1.Alternation and modulation of tumor necrosis factor-alpha and protein kinase C in celiac macrophage of mouse after serious scalding
Yong WANG ; Wenhua HUANG ; Daizhi PENG ; Xin ZHOU ; Jing LIU
Chinese Journal of Tissue Engineering Research 2005;9(10):249-251
BACKGROUND: Serious scalding leads to dysfunction of each aspect in immune system, and activated macrophage can secret many bioactive transmitters. The relationship between macrophage dysfunction and signal conduction after scalding is unclear at present.OBJECTIVE: To observe the alternation of tumor necrosis factor- alpha(TNF-α) at different time points after scalding and the activity of nuclear factor κB(NF-κB) and alternation of protein kinase C (PKC) after the application of specific modulator H-7 to explore whether PKC participates in the modulation of TNF-α in macrophage on signal conduction level for the clarification of some mechanisms of macrophage dysfunction.DESIGN: A randomized controlled experimental study by employing experimental animals as subjectsSETTING: An institute of burn research of a military medical university MATERIALS: The study was conducted in the Laboratory (state) of the Institute of Burn Research, Third Military Medical University of Chinese PLA between January and December 1999. Experimental animals were 32 healthy clean inbreeding Kunming white mice.METHODS: 15% Ⅲ scalding was created in mice for the establishment of routine scalding model. Mice were randomly divided into 6 groups according to different time points before or after scalding, I.e. 0(normal control group), 2, 6, 12, 24, or 48 hours group. Celiac macrophages were collected for the detection of TNF-α content by radioimmunoassay, NF-κB activity by electrophoretic mobility shift analysis (EMSA), and membrane or plasma PKC activity by isotope analysis.MAIN OUTCOME MEASURES: ① TNF-α content; ② NF-κB activity; ③Membrane or plasma PKC activity RESULTS: After scalding, macrophage excessively secreted TNF-α and reached its peak of (1 085.65 ± 122.99) ng/L at 12 hours, which was significantly higher than that of control group( t = 14.92, P < 0.01 ).Compared with control group, membrane PKC activity increased after scalding, which significantly heightened to(231.80 ± 31.66) nmol/min · g at 2hours( t = 7. 930, P < 0.01 ), slightly decreased to close to normal level of (174.29±16.80) nmol/min· gat 6hours(t=2.531, P <0.05), and rapidly elevated at 12 hours [512. 10 ±33.42) nmol/min · g] and 24 hours [ (454.70 ± 21.06) nmol/min · g] to reach its peak of(530.49 ± 28.54)nmol/min. G at 48 hours( t = 29.42, 28.03, 30. 19, P < 0. 01 ). Correlation analysis of the alternation between TNF-α and membrane PKC indicated a significant positive correlation( r = 0. 796 4, P < 0. 05) . As indicated by EMSA image, NF-κB activity significantly elevated after scalding. Twelve hours after scalding was set as modulation point, NF-κB activity was significantly inhibited by the application of H-7.CONCLUSION: The secretion of TNF-α and the activities of PKC and NF-κB are significantly activated in celiac macrophage after scalding, and PKC-NF-κB signal pathway participates in the modulation of TNF-α expression, which provide experimental data for the modulation of immune function and rehabilitative intervention during scalding.serious scalding.
