Epitopes are the basis of antigenicity and the smallest functional unit to induce immune response.Identification of B-cell epitopes is of great significance for the development of new vaccines and therapeutic drugs as well as diagnostic reagents.In this paper,the methods applied in the study of B-cell epitopes mapping in recent years are reviewed and their advantages and shortages are analyzed.

This study aims to construct C57/B6-L and A549 cell lines that stably overexpress circu-lar RNA LAMP3(circLAMP3),laying the foundation for further research on the biological func-tions of circLAMP3.Total RNA was extracted and reverse transcripted into cDNA from C57/B6-L and A549 cells to amplify the full-length sequence of circLAMP3.Then,the fragments of cir-cLAMP3 were ligated into pLC5-ciR vector to obtain pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 recombinant plasmids.The lentiviruses expressing circLAMP3 were packaged on tran-sient transfected HEK293T cells.C57/B6-L and A549 cells were infected with lentiviruses to gen-erate cell lines overexpressing circLAMP3 through puromycin screening.To verify the overexpres-sion efficiency of circLAMP3 of cell lines,we performed the fluorescence microscopy,PCR amplifi-cation,quantitative PCR(qPCR),and Sanger sequencing experiments.The results indicated that the overexpression plasmids of pLC5-Mouse-circLAMP3 and pLC5-Human-circLAMP3 were suc-cessfully constructed.Strong green fluorescence was observed under a fluorescence microscopy.C57/B6-L and A549 cell lines showed a significant increase in the expression of circLAMP3 by PCR and qPCR methods.Sanger sequencing results showed that the junction site of circLAMP3 was correct.This study successfully constructed C57/B6-L and A549 cell lines overexpressing circLAMP3,providing biomaterials for further exploration of the biological function of circLAMP3 in influenza virus replication.

Objective:To analyze the impact of post-dialysis blood pressure (Post-BP) on the long-term survival prognosis of maintenance hemodialysis (MHD) patients and the related risk factors.Methods:It was a retrospective cohort study. The data of patients who underwent their first hemodialysis (HD) from January 1, 2007, to June 30, 2021, as recorded in the dialysis registration system of the Kidney Disease Center, the First Affiliated Hospital, Zhejiang University School of Medicine was retrospectively analyzed. The mean Post-BP was calculated for each HD session 4-6 months after hemodialysis. According to the mean value of post-dialysis diastolic pressure (Post-DBP) at 4-6 months after dialysis, patients were divided into 3 groups (Post-DBP<80 mmHg, 80 mmHg≤Post-DBP<90 mmHg, Post-DBP≥90 mmHg). According to whether the mean value of post-dialysis systolic pressure (Post-SBP) was ≥140 mmHg and whether the mean value of Post-DBP was ≥80 mmHg, patients were divided into 4 groups (Post-SBP<140 mmHg, Post-DBP≥80 mmHg; Post-SBP≥140 mmHg, Post-DBP≥80 mmHg; Post-SBP<140 mmHg, Post-DBP<80 mmHg; Post-SBP≥140 mmHg, Post-DBP<80 mmHg). Patients' first dialysis time was used as the starting point of follow-up, and the end point of follow-up was death or conversion to peritoneal dialysis or kidney transplantation or up to December 31, 2021. Kaplan-Meier survival analysis, Log-rank test, and multivariate Cox regression model were used to analyze the relationship between Post-BP and survival rate and the related factors of prognosis in MHD patients.Results:According to inclusion criteria, a total of 1 213 patients were included. Kaplan-Meier survival curve showed that the long-term survival rate had statistically significant differences among Post-DBP<80 mmHg, 80 mmHg≤Post-DBP<90 mmHg and Post-DBP≥90 mmHg groups (Log-rank test, χ2=58.838, P<0.001), and Post-DBP<80 mmHg group was the lowest. Further comparing the cardiovascular diseases (CVD) mortality among the three groups, the curve showed a statistically significant difference (Log-rank test, χ2=27.926, P< 0.001), and the highest CVD mortality was found in the Post-DBP<80 mmHg group. Multivariate Cox regression model analysis showed that Post-DBP<80 mmHg was an independent associated factor for death in MHD patients (with Post-DBP mmHg≥90 group as reference, HR=4.197, 95% CI 1.452-12.197, P=0.008). When patients were divided into 4 groups according to whether the mean value of Post-SBP was ≥140 mmHg and whether the mean value of Post-DBP was ≥80 mmHg, Kaplan-Meier survival analysis showed a statistically significant difference in long-term survival rate among the four groups (Log-rank test, χ2=65.636, P<0.001), among which Post-SBP≥140 mmHg, Post-DBP<80 mmHg group had the lowest long-term survival rate. Further comparing the CVD mortality rate among the four groups, the curve showed a statistically significant difference (Log-rank test, χ2=29.784, P<0.001), and the highest CVD mortality rate was found in the Post-SBP≥140 mmHg, Post-DBP<80 mmHg group. Multivariate Cox regression analysis revealed that regardless of whether the average Post-SBP was ≥140 mmHg, Post-DBP<80 mmHg was an independent associated factor for death in MHD patients(with Post-SBP<140 mmHg, Post-DBP≥80 mmHg group as reference, Post-SBP≥140 mmHg, Post-DBP<80 mmHg group: HR=3.416, 95% CI 1.294-9.019, P=0.013; Post-SBP<140 mmHg, Post-DBP<80 mmHg group: HR=3.574, 95% CI 1.451-8.802, P=0.006). Conclusions:The long-term survival rate of the group with Post-SBP≥140 mmHg and Post-DBP<80 mmHg is significantly lower. Post-DBP<80 mmHg is an independent risk factor for death in MHD patients regardless of whether the average Post-SBP is ≥140 mmHg.

