1.Therapeutic effect of concentrated growth factors combined with self-curing calcium phosphate cement on periodontal intrabony defects: Clinical and radiographic evaluation.
Xinying WANG ; Xueyuan CHENG ; Yong ZHANG ; Fei LI ; Jinyu DUAN ; Jing QIAO
Journal of Peking University(Health Sciences) 2025;57(1):42-50
OBJECTIVE:
To clarify the role of concentrated growth factors (CGF) in the treatment of periodontal cement defects using calcium phosphate cement (CPC) with self-curing properties.
METHODS:
Thirty-six intrabony defects were randomly divided into two groups. The experimental group received CGF+CPC treatment (n=18), while the control group received CPC treatment alone (n=18). The probing depth, clinical attachment loss, and hard tissue filling as measured by cone beam CT (CBCT) were evaluated at baseline and 1 year postoperatively in both groups, and the levels of major growth factors in CGF and serum were compared [platelet-derived growth factor-BB (PDGF-BB), transforming growth factor-β1 (TGF-β1), insulin-like growth factor-1 (IGF-1), and vascular endothelial growth factor (VEGF)].
RESULTS:
At baseline, there were no statistically significant differences in probing depth, clinical attachment loss and CBCT measurements between the two groups (P>0.05). At 1 year postoperatively, significant improvements were observed in parameters mentioned above in both groups (P < 0.05). The CGF+CPC group seemed more effective compared with the CPC group in reduction of probing depth [(4.5±1.3) mm vs. (3.2±1.1) mm] and clinical attachment gain [(3.8±0.9) mm vs. (2.0±0.5) mm, P < 0.05]. Compared with the group treated with CPC alone, the hard tissue filling degree shown by CBCT in the CGF+CPC group was significantly increased [the reduction of the depth of the intrabony defects was (3.9±1.2) mm vs. (2.1±0.7) mm, respectively, P < 0.01]. At 1 year post-operatively, the volume of the intrabony defects shown by CBCT in the CGF+CPC group was reduced by (0.031 8±0.004 1) mL, which was significantly more than that in the CPC group [(0.019 7±0.001 2) mL, P < 0.05]. In addition, the concentration of the main growth factors (PDGF-BB, TGF-β1, IGF-1, and VEGF) in CGF were higher than those in serum (P < 0.001).
CONCLUSION
After 1 year of follow-up, the results of the present study indicated that CGF could significantly improve the clinical and radiological effects of CPC on the treatment of periodontal intrabony defects.
Humans
;
Calcium Phosphates/therapeutic use*
;
Male
;
Female
;
Bone Cements/therapeutic use*
;
Middle Aged
;
Cone-Beam Computed Tomography
;
Alveolar Bone Loss/therapy*
;
Becaplermin
;
Adult
;
Insulin-Like Growth Factor I
;
Intercellular Signaling Peptides and Proteins/therapeutic use*
;
Proto-Oncogene Proteins c-sis/blood*
;
Transforming Growth Factor beta1/blood*
;
Vascular Endothelial Growth Factor A/blood*
2.Dual rheumatoid factor and anti-cyclic citrullinated peptide antibody positivity affects the manifestations of rheumatoid arthritis.
Li Huan Angela Marie CHAN ; Khai Pang LEONG ; Justina Wei Lynn TAN ; Xiao GAO ; Wei Qiang SEE ; Ee Tzun KOH
Singapore medical journal 2025;66(9):486-491
INTRODUCTION:
Rheumatoid factor (RF) and anti-cyclic citrullinated peptide antibody (ACPA) are used in the diagnosis and prognostication of rheumatoid arthritis (RA). We wanted to determine the specific contributions of RF and ACPA to the biological nature of RA and whether they act synergistically.
METHODS:
We identified 731 patients from our prospective multi-ethnic RA cohort and categorised them into four groups: ACPA-positive, RF-positive, doubly positive and doubly negative. We compared the demographics, Disease Activity Score-28, Health Assessment Questionnaire score, quality of life using Short Form 36 and the use of prednisolone and disease-modifying antirheumatic drugs (DMARDs) of these patient groups.
