Objective:To accurately ascertain the whole genome sequencing and the composition of open reading frames (ORFs) of non-replicating Tiantan strain of vaccinia virus (NTV) using next-generation long-read sequencing technology.Methods:NTV, obtained from our laboratory stock, was amplified and purified on chicken embryo fibroblast cells(CEFs), and the full-length genomic nucleic acid of NTV was extracted. The PacBio HiFi sequencing platform was utilized for de novo assembly to obtain the complete genomic sequence of NTV. Using a homology annotation strategy, we identified its ORF composition and compared it with known non-replicating vaccinia virus strains. Results:The total length of NTV′s genome was 171 729 bp, with a GC content of 33%. Its unique inverted terminal repeat (ITR) region comprised hairpin structures, two tandem repeat regions, and three non-repeat regions. NTV contained 166 ORFs, with major differences observed in the ITR and its surrounding regions when compared to MVA-BN and NYVAC. These three strains shared a common set of 138 ORFs. NTV encoded six unique ORFs related to virus evasion of host antiviral response.Conclusions:This study accurately determines the whole genome sequencing and ORFs composition of NTV, and reveals its similarities and differences with other replication-deficient vaccinia virus strains, which pave a way for the development and application of the next generation of monkeypox vaccines and novel viral vectors.

Objective:To screen risk factors for carotid plaque vulnerability in patients with ischemic stroke and construct a risk prediction model.Methods:From November 2019 to January 2021, a total of 164 patients with ischemic stroke hospitalized in the Neurology Department of the Affiliated Hospital of Jining Medical University were selected as the study subjects by convenience sampling method. Color doppler ultrasound was used to measure carotid plaques in patients with ischemic stroke to determine whether they were vulnerable plaques. The patients were surveyed using the General Information Questionnaire and the Type D Personality Scale. Binary Logistic regression analysis was used to explore the risk factors of carotid plaque vulnerability in ischemic stroke patients, and based on this, a risk prediction model for carotid plaque vulnerability in ischemic stroke patients was constructed.Results:A total of 87 patients with ischemic stroke had vulnerable carotid plaques. The results of binary Logistic regression analysis showed that age ( OR=1.136, 95% CI: 1.052-1.226), total score of Type D Personality Scale ( OR=1.170, 95% CI: 1.043-1.312), smoking history ( OR=3.058, 95% CI: 1.054-8.875), homocysteine ( OR=1.400, 95% CI: 1.179-1.664), triglycerides ( OR=2.356, 95% CI: 1.534-3.619) were risk factors for carotid plaque vulnerability in stroke patients ( P<0.05). The results of risk prediction model based on risk factors show that, the area under the receiver operating characteristic of the subjects was 0.935, indicating good clinical predictive ability. Conclusions:Age, total score of Type D Personality Scale, smoking history, homocysteine and triglyceride are risk factors of carotid plaque vulnerability in stroke patients. The risk prediction model can early identify and screen high-risk factors for carotid plaque vulnerability in ischemic stroke patients, and is worthy of clinical promotion and practice.

This study aims to explore the molecular mechanism of Ganoderma against gastric cancer based on network pharmacology, molecular docking, and cell experiment. The active components and targets of Ganoderma were retrieved from Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP), and gastric cancer-related targets from GeneCards and Online Mendelian Inheritance in Man(OMIM). The protein-protein interaction(PPI) network of the common targets was constructed with STRING, followed by Gene Ontology(GO) term enrichment and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis of the common genes based on Bioconductor and R language. The medicinal-disease-component-target network and medicinal-disease-component-target-pathway network were established by Cytoscape. Molecular docking was performed between β-sitosterol(the key component in Ganoderma) and the top 15 targets in the PPI network. Cell experiment was performed to verify the findings. A total of 14 active components and 28 targets of Ganoderma were retrieved, and the medicinal and the disease shared 25 targets, including caspase-3(CASP3), caspase-8(CASP8), caspase-9(CASP9), and B-cell lymphoma-2(BCL2). The common targets involved 72 signaling pathways and apoptosis and p53 signaling pathway may play a crucial role in the effect of Ganoderma against gastric cancer. β-sitosterol had strong binding activity to the top 15 targets in the PPI network. The in vitro cell experiment demonstrated that β-sitosterol inhibited gastric cancer AGS cell proliferation by inducing cell apoptosis and cell cycle arrest in the S phase, which might be related to the regulation of the p53 pathway. This study shows the multi-component, multi-target, and multi-pathway characteristics of Ganoderma against gastric cancer, which lays a scientific basis for further research on the molecular mechanism.
Ganoderma ; Humans ; Medicine, Chinese Traditional ; Molecular Docking Simulation ; Network Pharmacology ; Stomach Neoplasms/genetics*