2.Changes and immune function mechanism of tumor necrosis factor-alpha in murine peritoneal macrophage after severe scald
Yong WANG ; Daizhi PENG ; Xin ZHOU ; Wenhua HUANG ; Jing LIU
Chinese Journal of Tissue Engineering Research 2005;9(6):233-235
BACKGROUND: Severe scald injury leads to a variety of disorders in the immune system. Activated macrophages are known to secrete many kinds of biologically active transmitter, but the relation between the functional disorder of the macrophages and signal transduction after burn injury has not been fully understood.OBJECTIVE: To observe the changes in nuclear factor(NF) -κκB activity and expressions of IκκB-α and tumor necrosis factor(TNF) -α in peritoneal macrophage of mice at different time points after severe scald injury and after the application of specific NF-κκB inhibitor pyrrolidine dithiocarbamate (PDTC), thereby to explore the mechanism of macrophage dysfunction in light of signal transduction.DESIGN: A randomized controlled experimental research.SETTING: State Key Laboratory of Trauma, Burn and Combined Injury.MATERIALS: The experiment was carried out in the State Key Laboratory of Institute of Burn Research, Third Military Medical University during the period from January to June, 1999, using 30 healthy clean-grade Kunming mice of inbred strain.INTERVENTIONS: Common scald injury models(with third degree burn of 15% total body surface area) were established in the mice, which were randomized into 6 groups according to different time points after the injury for observation, namely 0 hour(normal control group) and postburn 2, 6, 12,24 and 48 hours. Peritoneal macrophages were collected at these time points for examining TNF-α content using radio-immunoassay and NF-κκB activity by means of electrophoretic mobility shift assay(EMSA). The expressions of IκκB-α and TNF-α mRNA were determined by immunoblotting method and reverse transcription-PCR, respectively.MAIN OUTCOME MEASURES:Examinations of ① the content of TNF-α, ② NF-κκB activity,③ expression of IκB-α, and ④ expression of TNF-α mRNA.RESULTS: Macrophage secretion of TNF-α was enhanced postburn, reaching the peak level at 12 hours[(1085.65 ± 122.99) ng/L], which was significantly higher than that in the normal control group( t = 14.92, P < 0.01) .Postburn NF-κκB activity significantly increased after the injury, peaking at 2 hours[ (56. 8 ± 7.3)RDU], which occurred much earlier than the peak of TNF-α secretion( t=13. 31, P < 0.01 ). Compared with that in the normal control group, IκB-α expression decreased significantly 2 hours postburn ( t =4. 23, P < 0. 01) to 0. 632 ±0. 086, followed by gradual increase to the peak level to 1. 161 ± 0. 097 24 hours after the burn injury( t = 7.06, P< 0. 01) and then by slight decrease to 1. 149 ±0. 167 till 48 hours(t = 4. 82, P < 0.01) . Twelve hours after injury was the time point for intervention with PDTC application, when NF-κκB activity and TNF-α mRNA expression both decreased significantly( P < 0.01 ).CONCLUSION: NF-κB activity and TNF-αmRNA expression decrease significantly after severe scald. At high levels, IκB-α and NF-κκB maintain an interaction for their restriction. After burn injury, NF-κκB signal transduction pathway is involved in the modulation of TNF-α expression in mouse peritoneal macrophage.
3.The modulating of Qingguang’an II Formula on gut microbiota in mice with chronic high intraocular pressure by 16S rDNA sequencing
ZHOU Yasha ; GAO Wenyong ; HUANG Yu ; XIA Xin ; XIAO Li ; DENG Ying ; PENG Qinghua ; PENG Jun
Digital Chinese Medicine 2024;7(4):332-342
Methods:
A total of 10 specific pathogen free (SPF) grade female DBA/2J mice were randomly divided into model group and QGA II group (n = 5 for each group), while additional 5 SPF-grade female C57BL/6J mice were assigned to control group. Mice presented spontaneous high IOP and showed elevated approximately at the age of seven months. The high IOP was maintained until week 38, when gavage was initiated. Mice in control group underwent the same intragastric treatment, while those in QGA II group were gavaged with QGA II (9.67 g/kg), once a day for four weeks. Retinal morphology was examined using hematoxylin and eosin (HE) staining, with the number of retinal ganglion cells (RGCs) counted. The expression level of Brn3a protein, a specific marker for RGCs, was detected by immunofluorescence, with the mean optical density (OD) measured for quantitative analysis. In addition, 16S rDNA sequencing was leveraged to analyze changes in the diversity of gut microbiota, including their α-diversity (Chao1, Shannon, Pielou’s evenness, and observed species index) and β-diversity. Venn diagrams and linear discriminant analysis effect size (LEfSe) analysis was employed to investigate the number of amplicon sequence variants (ASVs), the abundance of differential gut microbiota species, and the classification of species at both the phylum and genus levels within the three groups of mice.
