Objective To develop a tumor vaccine containing broad-spectrum neoantigen tumor cell lysate(TCL)and CpG adjuvant,and to effectively deliver it to lymph node dendritic cells.Methods A novel polymer,9-fluorenylmethoxycarbonyl-polyethylene glycol-glycocholic acid(Fmoc-PEG-GCA),was employed to encapsulate the TCL and CpG through π-π stacking,resulting in high-density polyethylene glycol-modified glycocholic acid-decorated micelles TCL/CpG@GNP.The vaccine′s drug loading,encapsulation efficiency,particle size,polydispersity index(PDI),zeta potential,morphology,stability,cellular safety,uptake capability,immune stimulation effects on bone marrow-derived dendritic cells(BMDCs)and in vivo anti-tumor efficacy were evaluated.Results The vaccine TCL and CpG demonstrated a drug loading capacity of 6.26%and the encapsulation rate was 37.59%.The drug loading capacity of CpG was 7.05%,and the encapsulation efficiency was 56.86%.The particle size measured(139.26±27.23)nm,with a PDI of 0.249±0.015,indicating favorable dispersion properties.The zeta potential was recorded at(-21.23±0.36)mV.The TCL/CpG@GNP vaccine demonstrated good stability,cell safety and uptake ability,and can promote the activation and maturation of BMDCs.In tumor-bearing mouse models,TCL/CpG@GNP inhibited tumor growth,increased the proportion of T lymphocytes in peripheral blood,and elevated IFN-γ levels in spleen.Conclusion The TCL/CpG@GNP tumor vaccine can effectively activate BMDCs and induce strong anti-tumor immune memory in a mouse lung cancer model.

Objective To develop a tumor vaccine containing broad-spectrum neoantigen tumor cell lysate(TCL)and CpG adjuvant,and to effectively deliver it to lymph node dendritic cells.Methods A novel polymer,9-fluorenylmethoxycarbonyl-polyethylene glycol-glycocholic acid(Fmoc-PEG-GCA),was employed to encapsulate the TCL and CpG through π-π stacking,resulting in high-density polyethylene glycol-modified glycocholic acid-decorated micelles TCL/CpG@GNP.The vaccine′s drug loading,encapsulation efficiency,particle size,polydispersity index(PDI),zeta potential,morphology,stability,cellular safety,uptake capability,immune stimulation effects on bone marrow-derived dendritic cells(BMDCs)and in vivo anti-tumor efficacy were evaluated.Results The vaccine TCL and CpG demonstrated a drug loading capacity of 6.26%and the encapsulation rate was 37.59%.The drug loading capacity of CpG was 7.05%,and the encapsulation efficiency was 56.86%.The particle size measured(139.26±27.23)nm,with a PDI of 0.249±0.015,indicating favorable dispersion properties.The zeta potential was recorded at(-21.23±0.36)mV.The TCL/CpG@GNP vaccine demonstrated good stability,cell safety and uptake ability,and can promote the activation and maturation of BMDCs.In tumor-bearing mouse models,TCL/CpG@GNP inhibited tumor growth,increased the proportion of T lymphocytes in peripheral blood,and elevated IFN-γ levels in spleen.Conclusion The TCL/CpG@GNP tumor vaccine can effectively activate BMDCs and induce strong anti-tumor immune memory in a mouse lung cancer model.

Heart failure is a serious and end-stage status of various heart diseases, characterized by comparatively high rate of readmission and mortality, and has become an important public health issue. The risk of readmission and mortality following discharge of an index hospitalization are key indicators to evaluate the quality of medical care among patients with acute heart failure. Therefore, it is important to carry out risk prediction research for patients with acute heart failure, quantify the disease risk, perform risk stratification, optimize clinical decision-making, elevate patients' quality of life and prognosis, and comprehensively improve the medical quality of acute heart failure. During the past 20 years, foreign researchers have developed dozens of models to predict the risk of acute heart failure readmission and mortality, and Chinese researchers have also developed up to 10 models applicable to the Chinese population. However, there is no recommended risk prediction model for acute heart failure in current clinical guidelines across China. In this report, we aim to introduce the major models for predicting the risk of acute heart failure readmission and mortality from home and abroad, focus on putting forward limitations of established models, and initiating potential directions for future studies from the following aspects: integrate multi-source data, mine emerging biomarkers, establish polygenic risk scores, optimize machine learning methods, promote flexible adjustment, and broaden approaches that applicable for various scenarios. Accordingly, this study will help facilitate domestic research in predicting the risk of readmission and mortality among patients hospitalized for acute heart failure.

