1.Interaction between fibroblasts and keratinocytes in the wound edge skin tissue of a diabetic foot patient and the mechanism
Qiongfang RUAN ; Siyu ZHANG ; Maomao XI ; Jingjing RUAN ; Shuhua LIU ; Binghui LI ; Weiguo XIE
Chinese Journal of Burns 2024;40(8):762-771
Objective:To investigate the interaction between fibroblasts (Fb) and keratinocytes (KC) in the wound edge skin tissue of a diabetic foot patient and the mechanism.Methods:This was an experimental research. The wound edge skin tissue from a diabetic foot patient (male and 33 years old) admitted to the Department of Wound Repair of Liyuan Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology in August 2021 and from an acute foot injury patient (male and 50 years old) admitted to the Department of Hand Surgery of the hospital in September 2021 was collected. The single-cell transcriptome sequencing was performed to analyze the interaction between chemokine ligands of Fb subgroup and chemokine receptors of KC subgroup. The supernatant was collected after human foreskin fibroblast (HFF) was cultured routinely and with high concentration of glucose for 7 days as normal conditioned medium (CM) and high glucose CM, respectively. HaCaT cells were collected and divided into normal CM group cultured with normal CM and high glucose CM group cultured with high glucose CM, the scratch test was performed to calculate the cell migration rates at 24 and 48 h after scratch ( n=3). The content of cytokines in the two kinds of CM was detected by liquid suspension chip ( n=5). HFF was collected and divided into normal group cultured routinely and high glucose group cultured with high concentration of glucose for 7 days, and the mRNA expressions of C-X-C motif chemokine ligand 1 (CXCL1), CXCL2, CXCL8, and CXCL12 were detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction ( n=6). HaCaT cells in normal CM group and high glucose CM group were collected to detect the protein expressions of C-X-C motif chemokine receptor 4 (CXCR4) in cells cultured for 48 h by Western blotting ( n=3). HaCaT cells were collected and divided into normal CM group, high glucose CM group, normal CM+CXCL12 group, and high glucose CM+CXCL12 group. The first two groups of cells were treated as before, and the latter two groups of cells were cultured with normal CM and high glucose CM containing recombinant human CXCL12, respectively. Scratch test was performed, and cell migration rates were calculated at 24 and 48 h after scratch ( n=3); the protein expression of CXCR4 in cells cultured for 48 h was detected by Western blotting ( n=3). Results:Compared with those in the wound edge skin tissue of acute foot injury, the interactions between chemokine ligands (CXCL1, CXCL2, CXCL3, CXCL8, and CXCL12) of Fb subgroup and chemokine receptors (CXCR2 and CXCR4) of KC subgroup were significantly weakened in the wound edge skin tissue of diabetic foot. At 24 and 48 h after scratch, the migration rates of HaCaT cells in high glucose CM group were significantly lower than those in normal CM group (with t values of 23.50 and 15.65, respectively, P<0.05). Compared with that in normal CM, the content of CXCL1 in high glucose CM was significantly increased ( P<0.05), and the content of CXCL12 was significantly decreased ( P<0.05). After 7 days of culture, compared with those in normal group, the mRNA expressions of CXCL1, CXCL2, and CXCL8 in HFF in high glucose group were significantly increased (with t values of 4.25, 4.98, and 10.04, respectively, P<0.05), while the mRNA expression of CXCL12 was significantly decreased ( t=4.10, P<0.05). After 48 h of culture, the CXCR4 protein expression in HaCaT cells in high glucose CM group was significantly lower than that in normal CM group ( t= 5.13, P<0.05). At 24 and 48 h after scratch, the migration rates of HaCaT cells in high glucose CM group were significantly lower than those in normal CM group and high glucose CM+CXCL12 group (with P values all <0.05); at 24 h after scratch, the migration rate of HaCaT cells in normal CM+CXCL12 group was significantly lower than that in normal CM group ( P<0.05); at 48 h after scratch, the migration rate of HaCaT cells in normal CM+CXCL12 group was significantly higher than that in high glucose CM+CXCL12 group ( P<0.05). At 48 h of culture, the CXCR4 protein expression of HaCaT cells in high glucose CM+CXCL12 group was 0.446±0.050, which was significantly higher than 0.247±0.010 in high glucose CM group ( P<0.05) and similar to 0.522±0.082 in normal CM+CXCL12 group ( P>0.05); the CXCR4 protein expression in HaCaT cells in normal CM group was 0.509±0.055, which was significantly higher than that in high glucose CM group ( P<0.05). Conclusions:The interactions between chemokine ligands of Fb subgroup and chemokine receptors of KC subgroup were significantly weakened in the wound edge skin tissue of diabetic foot. High glucose can inhibit CXCL12 secretion of HFF, and the stimulation of its cell culture supernatant can decrease HaCaT cell migration ability and CXCR4 expression. Exogenous CXCL12 protein can increase the CXCR4 protein expression in HaCaT cells and enhance the cell migration ability.