Results:
HE staining revealed that compared with control group, model group showed significant reduction in the number of RGCs (P < 0.01), with intracellular vacuolar degeneration and nuclear pyknosis. After QGA II treatment, the number of RGCs was significantly increased compared with model group (P < 0.01), with notable improvements in intracellular vacuolar degeneration. Immunofluorescence analysis showed that the mean OD of Brn3a protein was significantly decreased in model group compared with control group (P < 0.01), while QGA II treatment significantly elevated its expression level (P < 0.01). Analysis of α-diversity showed that after QGA II intervention, the Chao1, Shannon, and Pielou’s evenness indices were significantly increased (P < 0.01), and the observed species index was elevated (P < 0.05). β-Diversity analysis demonstrated distinct clustering among the three groups, indicating relatively low similarity in bacterial community structures. ASV clustering identified a total of 14 061 ASVs across all groups, with 9 514 ASVs shared between model and QGA II groups. At the phylum level, the abundance of Bacteroidetes was significantly decreased in model group compared with control group (P < 0.01), while Firmicutes and the Firmicutes/Bacteroidetes (F/B) ratio were significantly increased (P < 0.01). QGA II treatment significantly reduced both Firmicutes abundance and the F/B ratio (P < 0.01). At the genus level, Lactobacillus was dominant across all groups, with its abundance significantly increased in model group (P < 0.01) and subsequently decreased following QGA II intervention (P < 0.05).
Conclusion
QGA II restructured the gut microbiota of DBA/2J mice with chronic high IOP, bringing changes in their diversity and abundance of components. Firmicutes, Bacteroidetes, Lactobacillus, along with their associated microorganisms, are likely critical components of the gut microbiota that contribute to the optic neuroprotective effects of QGA II on chronic high IOP mice.
4.Application of contrast-enhanced ultrasound in differential diagnosis of cervical lymph nodes
Xingyue HUANG ; Liao CHEN ; Peng LI ; Bin SUN ; Wei HU ; Xin HUANG ; Wenwei CHEN
Chinese Journal of Ultrasonography 2015;(12):1051-1055
Objective To evaluate the ability of contrast-enhanced ultrasonography (CEUS)in differentiate reactive lymph node,metastatic lymph node and lymphoma.Methods In a prospective study CEUS was performed in 129 patients with cervical lymph node enlargement.The entire process were recorded and preserved in DICOM format.The results were registered with Sonoliver.The receiver operating characteristic curve (ROC curve)analysis was performed to find the corresponding cutoff values. The selected node was removed surgically and submitted for histology.Results Of all the nodes,26 were reactive nodes,85 were metastases and 1 8 were lymphoma.Enhancement pattern was the most accurate way to characterize lymph nodes.The enhancement pattern of reactive lymph nodes was homogeneous and most of them were enhanced by lymphatic type while the metastatic lymph nodes were inhomogeneously enhanced or weakly enhanced by peripheral type.Lymph node lymphoma usually had no fixed enhancement pattern. Arrive time (AT),rise time (RT),time to peak (TTP),mean transit time (mTT),maximum intensity (IMAX),under the curve (AUC),rising slope(Kup),semi descending slope(Kdown)and perfusion index (PI)were significantly different in the three groups(P <0.05).RT,TTP and mTT of reactive lymph nodes were the shortest,which had significant difference compared with those of metastatic lymph nodes and lymphoma (P < 0.05 ).Kup,Kdown,IMAX%,AUC,PI in the reactive lymph nodes were significantly decreased compared with the metastatic group (P <0.05),but there was no significant difference compared with those in lymphoma (P >0.05 ).When TTP≥ 7.74 s,mTT≥26.54 s,metastatic lymph nodes were considered.When RT≥4.62 s,TTP ≥ 7.74 s,mTT ≥ 28.32 s,reactive lymph nodes were not considered. Conclusions Dynamic contrast-enhanced ultrasound image and enhancement pattern can distinguish neck lymph nodes while the optimal cut-off point time of the time-intensity curve parameters can further contribute to the identification of lymph nodes.