RESULTS:
Four hundred and ninety-one patients (67.2%) were ACPA+RF+, 54 (7.4%) were ACPA+RF-, 82 (11.2%) were ACPA-RF+ and 104 (14.2%) were ACPA-RF-. Mean disease duration before the study entry was not different in the four groups. Patients with older age of onset were less likely to be positive for RF and ACPA. Fewer ACPA+RF+ patients were in remission compared to those in the other groups ( P < 0.05). Erythrocyte sedimentation rate (ESR) was higher at study entry in the ACPA+RF+ group (40.4 mm/h vs. 30.6-30.9 mm/h, P < 0.05). Prednisolone and number of DMARDs used were higher in the ACPA+RF+ group compared to the doubly negative group. There were no differences in the functional status and quality of life.
CONCLUSIONS
RA patients who were positive for both ACPA and RF had lower remission rate, higher baseline ESR and required more corticosteroid and DMARD treatment compared to those who were singly positive or doubly negative. Being doubly positive confers a worse outcome to RA patients.
Humans
;
Arthritis, Rheumatoid/diagnosis*
;
Male
;
Female
;
Middle Aged
;
Rheumatoid Factor/blood*
;
Anti-Citrullinated Protein Antibodies/blood*
;
Adult
;
Quality of Life
;
Prospective Studies
;
Antirheumatic Agents/therapeutic use*
;
Aged
;
Peptides, Cyclic/immunology*
;
Prednisolone/therapeutic use*
;
Surveys and Questionnaires
;
Severity of Illness Index
;
Prognosis
3.Changes in circulating levels of calcium and bone metabolism biochemical markers in patients receiving denosumab treatment.
Yuancheng CHEN ; Wen WU ; Ling XU ; Haiou DENG ; Ruixue WANG ; Qianwen HUANG ; Liping XUAN ; Xueying CHEN ; Ximei ZHI
Journal of Southern Medical University 2025;45(4):760-764
OBJECTIVES:
To investigate the changes in blood levels of calcium and bone metabolism biochemical markers in patients with primary osteoporosis receiving treatment with denosumab.
METHODS:
Seventy-three patients with primary osteoporosis treated in our Department between December, 2021 and December 2023 were enrolled. All the patients were treated with calcium supplements, vitamin D and calcitriol in addition to regular denosumab treatment every 6 months. Blood calcium, parathyroid hormone (PTH), osteocalcin (OC), type I procollagen amino-terminal propeptide (PINP), and type I collagen carboxy-terminal telopeptide β special sequence (β‑CTX) data before and at 3, 6, 9, and 12 months after the first treatment were collected from each patient.
RESULTS:
Three months after the first denosumab treatment, the bone turnover markers (BTMs) OC, PINP, and β-CTX were significantly decreased compared to their baseline levels by 39.5% (P<0.001), 56.2% (P<0.001), and 81.8% (P<0.001), respectively. At 6, 9, and 12 months of treatment, OC, PINP, and β-CTX remained significantly lower than their baseline levels (P<0.001). Blood calcium level was decreased (P<0.05) and PTH level increased (P<0.05) significantly in these patients at months of denosumab treatment, but their levels were comparable to the baseline levels at 6, 9, and 12 months of the treatment (P>0.05).
CONCLUSIONS
Denosumab can suppress BTMs and has a good therapeutic effect in patients with primary osteoporosis, but reduction of blood calcium and elevation of PTH levels can occur during the first 3 months in spite of calcium supplementation. Blood calcium and PTH levels can recover the baseline levels as the treatment extended, suggesting the importance of monitoring blood calcium and PTH levels during denosumab treatment.