2.Risk factors of early myocardial injury and the impact of early myocardial injury on prognosis of patients with extensive burns
Nuo CHEN ; Maomao XI ; Qiongfang RUAN ; Zhigang CHU ; Wei ZHANG ; Jinli ZHANG ; Weiguo XIE
Chinese Journal of Burns 2023;39(5):417-423
Objective:To analyze the risk factors of early myocardial injury and the impact of early myocardial injury on prognosis of patients with extensive burns.Methods:A retrospective case series study was conducted. From January 2018 to August 2022, 361 patients with extensive burns who met the inclusion criteria were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital, including 231 males and 130 females, aged 50 (36, 58) years, with total burn area of 45% (35%, 60%) total body surface area. According to the highest level of creatine kinase isoenzyme-MB (CK-MB) within 72 h post injury, the patients were divided into early myocardial injury group (CK-MB≥75 U/L, 182 patients) and non-early myocardial injury group (CK-MB<75 U/L, 179 patients). The following data of patients in the 2 groups were collected and analyzed, including gender, age, total burn area, admission time post injury, combination with shock on admission, combination with inhalation injury on admission; the main blood test indexes such as myocardial enzyme spectrum, blood routine, liver and kidney function, and electrolytes within 72 h post injury; and treatment outcomes and fatality rate. Data were statistically analyzed with chi-square test, independent sample t test, or Mann-Whitney U test. The multivariate logistic regression analysis was conducted to screen the independent risk factors for early myocardial injury and for death in patients with extensive burns. Results:There were statistically significant differences in gender, combination with shock on admission, total burn area, and admission time post injury of patients between the two groups (with χ2 values of 6.40 and 6.10, Z values of 5.41 and 3.03, respectively, P<0.05). There were no statistically significant differences in age, combination with inhalation injury on admission of patients between the two groups ( P>0.05). The CK-MB, creatine kinase, lactate dehydrogenase, α-hydroxybutyrate dehydrogenase, white blood cell count, neutrophil-to-lymphocyte ratio (NLR), alanine aminotransferase (ALT), aspartate aminotransferase, potassium, and hemoglobin within 72 h post injury were significantly higher than those in non-early myocardial injury group (with Z values of 15.40, 6.26, 7.59, 7.02, 2.64, 4.53, 4.07, 6.32, and 4.12, t=2.34, respectively, P<0.05), while the level of calcium was significantly lower than that in non-early myocardial injury group ( Z=2.72, P<0.05). There were no statistically significant differences in other blood test indexes of patients between the two groups ( P>0.05). The total burn area, admission time post injury, NLR and ALT within 72 h post injury were the independent risk factors for early myocardial injury in patients with extensive burns (with odds ratios of 1.03, 1.07, 1.04, and 1.02, 95% confidence intervals of 1.02-1.05, 1.00-1.11, 1.02-1.07, and 1.00-1.03, respectively, P<0.05). The fatality rate of patients in early myocardial injury group was 8.8% (16/182), which was significantly higher than 2.8% (5/179) in non-early myocardial injury group ( χ2 =5.93, P<0.05). Early myocardial injury, age, combination with shock on admission, and combination with inhalation injury on admission were the independent risk factors for death in patients with extensive burns (with odds ratios of 3.60, 1.04, 6.53, and 3.14, 95% confidence intervals of 1.17-11.05, 1.01-1.07, 1.39-30.68, and 1.15-8.56, respectively, P<0.05). Conclusions:The total burn area, admission time post injury, NLR and ALT within 72 h post injury were the independent risk factors for early myocardial injury in patients with extensive burns. Patients with extensive burns with early myocardial injury have a higher fatality rate, and early myocardial injury is an independent risk factor for the patients' death.
