1.Role of imbalance of M1/M2 subsets of bone marrow macrophages in the pathogenesis of immune-mediated aplastic anemia in mice
Hui MU ; Hui JIA ; Zenghua LIN ; Honghui ZHENG ; Li WANG ; Hong LIU
Chinese Journal of Hematology 2021;42(11):945-951
Objective:To investigate the role of macrophages (M?) in the pathogenesis of modified immune-mediated aplastic anemia (AA) mice model.Methods:Before the establishment of the F1 AA mice model by total-body irradiation combined with allogeneic lymphocyte infusion, the mice of the CLO+AA group were treated with clodronate (CLO) liposomes to remove macrophages, and those of the PBS+AA group were treated with phosphate-buffered saline (PBS) liposomes and used as control. The severity of AA was observed by bone marrow (BM) pathological examination and peripheral blood cell count. Flow cytometry (FCM) was used to detect the CD4 +/CD8 + T lymphocyte subsets in the BM and M? subsets in the BM and spleen of each group. The levels of IFN-γ, TNF-α, G-CSF, GM-CSF, EPO, and TPO in the peripheral blood were detected using enzyme-linked immunosorbent assay. Finally, the relationships between inflammatory factors and M? subsets were analyzed. Results:The BM fatty conversion of mice in the CLO+AA group was significantly alleviated compared with the PBS+AA group. Hemoglobin counts were (91.50±31.63) and (110.65±24.15) g/L, respectively, and the platelet counts were (90.85±121.90) × 10 6/L and (461.13±483.45) ×10 6/L, respectively. The differences were all statistically significant (all P<0.05) . After removing macrophages, the proportions of CD4 + and CD8 + T lymphocytes in BM of mice in the CLO+AA group decreased, but the reduction of CD8 + T cells was more significant. The proportions of CD4 + T cells and CD8 + T cells in BM of the PBS+AA group were (18.5±10.17) % and (36.23±6.40) %, respectively, and in the CLO+AA group were (7.58±8.00) % and (6.67±5.78) %, respectively. Similarly, the percentage of macrophages in the spleen and BM in the CLO+AA group was significantly reduced compared with the PBS+AA group, most of which were M1 macrophages ( P<0.05) . The levels of IFN-γ in peripheral blood of the PBS+AA and CLO+AA groups were (602.37±104.62) ng/L and (303.01±87.22) ng/L, respectively, the levels of TNF-α were (34.46±1.42) ng/L and (23.25±4.21) ng/L, respectively, the levels of GM-CSF were (9.32 ± 2.00) ng/L and (64.85±12.25) ng/L, respectively, the levels of G-CSF were (5 891.78±2 632.39) ng/L and (17 784.16±488.36) ng/L, respectively, the levels of EPO were (9 667.31±4 501.95) ng/L and (2 078.02±897.56) ng/L, respectively, and the levels of TPO were (6.36±2.09) ng/L and (11.67±2.86) ng/L, respectively (all P<0.05) . Conclusions:This study confirmed that macrophages were involved in the pathogenesis of AA, and the degree of BM damage in AA mice was improved by removing macrophages in advance. The imbalance of M1/M2 macrophages and the changes of IFN-γ and TNF-α may be important mechanisms that eventually lead to AA.