Humans
;
Denosumab/therapeutic use*
;
Calcium/blood*
;
Parathyroid Hormone/blood*
;
Biomarkers/blood*
;
Osteoporosis/blood*
;
Osteocalcin/blood*
;
Procollagen/blood*
;
Female
;
Collagen Type I/blood*
;
Peptide Fragments/blood*
;
Bone Density Conservation Agents/therapeutic use*
;
Bone and Bones/metabolism*
;
Male
;
Middle Aged
;
Vitamin D
;
Peptides/blood*
;
Aged
4.Apelin promotes proliferation, migration, and angiogenesis in bladder cancer by activating the FGF2/FGFR1 pathway.
Wei SU ; Houhua LAI ; Xin TANG ; Qun ZHOU ; Yachun TANG ; Hao FU ; Xuancai CHEN
Journal of Southern Medical University 2025;45(6):1289-1296
OBJECTIVES:
To investigate the role of apelin in regulating proliferation, migration and angiogenesis of bladder cancer cells and the possible regulatory mechanism.
METHODS:
GEO database was used to screen the differentially expressed genes in bladder cancer tissues and cells. Bladder cancer and paired adjacent tissues were collected from 60 patients for analysis of apelin expressions in relation to clinicopathological parameters. In cultured bladder cancer J82 cells and human umbilical vein endothelial cells (HUVECs), the effects of transfection with an apelin-overexpressing plasmid or specific siRNAs targeting apelin, fibroblast growth factor 2 (FGF2) and fibroblast growth factor receptor 1 (FGFR1) on proliferation and migration of J82 cells and tube formation in HUVECs were examined using plate cloning assay, Transwell assay, and angiogenesis assay; the changes in FGF2 expression and FGFR1 phosphorylation were detected using Western blotting.
RESULTS:
The expression level of apelin was significantly higher in bladder cancer tissues than adjacent tissues, and bladder cancer cell lines (T24 and J82) also expressed higher mRNA and protein levels of apelin than SV-HUC-1 cells. Apelin expression level in bladder cancer tissues was correlated with tumor invasion, distant metastasis and advanced TNM stages. Apelin knockdown significantly suppressed proliferation and migration of J82 cells and decreased the total angiogenic length of HUVECs. In contrast, apelin overexpression significantly promoted proliferation and migration and enhanced FGFR1 phosphorylation in J82 cells, and increased the total angiogenesis length in HUVECs, but this effects were effectively mitigated by transfection of the cells with FGF2 siRNA or FGFR1 siRNA.
CONCLUSIONS
High expression of apelin promotes J82 cell proliferation and migration and HUVEC angiogenesis by promoting activation of the FGF2/FGFR1 pathway.
Humans
;
Urinary Bladder Neoplasms/blood supply*
;
Receptor, Fibroblast Growth Factor, Type 1/metabolism*
;
Cell Proliferation
;
Cell Movement
;
Fibroblast Growth Factor 2/metabolism*
;
Neovascularization, Pathologic
;
Human Umbilical Vein Endothelial Cells
;
Cell Line, Tumor
;
Signal Transduction
;
Apelin
;
Intercellular Signaling Peptides and Proteins/genetics*
;
Female
;
Male
;
Angiogenesis
5.Correlation between serum ghrelin and liver-expressed antimicrobial peptide-2 with idiopathic short stature in children.
Qing LIU ; Wei-Chun ZHANG ; Bo CHEN ; Ya-Wen SONG
Chinese Journal of Contemporary Pediatrics 2024;26(12):1261-1266
OBJECTIVES:
To investigate the expression levels of ghrelin and liver-expressed antimicrobial peptide-2 (LEAP-2) in children with idiopathic short stature (ISS) to provide reference for further understanding the etiology of short stature.
METHODS:
A prospective study was conducted from December 2021 to October 2023, involving 46 children diagnosed with ISS (ISS group) and 46 healthy children with normal height (control group) at the First Affiliated Hospital of Shihezi University. General data and serum levels of ghrelin and LEAP-2 were compared between the two groups. The predictive value of these two indicators for ISS was evaluated using receiver operating characteristic (ROC) curve analysis.
