Objective To investigate the differential expression of proteins in the serum of diabetic patients quantitative proteomics technologies, and to identify specific protein biomarkers. Methods Serum samples from three diabetic patients and three healthy controls in Shenzhen in 2023 were collected. Immunoprecipitation and nano liquid chromatography-quadrupole-electrostatic orbitrap-linear ion trap mass spectrometry (easy-nanoLC-Q-orbitrap-fusion) were employed to perform fraction and no-fraction-based quantitative analysis of the serum proteome in diabetic patients and healthy controls. The mass spectrometry data were analyzed by MaxQuant1.6.1.0, and the differential proteins identified by the fraction-based method were subjected to gene ontology (GO) functional clustering and kyoto encyclopedia of genes and genomes (KEGG) pathway analysis. Results A total of 251 reliable proteins were identified by the no-fraction-based method, among which 17 differentially expressed proteins were selected. In contrast, 494 reliable proteins were identified by the fraction-based method, and 74 differentially expressed proteins were selected. Among them, seven proteins showed consistent changes in both methods. Five proteins were downregulated, including pregnancy zone protein (PZP), carbonic anhydrase 1 (CAH1), mannose-binding protein C (MBL2), fibrinogen alpha chain (FIBA), and haptoglobin (HPT). Two proteins were upregulated, including apolipoprotein(a) (LPA) and complement factor H-related protein 4 (CFH4). GO analysis revealed that the differentially expressed proteins were mainly involved in molecular functions such as glycosaminoglycan binding and heparin binding. KEGG pathway analysis showed that the differentially expressed proteins were mainly associated with complement and coagulation cascades, cell adhesion, PI3K-Akt signaling pathway, and other pathways. Conclusions Fraction-based no-fractional quantitative proteomics analysis provides an effective approach for analyzing differential protein expression in the serum of diabetic patients, and offers a valuable technique for screening specific protein biomarkers of serum samples in large-scale cohorts of disease.

Abatract:Objective To study the relationship of the processes of chronic hepatitis B infection with the degree of depression and the level of serum Brain-derived neurotrophic factor (BDNF).Methods 126 patients with different processes of CHB infection were divided into 3 groups,which were asymptomatic HBV carriers (ASC group),liver cirrhosis (LC group)and hepatic cell carcinoma (HCC group).40 healthy volunteers were selected as normal control.Questionnaire was used to study demographic data and the degree of depression was assessed by using Hamilton Depression Rating Scale (HAMD-17).The levels of the serum BDNF were detected by ELISA.Results The ratio of divorced or widowed in LC and HCC groups was higher than that of the control (χ2 = 6.354,11.972;P < 0.01).Severity of depression in ASC,LC and HCC groups was higher than that of the control (χ2 = 16.151,42.150,49.636;P <0.01).Severity of depression in ASC and LC group was lower than that of HCC group (χ2 = 14.345,28.772;P <0.01).The role of BDNF (x ±s,ng/ml)in ASC,LC,HCC and control group were 11.10±3.26,8.66 ± 3.11,7.39 ±2.52 and 12.18±2.59.The levels of serum BDNF in LC and HCC group was lower than ASC group and the control (P <0.01).The process of CHB infection had positive correlation with the score of HAMD (r=0.719,P <0.01),but had negative correlation with thelevels of serum BDNF (r=-0.504,P <0.01). There was negative correlation relationship between the score of HAMD and the levels of serum BDNF (r=-0.526,P <0.01).Conclusion The process of CHB infection can obviously aggravate the degree of depression,and reduce the levels of BDNF gradually.The levels of serum BDNF can be suggested as a psychological laboratory reference basis for the CHB pa-tients with depression.It is expected to improve the level of clinical diagnosis and treatment in CHB with depression.

Objective To clone and express Mycobacterium tuberculosis fusion antigen ESAT-6 and CFP-10 ( EC) and to evaluate the biological activity of the fusion antigen EC in inducing specific cytokines secretion from THP -1 cells. Methods The fusion antigen EC gene was cloned into pET-30 a prokaryotic expression vector and expressed highly in E.coli BL21.Then, the THP-1 cells were stimulated with purified fusion antigen EC of different concentrations (10 and 20 μg/ml).Culture supernatants were collected after 12 h and 24 h, respectively.The secretion levels of IL-2, IL-4, IL-6, IL-8, IL-10, GM-CSF, TNF-αand IFN-γin THP-1 cell culture supernatants were detected using Bio-PlexProTM Assays kit.Results The M.tuberculosis fusion antigen EC was cloned and expressed successfully .The secretion levels of IL-6, IL-8 and TNF-αin EC infected THP-1 cells were significantly higher than those in THP-1 cells (P<0.05).The secretion levels of other cytokines did not change significantly .Conclusion The obtained M.tuberculosis fusion antigen EC has biological activity in inducing the THP-1 cells to secrete a higher level of IL-6,IL-8 and TNF-α.