3.Pan-cancer analysis of ubiquitin-specific protease 7 and its expression changes in the carcinogenesis of scar ulcer
Siyu ZHANG ; Jingjing RUAN ; Dongmei JIN ; Nuo CHEN ; Weiguo XIE ; Qiongfang RUAN
Chinese Journal of Burns 2023;39(6):518-526
Objective:To explore the biological role and clinical significance of ubiquitin-specific protease 7 (USP7) in the carcinogenesis of scar ulcer.Methods:A retrospective observational study combined with bioinformatics analysis was used. The RNA expression profile data of USP7 in tumor and/or its corresponding paracancular normal tissue were obtained from The Cancer Genome Atlas (TCGA) database and the Gene Expression Omnibus database, and the RNA sequencing data were transformed by log 2. The variations of USP7 gene were analyzed by cBioPortal database. The USP7 mRNA expression in tumor and adjacent normal tissue in TCGA database were obtained by using the "Gene_DE" module in TIMER 2.0 database. The survival rates of patients with high and low USP7 expression in cutaneous melanoma (SKCM), cervical squamous cell carcinoma (CESC), lung squamous cell carcinoma (LUSC), and head and neck squamous cell carcinoma (HNSC) were analyzed using the Gene Expression Profile Interactive Analysis 2 (GEPIA2) database, and the Kaplan-Meier survival curves were drawn. Sangerbox database was used to analyze the correlation of USP7 expression in pan-cancer with microsatellite instability (MSI) or tumor mutation burden (TMB) pan-cancer. Through the "correlation analysis" module in the GEPIA2 database, the correlation of USP7 expression in pan-cancer with the expression levels of five DNA mismatch repair genes ( MLH1, MSH2, MSH6, PMS2, and EPCAM) and three essential DNA methyltransferases (DNMT)--DNMT1, DNMT3A, and DNMT3B were evaluated. The USP7 expression in CESC, HNSC, LUSC, and SKCM and its correlation with infiltration of immune cells (B cells, CD4 + T cells, CD8 + T cells, neutrophils, macrophages, and dendritic cells) were analyzed by the "Immune-Gene" module in TIMER 2.0 database. The "Similar Genes Detection" module of GEPIA2 database was used to obtain the top 100 protein sets with similar expression patterns to USP7. Intersection analysis was performed between the aforementioned protein sets and the top 50 protein sets that were directly physically bound to USP7 obtained by using the STRING database. Kyoto Encyclopedia of Genes and Genomes (KEGG) and Gene Ontology (GO) enrichment analysis were performed for the two protein sets mentioned above using the DAVID database. The samples of normal skin, hypertrophic scar, scar ulcer, and scar carcinoma with corresponding clinicopathologic features were collected from the Department of Pathology of Tongren Hospital of Wuhan University & Wuhan Third Hospital from October 2018 to October 2022, and the USP7 expression in tissue was detected by immunohistochemical method, with the number of samples of 6. Data were statistically analyzed with Log-rank test, one-way analysis of variance, and Bonferroni test. Results:In pan-cancer, the main gene variations of USP7 were mutation and amplification, and the top 3 tumors with the highest variation frequency (>6%) were bladder urothelial carcinoma, SKCM, and endometrial carcinoma. The main mutation of USP7 gene in pan-cancer was missense mutation. In SKCM with the highest mutation frequency, the main type of mutation was missense mutation in USP7_ICP0_bdg domain. USP7 mRNA expression in breast invasive carcinoma, bile duct carcinoma, colon carcinoma, esophageal carcinoma, HNSC, renal chromophobe cell carcinoma, hepatocellular carcinoma, lung adenocarcinoma, LUSC, prostate carcinoma, and gastric carcinoma was significantly higher than that in corresponding paracancer normal tissue ( P<0.05). USP7 mRNA expression in glioblastoma multiforme, renal clear cell carcinoma, renal papillary cell carcinoma, and thyroid carcinoma was significantly lower than that in corresponding paracancular normal tissue ( P<0.05). In addition, USP7 mRNA expression in SKCM metastases was much higher than that in primary tumor tissue ( P<0.05). Survival curves showed no significant difference in survival rate between patients with high USP7 expression and patients with low USP7 expression in CESC, HNSC, LUSC, and SKCM (Log-rank P>0.05, with hazard ratios of 1.00, 0.99, 1.00, and 1.30, respectively). USP7 expression in colon cancer, colorectal cancer, thymic cancer, and thyroid cancer was negatively correlated with TMB (with Pearson correlation coefficients of -0.26, -0.19, -0.19, and 0.11, respectively, P<0.05). USP7 expression in glioma, CESC, lung adenocarcinoma, mixed renal carcinoma, and LUSC was positively correlated with MSI expression (with Pearson correlation coefficients of 0.22, 0.14, 0.15, 0.08, and 0.14, respectively, P<0.05), and USP7 expression in colon cancer, colorectal cancer, invasive breast cancer, prostate cancer, HNSC, thyroid cancer, and diffuse large B-cell lymphoma were significantly negatively correlated with MSI expression (with Pearson correlation coefficients of -0.31, -0.27, -0.13, -0.19, -0.16, -0.18, and -0.53, respectively, P<0.05). The expression of USP7 in CESC was positively correlated with that of both MSH2 and MSH6 (with Spearman correlation coefficients of 0.51 and 0.44, respectively, P<0.05), and the expression of USP7 in HNSC was positively correlated with the expression of EPCAM, MLH1, MSH2, MSH6, and PMS2 (with Spearman correlation coefficients of 0.39, 0.14, 0.49, 0.54, and 0.41, respectively, P<0.05), and the expression of USP7 in LUSC was positively correlated with the expression of EPCAM, MSH2, MSH6, and PMS2 (with Spearman correlation coefficients of 0.20, 0.36, 0.40, and 0.34, respectively, P<0.05), and the expression of USP7 in SKCM was positively correlated with the expression of EPCAM, MLH1, MSH2, MSH6, and PMS2 (with Spearman correlation coefficients of 0.11, 0.33, 0.42, 0.55, and 0.34, respectively, P<0.05). The expression of USP7 in CESC, HNSC, LUSC, and SKCM was significantly positively correlated with the expression of DNMT1, DNMT3A, and DNMT3B (with Spearman correlation coefficients of 0.42, 0.34, 0.22, 0.45, 0.52, 0.22, 0.36, 0.36, 0.22, 0.38, 0.46, and 0.21, respectively, P<0.05). The expression of USP7 in CESC, HNSC, LUSC, and SKCM was positively correlated with CD4 + T cell infiltration (with Partial correlation coefficients of 0.14, 0.22, 0.13, and 0.16, respectively, P<0.05). Being similar to the pattern of USP7 expression and ranked among top 100 protein sets, the top 5 proteins were C16orf72, BCLAF1, UBN, GSPT1, ERI2 (with Spearman correlation coefficients of 0.83, 0.74, 0.73, and 0.72, respectively, all P values<0.05). The top 50 protein sets that directly physically bind to USP7 overlapped with the aforementioned protein set by only one protein, thyroid hormone receptor interaction factor 12. KEGG enrichment analysis showed that USP7 related genes were involved in cell cycle, spliceosome, cell senescence, and p53 signal pathway. GO enrichment analysis showed that USP7 related genes were involved in transcriptional regulation, protein ubiquitination, DNA repair, and cytoplasmic pattern recognition receptor signal pathways. Analysis of clinical samples showed that USP7 expression was significantly higher in hypertrophic scars (0.35±0.05), scar ulcers (0.43±0.04), and scar cancers (0.61±0.03) than in normal skin (0.18±0.04), P<0.05. Conclusions:USP7 may be a clinical biomarker for the progression of cicatricial ulcer cancer.