RESULTS:
The serum level of ghrelin in the ISS group was higher than that in the control group, while the level of LEAP-2 was lower (P<0.05). The ratio of LEAP-2 to ghrelin was lower in the ISS group compared to the control group (P<0.05). Multivariate logistic regression analysis showed that HtSDS, IGF-1, ghrelin, LEAP-2, and the ratio of LEAP-2/ghrelin were independently associated with the occurrence of ISS (P<0.05). ROC curve analysis indicated that the AUCs for ghrelin, LEAP-2, the ratio of ghrelin to LEAP-2, and their combination in predicting ISS were all >0.8. The optimal cutoff values for ghrelin, LEAP-2, and the LEAP-2/ghrelin ratio were 5 607 pg/mL, 1 155 pg/mL, and 0.212, respectively. In children with ISS, ghrelin showed a negative correlation with chronological age, LEAP-2, and the LEAP-2/ghrelin ratio (P<0.05), while it was positively correlated with growth rate and peak growth hormone levels (P<0.05). LEAP-2 was negatively correlated with growth rate, peak growth hormone levels, and ghrelin (P<0.05), but positively correlated with chronological age and the LEAP-2/ghrelin ratio (P<0.05).
CONCLUSIONS
Ghrelin and LEAP-2 are correlated with the occurrence of ISS, which may provide references for the diagnosis and etiological analysis of children with ISS.
Humans
;
Ghrelin/blood*
;
Male
;
Child
;
Female
;
Prospective Studies
;
Child, Preschool
;
Growth Disorders/etiology*
;
Antimicrobial Cationic Peptides/blood*
;
Body Height
;
Adolescent
;
Insulin-Like Growth Factor I/analysis*
;
Blood Proteins
6.Predictive value of plasma heparin-binding protein combined with albumin for 28-day mortality in patients with sepsis.
Jiangping LIU ; Yajun LI ; Yawen ZHENG ; Cuijie ZHANG ; Lihua HUANG ; Xiaopeng NING ; Wenfei WANG ; Qingli DOU
Chinese Critical Care Medicine 2024;36(12):1233-1237
OBJECTIVE:
To evaluate the predictive value of plasma heparin-binding protein (HBP) combined with albumin (Alb) for predicting 28-day mortality in patients with sepsis.
METHODS:
The clinical data of patients with sepsis admitted to the emergency intensive care unit (EICU) of the People's Hospital of Shenzhen Baoan District from March 2020 to March 2024 were retrospectively analyzed. The study began at the time of the first diagnosis of sepsis upon EICU admission and ended upon patient death or at 28 days. The gender, age, length of stay in EICU, underlying diseases, and infection sites were recorded. Within 24 hours of sepsis diagnosis, blood culture results, white blood cell count (WBC), C-reactive protein (CRP), procalcitonin (PCT), blood lactate acid (Lac), HBP, Alb, sequential organ failure assessment (SOFA), acute physiology and chronic health evaluation II (APACHE II), mortality in emergency department sepsis score (MEDS), modified early warning score (MEWS), number of organ failures, use of vasopressors, application of mechanical ventilation, renal replacement therapy, and 28-day prognosis were recorded, the differences in these indicators between two groups were compared. Univariate and multivariate Logistic regression analyses were used to analyze the risk factors of 28-day mortality in patients with sepsis. Receiver operator characteristic curve (ROC curve) was drawn, and the area under the ROC curve (AUC) was calculated to evaluate the early predictive value of various risk factors for 28-day mortality in patients with sepsis.