Objective To investigate the related factors of percutaneous balloon compression (PBC) for the treatment of trigeminal neuralgia complicating cerebrovascular disease in order to reduce the incidence of surgical complications. Methods The clinical data in patients with trigeminal neuralgia complicating cerebrovascular disease treated with PBC in the People's Hospital of Liaoning Province from December 2000 to December 2010 were analyzed retrospectively. The reasons for its occurrence were investigated. Results Of the 6 759 patients with trigeminal neuralgia who received PBC treatment, 10 (0. 15％ ) developed cerebrovascular disease,and 2 of them had arteriovenous fistula; 5 patients developed intracranial hemorrhage: 2 acute subdural hemorrhage, 2 intraventricular hemorrhage with subarachnoid hemorrhage, and 1 small amount of bleeding in brain stem; 1 patient developed internal carotid hemorrhage during the intraoperative puncture and the operation was stopped temporarily, and he was reoperated successfully after 2 weeks; 2 patients had cerebral infarction, they were all in the risk population of cerebrovascular disease, and 3 weeks after treatment, their disease conditions were improved. One of them left severe paralysis of limbs at discharge. Condusions The treatment of trigeminal neuralgia with PBC is safe and the incidence of cerebrovascular complications is lower. The surgical operations and blood pressure changes are the important causes of cerebrovascular complications.

Objective To explore the mechanism of Zishen Jianpi Huayu Tablets(Corni Fructus,Notoginseng Radix et Rhizoma,Astragali Radix,Puerariae Lobatae Radix,Spatholobi Caulis,Rehmanniae Radix)in the treatment of diabetic retinopathy(DR)by means of network pharmacology and molecular docking technique,and verified by in vitro experiments.Methods The active components of Zishen Jianpi Huayu Tablets and their corresponding target proteins were screened using the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)and the BATMAN-TCM database.Drug target proteins were converted to their corresponding gene names through the UniProt database.DR-related targets were searched using"diabetic retinopathy"as a keyword in GeneCards,DrugBank,OMIM,and TTD databases.Common targets between the disease and the drug were identified using the Venny tool.These common targets were analyzed using the String database,a protein-protein interaction(PPI)network was constructed.Topological heterogeneity analysis was performed using Cytoscape 3.9.1 to select core targets and create a PPI network diagram.These common targets were entered into the Metascape database for Gene Ontology(GO)function analysis and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analysis to identify potential action pathways.Molecular docking of the main active components and core targets was performed using Auto Dock tools software,followed by further experimental validation.The CCK-8 assay was used to assess the effect of Zishen Jianpi Huayu Tablet medicated serum on the cell viability of Human Retinal Microvascular Endothelial Cells(HRmECs)under high glucose conditions,and RT-qPCR was used to measure the expression of IL-1β,AKT1,VEGFA,and TP53 mRNA in HRmECs.Results(1)The effective components and corresponding target proteins of Zishen Jianpi Huayu Tablets were screened by Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP)and BATMAN-TCM database.The disease-related targets of DR were searched by GeneCards,OMIM and TTD databases.The use of VENNY platform for drug active components target and DR disease-related target to take intersection(common target),that is,Zishen Jianpi Huayu Tablets in the treatment of DR potential target.The network of"drugs-active components-common targets"was constructed to screen out the key active components of Zishen Jianpi Huayu Tablets in the treatment of DR.Import the common target into STRING database,obtain the PPI network relationship,and screen out the core target.Metascape platform was used to analyze the GO function and KEGG pathway enrichment of the common targets.The key active components and core targets were verified by Autodock 4 software for molecular docking.(2)The drug-containing serum and blank serum of Zishen Jianpi Huayu Tablets was prepared.Human retinal microvascular endothelial cells(HRmECs)were randomly divided into 5 groups:the control group(low-sugar DMEM medium+10%blank serum),high-glucose group(high-sugar DMEM medium+10%blank serum)and Zishen Jianpi Huayu Tablets containing low-,medium-and high-dose serum(high-sugar DMEM medium+10%low-,medium-and high-dose drug containing serum)were detected after 48 hours of culture.The proliferative activity of HRmECs cells was detected by CCK-8 method,and the mRNA expressions of IL-1β,AKT1,VEGFA and TP53 in HRmECs cells were detected by RT-qPCR method.Conclusion Zishen Jianpi Huayu Tablets may act on core targets such as IL-1β,IL-6 and VEGFA,as well as key pathways such as NF-κB signaling pathway,AGE-RAGE signaling pathway and PI3K-AKT pathway through various active components such as quercetin,kaempferol and rehmannia flavonoids,so as to play a therapeutic role in DR.