Objective:To construct a bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 and to evaluate its immunogenicity in mice.Methods:The coding sequences for spike 1 (S1) protein of SARS-CoV-2 Beta variant and hemagglutinin (HA) of influenza A virus Cambodia (H3N2) strain were codon-optimized and synthesized. The two coding genes were ligated by the self-cleaving 2A peptide using over-lapping PCR to construct S1-2A-HA fragment, which was inserted into pVRC vector to construct the bivalent DNA vaccine, named as pVRC-S1-2A-HA. Indirect immunofluorescence assay (IFA) and Western blot were performed to detect the expression of S1 and HA proteins. BALB/c mice were immunized with pVRC-S1-2A-HA by intramuscular injection and electroporation. The humoral immune responses induced in mice were detected by indirect ELISA, pseudovirus neutralization assay and hemagglutination inhibition assay. Cellular immune responses were detected by IFN-γ ELISPOT, intracellular cytokine staining (ICS) and cytometric bead array (CBA).Results:The bivalent DNA vaccine pVRC-S1-2A-HA could express S1 and HA proteins in vitro. Specific cellular immune responses against S1 protein and specific IgG antibody against HA protein were significantly induced in mice with single-dose immunization. The antigen-specific immunity was significantly enhanced after booster immunization. The geometric mean titer (GMT) of specific IgG antibody increased to 3 251 for S1 protein and 45 407 for HA protein after two-dose immunization. Moreover, the S1-specific T cells increased to 1 238 SFC/10 6 cells. ICS results indicated that the booster vaccination induced CD4 + T and CD8 + T cells to produce IL-2, IFN-γ and TNF-α in mice. The secretion of various cytokines including IL-2, IL- 4, IL-6, IL-10 and IFN-γ in mouse splenocytes was induced after single-dose immunization. Conclusions:A bivalent DNA vaccine against SARS-CoV-2 and influenza A virus H3N2 was constructed and could induce S1- and HA-specific humoral and cellular immune responses in mice, suggesting the great potential of it for further development and application.