2.Efficacy of ultrasound-guided bilateral erector spinae plane block for pediatric patients undergoing Nuss procedure with general anesthesia
Zenghua XU ; Jianmin ZHANG ; Chao ZHENG ; Jingjing CAI ; Tiehua ZHENG ; Lijing LI ; Fang WANG
Chinese Journal of Anesthesiology 2020;40(2):186-189
Objective:To evaluate the efficacy of ultrasound-guided bilateral erector spinae plane (ESP) block for pediatric patients undergoing Nuss procedure with general anesthesia.Methods:Thirty-two American Society of Anesthesiologists physical status Ⅰ or Ⅱ patients, aged 4-15 yr, undergoing Nuss procedure with general anesthesia, were divided into ESP block plus general anesthesia group (group EG, n=16) and general anesthesia group (group G, n=16) using a random number table method.Bilateral ESP blocks were performed after anesthesia induction in group EG.Intravenous analgesia was performed postoperatively in both groups.Face Legs Activity Cry Consolability (FLACC) scores both at rest and during coughing were recorded on admission to postanesthesia care unit (PACU) and at 1, 6, 12, 24 and 48 h after operation.When FLACC score at rest >4, ibuprofen 5.0-7.5 mg/kg was taken orally according to the requirements of pediatric patients and their parents.The intraoperative amount of propofol and remifentanil used and postoperative requirement for fentanyl and ibuprofen were recorded.The development of postoperative hypoxia (SpO 2 <90%), nausea and vomiting, and urinary retention was recorded.The development of puncture site infection, hematoma and subcutaneous emphysema was recorded.The tracheal extubation time, duration in PACU and length of postoperative hospital stay were also recorded. Results:Compared with group G, FLACC scores at rest within 12 h after operation and during coughing within 6 h after operation were significantly decreased, the intraoperative consumption of remifentanil was reduced, the postoperative requirement for fentanyl and ibuprofen was decreased, the incidence of hypoxemia was decreased, the tracheal extubation time and duration in PACU were shortened ( P<0.05), and no significant change was found in the consumption of propofol, incidence of nausea and vomiting, or length of postoperative hospital stay in group EG ( P>0.05). Conclusion:Ultrasound-guided bilateral ESP block can reduce the consumption of opioids in the perioperative period and enhance the analgesic efficacy with a higher safety, which is helpful in promoting short-term outcomes for pediatric patients undergoing Nuss procedure with general anesthesia.
3.Expression profile of circular RNA in inflammatory response in human bronchial epithelial cells induced by carbon black nanoparticles
Jiahao ZHANG ; Sijia ZHOU ; Zheng KUANG ; Zenghua QIN ; Liwen TAN ; Yueting SHAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(8):576-581
Objective:To explore the toxic effect of carbon black nanoparticles on human bronchial epithelial cells, and identify the differentially expressed circular RNA based on the full transcriptome high-throughput sequencing, so as to provide evidence for the development of biomarkers exposed to carbon black nanoparticles and their application on epigenetic toxicology.Methods:In June 2020, 16 HBE cells were treated with carbon black nanoparticles at concentrations of 20, 40 and 80 μg/ml, and 16 HBE cells without any intervention were used as the control group. The cytotoxicity of carbon black nanoparticles was detected by CCK8 and LDH experiments. Real-time quantitative fluorescent PCR (qRT-PCR) and ELISA were used to detect the changes of interleukin-6 (IL-6) and interleukin-8 (IL-6, IL-8) mRNA and protein levels of carbon black nanoparticles with concentration gradient after 72 h exposure. Western blot analysis was conducted to detect the expression levels of toll-like receptor 4 (TLR4), phosphorylated nuclear factor-κB (P-NF-κB), apoptosis-related speckled protein (ASC) and Caspase-1 associated with nuclear factor-κB. According to high-throughput sequencing results, differentially expressed Circrnas were screened and identified by qRT-PCR, and those with stable differentially expressed circrnas and the strongest association with the NF-κB pathway were selected for ring performance identification.Results:After being exposed to carbon black nanoparticles for 72 h, the activity of 16HBE cells decreased significantly ( P<0.05), and the release of lactate dehydrogenase increased significantly ( P<0.05). Compared with control group, mRNA expression levels of IL-6 and IL-8, protein levels of IL-6 and IL-8 were increased, and protein levels of TLR4, p-NF-κB, ASC and Caspase-1 were significantly up-regulated in 16 HBE cells of different concentrations, with statistical significance ( P<0.05). Compared with the control group, a total of 492 differentially expressed circular Rnas (|log2 FC|>1) were detected. Among the 5 differentially expressed ( P<0.05) circular Rnas, circ_002642 was selected as the object of subsequent research on circular Rnas, affter 72 hours of exposure to 80 μg/ml CBNPs, 16HBE cells showed signlficantly higher expression of circ_002642 ( P<0.05) . Conclusion:Carbon black nanoparticles can induce differentially expressed circular RNAs associated with inflammatory response in human bronchial epithelial cells.