Objective To investigate the effects of long non-coding RNA (LncRNA) solute carrier organic anion transporter family member 4A1 (SLCO4A1-AS1) targeting microRNA-615-5p (miR-615-5p) in esophageal cancer cells on cell proliferation, apoptosis, and expression of inflammatory factors. Methods Real-time quantitative polymerase chain reaction (RT-qPCR) was used to analyze the expression of LncRNA SLCO4A1-AS1 and miR-615-5p in esophageal cancer tissues and cell lines. Eca109 cells were transfected with si-NC, si-LncRNA SLCO4A1-AS1, miR-NC, miR-615-5p mimic, pcDNA, pcDNA-LncRNA SLCO4A1-AS1, si-LncRNA SLCO4A1-AS1+anti-miR-NC, and si-LncRNA SLCO4A1-AS1+anti-miR-615-5p. Cell viability and apoptosis rate were measured by CCK-8 and flow cytometry, respectively; cell proliferation was determined by plate cloning assay; and levels of interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-α) in the culture medium were measured using an enzyme-linked immunosorbent assay (ELISA) kit. A dual luciferase reporter gene assay was used to determine the relationship between LncRNA SLCO4A1-AS1 and miR-615-5p. Results Expression of LncRNA SLCO4A1-AS1 was upregulated, and miR-615-5p expression was downregulated in esophageal cancer tissues and cell lines. After inhibiting LncRNA SLCO4A1-AS1 expression, cell viability, the number of cell clones, and levels of IL-6 and TNF-α in the culture medium decreased, while miR-615-5p expression and apoptosis rate increased (P < 0.05). Compared with the miR-NC group, the miR-615-5p group showed increased miR-615-5p expression, levels of Cleaved-caspase-3 protein, and apoptosis rate, and decreased cell viability, the number of cell clones, and levels of IL-6 and TNF-α in the culture medium (P < 0.05). Compared with the si-LncRNA SLCO4A1-AS1+anti-miR-NC group, the si-LncRNA SLCO4A1-AS1+anti-miR-615-5p group showed decreased miR-615-5p expression, levels of Cleaved-caspase-3 protein, and apoptosis rate, and increased cell viability, the number of cell clones, and levels of IL-6 and TNF-α in the culture medium (P < 0.05). Conclusion LncRNA SLCO4A1-AS1 can promote the occurrence and development of esophageal cancer. Inhibiting LncRNA SLCO4A1-AS1 can reduce the proliferation of esophageal cancer cells, decrease the expression of inflammatory factors, and induce apoptosis.