Objective Investigate the expression level and clinical significance of microRNA-223-3p(miR-223-3p)in the serum of pregnant women with preterm premature rupture of membranes(PPROM).Methods A total of 91 pregnant women who underwent cesarean delivery at the Xuzhou Maternal and Child Health Hospital affiliated with Xuzhou Medical University between April and September 2023 were selected for the study.This included 60 cases of PPROM and 31 cases of term normal pregnant women as the research subjects.The study group was divided based on the pathological results of the membranes into:PPROM group without histological chorioamnionitis(HCA)(PPROM group,n = 37),and PPROM group with HCA(PPROM+HCA group,n = 23).Serum samples from pregnant women were collected before hospital treatment,and the expression level of miR-223-3p in the serum was detected by quantitative real-time PCR(qRT-PCR).The levels of inflammatory factors such as IL-1β,IL-6,IL-8,and TNF-α in the serum were measured by enzyme-linked immunosorbent assay(ELISA).Results The expression level of miR-223-3p in the serum of the PPROM+HCA group was significantly higher than that of the PPROM group and the normal group(P＜0.05).The expression levels of IL-1β,IL-6,IL-8,and TNF-α in the serum of the PPROM+HCA group were also significantly higher than those of the PPROM group and the normal group(P＜0.05).There was a positive correlation between the expression level of miR-223-3p in the serum of the PPROM+HCA group and the expression levels of IL-1β,IL-6,IL-8,and TNF-α(r = 0.553,0.505,0438,0.656,P＜0.05).Conclusion The upregulation of miR-223-3p in the serum of pregnant women with PPROM+HCA is associated with the severity of inflammation in PPROM.

OBJECTIVETo assess the association of a disintegrin and metallo-proteinase with thrombospondin type 1 motifs (ADAMTS-1) gene polymorphism and ischemic stroke caused by large artery atherosclerosis (LAA).

METHODSIn total 767 patients and 506 controls were recruited. Single nucleotide polymorphisms (SNPs) rs416905 (T/C) and rs402007 (G/C) of the ADAMTS-1 gene were genotyped by polymerase chain reaction and DNA sequencing.

RESULTSFrequencies of the rs402007 GC+CC genotype and the C allele were significantly different between the two groups (68.84% vs. 60.67%, χ2=9.012, P=0.003, OR=1.432; 45.24% vs. 38.54%, χ2=11.208, P=0.001, OR=1.318). Binary logistic regression has confirmed that the above difference was significant (P=0.001, OR=1.521, 95%CI: 1.183-1.955). The frequencies of TC+CC and GC+CC genotypes were similar between the two groups, and so was it with the C allele. The two SNPs had been in complete linkage disequilibrium (D'=1.0, r2=1.0).

CONCLUSIONThe rs416905 and rs402007 polymorphisms of the ADAMTS-1 gene may be associated with ischemic stroke caused by LAA. The C allele of the rs402007 locus may be a susceptibility factor for this subtype of stroke.

ADAM Proteins ; genetics ; ADAMTS1 Protein ; Aged ; Alleles ; Atherosclerosis ; complications ; Base Sequence ; Blood Glucose ; metabolism ; Brain Ischemia ; complications ; Fasting ; blood ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Logistic Models ; Male ; Middle Aged ; Polymerase Chain Reaction ; Polymorphism, Single Nucleotide ; Risk Factors ; Sequence Analysis, DNA ; Smoking ; Stroke ; blood ; etiology ; genetics