4.Expression profile of circular RNA in inflammatory response in human bronchial epithelial cells induced by carbon black nanoparticles
Jiahao ZHANG ; Sijia ZHOU ; Zheng KUANG ; Zenghua QIN ; Liwen TAN ; Yueting SHAO
Chinese Journal of Industrial Hygiene and Occupational Diseases 2023;41(8):576-581
Objective:To explore the toxic effect of carbon black nanoparticles on human bronchial epithelial cells, and identify the differentially expressed circular RNA based on the full transcriptome high-throughput sequencing, so as to provide evidence for the development of biomarkers exposed to carbon black nanoparticles and their application on epigenetic toxicology.Methods:In June 2020, 16 HBE cells were treated with carbon black nanoparticles at concentrations of 20, 40 and 80 μg/ml, and 16 HBE cells without any intervention were used as the control group. The cytotoxicity of carbon black nanoparticles was detected by CCK8 and LDH experiments. Real-time quantitative fluorescent PCR (qRT-PCR) and ELISA were used to detect the changes of interleukin-6 (IL-6) and interleukin-8 (IL-6, IL-8) mRNA and protein levels of carbon black nanoparticles with concentration gradient after 72 h exposure. Western blot analysis was conducted to detect the expression levels of toll-like receptor 4 (TLR4), phosphorylated nuclear factor-κB (P-NF-κB), apoptosis-related speckled protein (ASC) and Caspase-1 associated with nuclear factor-κB. According to high-throughput sequencing results, differentially expressed Circrnas were screened and identified by qRT-PCR, and those with stable differentially expressed circrnas and the strongest association with the NF-κB pathway were selected for ring performance identification.Results:After being exposed to carbon black nanoparticles for 72 h, the activity of 16HBE cells decreased significantly ( P<0.05), and the release of lactate dehydrogenase increased significantly ( P<0.05). Compared with control group, mRNA expression levels of IL-6 and IL-8, protein levels of IL-6 and IL-8 were increased, and protein levels of TLR4, p-NF-κB, ASC and Caspase-1 were significantly up-regulated in 16 HBE cells of different concentrations, with statistical significance ( P<0.05). Compared with the control group, a total of 492 differentially expressed circular Rnas (|log2 FC|>1) were detected. Among the 5 differentially expressed ( P<0.05) circular Rnas, circ_002642 was selected as the object of subsequent research on circular Rnas, affter 72 hours of exposure to 80 μg/ml CBNPs, 16HBE cells showed signlficantly higher expression of circ_002642 ( P<0.05) . Conclusion:Carbon black nanoparticles can induce differentially expressed circular RNAs associated with inflammatory response in human bronchial epithelial cells.
5.Assessment of lymph node metastasis in pancreatic ductal adenocarcinoma using extracellular volume fraction
Ruru GAO ; Zenghua ZHENG ; Xuan QIN ; Jianbing MA ; Jun HAN ; Jian DING ; Xiang ZHU
China Modern Doctor 2024;62(26):38-40,45
Objective To assess the diagnostic efficacy of extracellular volume fraction(fECV)based on multi-slice spiral CT in evaluating lymph node metastasis of pancreatic ductal adenocarcinoma.Methods A total of 74 patients diagnosed as pancreatic ductal adenocarcinoma from March 2018 to March 2023 in the First Hospital of Jiaxing were retrospectively collected.All patients underwent enhanced multi-slice spiral CT examination.Hematocrit was collected before examination.CT values at the plain phase,portal phase,the equilibrium phase of the lesion and CT values of abdominal aorta at the same level were respectively measured after examination,and fECV was calculated.Using postoperative pathological results as the"gold standard",the efficacy of fECV in diagnosing lymph node metastases of pancreatic ductal adenocarcinoma was evaluated by receiver operating characteristic curve.Results There were 33 cases with lymph node metastasis in positive group and 41 cases in negative group.There was no statistically significant difference in fECV at the portal phase between lymph node metastasis positive group and negative group(t=0.80,P>0.05).There was statistically significant difference in fECV at the equilibrium phase between positive and negative lymph node metastasis group(t=2.84,P<0.001).The area under the curve of lymph node metastasis in portal phase and equilibrium phase were 0.517 and 0.870,respectively.Conclusion The fECV based on multi-slice spiral CT equilibrium phase can evaluate lymph node metastasis of pancreatic ductal adenocarcinoma.