4.Histopathological and genetical diagnosis of one case of neonatal ectodermal dysplasia/skin fragility syndrome
Qiongfang RUAN ; Chen XIA ; Weiguo XIE
Chinese Journal of Burns 2020;36(6):500-502
On August 6, 2015, a male infant with ectodermal dysplasia/skin fragility syndrome at 6 hours of birth was admitted to the Burn Department of Tongren Hospital of Wuhan University & Wuhan Third Hospital. The ulcerous skin tissue in thoracic area was harvested. The histopathological change of wound tissue was observed with hematoxylin-eosin staining. The result showed that the epidermal muscle cell layer was slightly released, there were bullae under the epidermis, the dermal papilla layer disappeared, and a small amount of inflammatory cells infiltrated in the dermis. The expression of plakophilin 1 (PKP1) in wound tissue was observed with immunohistochemical staining. The result showed that the PKP1 expression was completely absent. The PKP1 gene mutation site was identified by target sequencing. The result showed that the PKP1 gene had a homozygous mutation at intron ( PKP1: c.203-1G>A). Most of the wounds of the pediatric patient healed after 35 days of treatment, with many scattered residual wounds visible, and new blisters and skin lesions continue to appear.
5.Analysis of causes of death and etiological characteristics of skin tissue donors
Ziqing YE ; Weiguo XIE ; Xiongbing LU ; Xin CHEN ; Chaoli ZHAO ; Qiongfang RUAN ; Wei ZHANG
Chinese Journal of Burns 2020;36(6):446-450
Objective:To investigate the causes of death and etiological characteristics of skin tissue donors, and to provide reference for allogeneic skin transplantation.Methods:From October 2008 to October 2018, 49 skin tissue donors accepted by the Burn Department of Wuhan Third Hospital met the inclusion criteria of this study, and a cross-sectional study was conducted. According to the cause of death, the donors were divided into accidental death group (19 cases) and non-accidental death group (30 cases). The sex and death age of 49 donors were recorded, and the death age between different sex donors and that of donors between accidental death group and non-accidental death group were compared. Diseases or circumstances that caused the death of donors, hepatitis B, hepatitis C, acquired immunodeficiency syndrome, syphilis virus carrying status, and peripheral blood microbial culture results of 49 donors were recorded, and the detection of blood-borne infectious risk factors of donors between accidental death group and non-accidental death group was compared. Abnormal skin tissue was also selected during allogenic skin graft preparing for pathological examination. Data were statistically analyzed with Mann-Whitney U test and continuity correction chi-square test. Results:(1) Out of the 49 donors in this group, 38 were male (77.55%) and 11 were female (22.45%). The death age was 42.00 (24.00, 55.00) years, and the death age of male donors was similar to that of female donors ( Z=0.120, P>0.05). The death age of donors in accidental death group was lower than that in non-accidental death group, but the difference was not statistically significant ( Z=-1.581, P>0.05). (2) Among the causes and circumstances of the 49 donors in this group, there were 19 cases (38.78%) of injury, poisoning, and some other consequences of external causes, 11 cases (22.45%) of circulatory system diseases, 9 cases (18.37%) of tumors, 3 cases (6.12%) of nervous system diseases, 2 cases (4.08%) of respiratory system diseases, and 2 cases (4.08%) of congenital malformation, deformation, and chromosome abnormality, 1 case (2.04%) of blood and hematopoietic organ diseases and some diseases related to immune mechanism, 1 case (2.04%) of digestive system disease, and 1 case (2.04%) of genitourinary system disease. (3) There were 9 donors (18.37%) with blood-borne infectious risk factors among the 49 donors in this group, including 8 cases (16.33%) of blood-borne infectious diseases, which were 5 cases (10.20%) of hepatitis B, 2 cases (4.08%) of syphilis, and 1 case (2.04%) of hepatitis C, respectively. Blood microorganism culture was positive in 1 case (2.04%), in which multi-drug resistant Pseudomonas aeruginosa was detected. Risk factors of blood-borne infection were detected in 2 donors in accidental death group, with detection ratio lower than that in non-accidental death group (7 cases), but the difference was not statistically significant ( χ2=0.562, P>0.05). (4) A total of 8 donors′ abnormal skin tissue were selected, including 4 cases of intradermal pigmented nevus, 1 case of scar, 1 case of pseudoepithelioma hyperplasia, 1 case of epidermal verrucous hyperplasia, and 1 case of large amount of pigment granules in dermis. Conclusions:Non-accidental death caused by diseases is the main cause of death of skin tissue donors, and the risk of donor-derived infection of non-accidentally dead donors is slightly higher than that of accidentally dead donors. Before the allogeneic skin is obtained and transplanted, the cause of death of the donor should be carefully investigated, and the health status should be evaluated, so as to avoid the occurrence of donor-derived infection.