Objective:To investigate the expression levels of microRNA-124 (miR-124) and microRNA-494(miR-494) in the serum of elderly patients with Parkinson disease (PD) and its clinical significance.Methods:Ninety PD patients (PD group) who were hospitalized in Zhengzhou Central Hospital Affiliated to Zhengzhou University from March 2018 to April 2020 were selected.At the same time, 100 non-PD elderly people examined in the physical examination center of the same hospital who matched with age and gender of PD patients were selected as the control group.After 12 hours of fasting, 4 ml of venous blood was taken from all subjects.All PD patients were graded by unified Parkinson disease rating scale(UPDRS) from the aspects of mental state, behavior and emotion, quality of life and motor examination, and graded by the Hoehn-Yahr rating scale for Parkinson disease.The expression levels of miR-124 and miR-494 in serum were detected by real-time fluorescent quantitative PCR (qRT-PCR), and the diagnostic values of miR-124 and miR-494 in PD patients were evaluated by ROC curve.Results:Hoehn-Yahr grade of PD patients with UPDRS≤60 points was significantly lower than that of patients with UPDRS >60 points((2.47±0.43) vs (3.42±0.47))( t=9.055, P<0.001), and there was no significant difference in serum miR-124 and miR-494 expression levels((0.72±0.14) vs (0.70±0.12), (1.17±0.19) vs (1.18±0.22)) ( t=0.633, 0.230, P=0.529, 0.819). Compared with that in control group, the expression of miR-124 in PD group was down-regulated ((0.71±0.20) vs (1.05±0.24)), and the expression of miR-494 was up-regulated((1.18±0.26) vs (0.96±0.22)) ( t=10.542, 6.315, P<0.001). The results of ROC showed that the area under curve (AUC) of serum miR-124 and miR-494 in the diagnosis of PD were 0.847 and 0.760 respectively, the cutoff values were 0.901 and 1.126, respectively, the sensitivities were 86.67% and 61.11% respectively, and the specificities were 75.03% and 79.00% respectively. The AUC of the combined diagnosis of PD was 0.898, and the sensitivity and specificity were 85.56% and 85.00% respectively. Conclusions:The expression of miR-124 is low in PD patients, while the expression of miR-494 is high, which suggests that the changes of the two miRNA levels may be related with the occurrence and development of PD.Both of them have a certain diagnostic value for PD, and the value of combined diagnosis is higher.

Objective　To analyze the relationship between single nucleotide polymorphism （SNP） of microRNA-146a （miR-146a） gene and Parkinson’s disease （PD）. Methods　In a case-control study，446 patients （PD group） with primary PD were selected from our hospital from October 2015 to August 2019，and 450 healthy people in the health examination center of the same period were selected as the control group，the SNP of rs2910146 and rs5709329 loci in miR-146a were detected by DNA PCR sequencing technology；and the risk factors of PD were analyzed by logistic regression. Results　The genotypes of rs2910146 and rs5709329 in miR-146a in PD group and control group were consistent with Hardy-Weinberg equilibrium law （P>0.05）. There was no significant difference in gender，age，BMI and education between PD group and control group （P>0.05）. Compared with the control group，the G allele and GG genotype frequency of rs2910146 locus in PD group were significantly lower （P<0.05）. There was no significant difference in allele frequency and genotype distribution of rs5709329 locus between PD group and control group （P>0.05）. SNP at rs2910146 in miR-146a was related to cognitive dysfunction and age of PD patients （P<0.05），but not to gender，PD subtype and first-onset limb （P>0.05）. Carrying GG genotype was an independent protective factor for PD （P<0.05）.Conclusion　SNP at rs2910146 locus in miR-146a is related to age and cognitive impairment in PD patients，and carrying GG genotype may reduce the risk of PD.