6.Anesthesia management of children undergoing resection of pheochromocytoma and paraganglioma
Zenghua XU ; Jianmin ZHANG ; Nan ZOU ; Tiehua ZHENG ; Lianghong HUO ; Lijing LI ; Fang WANG
Chinese Journal of Anesthesiology 2024;44(2):209-213
Objective:To retrospectively analyze the anesthetic management characteristics of children undergoing resection of pheochromocytoma and paraganglioma (PPGL).Methods:The clinical data from patients undergoing resection of PPGL and confirmed histologically from January 1, 2010 to June 30, 2023 were retrospectively collected. The baseline characteristics, intraoperative conditions and postoperative complications were recorded.Results:The clinical data from 47 pediatric patients were analyzed. The overall incidence of hemodynamic instability events was 68% (32 cases). Lowering preoperative blood pressure to normal levels and the maximum diameter of tumor≥6 cm was helpful in reducing the incidence of the intraoperative hemodynamic instability events ( P<0.05). Postoperative hypotension developed in 7 cases, acute left heart failure in 1 case, arrhythmia in 1 case, adrenocortical insufficiency in 4 cases, and pulmonary infection in 13 cases. Conclusions:Thorough preoperative preparation, evidence-based anesthetic management, and meticulous postoperative vital sign monitoring can increase the perioperative safety for children undergoing resection of PPGL, thereby reducing the incidence of complications.
7.Optimization strategies of anesthesia for pediatric patients with pectus excavatum undergoing Nuss procedure: ultrasound-guided erector spinae plane block combined with general anesthesia
Yi REN ; Lei HUA ; Fuzhou ZHANG ; Yangwei MA ; Tiehua ZHENG ; Zenghua XU ; Jianmin ZHANG
Chinese Journal of Anesthesiology 2024;44(8):972-976
Objective:To evaluate the optimization effect of ultrasound-guided erector spinae plane block (ESPB)combined with general anesthesia on Nuss procedure in pediatric patients with pectus excavatum undergoing Nuss procedure.Methods:One hundred and eight pediatric patients of either sex, aged 7-18 yr, with body mass index within the normal range, of American Society of Anesthesiologists Physical Status classification Ⅰ or Ⅱ, undergoing elective Nuss procedure, were randomized into 3 groups ( n=36 each) using a random number table method: ultrasound-guided ESPB combined with general anesthesia group (group E), thoracic epidural anesthesia plus general analgesia group (group T), and general analgesia group (group G). Pediatric patients received ultrasound-guided bilateral ESPB (0.3% ropivacaine was injected, 0.5 ml/kg bilateral total volume [maximum 20 ml]) or thoracic epidural anesthesia (0.3% ropivacaine 0.5 ml/kg [maximum 20 ml/side]) after induction of anesthesia in group E and group T, respectively. All the three groups underwent general anesthesia, anesthesia was induced with propofol 2-3 mg/kg, sufentanil 0.3-0.5 μg/kg and cis-atracurium 0.1-0.2 mg/kg and maintained with propofol 6-10 mg·kg -1·h -1 and remifentanil 0.1-0.3 μg·kg -1·min -1. The patients were endotracheally intubated and mechanically ventilated after anesthesia induction. The parent-controlled intravenous analgesia (PCIA) was used for patients in all the three groups postoperatively. The consumption of intraoperative remifentanil and postoperative consumption of sufentanil for PCIA in each period after surgery were recorded. The time to the first rescue analgesia, consumption of ibuprofen and postoperative complications were also recorded. Results:Compared with group G, the intraoperative consumption of remifentanil and postoperative consumption of sufentanil for PCIA in each period were significantly decreased, the time to the first rescue analgesia was prolonged, and the consumption of ibuprofen was decreased in group E ( P<0.05). There was no significant difference among the three groups ( P>0.05). Conclusions:ESPB combined with general analgesia is helpful in reducing postoperative hyperalgesia in pediatric patients undergoing the Nuss procedure.