6.A Meta-analysis of the risk factors of delirium in ICU
Jie HUANG ; Qian XIAO ; Ying WU ; Chunli WANG ; Peng YUE ; Xin YANG
Chinese Journal of Nursing 2010;45(1):6-9
Objective To identify the risk factors of delirium in ICU by Meta-analysis. Methods Quality of the studies was assessed in terms of study design, definitions of main variables, statistics, and bias control. Analysis of sensitivity and heterogeneity were performed and cumulative effects were calculated using either fixed or random effects models by RevMan 4.2.Results Ten studies met all inclusive and exclusive criteria. Simple sizes range from 100 to 3308. Twenty-one risk factors of delirium in ICU were involved, but only alcohol abuse, respiratory disease, infection, APACHE Ⅱ,elevated level of serum urea nitrogen, hyponatremia, hyperbilirubinemia and using sedatives were identified as having a cumulative effect on delirium in ICU. Conclusions Infection, abnormality or disturbance of metabolism and intoxication or acute withdrawal from drug or alcohol are independent predictors of delirium in ICU, while advanced age and hypoxemia, which are commonly considered as independent risk factors of delirium in ICU, are still inconclusive.
7.Adeno-associated viral vector mediated and cardiac-specific delivery of CD151 gene in ischemic rat hearts.
Quan, WEI ; Zhaoyu, LIU ; Yujie, FEI ; Dan, PENG ; Houjuan, ZUO ; Xiaolin, HUANG ; Zhengxiang, LIU ; Xin, ZHANG
Journal of Huazhong University of Science and Technology (Medical Sciences) 2011;31(1):46-51
Our previous studies demonstrated that CD151 gene promoted neovascularization in ischemic heart model. To improve the delivery efficacy and target specificity of CD151 gene to ischemic heart, we generated an adeno-associated virus (AAV) vector in which CD151 expression was controlled by the myosin light chain (MLC-2v) promoter to achieve the cardiac-specific expression of CD151 gene in ischemic myocardium and to limit unwanted CD151 expression in extracardiac organs. The function of this vector was examined in rat ischemic myocardium model. The protein expression of CD151 in the ischemic myocardium areas, liver and kidney was confirmed by using Western blot, while the microvessels within ischemic myocardium areas were detected by using immunohistochemistry. The results showed that MLC-2v significantly enhanced the expression of CD151 in ischemic myocardium, but attenuated its expression in other organs. The forced CD151 expression could increase the number of microvessels in the ischemic myocardium. This study demonstrates the AAV-mediated and MLC-2v regulated CD151 gene is highly expressed in the ischemic myocardium and cardiac-specific delivery that is more efficiently targets CD151 to the ischemia myocardium after myocardial infarction.
8.Clinical effect of head and neck reconstruction using microsurgical free flap transfer techniques
Chi MAO ; Guang-Yan YU ; Xin PENG ; Lei ZHANG ; Chuan-Bin GUO ; Min-Xian HUANG
Chinese Journal of Microsurgery 2000;0(03):-
Objective To analyze our clinical results of head and neck reconstruction using microsur- gical free flap transfer techniques.Methods The free flap donor sites with long vascular pedicle and large diameter of vessel were routinely chosed,and chose receipt vessels with large diameter and proper position, and perform vessel ananstomosis under surgical loups instead of microscope.The un-buried free flap with a mo- nitoring window were harvest,and do double venous anastomoses in some flaps to ensure adequate venous out- flow.Results From May 1999 to March 2005,1066 consecutive free flap transfers were used to reconstruct head and neck defects.The overall success rate of free flap was 98.3%.The vessel thrombosis rate was 3.1%,and the flap salvage rate was 45.5%.Conclusion Head and neck reconstruetion using microsurgi- cal free flap transfer technique is safe and reliable,and good clinical results can be obtained.