OBJECTIVE: To explore the correlation between special A/O genotype and the O phenotype. METHODS: Group O samples with partially reduced or lack of isoagglutinins were collected to determinate the ABO genotype with a PCR-sequence specific primer (PCR-SSP) assay. Seven samples with A/O genotype were selected for further study. Serological tests including forward and reverse typing, H antigen determination and adsorption/elution were carried out with a tube method. Genomic DNA was genotyped by amplifying and sequencing of the coding regions of exons 1 to 7 of the ABO gene. RESULTS: Seven samples were serotyped as group O by the forward typing test. However, reduced anti-A activity was found in 5 samples by the reverse typing test, reduced anti-A and anti-B activities were found in 1 sample, and no anti-A isoagglutinin activity was found with 1 sample. H antigen was determined in all samples by routine serologic method. Neither anti-A nor anti-B was eluted from red cells derived from all samples. Three samples were genotyped as Ael02/O02, whilst the remainders were Ael02/O13, Ael02/O65, Am04/O75, Ael06/O02, respectively. CONCLUSION Special A/O genotype may not express the A antigen, leading to the generation of group O red cells. Reduced or missed anti-A activity is the typical serological feature of this special group of O phenotype, for which ABO*Ael02 and ABO*O02 are the major alleles. Group O individuals with isoagglutinin detection problem should be grouped by serological tests and genomic DNA analysis.
ABO Blood-Group System ; Alleles ; Exons ; Genotype ; Humans ; Phenotype

Objective To improve the quality of clinical biochemistry laboratory by quality indicators of pre-analytical,analytical,post-analytical phase and the whole process.Methods Analytical Phase:The Sigma values of items were calculated,applying the equation Sigma =(TEa％-Bias％)/CV％.Total allowable error (TEa) is from analyticalal specification defined in WS/T403-2012 of China,Bias％ is from the evaluation results of National Center for Clinical Laboratory (NCCL) trueness verification PT series and CV％ is from internal quality control data during the last 6 months in our lab.Normalized Sigma metrics plot was made to evaluate the analysis performance and the quality control strategies were designed accordingly.The quality goal indexes (QGI) were also calculated to propose improvement measures for items below 6 Sigma.Quality indicators of pre-,post-analytical and whole analytical phase,such as quality of specimen,critical value notification,critical value notification in time,TAT of hs-cTnT,TAT of emergency biochemical items,rewrite of laboratory reports and unacceptable performance in EQA-PT were measured in Sigma metrics too.The Sigma metrics changes before and after taking improvement measures were compared to conform the effectiveness.Results The average Sigma value of 17 biochemical tests was 5.29,of which 8 items (UA,K,ALP,CK,AMY,AST,TG,Na) achieved excellent to world class level (≥ 5 Sigma),6 items (LDH,Cre,TC,ALT,Mg,Glu) achieved marginal to good level (5 ＞ Sigma ≥ 3),BUN performed poorly (3 ＞ Sigma ≥ 2),Ca,TP performed unacceptably (Sigma ＜ 2) with serious quality defects.The Sigma values of unacceptable specimen,critical value notification,critical value notification in time,unacceptable turn around time (TAT) of hs-cTnT,unacceptable turn around time (TAT) of emergency biochemical items,rewrite of laboratory reports,unacceptable performance in EQA-PT were 4.17,3.60,2.75,1.72,3.27,4.52,3.33 respectively,rising to 4.30,4.30,2.90,2.45,3.75,4.80,3.60 accordingly after improvement.Conclusions Sigma metrics is potentially an ideal approach for clinical biochemistry laboratories management,which is helpful to find out problems,put forward improvement measures,and confirm the effectiveness,so as to achieve the purpose of continuous quality improvement.

