1.Comparison of different kinds of combination of laparoscopic operation in treatment of acute appendicitis complicated with gallbladder stones
Hai XIANG ; Binggai XIANG ; Zhenliang LIN ; Zhangcheng ZHAO ; Jiangtao WEN
China Journal of Endoscopy 2017;23(1):65-69
Objective To investigate the clinical effects differences of three-port and single-port invasively combination laparoscopic cholecystectomy and appendectomy in the treatment of acute appendicitis complicated with gallbladder stone. Methods 110 patients with acute appendicitis complicated with gallbladder stones from August 2012 to August 2015 were randomly divided into control group (55 patients) with three-port laparoscopic operation and observation group (55 patients) with single-port laparoscopic operation;and the clinical indexes for operation related, operation overall satisfaction score of Brown, the VAS score of depression and anxiety before and after operation and postoperative complications of both groups were compared. Results The incision length of observation group was significantly shorter than control group (P< 0.05). The operative time of observation group was significantly longer than control group (P < 0.05). The operation overall satisfaction of Brown of observation group was significantly higher than control group (P < 0.05). The VAS score of depression and anxiety after operation of observation group was significantly better than control group and before operation (P<0.05). There was no signiifcant difference in the incidence of postoperative complications between 2 groups (P < 0.05). Conclusion Compared with three-port laparoscopic operation, single-port invasively combination laparoscopic cholecystectomy and appendectomy in the treatment of acute appendicitis complicated with gallbladder stone can efifciently decrease the incision length, improve the aesthetic degree and postoperative negative emotions and not lead to increased risk of postoperative complications.
2.Role of group 3 innate lymphoid cells in intestinal barrier
Zhenliang WEN ; Dechang CHEN ; Jinjun BIAN
Chinese Critical Care Medicine 2019;31(2):252-256
Intestinal?barrier?act?as?the?crucial?defender?against?pathogen?invasion,?and?is?indispensable?in?maintaining?tissue?homeostasis?both?locally?and?systemically.?Severe?disease?can?lead?to?impaired?intestinal?barrier.?In?addition?to?cause?a?variety?of?gastrointestinal?diseases,?intestinal?barrier?damage?can?also?worsen?the?disease?progression?in?critically?ill?patients.?Innate?lymphoid?cells?(ILCs)?is?a?group?of?newly?defined?innate?immune?cells?which?have?some?characteristics?as?adaptive?immune?cells.?Group?3?innate?lymphoid?cells?(ILC3),?which?mainly?reside?at?gut?associate?mucosal?tissue,?have?been?reported?to?play?a?critical?role?in?maintaining?intestinal?barrier?function.?After?a?brief?introduction?about?its?origination?and?classification,?we?will?focus?on?function?of?ILC3?physiologically?and?pathologically,?and?provide?a?new?theoretical?basis?for?maintaining?intestinal?barrier?function?under?pathological?conditions?in??this?review.
3.Ulinastatin via stabilizing endothelial junction protein to ameliorating hyper-permeability of vascular endothelial cell induced by matrix metalloproteinase-9
Zhenliang WEN ; Xiangwei WU ; Dechang CHEN ; Jinjun BIAN
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2019;26(1):16-20
Objective To investigate the role and mechanism of ulinastatin on the hyper-permeability of vascular endothelial cell induced by matrix metalloproteinase-9 (MMP-9). Methods Human umbilical vein endothelial cells (HUVEC) were cultured in vitro to establish a complete monolayer vascular endothelial cell model. The monolayer vascular endothelial cells were randomly divided into three groups: blank control group [phosphate buffered saline (PBS) added], MMP-9 model group (1 mg/L MMP-9 added) and ulinastatin group (1 mg/L MMP-9 and 1 000 kU/L ulinastatin added). The permeability of monolayer vascular endothelial cells was measured by fluorescein isothiocyanate (FITC)-labeled dextran (FD40) leaking method; the soluble vascular endothelial cells calcium dependent adherin (VE-cadherin) concentration in culture solution was determined by enzyme linked immunosorbent assay (ELISA);the protein expression levels of zonular occlusion protein-1 or tight junction (ZO-1), VE-cadherin, claudin-5 were detected by Western Blot and immunofluorescence methods. Results Compared with the blank control group, the permeability of vascular