Objective:To determine the impact of early serum sodium concentrations on the survival prognosis in maintenance hemodialysis (MHD) patients.Methods:It was a retrospective cohort study. The newly admitted hemodialysis patients who were included in the registration system of Zhejiang Province Dialysis Quality Control Center from January 1, 2010 to December 31, 2019 were identified. Follow-up was conducted until December 31, 2020. Baseline data were collected for the first three months of dialysis, in which the mean level of serum sodium was defined as early serum sodium. Patients were divided into five groups based on early serum sodium level. Restricted cubic spline (RCS) was used to fit the relationship between long-term serum sodium level and risk of death. Kaplan-Meier model and Log-rank test were used to compare the survival rates of different groups. Multivariable Cox regression was used to analyze the correlation between early serum sodium level and death.Results:A total of 26 309 MHD patients were included in this study, and their ages were (59.07±15.41) years (ranging from 18 to 100 years). Among them, 13 643 (51.9%) were over 60 years old and 15 843 (60.2%) were males. Among the primary diseases of chronic renal failure, chronic glomerulonephritis was the first [13 703 cases (52.1%)], followed by diabetic nephropathy [6 460 cases (24.6%)], hypertensive nephropathy [1 293 cases (4.9%)], polycystic kidney disease [1 164 cases (4.4%)], etc. According to early serum sodium level, 12 883 patients (49.0%) had hyponatremia (serum sodium <135 mmol/L), of which 4 001 patients (15.2%) had serum sodium ≤130 mmol/L; 1 529 patients (5.8%) had hypernatremia (serum sodium >145 mmol/L). Patients were divided into the following 5 groups: 4 001 cases (15.2%) in group 1 (serum sodium ≤130 mmol/L), 8 882 cases (33.8%) in group 2 (130 145 mmol/L). Among them, patients in the Na≤130 mmol/L group had a slightly older age, a higher proportion of diabetes and cardiovascular disease, a lower level of blood uric acid, albumin, hemoglobin, and a higher level of alkaline phosphatase and leukocytes, while patients in the Na >145 mmol/L group had an older age and a higher proportion of cardiovascular disease. After follow-up of (55.67±33.58) months, a total of 4 954 patients (18.8%) died, 1 537 patients (5.8%) underwent kidney transplantation, 128 patients (0.5%) were converted to peritoneal dialysis. Of the deaths, 990 (20.0%) were due to cardiovascular diseases, 498 (10.1%) to cerebrovascular diseases and 400 (8.1%) to infections, and cardiovascular disease was the main cause of death. RCS curve fitting of the relationship between serum sodium level and risk of death found that the all-cause mortality hazard ratio ( HR) increased with decreasing or increasing serum sodium, and the optimal serum sodium was between 135 mmol/L and 140 mmol/L. Kaplan-Meier survival curve showed that the risk of all-cause death (Log-rank test, χ2=66.5, P<0.001), the risk of cardiovascular death (Log-rank test, χ2=31.5, P<0.001) and the risk of infection death (Log-rank test, χ 2=28.6, P<0.001) were significantly different among five groups. The 10-year cumulative survival rate was 63.0%, 71.5%, 72.5%, 67.8% and 61.4% in groups with different serum sodium levels from low to high, and the 10-year cumulative cardiovascular mortality rate was 9.6%, 6.2%, 5.5%, 7.3% and 11.7%, and the 10-year cumulative infection mortality rate was 4.9%, 3.2%, 1.7%, 2.8% and 3.9%. Multivariable Cox regression showed early serum sodium level >145 mmol/L was an independent relevant factor for all-cause death in MHD patients ( HR=1.237, 95% CI 1.045-1.465, P=0.013). Conclusions:MDH patients are more likely to develop hyponatremia in the early stage of dialysis. The cumulative survival rate of all-cause death, cardiovascular death and infection death in patients with predialysis serum sodium ≤130 mmol/L and >145 mmol/L within three months after initiation of dialysis is significantly lower than those in other levels. Early serum sodium >145 mmol/L is associated with higher mortality in MHD patients.