Objective:To determine the clinical, pathological and imaging phenotypes of pediatric patients with anti-3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR) myopathy to explore its diagnostic strategies.Methods:The clinical features of 10 pediatric patients with anti-HMGCR myopathy in the Department of Neurology, Peking University First Hospital from July 2014 to July 2021 were collected. Muscle biopsies were performed in all patients, with histological, enzymatic histochemical and immunohistochemical staining.Results:The male to female ratio was 6∶4, the age of onset was 3-16 (8.3±3.7) years, 2 cases had subacute onset, and 8 cases experienced chronic progressive onset. All patients presented with neck and proximal muscular weakness of all limbs. Skin rash was observed in 2 cases. Serum creatine kinase was 998-27 981 U/L. The electromyography results were available from 6 cases, who experienced myogenic changes. The muscle magnetic resonance imaging was performed in 5 cases and revealed muscle edema predominantly in posterior compartment of thigh, with mild fatty infiltrate in 2 cases. An initial diagnosis was limb-girdle muscular dystrophy in 7 cases, but with subsequently negative genetic testing. Muscle biopsies revealed scattered necrotic fibers and regenerating fibers, complement deposition in sarcolemma basement-membrane areas of non-necrotic fibers and a few of lymphocyte infiltrate in all specimens. Moreover, a high frequency of major histocompatibility complex Ⅰ expression in muscle fibers was observed in 9 cases, proliferation of connective tissue of endomysium in 8 cases, muscle fiber hypertrophy in 4 cases and vacuoles in 2 cases.Conclusions:Pediatric anti-HMGCR myopathy is frequently misdiagnosed as muscular dystrophy. Systematic consideration of anti-HMGCR myopathy and testing for myositis specific antibody in children with genetically unconfirmed muscular dystrophy may help the differential diagnosis.

Purpose: Anterior gradient 3 (AGR3) belongs to human anterior gradient (AGR) family. The function of AGR3 on cancer remains unknown. This research aimed to investigate if AGR3 had prognostic values in invasive ductal carcinoma (IDC) of breast cancer and could promote tumor progression. Materials and Methods: AGR3 expression was detected in breast benign lesions, ductal carcinoma in situ and IDC by immunohistochemistry analysis. AGR3’s correlations with clinicopathological features and prognosis of IDC patients were analyzed. By cell function experiments, collagen gel droplet-embedded culture drug sensitivity test and cytotoxic analysis, AGR3’s impacts on proliferation, invasion ability, and chemotherapeutic drug sensitivity of breast cancer cells were also detected. Results: AGR3 was up-regulated in luminal subtype of histological grade I-II of IDC patients and positively correlated with high risks of recurrence and distant metastasis. AGR3 high expression could lead to bone or liver metastasis and predict poor prognosis of luminal B. In cell lines, AGR3 could promote proliferation and invasion ability of breast cancer cells which were consistent with clinical analysis. Besides, AGR3 could indicate poor prognosis of breast cancer patients treated with taxane but a favorable prognosis with 5-fluoropyrimidines. And breast cancer cells with AGR3 high expression were resistant to taxane but sensitive to 5-fluoropyrimidines. Conclusion AGR3 might be a potential prognostic indicator in luminal B subtype of IDC patients of histological grade I-II. And patients with AGR3 high expression should be treated with chemotherapy regimens consisting of 5-fluoropyrimidines but no taxane.

Objective:To assess the impact of vaccination at school and influenza vaccination rates among school-age children on school absenteeism in Shenzhen.Methods:The study subjects were primary school students in Shenzhen. School absenteeism panel database from December 2017 to June 2020 of 286 primary schools in Shenzhen was merged with vaccination rates and organizational patterns ( i.e., vaccination at school vs. non-school) data of 9 districts in Shenzhen after influenza vaccination for children. The outcome was the number of school absenteeism. The treatment and control groups were distinguished by organizational patterns and district vaccination rates. Difference-in-Difference (DiD) Poisson regressions were used to analyze the effectiveness of vaccination at school and higher vaccination rates. Besides, a robustness test was performed on the regression results. Results:Poisson regression analysis and robustness test of regression results showed that vaccination at school and higher vaccination rates effectively reduced the risk of school absenteeism, with effectiveness against absenteeism of 32.6% (95% CI: 17.0%-45.3%, P<0.01) and 53.0% (95% CI: 42.1%-61.8%, P<0.01), respectively. Conclusion:A free influenza vaccination program for school-age children in Shenzhen and prioritizing school-based vaccination may be an effective measure to reduce the risk of school absenteeism.