RESULTS:
A total of 300 patients with sepsis were included, with 16 excluded, resulting in 284 patients being analyzed. Among them, 191 survived and 93 died within 28 days. There were no statistically significant differences between the two groups in terms of gender, age, underlying diseases, infection sites, blood culture positivity rate, number of organ failures, and length of stay in EICU. Univariate analysis showed that the rate of vasopressor use, the rate of mechanical ventilation, HBP, PCT, CRP, Lac, SOFA score, APACHE II score, MEDS score, and MEWS score were significantly higher in the death group than those in the survival group, while Alb was significantly lower in the death group than that in the survival group. Multivariate Logistic regression analysis showed that HBP and Alb were independent risk factors for predicting 28-day mortality in patients with sepsis [odds ratio (OR) and 95% confidence interval (95%CI) were 1.093 (0.989-1.128) and 1.174 (1.095-1.259), both P < 0.05]. ROC curve analysis showed that both HBP and Alb had certain predictive value for 28-day mortality in patients with sepsis [AUC and 95%CI were 0.820 (0.717-0.923) and 0.786 (0.682-0.890), both P < 0.05]. When the critical value of HBP was 117.50 μg/L, the sensitivity was 85.90%, and the specificity was 70.50%. When the critical value of Alb was 28.30 g/L, the sensitivity was 69.30%, and the specificity was 81.20%. When the two indexes were combined for diagnosis, the AUC was 0.881 (95%CI was 0.817-0.945, P < 0.001), the sensitivity was 92.70%, and the specificity was 76.80%.
CONCLUSIONS
HBP and Alb are independent risk factors for predicting 28-day mortality in patients with sepsis. The combined prediction efficiency of HBP and Alb for 28-day mortality in patients with sepsis is superior to a single indicator.
Humans
;
Sepsis/diagnosis*
;
Retrospective Studies
;
Predictive Value of Tests
;
Intensive Care Units
;
Blood Proteins/analysis*
;
Prognosis
;
Antimicrobial Cationic Peptides/blood*
;
APACHE
;
Male
;
Female
;
Organ Dysfunction Scores
;
ROC Curve
;
Middle Aged
;
C-Reactive Protein/analysis*
;
Emergency Service, Hospital
;
Aged
;
Hospital Mortality
;
Serum Albumin/analysis*
7.Electroacupuncture Improves Blood-Brain Barrier and Hippocampal Neuroinflammation in SAMP8 Mice by Inhibiting HMGB1/TLR4 and RAGE/NADPH Signaling Pathways.
Yuan WANG ; Qiang WANG ; Di LUO ; Pu ZHAO ; Sha-Sha ZHONG ; Biao DAI ; Jia-Jyu WANG ; Yi-Tong WAN ; Zhi-Bin LIU ; Huan YANG
Chinese journal of integrative medicine 2023;29(5):448-458
OBJECTIVE:
To investigate the molecular mechanisms underlying the beneficial effect of electroacupuncture (EA) in experimental models of Alzheimer's disease (AD) in vivo.
METHODS:
Senescence-accelerated mouse prone 8 (SAMP8) mice were used as AD models and received EA at Yingxiang (LI 20, bilateral) and Yintang (GV 29) points for 20 days. For certain experiments, SAMP8 mice were injected intravenously with human fibrin (2 mg). The Morris water maze test was used to assess cognitive and memory abilities. The changes of tight junctions of blood-brain barrier (BBB) in mice were observed by transmission electron microscope. The expressions of fibrin, amyloid- β (Aβ), and ionized calcium-binding adapter molecule 1 (IBa-1) in mouse hippocampus (CA1/CA3) were detected by reverse transcription-quantitative polymerase chain reaction (qRT-PCR), Western blot or immunohistochemical staining. The expression of fibrin in mouse plasma was detected by enzyme-linked immunosorbent assay. The expressions of tight junction proteins zonula occludens-1 and claudin-5 in hippocampus were detected by qRT-PCR and immunofluorescence staining. Apoptosis of hippocampal neurons was detected by terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining.