BACKGROUND: This study aimed to evaluate the correlation between long non-coding RNA (lncRNA)-related single nucleotide polymorphisms (SNPs) and susceptibility to silicosis. METHODS: First, RNA-sequencing (RNA-seq) data were comprehensively analyzed in the peripheral blood lymphocytes of eight participants (four silicosis cases and four healthy controls) exposed to silica dust to identify differentially expressed lncRNAs (DE-lncRNAs). The functional SNPs in the identified DE-lncRNAs were then identified using several databases. Finally, the association between functional SNPs and susceptibility to silicosis was evaluated by a two-stage case-control study. The SNPs of 155 silicosis cases and 141 healthy silica-exposed controls were screened by genome-wide association study (GWAS), and the candidate SNPs of 194 silicosis cases and 235 healthy silica-exposed controls were validated by genotyping using the improved Mutiligase Detection Reaction (iMLDR) system. RESULTS: A total of 76 DE-lncRNAs were identified by RNA-seq data analysis (cut-offs: fold change > 2 or fold change < 0.5, P < 0.05), while 127 functional SNPs among those 76 DE-lncRNAs were identified through multiple public databases. Furthermore, five SNPs were found to be significantly correlated with the risk of silicosis by GWAS screening (P < 0.05), while the results of GWAS and iMLDR validation indicated that the variant A allele of rs1814521 was associated with a reduced risk of silicosis (OR = 0.76, 95% CI = 0.62-0.94, P = 0.011). CONCLUSION The presence of the SNP rs1814521 in the lncRNA ADGRG3 is associated with susceptibility to silicosis. Moreover, ADGRG3 was found to be lowly expressed in silicosis cases. The underlying biological mechanisms by which lncRNA ADGRG3 and rs1814521 regulate the development of silicosis need further study.
Case-Control Studies ; Genetic Predisposition to Disease ; Genome-Wide Association Study ; Humans ; Polymorphism, Single Nucleotide ; RNA, Long Noncoding/genetics* ; Silicosis/genetics*

Objective:To investigate the clinical characteristics and prognosis of IL3-IGH fusion gene-positive pediatric acute lymphoblastic leukemia (ALL) with hypereosinophilia as the first presentation.Methods:The clinical data of 1 pediatric IL3-IGH fusion gene-positive ALL patient with hypereosinophilia as the first presentation in January 2021 in Fujian Medical University Union Hospital was retrospectively analyzed and relevant literature was reviewed.Results:This 11-year-old male patient underwent bone marrow examination, and results showed that the proportion of eosinophils was increased; immunophenotyping disclosed that there were about 49.4% abnormal naive B lymphocytes in bone marrow; 43 leukemia fusion genes showed all negative; the whole transcriptome sequencing showed IL3-IGH fusion gene-positive. The patient was finally diagnosed as B-ALL with IL3-IGH fusion gene. According to the Chinese Children Cancer Group (CCCG)-ALL 2020 regimen, eosinophils returned to normal after induction therapy. Bone marrow examination on day 19 of induction showed that the proportion of promyelocytes was 0.005, the proportion of eosinophils was 0.05, and the minimal residual disease (MRD) was 23.02%. Bone marrow examination on day 46 of induction showed remission, and MRD was 0.18%. Consolidation chemotherapy used CAT (cyclophosphamide 1 g/m 2 once; cytarabine 50 mg/m 2, 12 h once, 7 days in total; mercaptopurine 40 mg/m 2, once per night, 7 days in total) regimen. Then the patient was added with lusotinib (75 mg 12 h once) orally and continued to receive high-dose methotrexate (5 g/m 2) regimen chemotherapy for 2 courses, the MRD was 0.20%. Chimeric antigen receptor T-cell (CAR-T) regimen was administered, followed by negative MRD. Conclusions:IL3-IGH fusion gene ALL is more frequently found in males, and more common in older children and young adults. It is prone to organ infiltration damage, and it has a high rate of induction failure and recurrence as well as poor prognosis.

To present the clinical characteristics and the misdiagnosis rate of acute coronary syndrome manifested primarily as throat discomfort, we conducted a multicentric and retrospective study in the cardiology and otorhinolaryngology departments. Records of patients with primary complaint of throat discomfort, absence of chest pain at onset, and an ultimate diagnosis of acute coronary syndrome, as well as patients with pharyngitis (as controls) were collected from May 2015 to April 2016. The patients' main manifestations were compared. Logistic regression results showed that chest tightness, dyspnea, perspiring, and exertional throat symptoms were significantly associated with acute coronary syndrome, with odds ratios of 8.3 (95% CI 2.2-31.5), 10.9 (95% CI 1.8-66.9), 25.4 (95% CI 3.6-179.9), and 81.2 (95% CI 13.0-506.7). A total of 25 (56.82%) out of 44 acute coronary syndrome patients, who were first admitted to the otorhinolaryngology department, were misdiagnosed, with a 12% (3/25) mortality rate. Throat discomfort can be the principal manifestation of acute coronary syndrome. Such patients exhibit high misdiagnosis and mortality rates. Exertional throat symptoms, chest tightness, perspiring, and dyspnea were important indicators of acute coronary syndrome in patients whose main complaint was throat discomfort. The awareness of this condition will result in prompt diagnosis and reduce morbidity and mortality.
Acute Coronary Syndrome/etiology* ; Dyspnea/etiology* ; Humans ; Pharyngitis/diagnosis* ; Pharynx ; Retrospective Studies