6.Changes in serum osteoprotegerin/receptor activator of nuclear factor-κB ligand and related indexes of calcium and phosphorus in the early stage of severe burn patients
Xiang GONG ; Ziqing YE ; Wei ZHANG ; Gang YU ; Qiongfang RUAN ; Weiguo XIE
Chinese Journal of Burns 2020;36(8):704-709
Objective:To investigate the early changes in serum osteoprotegerin/receptor activator of nuclear factor-κB ligand (RANKL) and related indexes of calcium and phosphorus in severe burn patients.Methods:Thirty severe burn patients who met the inclusion criteria and were admitted to Tongren Hospital of Wuhan University & Wuhan Third Hospital within 8 hours post injury from June 2017 to December 2018 were recruited into severe burn group (24 males and 6 females, aged (38±13) years). Ten healthy volunteers with normal physical examination results in the Physical Examination Center of the same hospital in the same period of time were recruited into healthy control group (7 males and 3 females, aged (37±8) years). A prospective controlled study was conducted. The fasting venous blood of 5 mL was taken from each patient in severe burn group on post injury day (PID) 1, 7, 14, 21, and 28, respectively, and the fasting venous blood of 5 mL was taken from each volunteer in healthy control group. The serum osteoprotegerin, RANKL, 25 hydroxyvitamin D, and parathyroid hormone (PTH) levels were determined by enzyme-linked immunosorbent assay, and the RANKL/osteoprotegerin ratio was calculated. Serum albumin, serum calcium, and serum phosphorus levels were determined by bromocresol green method, methylthymol blue method, and phosphomolybdic acid method, respectively. Data were statistically analyzed with Fisher′s exact probability test, analysis of variance for repeated measurement, Mann-Whitney U test, independent sample t test, and Bonferroni correction. Results:(1) The serum osteoprotegerin levels of patients in severe burn group on PID 1, 7, 14, 21, and 28 were 155.11 (102.91, 187.02), 170.07 (84.60, 196.86), 174.95 (59.09, 208.35), 190.01 (47.08, 214.52), and 188.85 (58.73, 223.13) pg/mL, respectively, which were significantly higher than 33.34 (28.59, 45.68) pg/mL of volunteers in healthy control group, Z=-3.436, -4.311, -3.248, -2.811, -4.217, P<0.01. The serum levels of RANKL of patients in severe burn group on PID 1, 7, 14, 21, and 28 were (1 869±791), (1 746±857), (1 781±713), (2 015±825), and (2 272±583) pg/mL, respectively, significantly higher than (49±16) pg/mL of volunteers in healthy control group, t=12.600, 10.844, 13.294, 13.041, 20.880, P<0.01. The ratios of RANKL/osteoprotegerin of patients in severe burn group on PID 1, 7, 14, 21, and 28 were 12.23 (8.10, 24.73), 11.40 (8.25, 16.96), 11.15 (6.91, 38.32), 12.98 (9.22, 49.68), and 13.91 (10.29, 40.68), respectively, which were significantly higher than 1.17 (0.91, 1.74) of volunteers in healthy control group, Z=-4.560, -4.529, -4.529, -4.560, -4.623, P<0.01. (2) The serum level of 25 hydroxyvitamin D of patients in severe burn group on PID 1 was significantly lower than that of volunteers in healthy control group ( Z=-2.749, P<0.01). Compared with those of volunteers in healthy control group, the serum levels of albumin of patients in severe burn group on PID 1, 7, 14, 21, and 28 were significantly lower ( t=-4.374, -7.689, -8.257, -7.651, -6.259, P<0.01), the serum levels of PTH were significantly elevated ( Z=-4.685, -4.685, -4.685, -4.654, -4.685, P<0.01), and the serum levels of phosphorus were not changed significantly. The serum levels of calcium of patients in severe burn group on PID 1, 7, 14, and 21 were significantly lower than the level of volunteers in healthy control group ( Z=-2.375, -3.455, -2.442, -2.016, P<0.05 or P<0.01). Conclusions:The serum osteoprotegerin, RANKL, RANKL/osteoprotegerin ratio, and PTH are increased, and the serum 25 hydroxyvitamin D, albumin, and calcium are decreased in the early stage of severe burn patients, which may be the mechanism leading to bone loss in patients.