RESULTS:
Fibrin was time-dependently deposited in the hippocampus of SAMP8 mice and this was inhibited by EA treatment (P<0.05 or P<0.01). Furthermore, EA treatment suppressed the accumulation of Aβ in the hippocampus of SAMP8 mice (P<0.01), which was reversed by fibrin injection (P<0.05 or P<0.01). EA improved SAMP8 mice cognitive impairment and BBB permeability (P<0.05 or P<0.01). Moreover, EA decreased reactive oxygen species levels and neuroinflammation in the hippocampus of SAMP8 mice, which was reversed by fibrin injection (P<0.05 or P<0.01). Mechanistically, EA inhibited the promoting effect of fibrin on the high mobility group box protein 1 (HMGB1)/toll-like receptor 4 (TLR4) and receptor for advanced glycation end products (RAGE)/nicotinamide adenine dinucleotide phosphate (NADPH) signaling pathways (P<0.01).
CONCLUSION
EA may potentially improve cognitive impairment in AD via inhibition of fibrin/A β deposition and deactivation of the HMGB1/TLR4 and RAGE/NADPH signaling pathways.
Mice
;
Humans
;
Animals
;
NADP/metabolism*
;
Toll-Like Receptor 4
;
HMGB1 Protein/metabolism*
;
Receptor for Advanced Glycation End Products/metabolism*
;
Blood-Brain Barrier/metabolism*
;
Neuroinflammatory Diseases
;
Electroacupuncture
;
Alzheimer Disease/therapy*
;
Hippocampus/metabolism*
;
Amyloid beta-Peptides/metabolism*
8.Value of heparin-binding protein in the diagnosis of severe infection in children: a prospective study.
Jun-Chao DENG ; Fang-Li ZHAO ; Li-Na QIAO
Chinese Journal of Contemporary Pediatrics 2022;24(1):85-89
OBJECTIVES:
To study the value of heparin-binding protein (HBP) in the diagnosis of severe infection in children.
METHODS:
This study was a prospective observational study. The medical data of children who were admitted to the pediatric intensive care unit due to infection from January 2019 to January 2020 were collected. According to the diagnostic criteria for severe sepsis and sepsis, the children were divided into a severe sepsis group with 49 children, a sepsis group with 82 children, and a non-severe infection group with 33 children. The three groups were compared in terms of related biomarkers such as plasma HBP, serum C-reactive protein, serum procalcitonin, and platelet count. The receiver operating characteristic (ROC) curve was plotted to investigate the value of plasma HBP level in the diagnosis of severe infection (including severe sepsis and sepsis).
RESULTS:
The severe sepsis and sepsis groups had a significantly higher plasma HBP level on admission than the non-severe infection group (P<0.05). Compared with the sepsis and non-severe groups, the severe sepsis group had significantly higher serum levels of C-reactive protein and procalcitonin and a significantly lower platelet count (P<0.05). Plasma HBP level had an area under the ROC curve of 0.590 in determining severe infection, with a sensitivity of 38.0% and a specificity of 82.4% (P<0.05).
CONCLUSIONS
There is an increase in plasma HBP level in children with severe infection, and plasma HBP level has a lower sensitivity but a higher specificity in the diagnosis of severe infection and can thus be used as one of the markers for the judgment of severe infection in children.
Antimicrobial Cationic Peptides
;
Biomarkers
;
Blood Proteins
;
C-Reactive Protein/analysis*
;
Child
;
Humans
;
Procalcitonin
;
Prospective Studies
;
ROC Curve
;
Sepsis/diagnosis*
9.Serum vitamin K2 level and its association with bone metabolism markers in 1 732 children.
Chinese Journal of Contemporary Pediatrics 2022;24(10):1130-1135
OBJECTIVES:
To study the level of serum vitamin K2 (VitK2) and its association with bone metabolism markers osteocalcin (OC), type I procollagen amino-terminal peptide (PINP), and type I collagen carboxy-terminal peptide (CTX) in children.