Objective:To compare the clinical efficacy of endoscopic resection and laparoscopic surgery in the treatment of gastric gastrointestinal stromal tumor (GIST) with a maximum diameter of 2 to 5 cm, and to analyze the influence of factors such as tumor surface, growth pattern and lesion origin on the choice of resection method, so as to provide a safer and more effective treatment for patients with gastric GIST.Methods:From January 2012 to November 2019, at the First Affiliated Hospital of Zhengzhou University, the clinical data of 301 patients with gastric GIST who underwent endoscopic resection (137 cases in the endoscopic resection group) or laparoscopic surgery (164 cases in the laparoscopic surgery group) were retrospectively analyzed, including age, gender, whether there was depression on the tumor surface (the local subsidence depth of the mucosa on the tumor surface was >5 mm), whether the tumor surface was irregular (non-hemispherical or non-elliptical tumor surface), whether there was combined ulcer, location, shape, origin of the lesion, growth pattern (intralumina growth or combined intraluminal and extraluminal growth), risk classification (very low risk, low risk, medium risk, high risk), whether the tumor was en bloc resection, operation time, whether bleeding or not, fasting time, indwelling time of gastric tube, time of hospitalization, time of postoperative hospital stay, postoperative complications and follow-up. Independent sample t test, chi-square test or Fisher′s exact test and Wilcoxon rank sum test were used for statistical analysis. Results:Among the 137 patients with gastric GIST in the endoscopic resection group, 85 cases (62.0%) underwent endoscopic submucosal dissection, 9 cases (6.6%) underwent endoscopic submucosal excavation, 42 cases (30.7%) underwent endoscopic full-thickness resection, and 1 case (0.7%) underwent submucosal tunnel endoscopic resection. There were no significant differences in gender, age, lesion location, tumor size, and risk classification between the endoscopic resection group and the laparoscopic surgery group (all P>0.05). The tumor surface was depressed, with ulcer or irregular in 1, 49, 26, and 2 cases of patients with gastric GIST of very low risk, low risk, medium risk and high risk, respectively. There was statistically significant difference in the proportion of depression, irregularity and ulcer on the tumor surface at different risk levels ( Z=-2.55, P=0.011). The complete tumor resection rate of the endoscopic resection group was lower than that of the laparoscopic surgery group (86.1%, 118/137 vs. 100.0%, 164/164), and the difference was statistically significant ( χ2=24.28, P<0.001). However the operation time, fasting time, the indwelling time of gastric tube, time of hospitalization, and the time of postoperative hospital stay of the endoscopic resection group were shorter than those of the laparoscopic surgery group, and the total hospitalization cost was lower than that of the laparoscopic surgery group (90.0 min (62.5 min, 150.0 min) vs. 119.5 min, (80.0 min, 154.2 min); 3 d (3 d, 4 d) vs. 5 d (4 d, 7 d); 3 d (2 d, 4 d) vs. 4 d (2 d, 6 d); 11 d (10 d, 14 d) vs. 16 d (12 d, 20 d); 7 d (6 d, 9 d) vs. 9 d (7 d, 11 d); (38 211.6±10 221.0) yuan vs. (59 926.1±17 786.1) yuan), and the differences were statistically significant ( Z=-2.46, -7.12, -4.44, -6.89 and -5.92, t=-13.24; all P<0.05). The incidence of postoperative abdominal pain and other severe postoperative complications (including shock, respiratory failure, pulmonary embolism, gastroparesis, etc.) of the endoscopic resection group were all lower than those of the laparoscopic surgery group (16.8%, 23/137 vs. 27.4%, 45/164; 0.7%, 1/137 vs. 4.9%, 8/164), and the differences were statistically significant ( χ2=4.84, Fisher′s exact test, P=0.028 and 0.043). There were no significant differences in the incidence of intraoperative bleeding, postoperative bleeding, fever and perforation between the two groups (all P>0.05). The incidence of operation-related complications of lesions with intraluminal growth and originating from muscularis propria in the endoscopic resection group were lower than those of the laparoscopic surgery group (19.5%, 25/128 vs. 32.6%, 45/138; 12.6%, 12/95 vs. 31.4%, 37/118), and the differences were statistically significant ( χ2=5.86 and 10.42, P=0.016 and 0.001). There was no significant difference in the postoperative tumor recurrent rate between the endoscopic resection group and the laparoscopic surgery group (0, 0/137 vs. 2.4%, 4/164; Fisher’s exact test, P=0.129). Conclusions:Endoscopic treatment is safe and effective for gastric GIST with a maximum diameter of 2 to 5 cm, which is superior to laparoscopic surgery. However, laparoscopic surgery is recommended for tumor with depressed, ulcerative, or irregular surface and combined intraluminal and extraluminal growth.

Background/Aims: Nonalcoholic fatty liver disease (NAFLD) is closely associated with diabetes. The cumulative impact of both diseases synergistically increases risk of adverse events. However, present population analysis is predominantly conducted with reference to non-NAFLD individuals and has not yet examined the impact of prediabetes. Hence, we sought to conduct a retrospective analysis on the impact of diabetic status in NAFLD patients, referencing non-diabetic NAFLD individuals. Methods: Data from the National Health and Nutrition Examination Survey 1999–2018 was used. Hepatic steatosis was defined with United States Fatty Liver Index (US-FLI) and FLI at a cut-off of 30 and 60 respectively, in absence of substantial alcohol use. A multivariate generalized linear model was used for risk ratios of binary outcomes while survival analysis was conducted with Cox regression and Fine Gray model for competing risk. Results: Of 32,234 patients, 28.92% were identified to have NAFLD. 36.04%, 38.32% and 25.63% were non-diabetic, prediabetic and diabetic respectively. Diabetic NAFLD significantly increased risk of cardiovascular disease (CVD), stroke, chronic kidney disease, all-cause and CVD mortality compared to non-diabetic NAFLD. However, prediabetic NAFLD only significantly increased the risk of CVD and did not result in a higher risk of mortality. Conclusions Given the increased risk of adverse outcomes, this study highlights the importance of regular diabetes screening in NAFLD and adoption of prompt lifestyle modifications to reduce disease progression. Facing high cardiovascular burden, prediabetic and diabetic NAFLD individuals can benefit from early cardiovascular referrals to reduce risk of CVD events and mortality.

