage methods, the impacttovarious other tissues, as well as the effects on intervertebral discs and their mechanisms were elaborated in order to provide a basis for the study of intervertebral disc degenerative diseases.

Objective To compare the CT findings of poor-differentiated gastric neuroendocrine carcinomas (PD-NECs) and poordifferentiated gastric adenocarcinomas (PD-ADCs),and to increase the diagnostic accuracy rate.Methods In this retrospective study that enrolled 29 gastric PD-NECs and 29 gastric PD-ADCs,whose diagnoses were proven pathologically,we assessed their clinical characteristics and CT findings and then used univariate and multivariate analyses for statistical comparisons.Results The univariate analysis revealed that gastric PD-NECs were significantly smaller (P =0.002),more often showed well-defined margins (P＜0.001),and more frequently accompanied with intact overlying mucosa on CT when compared with gastric PD-ADCs (P ＜0.001).In the multivariate analysis,the presence of intact overlying mucosa (OR=0.028,95 ％ CI:0.001-0.863,P =0.041) and tumor-to-mucosa HU ratio on the arterial phase (OR=0.015,95 ％ CI:0.000-0.495,P =0.019) were two independent factors affecting the identification of gastric PD-NECs and gastric PD-ADCs.Receiver operating characteristic analysis (ROC) showed that the value of tumor-to-mucosa H U ratio on the arterial phase in differentiating them was moderate (AUC=0.72).Conclusion Gastric PD-NECs have better-defined margin,more intact overlying mucosa and higher tumor-to-mucosa HU ratio on the arterial phase than gastric PD-ADCs.

Objective:To evaluate the distribution characteristics of choroidal vascularity index (CVI) in macula among normal children using swept-source optical coherence tomography (SS-OCT), and to investigate the influencing factors.Methods:A cross-sectional study was conducted.Sixty-three children aged 6 to 12 years were enrolled in The First Affiliated Hospital of Zhengzhou University from May 2021 to November 2021.Spherical equivalent refraction, axial length (AL) and other ocular biological parameters were measured.Macula-centered CVI and choroidal thickness (ChT) were measured by SS-OCT angiography.According to the ETDRS partition, the obtained image was divided into macular central fovea (0-1 mm diameter), inner ring (1-3 mm diameter) and outer ring (3-6 mm diameter) zones.Data from the right eye were selected for statistical analysis.CVI in the three rings and four zones (superior, temporal, inferior and nasal zones) of the inner and outer rings were compared.Univariate and multiple linear regression analyses were used to analyze the correlation between CVI and sex, AL, anterior chamber depth (ACD), ChT and pupil diameter.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of The First Affiliated Hospital of Zhengzhou University (No.2021-KY-0399-003). Written informed consent was obtained from each guardian.Results:The average CVI in the macular central fovea, inner ring and outer ring were 0.35±0.12, 0.32±0.10 and 0.27±0.08, respectively, with a significant difference ( F=10.96, P<0.001), and significant differences in CVI were found in pairwise comparisons of the three ring zones (all at P<0.05). Significant differences in CVI were found among superior, temporal, inferior and nasal zones in inner and outer rings ( Fring=11.60, P=0.001; Fzone=12.02, P<0.05). The CVI was smaller in superior and nasal zones in inner ring than in temporal zone, greater in superior, temporal and inferior zones in outer ring than in nasal zone, smaller in superior zone of outer ring than that of the inner ring, and the differences were statistically significant (all at P<0.001). The single factor linear regression analysis showed that ChT in the fovea, inner ring and outer ring were the influencing factors of CVI in the three ring zones (all at β=0.001, P<0.001). Pupil diameter ( β=0.034, P=0.038; β=0.040, P=0.003; β=0.024, P=0.011) and ACD ( β=0.097, P=0.034; β=0.097, P=0.013; β=0.061, P=0.032) were the influencing factors of CVI in fovea, inner ring and outer ring.After multiple linear regression analysis, the regression equations were established as follows: CVI in the macular fovea=0.001×ChT in the macular fovea + 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring-0.301 ( R2=0.514, F=6.875, P<0.001); CVI in the inner ring=0.001×ChT in the macular fovea+ 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring+ 0.088×AL-0.307 ( R2=0.603, F=9.870, P<0.001); CVI in the outer ring=0.001×ChT in the macular fovea+ 0.001×ChT in the inner ring+ 0.001×ChT in the outer ring-0.135 ( R2=0.601, F=9.781, P<0.001). Conclusions:In children aged 6-12 years old, the CVI is higher in the macular central fovea than in inner and outer rings, and the CVI in nasal zone is the smallest in both inner and outer rings.The thicker the ChT, the higher the CVI in all zones in the macular area; the deeper the anterior chamber, the higher the CVI in the inner ring.

Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016–2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.