OBJECTIVE To investigate the antibacterial activities of flavaspidic acid BB against the main pathogens of skin and soft tissue infections such as methicillin -resistant Staphylococcus aureus （MRSA）and methicillin -sensitive Staphylococcus aureus （MSSA） in vitro and in vivo . METHODS Microdilution method was used to determine the minimum inhibitory concentration（MIC）and minimum bactericidal concentration （MBC）of flavaspidic acid BB on 10 strains of MRSA and 13 strains of MSSA ；time-kill curves of MRSA 11 and MSSA 23 strains were drawn by plate method ；the checkerboard dilution method was used to determine the combined antibacterial effect of flavaspidic acid BB and mupirocin ；the skin abscess model of mice was established to evaluate the antibacterial effects of 0.5%，1%，2% flavaspidic acid BB cream on MRSA 11 or MSSA 23 strains in vivo；scanning electron microscopy was used to observe the effects of flavaspidic acid BB on the morphology of MRSA 11 or MSSA23 strains. The contents of nucleic acid leakage and extracellular protein were also determined . RESULTS MIC and MBC ranges of flavaspidic acid BB against 10 strains of MRSA were 6.30-80.00 μg/mL and 40.00-640.00 μg/mL；MIC and MBC ranges of 13 strains of MSSA were 40.00-80.00 μg/mL and 63.50-126.99 μg/mL，respectively. The bactericidal rate of MRSA 11 and MSSA23 strains were 99.9% after interfered with plavaspidic acid BB for 36 h. The combination of flavaspidic acid BB and mupirocin showed an additive effect on MRSA strain and an additive or synergistic effect on MSSA strain . The 0.5%，1% and 2% flavaspidic acid BB cream could reduce the volume of abscess and the bacterial load of skin abscess model mice to some extent （P＜0.05），and improved the pathological changes of skin inflammation in skin abscess site of mice . Flavaspidic acid BB could make MRSA 11 and MSSA 23 cells shrinkage and rupture ，and significantly increase the contents of nucleic acid leakage and extracellular protein （P＜0.01）. CONCLUSIONS Flavaspidicacid BB shows good antibacterial effect on MRSA and MSSAE-mail：tchp66@163.com in vitro and in vivo ，the mechanism of which may be related to the change of membrane permeability .

OBJECTIVE To establish an analytical method based on ultra-performance liquid chromatography-quadrupole-time- of-flight mass spectrometry （UPLC-Q-TOF/MS） combined with UNIFI platform for rapid identification of the chemical constituents in Abrus cantoniensis Hance. METHODS The chromatographic separation was performed on ACQUITY PRM HSS T3 FIT column for gradient elution with the mobile phase consisted of 0.1% formic acid aqueous solution-acetonitrile. The flow rate was 0.3 mL/min， and injection volume was 2 μL. Electrospray ionization was used to collect the mass spectrometry data of the chemical constituents of A. cantoniensis Hance with full information tandem mass spectrometry technology in positive and negative ion modes. The chemical constituent database of A. cantoniensis Hance was established. Targeted and non-targeted analyses were conducted based on UNIFI platform， and chemical constituents were further identified in combination with accurate molecular mass， secondary fragment ion information and equivalence with reference substances and literature data， etc. RESULTS Totally 46 compounds of A. cantoniensis Hance were successfully identified， including 19 flavonoids， 8 triterpenoids， 3 alkaloids， 5 organic acids and 11 other components. Among them， 11 compounds were firstly found in A. cantoniensis Hance， and 9 compounds were confirmed by reference substance. CONCLUSIONS The analytical method based on UPLC-Q-TOF/MS combined with UNIFI platform can quickly identify the chemical constituents of A. cantoniensis Hance. Flavonoids and triterpenes are the main components in A. cantoniensis Hance.