METHODS:
A prospective analysis was performed on 1 732 children who underwent routine physical examination from October 2020 to October 2021. The serum levels of VitK2 and 25-hydroxy vitamin D [25(OH)D] were measured. According to age, they were divided into four groups: <1 year, 1-3 years group, >3-6 years group, and >6-14 years. A total of 309 children with 25(OH)D≥50 nmol/L were screened out, and serum levels of OC, PINP, and CTX were measured to investigate the correlation of the serum levels of OC, PINP, and CTX with serum VitK2 levels in different age groups.
RESULTS:
The prevalence rate of serum VitK2 deficiency was 52.31% (906/1 732). The VitK2 deficiency group had higher prevalence rates of overweight/obesity and growth pain (≥3 years of age) than the normal VitK2 group (P<0.05). There were differences in the prevalence rate of serum VitK2 deficiency (P<0.0083) and the serum level of VitK2 (P<0.05) between the 1-3 years group and the >6-14 years group. The <1 year group had a higher serum level of CTX and a lower serum level of PINP than the >3-6 years group and the >6-14 years group (P<0.05). The <1 year group had a lower serum level of OC than the >6-14 years group (P<0.05). Serum VitK2 level was positively correlated with OC level (rs=0.347, P<0.01), and CTX level was negatively correlated with PINP level (rs=-0.317, P<0.01).
CONCLUSIONS
Serum VitK2 deficiency may be associated with overweight/obesity. Serum VitK2 may affect the level of OC and even bone health.
Child
;
Humans
;
Infant
;
Biomarkers/metabolism*
;
Collagen Type I/metabolism*
;
Obesity/complications*
;
Osteocalcin/metabolism*
;
Overweight/complications*
;
Peptide Fragments/metabolism*
;
Peptides/metabolism*
;
Procollagen/metabolism*
;
Vitamin K/blood*
;
Child, Preschool
;
Adolescent
;
Bone and Bones/metabolism*
10.Changes of heparin-binding protein in severe burn patients during shock stage and its effects on human umbilical vein endothelial cells and neutrophils.
Xin Xin QI ; Lu LIU ; Yun Xi YANG ; Jia Min HUANG ; Bing Wei SUN
Chinese Journal of Burns 2022;38(2):147-155
Objective: To investigate the changes of heparin-binding protein (HBP) in severe burn patients during shock stage and its effects on human umbilical vein endothelial cells (HUVECs) and neutrophils in vitro. Methods: Prospective observational and experimental research methods were used. Twenty severe burn patients who met the inclusion criteria and were admitted to the Department of Burns and Plastic Surgery of Affiliated Suzhou Hospital of Nanjing Medical University from August to November 2020 were included in severe burn group (12 males and 8 females, aged 44.5 (31.0, 58.0) years). During the same period, 20 healthy volunteers with normal physical examination results in the unit's Physical Examination Center were recruited into healthy control group (13 males and 7 females, aged 39.5 (26.0, 53.0) years). Enzyme-linked immunosorbent assay (ELISA) method was used to detect the protein expression levels of HBP and tissue inhibitor of metalloproteinase 1 (TIMP-1) in plasma of patients within 48 hours after injury in severe burn group and in plasma of volunteers in healthy control group. The correlation between protein expression of HBP and that of TIMP-1 in the plasma in the two groups was analyzed by Pearson correlation analysis. The fourth passage of HUVECs in logarithmic growth phase were used for the experiment. The HUVECs were divided into normal control group with routine culture (the same treatment below) and recombinant HBP (rHBP)-treated 12 h group, rHBP-treated 24 h group, and rHBP-treated 48 h group with corresponding treatment according to the random number table (the same grouping method below), and the mRNA expression of TIMP-1 in cells was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction. The HUVECs were divided into normal control group and rHBP-treated 48 h group with corresponding treatment, and the protein expression of TIMP-1 in the cells was detected by Western blotting. The HUVECs were divided into normal control group, rHBP alone group, aprotinin alone group, and rHBP+aprotinin group treated with the corresponding reagents (with the final molarity of rHBP being 200 nmol/L and the final concentration of aprotinin