9.Surgical exploration versus conservative treatment for refractory arterial crisis during the hypersensitive period after digital replantation
Peng WEI ; Hong CHEN ; Xin WANG ; Weiwen ZHANG ; Xiaofeng WANG ; Jianbo XUE ; Liming ZHOU ; Jian HUANG ;
Chinese Journal of Orthopaedic Trauma 2004;0(06):-
Objective To evaluate the significance of surgical exploration for the refractory arterial crisis daring the hypersensitive period (48 h to 96 h) after replantation of severed fingers.Methods One hundred and seventy-one patients experienced refractory arterial crisis during the hypersensitive period after replantation of the proximal thumb from February 1995 to February,2005 in our department.Eighty-seven of them were managed with surgical exploration,including incision injury (n=6),saw injury (n=17),rotation and avulsion injury (n=30), and crush injury (n=34).Eighty-four cases received conservative treatment,including incision injury (n=6),saw injury (n=16).rotation and avulsion injury (n=29),and crush injury (n=33).In the surgery group,the e- mergent explorations were performed as soon as the refractory arterial crisis arose,If arterial spasm or/and thrombosis were found,the involved parts were resected before the artery ends were anastomosed or the finger artery was repaired by cubital vein graft.In the other group,conservative managements were carried out by using intramuscular injection of 30 mg Papaverine and intravenous injection of 20,000-unit Urokinase in 20 mL normal saline.If symptums were not alleviated after half an hour,the procedures were repeated.The conservative managements also included abirritative antipsychotics and analgesia of anodyne.Meanwhile,the survival state of all the digital replants was observed. Results In the surgery group,78 fingers survived,the surviving rate being 89.7%.In the conservative group,41 fingers survived with a surviving rate of 48.8%.The difference was statistically significant (P<0.01).No obvious complications happened in the two groups.Conclusion Since surgical exploration is crucial to management of refractory arterial crisis during the hypersensitive period after replantation of severed fingers,it should not be readily abandoned.
10.Primary clinical study of hippocampal-sparing prophylactic cranial irradiation in limited-stage small-cell lung cancer
Xin DONG ; Zongmei ZHOU ; Junjie MIAO ; Xinyuan CHEN ; Zhihui HU ; Peng HUANG ; Yin ZHANG
Chinese Journal of Radiation Oncology 2015;24(2):131-136
Objective To preliminarily observe the clinical efficacy of hippocampal-sparing prophylactic cranial irradiation (HS-PCI) using helical tomotherapy (HT) in patients with limited-stage small-cell lung cancer (LS-SCLC) after chemoradiotherapy,and compare HT with intensity-modulated radiotherapy (IMRT) and volumetric modulated arc therapy (VMAT) in dose distribution.Methods From April to June,2014,six patients with LS-SCLC who had achieved a complete remission after chemoradiotherapy were assigned to HS-PCI using HT within a month after brain metastasis was ruled out using brain magnetic resonance imaging (MRI).After fusing CT images and MRI images,the hippocampus was contoured in the fusion images and hippocampal avoidance regions were created using a volumetric expansion of 3 mm around the hippocampus.A dose of 25 Gy in 10 fractions to 95% of planning target volume (PTV) was prescribed in HT,IMRT,and VMAT.The clinical efficacy,adverse reactions,neurocognitive function,and brain metastasis were evaluated for HT.The dose distribution in PTV and hippocampus were compared between HT,IMRT,and VMAT.Results There were one patient with abdominal wall and abdominal lymph node metastases,one patient with local recurrence,and no patient with brain metastasis during the observation period.The numbers of patients with grade 1 and grade 2 headache,dizziness,and hair loss reactions were 3 and 1,3 and 1,and 4 and 2,respectively.There were no significant differences in the average score of the Mini-Mental State Examination before treatment and at 3 and 6 months after treatment (29.7,29.2,and 29.3 ; P =0.083,0.317,and 0.157).The mean dose to the hippocampus was 16.85 Gy for IMRT and 17.59 Gy for VMAT.For HT,the mean doses to the hippocampus and avoidance regions were reduced to 5.26 Gy and 6.21 Gy,respectively.The prescribed dose for HT was reduced by 79% and 71% compared with IMRT and VMAT,respectively.The average coverage rate of the prescribed dose was 94.48% for HT.Conclusions HT achieves promising dose distribution and target coverage in sparing of the hippocampus.Moreover,HT dose not increase the incidence of adverse reactions.The change in neurocognitive function needs to be further studied with longterm observation and large-scale sampling.