Objective We aim to evaluate the value of the current serum creatinine reference interval ( RI ) provided by Industry Standard WS /T 404.5 in clinical practice.Methods The first time serum creatinine levels and urinary albumin /creatinine ratio were obtained from 67 605 adult patients ( <60 years old) who were treated in the First Hospital of China Medical University between October 1, 2014 and September 30, 2015 in this cross-sectional study.Estimated glomerular filtration rate ( eGFR ) calculated by chronic kidney disease epidemiology collaboration (CKD-EPI) equation and urinary albumin /creatinine ratio (ACR) were used to evaluate the clinical practical significance of current and old serum creatinine RIs as the criteria.Results 4.3% of normal subjects based on current RI were showed decreased eGFR, 98% of abnormal subjects based on the current RI were founded to have decreased eGFR . 1378 subjects were evaluated as increased based on current RI but as normal based on old RI , and 93.5% of these subjects were showed decreased eGFR .In addition, ACR was measured in 26 cases, and 18 out of 26 cases (69.2%) were confirmed to have elevated ACR (≥30 mg/g) and proteinuria.On the other hand of analysis, screening positive rates of declined eGFR were 43.6% by old RI and 61.9% by current RI in the subjects with eGFR under 90 ml(min ×1.73 m 2 ), and the performance of the current RI was obviously improved(χ2 =212.648,P <0.001).Conclusions The current reference interval of serum creatinine is favorable for the detection of renal dysfunction in patients .It is recommended that the current reference interval can be applied in the clinical laboratories as early as possible .

Aim To study the synergistic anti-depres-sion effect of 3 , 6-disinapoyl sucrose ( DISS ) and tenuifoliside A ( TFSA ) from Radix Polygalae and the preliminary mechanism . Methods Using the classical behavioral despair and depression model of mouse tail suspension test, 120 mice were divided into control group, positive group, DISS 5 mg·kg-1 group,DISS 10 mg·kg-1 group,TFSA 5 mg·kg-1 group,TFSA 10 mg· kg-1 group, DISS 5 mg · kg-1 +TFSA 5 mg · kg-1 group,DISS 5 mg·kg-1 +TFSA 10 mg·kg-1 group,DISS 10 mg·kg-1 +TFSA 5 mg·kg-1 group and DISS 10 mg · kg-1 +TFSA 10 mg · kg-1 group randomly. They were given intragastric injection for 7 days continuously, to observe the effect of DISS and TFSA monomer and its combination on the time of mouse tail suspension. Expression of BDNF in the hip-pocampus of mice was detected by immunohistochemis-try. The expressions of CREB, pCREB, CRTC1 and BDNF in the hippocampus of mice were detected by Western blot method. Results The administration of DISS and TFSA could shorten the immobility time of mice subjected to the tail. DISS ( 10 mg · kg-1 ) and TFSA( 10 mg · kg-1 ) group were significantly lower than single dose drug group(P<0. 05). DISS and TF-SA and the combination groups could increase the ex-pression of BDNF in hippocampus and cortex by immu-nohistochemistry(P <0. 05). At the same time, the contents of CREB, CRTC1, pCREB and BDNF protein in the hippocampus were increased by DISS and TF-SA, and the combination group was significantly higher than the single drug group ( P<0. 05 ) . Conclusion The administration of DISS and TFSA are used to acti-vate CREB transcription factor CRTC1 , and activate the phosphorylation of CREB in the hippocampus, and then increase the expression of BDNF in the hippocam-pus and plays a synergistic antidepressant effect.

Objective To investigate the situation of human papillomavirus (HPV) infection and the distribution of HPV genotypes among women attending hospital in Meizhou,Guangdong province.Methods Flow-through hybridization and gene chip technique was used to detect HPV in cervical exfoliated cell specimens collected from 24 450 women for HPV screening in Department of Gynecology,Meizhou People's Hospital,Meizhou Hospital Affiliated to Sun Yat-sen University,and then the situation of HPV infection and distribution of HPV genotypes were analyzed.Results Among 24 450 female exfoliated cell specimens,3 922 were found to be positive for HPV infection,with the total infection rate of 16.04％.Among 3 922 samples,the top three high-risk subtypes of HPV were HPV-16 (30.37％),HPV-52 (17.77％) and HPV-58 (15.53％),the majority of low-risk HPV was HPV-CP8304 (5.61％).The positivity of various HPV types peaked among 30-50 years old.The differences of the HPV positive rates in different age groups was statistical significance (P ＜ 0.001).Conclusions The majority of women attending hospital detected with HPV-16 in Meizhou and the positivity of various HPV types peaked among women aged 30-50 years.Genotyping of HPV was meaningful for preventing and treating cervical cancer.