endothelial cells in MMP-9 model group was significantly increased [(cm2/h, ×10-2):3.35±0.56 vs. 0.94±0.06, P < 0.05]; the concentrations of soluble VE-cadherin in the Transwell upper and lower chambers were increased significantly [upper chamber (μg/L): 5.02±0.40 vs. 3.83±0.42, lower chamber (μg/L):4.92±1.05 vs. 3.24±1.24, both P < 0.05]; the protein expression levels of ZO-1, VE-cadherin and claudin-5 were significantly decreased [ZO-1/β-actin: 0.152±0.067 vs. 0.262±0.090, VE-cadherin/β-actin: 0.137±0.048 vs. 0.246±0.094, claudin-5/β-actin: 0.148±0.062 vs. 0.336±0.119, all P < 0.05], and obvious rupture sites appeared in their fluorescent patterns, and fluorescent particles were significantly reduced; compared with MMP-9 model group, the permeability of vascular endothelial cells in ulinastatin group was significantly decreased [(cm2/h, ×10-2): 1.80±0.34 vs. 3.35±0.56, P < 0.05]; the soluble VE-cadherin concentrations were significantly reduced in upper and lower chambers than those in the MMP-9 model group [upper chamber (μg/L): 4.41±0.37 vs. 5.02±0.40, lower chamber (μg/L):3.85±1.04 vs. 4.92±1.05, both P < 0.05], the expressions of endothelial junction protein were significantly increased in ulinastatin group (ZO-1/β-actin: 0.229±0.097 vs. 0.152±0.067, VE-cadherin/β-actin: 0.236±0.089 vs. 0.137±0.048, claudin-5/β-actin: 0.262±0.101 vs. 0.148±0.062, all P < 0.05], and the continuity of their fluorescent patterns and fluorescent particles were both increased. Conclusion The in vitro experiment showed that the hyper-permeability of vascular endothelial cells induced by MMP-9 can be attenuated by ulinastatin through decreasing the destruction of VE-cadherin and maintaining the protein expression levels of ZO-1, VE-cadherin and claudin-5 in vascular endothelial cells.
4.Lipopolysaccharide induced intestinal epithelial injury: a novel organoids-based model for sepsis in vitro.
Sisi HUANG ; Sheng ZHANG ; Limin CHEN ; Xiaojun PAN ; Zhenliang WEN ; Yizhu CHEN ; Lidi ZHANG ; Jiao LIU ; Dechang CHEN
Chinese Medical Journal 2022;135(18):2232-2239
BACKGROUND:
Advances in organoid culture technology have provided a greater understanding of disease pathogenesis, which has been rarely studied in sepsis before. We aim to establish a suitable organoids-based intestinal injury model for sepsis.
METHODS:
Stable passaged organoids were constructed and pre-treated with lipopolysaccharide (LPS) to mimic sepsis-induced intestinal injury. The LPS-induced sepsis model was used as a reference. We used quantitative real-time polymerase chain reaction to evaluate the RNA levels of inflammatory factors and antimicrobial peptides. Enzyme-linked immunosorbent assay was used to evaluate the protein levels, hematoxylin and eosin staining was used to evaluate the pathology of the small intestine of mice, and immunohistochemistry and immunofluorescence were used to evaluate the intestinal epithelial barrier function. Perkin Elmer Operetta™ was used to obtain high-resolution images of three-dimensional organoids.
RESULTS:
An LPS concentration >150 μg/mL after 24 h was identified to cause organoid growth restriction. The fluorescence intensity of zonula occludens-1 and occludins at LPS concentrations >100 μg/mL decreased significantly after 24 h. After LPS stimulation for 8 h, the RNA expression levels of interleukin (IL)-1α, tumor necrosis factor alpha, granulocyte-macrophage colony-stimulating factor, IL-6, and regenerating islet-derived protein 3 alpha, beta, and gamma increased. These results resembled those of intestinal epithelial layer alterations in a mouse sepsis model. For IL-10, the RNA expression level increased only when the LPS level >200 μg/mL for 24 h.
CONCLUSIONS
This study provides the primary intestinal in vitro model to study the effects of LPS-induced intestinal injury resembling sepsis. This model provides a platform for immune associated mechanism exploration and effective drug screening.
Mice
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Animals
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Lipopolysaccharides/toxicity*
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Sepsis
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Intestinal Diseases
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Tumor Necrosis Factor-alpha
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Disease Models, Animal
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Organoids
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RNA