being 20 μg/mL, respectively), cultured for 48 h, and ELISA was used to detect the protein expression of TIMP-1 in the culture supernatant of cells. The neutrophils were isolated from the peripheral venous blood of the aforementioned 10 healthy volunteers by immunomagnetic bead sorting, and the cells were divided into normal control group, recombinant TIMP-1 (rTIMP-1) alone group, phorbol acetate (PMA) alone group, and rTIMP-1+PMA group treated with corresponding reagents (with the final concentration of rTIMP-1 being 500 ng/mL and the final molarity of PMA being 10 nmol/L, respectively). After being cultured for 1 h, the expression of CD63 protein in cells was detected by immunofluorescence method, the positive expression rate of CD63 protein in cells was detected by flow cytometry, and the protein expression levels of HBP and myeloperoxidase (MPO) in the culture supernatant of cells were detected by ELISA. The normal control group underwent the above-mentioned related tests at appropriate time points. The number of samples was 3 in each group of cell experiment. Data were statistically analyzed with chi-square test, Mann-Whitney U test, Kruskal-Wallis H test, and Tamhane's T2 test. Results: The protein expression levels of HBP and TIMP-1 in the plasma of patients in severe burn group were 404.9 (283.1, 653.2) and 262.1 (240.6, 317.4) ng/mL, respectively, which were both significantly higher than 61.6 (45.0, 68.9) and 81.0 (66.3, 90.0) ng/mL of volunteers in healthy control group (with Z values of -5.41 and -5.21, respectively, P<0.01). The correlation between the protein expression of HBP and that of TIMP-1 in the plasma of volunteers in healthy control group was not strong (P>0.05). The protein expression of HBP was significantly positively correlated with that of TIMP-1 in the plasma of patients in severe burn group (r=0.64, P<0.01). Compared with that in normal control group, the mRNA expression of TIMP-1 in HUVECs was significantly increased in rHBP-treated 12 h group, rHBP-treated 24 h group, and rHBP-treated 48 h group (with t values of -3.58, -2.25, and -1.26, respectively, P<0.05). Western blotting detection showed that compared with that in normal control group, the protein expression of TIMP-1 in HUVECs in rHBP-treated 48 h group was significantly enhanced. After 48 h of culture, compared with that in normal control group, the protein expression level of TIMP-1 in the culture supernatant of HUVECs in rHBP alone group was significantly increased (t=9.43, P<0.05), while the protein expression level of TIMP-1 in the culture supernatant of HUVECs didn't change significantly in aprotinin alone group or rHBP+aprotinin group (P>0.05); compared with that in rHBP alone group, the protein expression level of TIMP-1 in the culture supernatant of HUVECs in rHBP+aprotinin group was significantly decreased (t=4.76, P<0.01). After 1 h of culture, the trend of CD63 protein expression in neutrophils detected by immunofluorescence method and that by flow cytometry were consistent in each group. After 1 h of culture, compared with that in normal control group, the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells in rTIMP-1 alone group all had no significant changes (P>0.05), while the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells were all significantly increased in PMA alone group and rTIMP-1+PMA group (with t values of 2.41, 3.82, 5.73, 1.05, 4.16, and 1.08, respectively, P<0.05 or P<0.01); compared with that in PMA alone group, the positive expression rate of CD63 protein in the neutrophils and the protein expression levels of HBP and MPO in the culture supernatant of cells in rTIMP-1+PMA group were all significantly decreased (with t values of 5.26, 2.83, and 1.26, respectively, P<0.05 or P<0.01). Conclusions: The expression level of HBP in the plasma of severe burn patients is increased during shock stage. HBP can induce HUVECs to secrete TIMP-1 in vitro, and TIMP-1 can reduce the expression of CD63 molecule in human neutrophils.
Adult
;
Antimicrobial Cationic Peptides
;
Blood Proteins
;
Burns
;
Female
;
Human Umbilical Vein Endothelial Cells
;
Humans
;
Male
;
Neutrophils
;
Tissue Inhibitor of Metalloproteinase-1

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