Objective To investigate the change patterns of functional brain networks and their relations with cognitive function in patients with end-stage renal disease (ESRD).Methods Sixty-two patients with ESRD (ESRD group),admitted to our hospital from July 2018 to June 2019,and 36 age-,gender-,and education level-matched healthy controls (HC group) were enrolled.Mini-mental State Examination (MMSE),Montreal Cognitive Assessment (MoCA),Trail Making Test A (TMT-A),TMT-B and Symbol Digit Modalities Test (SDMT) were used to evaluate the cognitive function for all subjects.Resting-state functional magnetic resonance imaging data were acquired;after data preprocessing,the brain functional networks were constructed and the topological parameters were calculated.Statistical methods were used to compare the differences of cognitive function scores and topological parameters between the two groups,and to analyze the correlations between these topological parameters and cognitive function scores in the ESRD group.Results The MMSE,MoCA and SDMT scores of the ESRD group were significantly lower than those of the HC group (P＜0.05),and the ESRD group took significantly longer time to complete TMT-A and TMT-B than the HC group (P＜0.05).The ESRD group had significantly lower normalized clustering coefficient (γ),small-worldness (σ) and local efficiency (Elocal) values than the HC group (P＜0.05).Patients in the ESRD group exhibited significantly decreased nodal efficiency in the paralimbic-limbic network (including the bilateral insula,median cingulate and paracingulate gyri,hippocampus,parahippocampal gyrus,amygdala,temporal pole:superior temporal gyrus,and temporal pole:middle temporal gyrus),right heschl gyrus and left superior temporal gyrus,and exhibited significantly increased nodal efficiency in the visual network (including the right distal-shaped gyrus,bilateral wedge,and left superior and middle occipital gyrus) as compared with the HC group (P＜0.05).In the ESRD group,the area under the curve (AUC) ofγ and σ was positively correlated with MoCA scores (r=0.698,P=0.000;r=0.661,P=0.000),and the AUC of Elocal showed positive correlation with MMSE scores (r=0.407,P=0.003).Conclusion Abnormal topological organization of the functional brain networks is revealed in patients with ESRD,which affects the cognitive function of these patients.

Objective To explore the association between frailty and muscle performances of hospitalized elder adults with coronary artery disease.Methods A total of 122 hospitalized patients aged 65-85 years old with coronary artery disease from Department of Geriatrics and Cardiology,Peking Union Medical College Hospital between December 2017 and March 2018 were enrolled in the study.A comprehensive geriatric assessment was performed to evaluate existing comorbidity and geriatric syndromes of the patients.Frailty was assessed using the Clinical Fraity Scale.The patients were classified as frail and non-frail,according to the scale.Muscle performances were assessed using grip strength,gait speed,etc.Whole body and appendicular skeletal muscle mass was detected with bioelectrical impedance analysis in patients with reduced grip strength or slowed gait speed.Appendicular skeletal muscle index (ASMI) was calculated.Results Among all subjects,28 were with frailty (23.0％) and 94 were without (77.0％).The frail patients were older [(76.7±5.4) years vs.(72.2±5.6)years],had higher Charlson comorbidity index [2.0(1.0,2.75)vs.1.0(0,2.0)],and higher proportion of malnutrition (14.29％ vs.1.06％),urinary incontinence (39.29％ vs.15.96％),using walking-aid (28.57％ vs.6.38％),and more kinds of taken drugs (8.1±3.0 vs.6.6±2.7),than the non-frail patients.Prealbumin levels [(207.8±60.0)mg/L vs.(234.3±45.4)mg/L] were lower,and highly sensitive C-reactive protein levels [(5.89±9.57)mg/L vs.(1.89±2.49)mg/L] were higher in the frail patients than in the non-frail patients (all P＜0.05).Compared with non-frail patients,the frail patients had poorer grip strength [(19.67±7)kg vs.(29.23±8.29)kg] and slower gait speed [(0.54±0.2)m/s vs.(0.91±0.22)m/s](all P＜0.001).Spearman rank correlation analyses showed that grip strength was positively correlated with the appendicular skeletal muscle mass(r =0.811),whole body skeletal muscle mass(r =0.74) and the ASMI (r =0.783),respectively.Conclusions The incidence of frailty among hospitalized older adults with coronary artery disease is high.Poor muscle performances were common in these patients.Assessment of frailty and muscle performances can help to evaluate the overall function of older adults with cardiovascular disease in a comprehensive way.