7. Observation on clinical effects of recombinant human growth hormone on the treatment of children with severe burn
Zhigang CHU ; Ze LI ; Aihua WANG ; Qiongfang RUAN ; Hong WU ; Jingjing RUAN ; Weiguo XIE
Chinese Journal of Burns 2018;34(8):522-525
Objective:
To observe the clinical effects of recombinant human growth hormone (rhGH) on children with severe burn.
Methods:
Clinical data of 94 children with severe burn, hospitalized in our burn unit from April 2012 to December 2016, conforming to the study criteria, were retrospectively analyzed. According to the use of rhGH, children were divided into rhGH group (
8. Analysis of microRNA expression profile in serum of patients with electrical burn or thermal burn
Qiongfang RUAN ; Meijun JIANG ; Ziqing YE ; Chaoli ZHAO ; Weiguo XIE
Chinese Journal of Burns 2017;33(1):37-42
Objective:
To explore the differential expression of microRNAs in the serum among patients with electrical burn or thermal burn and healthy persons and to explore the significance.
Methods:
In this study we included three patients with electrical burn and three patients with thermal burn, conforming to the inclusion criteria and hospitalized in our burn ward from June to August 2015, and three healthy adult volunteers. Their serum samples were separated from whole blood and divided into electrical burn group, thermal burn group, and normal control group. Total RNA was extracted from their serum samples using Trizol method. The differentially expressed microRNAs (with differential ratio larger than or equal to 2.000, less than or equal to 0.500) among the three groups were screened by microRNA chip technique. Then cluster and Venn diagram analysis of the differentially expressed microRNAs were performed. Enrichment analysis of Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway was performed on the distinctly changed microRNAs (with differential ratio larger than or equal to 5.000, less than or equal to 0.500).
Results:
There were 220 differentially expressed microRNAs among serum of the three groups. MicroRNA expression profiles in serum of electrical burn and thermal burn groups were different from that in serum of normal control group. Compared with those in serum of normal control group, the expressions of 59 microRNAs changed more than 2.000 times in serum of electrical burn group, with 50 up-regulated microRNAs and 9 down-regulated microRNAs; the expressions of 40 microRNAs changed more than 2.000 times in serum of thermal burn group, with 21 up-regulated microRNAs and 19 down-regulated microRNAs. Compared with those in serum of thermal burn group, the expressions of 167 microRNAs changed more than 2.000 times in serum of electrical burn group. There were 17 exclusively expressed microRNAs in serum of thermal burn group and 26 exclusively expressed microRNAs in serum of electrical burn group, compared with those in serum of normal control group. Enrichment analysis of KEGG signaling pathway showed that compared with those in serum of normal control group, microRNAs which changed distinctly in serum of electrical burn group took part in the insulin secretion signaling pathway, arrhythmogenic right ventricular cardiomyopathy signaling pathway, hypertrophic cardiomyopathy signaling pathway, glutamatergic synapse signaling pathway, calcium signaling pathway, cyclic adenosine monophosphate signaling pathway, glycerophospholipid metabolism, pyrimidine metabolism, serotonergic synapse signaling pathway, etc, while microRNAs which changed distinctly in serum of thermal burn group took part in the tumor transcription misregulation signaling pathway, proteoglycans in tumor signaling pathway, microRNAs in tumor signaling pathway, long-term potentiation signaling pathway, citrate cycle signaling pathway, tumor necrosis factor signaling pathway, focal adhesion signaling pathway, endocytosis signaling pathway, insulin secretion signaling pathway, p53 signaling pathway, and estrogen signaling pathway, etc.