Objective: To conduct a follow-up investigation of structural changes of the corpus callosum fibers of toddlers (2 to 5 years of age) with autism spectrum disorder(ASD) and to explore the associations with clinical symptoms. Method: In this prospective randomized controlled study, ASD children who were diagnosed in the Child Mental Health Research Center, Nanjing Brain Hospital Affiliated to Nanjing Medical University from May 2011 to November 2012 were included in the ASD group, and developmentally delayed children were included in the control group (DD group). Diffusion tensor imaging (DTI) data from the two groups were obtained at two age levels: 2-3 years of age, and 4-5 years of age. Region of interest analysis was applied to assess characteristic values of total area and sub-regions of corpus callosum: the fraction anisotropy (FA), the mean diffusivity (MD), the radial diffusivity (RD) and the axial diffusivity (AD). All children were assessed using the Autism Diagnostic Interview-Revised (ADI-R) and Autism Treatment Evaluation Checklist (ATEC). The characteristic values of total area and sub-regions of corpus callosum of ASD group at two age levels were analyzed by paired sample t test; the characteristic values of total area and sub-regions of corpus callosum of ASD group and DD group were analyzed by independent-sample t test; the correlations between FA values of the total area and sub-regions of corpus callosum and ADI-R or ATEC scores were analyzed by Pearson correlation analysis. Result: Forty cases meeting inclusion criteria were enrolled in ASD group, and 31 eligible cases were enrolled in the control group. Four children in the ASD group were lost to follow-up, and 5 children in the control group were lost to follow-up. Longitudinal comparison between the two age subgroups of ASD patients showed that the FA values of the total corpus callosum increased (0.499 55±0.027 59 vs. 0.505 83±0.086 64, t=4.88, P<0.05), but MD values, RD values and AD values of the total corpus callosum area decreased (0.000 89±0.000 03 vs. 0.000 81±0.000 14, 0.000 61±0.000 04 vs. 0.000 55±0.000 09, 0.001 43±0.000 03 vs. 0.001 38±0.000 03, t=9.31, 7.90, 8.66, P<0.05 for all comparisons). In the area of corpus callosum genu, FA and AD values increased (t=5.59, 8.48, P<0.05 for both comparisons), but MD and RD values decreased (t=12.67, 11.28, P<0.05 for both comparisns). In the area of corpus callosum body, FA and RD values increased(t=5.46, 8.48, P<0.05 for both comparisons), but MD and AD values decreased (t=8.08, 6.22, P<0.05 for both comparisons). In the area of corpus callosum splenium, MD, RD and AD values decreased (t=6.81, 4.44, 5.51, P < 0.05 for all comparisons). Among the participants 2 to 3 years of age, there were no significantly differences in FA values of total area and sub-regions of corpus callosum between ASD group and the DD group (P > 0.05 for all comparisons); as compared with the DD group, ASD group had higher AD values of total area and splenium of corpus callosum (0.001 43±0.000 03 vs. 0.001 40±0.000 04, 0.001 34±0.000 03 vs. 0.001 32±0.000 04, t=1.56, 1.14, P < 0.05 for both comparisons); ASD group had lower AD values but higher RD and MD values of corpus callosum genu (t=0.07, 0.55, 0.07, P < 0.05 for all comparisons); ASD group had lower RD values of corpus callosum body (t=0.07, P < 0.05). Among the participants 4 to 5 years of age, as compared with the DD group, ASD group had higher FA value of total corpus callosum area(0.505 83±0.086 64 vs. 0.483 77±0.099 30, t=8.56, P < 0.05), lower RD value of total corpus callosum(0.000 55±0.000 09 vs. 0.000 56±0.000 12, t=14.44, P < 0.05), lower RD values of corpus callosum body (t=2.20, P < 0.05), higher FA values (t=3.35, P < 0.05) but lower AD values of corpus callosum splenium (t=2.20, P < 0.05). A correlation analysis between FA values of total area and sub-regions of corpus callosum and clinical variables showed that the FA values of total area and splenium of corpus callosum in ASD group at 2 to 3 years of age were negatively correlated with the scores of language skills in ATEC (r=-0.35,-0.36, P < 0.05 for both comparisons). And after two years, FA values of total corpus callosum were positively correlated with the scores of social communication in ATEC (r=0.34, P < 0.05). There was no significant correlation between FA values of sub-regions of corpus callosum and the scores of ATEC (P > 0.05 for all comparisons). There was no significant correlation between FA values of total area and sub-regions of corpus callosum and the scores of ADI-R (P > 0.05 for all comparisons). Conclusion The fiber structure of corpus callosum was still in the process of maturing during the age of 2 to 5 years; however, compared with DD group, ASD group had more extensive structural abnormalities of the corpus callosum fibers as age increased, and the structural abnormalities had correlation with the core symptoms of ASD. Trial registration Chinese Clinical Trial Registry, ChiCTR-OPC-17011995.

Background: The H5 avian influenza viruses (AIVs) of clade 2.3.4.4 circulate in wild and domestic birds worldwide. In 2017, nine strains of H5N6 AIVs were isolated from aquatic poultry in Xinjiang, Northwest China. Objectives: This study aimed to analyze the origin, reassortment, and mutations of the AIV isolates. Methods: AIVs were isolated from oropharyngeal and cloacal swabs of poultry. Identification was accomplished by inoculating isolates into embryonated chicken eggs and performing hemagglutination tests and reverse transcription polymerase chain reaction (RT-PCR). The viral genomes were amplified with RT-PCR and then sequenced. The sequence alignment, phylogenetic, and molecular characteristic analyses were performed by using bioinformatic software. Results: Nine isolates originated from the same ancestor. The viral HA gene belonged to clade 2.3.4.4B, while the NA gene had a close phylogenetic relationship with the 2.3.4.4C H5N6 highly pathogenic avian influenza viruses (HPAIVs) isolated from shoveler ducks in Ningxia in 2015. The NP gene was grouped into an independent subcluster within the 2.3.4.4B H5N8 AIVs, and the remaining six genes all had close phylogenetic relationships with the 2.3.4.4B H5N8 HPAIVs isolated from the wild birds in China, Egypt, Uganda, Cameroon, and India in 2016–2017, Multiple basic amino acid residues associated with HPAIVs were located adjacent to the cleavage site of the HA protein. The nine isolates comprised reassortant 2.3.4.4B HPAIVs originating from 2.3.4.4B H5N8 and 2.3.4.4C H5N6 viruses in wild birds. Conclusions These results suggest that the Northern Tianshan Mountain wetlands in Xinjiang may have a key role in AIVs disseminating from Central China to the Eurasian continent and East African.