Leukocyte differentiation antigens (LDAs) play important roles in the immune system, by serving as surface markers and participating in multiple biological activities, such as recognizing pathogens, mediating membrane signals, interacting with other cells or systems, and regulating cell differentiation and activation. Data mining is a powerful tool used to identify novel LDAs from whole genome. LRRC25 (leucine rich repeat-containing 25) was predicted to have a role in the function of myeloid cells by a large-scale "omics" data analysis. Further experimental validation showed that LRRC25 is highly expressed in primary myeloid cells, such as granulocytes and monocytes, and lowly/intermediately expressed in B cells, but not in T cells and almost all NK cells. It was down-regulated in multiple acute myeloid leukemia (AML) cell lines and bone marrow cells of AML patients and up-regulated after all-trans retinoic acid (ATRA)-mediated granulocytic differentiation in AML cell lines and acute promyelocytic leukemia (APL; AML-M3, FAB classification) cells. Localization analysis showed that LRRC25 is a type I transmembrane molecule. Although ectopic LRRC25 did not promote spontaneous differentiation of NB4 cells, knockdown of LRRC25 by siRNA or shRNA and knockout of LRRC25 by the CRISPR-Cas9 system attenuated ATRA-induced terminal granulocytic differentiation, and restoration of LRRC25 in knockout cells could rescue ATRA-induced granulocytic differentiation. Therefore, LRRC25, a potential leukocyte differentiation antigen, is a key regulator of ATRA-induced granulocytic differentiation.
Antigens, Differentiation ; immunology ; metabolism ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Granulocytes ; cytology ; drug effects ; immunology ; metabolism ; Humans ; Leukocytes ; cytology ; drug effects ; immunology ; metabolism ; Membrane Proteins ; antagonists & inhibitors ; immunology ; metabolism ; RNA, Small Interfering ; pharmacology ; Tretinoin ; pharmacology

Background: CKLF-like MARVEL transmembrane domain containing (CMTM) superfamily is involved in the occurrence and development of inflammation, cancer and a variety of diseases.Human CMTM3 has been proposed as a putative tumor suppressor gene.Aims: To investigate the expression of CMTM3 in Helicobacter pylori (Hp) infection-related chronic gastritis and its significance.Methods: Sixty cases of outpatients with chronic gastritis (30 Hp-positive and 30 Hp-negative) were enrolled for detection of CMTM3 and interleukin-6 (IL-6) expressions in gastric mucosa by immunohistochemistry.The correlation of expressions of CMTM3 and IL-6 was analyzed.Results: The positivity rates of CMTM3 and IL-6 in gastric mucosa were significantly higher in Hp-positive chronic gastritis than in Hp-negative ones (CMTM3: 63.3% vs.30.0%, P<0.05;IL-6: 73.3% vs.13.3%, P<0.01).In patients with Hp-positive chronic gastritis, CMTM3 and IL-6 were co-expressed in 53.3% (16/30) of the patients and localized in the same position.Expression of CMTM3 was positively correlated with IL-6 expression in Hp-positive chronic gastritis patients (r=0.58, P<0.05).Conclusions: CMTM3 is highly expressed in chronic gastritis patients with Hp infection.It may participate in the occurrence and development of Hp infection-related chronic gastritis with inflammatory cytokines such as IL-6.Hp infection might be one of the mechanisms involved in CMTM3 up-regulation.