Conclusions
MicroRNA expression profiles in serum of electrical and thermal burn are different from that in serum of healthy adult. The signaling pathways enriched with target genes which are regulated by the differentially expressed microRNAs are related to the pathological changes and clinical manifestations after electrical or thermal burn.
9. Differential expression of microRNAs in serum of severe burn patients and analysis of the signaling pathway at early stage
Shan WANG ; Qiongfang RUAN ; Weiguo XIE ; Lan CHEN ; Meijun JIANG ; Jingjing RUAN ; Ziqing YE
Chinese Journal of Burns 2017;33(10):639-643
Objective:
To explore differential expression of microRNAs in serum of patients with severe burn and analysis of the signaling pathway at early stage.
Methods:
In this study, we included three healthy adult volunteers and three patients with severe burn, conforming to the inclusion criteria and hospitalized in Tongren Hospital of Wuhan University & Wuhan Third Hospital in July 2015. Venous whole blood of 6 mL of each burn patient and healthy volunteer was collected at 24 to 48 h post injury of burn patients. The whole blood was divided into burn group and healthy control group. Whole blood of 2 mL of each one was used to determine white blood cell count and neutrophile granulocyte content. Serum was separated from the other whole blood of 4 mL of each one. Half of serum was used to determine content of blood glucose, total protein, and albumin; another half of serum was used to extract total RNA with Trizol method. The differentially expressed microRNA, with differential expression ratio larger than or equal to 1.500 between 2 groups, were screened by microRNA chip technique. Then cluster analysis and functional enrichment analysis of Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway were performed on the differentially expressed microRNAs. Data were processed with
10.Effect of monocyte-secreted VEGF induced by electrical burn serum on monocyte-endothelial cell adhesion
Qiongfang RUAN ; Chaoli ZHAO ; Ziqing YE ; Qionghui XIE ; Weiguo XIE
Chinese Journal of Pathophysiology 2015;33(4):755-758
AIM: To observe the level of vascular endothelial growth factor (VEGF) secreted by monocytes cultured with electrical burn serum, and to explore the effect of VEGF on monocyte-endothelial cell adhesion.METH-ODS:The electrical burn serum of the rat was prepared.The normal serum from the rats without treating electric current was also collected for control.The contents of VEGF and its soluble receptor sFlt-1 in electrical burn group were determined by double-antibody sandwich ELISA.THP-1 cells were randomly divided into normal serum group and electrical burn serum group.The contents of VEGF and sFlt-1 in the culture supernatants were measured by double-antibody sandwich ELISA. THP-1 cells were also randomly divided into another 4 groups:normal serum group, electrical burn serum group, normal serum +inhibitor group and electrical burn serum +inhibitor group.THP-1 cells, which were incubated with the serum for 3 h and 6 h, were labeled with calcein-AM and then were added into the well with monolayer of endothelial cell line EA.hy926 to detect monocyte-endothelial cell adhesion.RESULTS:The levels of serum VEGF of the rats with electrical burns were significantly increased, the levels of serum sFlt-1 were significantly decreased as compared with the controls. The levels of VEGF secreted by THP-1 cells cultured with electrical burn serum were significantly increased, the levels of sFlt-1 were decreased correspondingly.Electrical burn serum enhanced monocyte-endothelial cell adhesion, sFlt-1 inhibi-ted the adhesion between monocytes and endothelial cells.CONCLUSION:The monocytes exposed to the electrical burn serum secrete VEGF, which enhance the adhesion between monocytes and endothelial cells.Blockage of VEGF activity may effectively inhibit monocyte-endothelial cell adhesion.

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