1.Efficacy of robot-assisted surgery and laparoscopic surgery for choledochal cyst: a Meta-analysis
Tianhui GUO ; Qihui HU ; Cong CHEN ; Rui TAO ; Jintong HE ; Jixing WANG ; Zhenhao HUANG
Chinese Journal of Digestive Surgery 2024;23(2):289-296
		                        		
		                        			
		                        			The Choledochal cyst is an extremely rare congenital anomaly of the bile duct. Early cyst resection and Roux-en-Y hepatojejunostomy are the primary surgical methods for treating choledochal cyst. With the emergence of enhanced recovery after surgery, laparoscopic surgery has effectively reduced the incidence of biliary complications and wound infections, but it still does not meet people's requirements for minimally invasive surgery. Robotic surgery system has the potential to enhance surgical precision and the maneuverability of surgeons due to clear surgical visualization and flexible mechanical arms. The authors review the relevant literatures and conduct a Meta-analysis to evaluate the efficacy of robot-assisted surgery and laparoscopic surgery for choledochal cyst.
		                        		
		                        		
		                        		
		                        	
2.Efficacy and safety of ixekizumab in Chinese patients with plaque psoriasis.
He HUANG ; Min CHEN ; Wenjuan WU ; Tianhui YANG ; Hao LIU ; Zhengwei ZHU ; Wenjun WANG ; Sen YANG ; Xian DING ; Hui WANG ; Yujun SHENG ; Yaohua ZHANG ; Min LI ; Xuejun ZHANG
Chinese Medical Journal 2023;136(3):360-361
3.The effect of adeno⁃associated virus delivery of shRNA against EP3 receptors in the bilateral lateral parabrachial nucleus of rats on fever
Tianhui He ; Nanping Wang ; Sihao Wu ; Yanlin Wei ; Jianhui Xu ; Jie Zhang
Acta Universitatis Medicinalis Anhui 2023;58(11):1872-1877
		                        		
		                        			Objective  :
		                        			To investigate the effect of adeno⁃associated virus ( AAV) delivery of short hairpin RNA ( shRNA) against the Ptger3 gene in the lateral parabrachial nucleus (LPB) on the fever induced by microinjection of prostaglandin E2 (PGE2 ) into the LPB and the intraperitoneal injection of lipopolysaccharide (LPS) .
		                        		
		                        			Methods:
		                        			AAV2⁃shRNA⁃Ptger3(EP3) ⅣEGFP ( shRNA⁃EP3) and AAV2⁃ CMV⁃ EGFP ( shRNA⁃control) viruses were constructed and transfected the rat LPB by stereotaxic injection.  Four weeks later, the transfection efficiency of AAV viruses was observed by fluorescence microscopy , and the knockdown efficiency was determined by real⁃time PCR of EP3 receptor mRNA on the LPB. The effects of microinjection of saline or PGE2 in the LPB or intraperitoneal injection of LPS on body temperature (Tcore ) and energy expenditure (EE) of shRNA⁃control group and shRNA⁃EP3 group were monitored using an animal monitoring system with temperature telemetry.
		                        		
		                        			Results :
		                        			 AAV virus transfecnificant difference in basal body temperature between shRNA⁃control group and shRNA⁃EP3 group.  Tcore and EE were briefly and slightly increased after microinjection of saline in the LPB , but there was no significant difference between the two groups. Compared with the shRNA⁃control group , the febrile response induced by LPB PGE2 was attenuated in the shRNA⁃EP3 group (P < 0. 05) . Furthermore , the knockdown of EP3 receptor of LPB also attenuated the LPS⁃induced fever, and the Tcore 5. 5 h post⁃LPS in the shRNA⁃EP3 rats increased compared with the baseline (P < 0. 05) , which was lower than that in the shRNA⁃control rats ( P < 0. 01) .  
		                        		
		                        			Conclusion   
		                        			EP3 receptor knockdown in LPB attenuates the febrile response induced by microinjection of PGE2 in the LPB and intraperitoneal injection of LPS , suggesting that EP3 receptors of LPB mediate the pyrogenic action of LPB PGE2 and partly participate in LPS⁃induced fever.
		                        		
		                        		
		                        		
		                        	
4.Excitory effect of arginine vasopressin on median preoptic glutamatergic neurons and its mechanism
Xiaoyu Hou ; Yi' ; an Song ; Tianhui He ; Wenmin Gao ; Jie Zhang ; Jianhui Xu
Acta Universitatis Medicinalis Anhui 2023;58(3):418-422,428
		                        		
		                        			Objective :
		                        			To investigate the effect of Arginine Vasopressin (AVP) on the median preoptic glutamatergic  (MnPOVglut2 ) neurons and its mechanism.
		                        		
		                        			Methods :
		                        			Brain  slices  were prepared from  male Vglut2-tdTomato mice.MnPOVglut2  neurons expressing red fluorescent protein were located by using fluorescence microscope.Wholecell patch clamp technique was used to observe the effect of AVP on the firing frequency of MnPOVglut2  neurons,the  effect of synaptic transmission blockers  ( STBs) on the AVP-induced change in the firing frequency of MnPOVglut2   neurons,and the effect of AVP V1a receptor antagonist on the AVP-induced change in the firing frequency of MnPOVglut2  neurons.  
		                        		
		                        			Results:
		                        			The mean firing frequency of MnPOVglut2  neurons increased during perfusion with artificial  cerebrospinal fluid  (ACSF) and AVP compared with that during perfusion with ACSF  (P<0. 01) ,indicating that  AVP excited the MnPOVglut2  neurons.The mean firing frequency of MnPOVglut2  neurons still increased during perfusion with ACSF,STBs,and AVP compared with that during perfusion with ACSF and STBs  (P<0. 001) ; moreover,the magnitude of AVP-induced increase in firing frequency didn't change significantly during perfusion with  ACSF,STBs,and AVP compared with that during perfusion with ACSF and AVP  (P >0. 05 ) ,suggesting  that  AVP excited the MnPOVglut2  neurons directly in a postsynaptic manner.The magnitude of AVP-induced increase in  the firing frequency of MnPOVglut2  neurons declined during perfusion with ACSF,STBs,AVP,and V1areceptor antagonist compared with that during perfusion with ACSF,STBs,and AVP  (P<0. 01) ,suggesting that AVP excited  MnPOVglut2  neurons directly via V1a receptor.
		                        		
		                        			Conclusion 
		                        			AVP can excite MnPOVglut2  neurons via V1areceptor directly in a postsynaptic manner.This study reveals the molecular marker of MnPO neurons which AVP act on.
		                        		
		                        		
		                        		
		                        	
5.Analysis of risk factors for prolonged mechanical ventilation in patients with sepsis after abdominal surgery
Tianhui HE ; Feng SHEN ; Shuwen LI ; Bo LIU
Chinese Critical Care Medicine 2022;34(7):693-698
		                        		
		                        			
		                        			Objective:To analyze the risk factors of prolonged mechanical ventilation (PMV) in patients with sepsis complicated by abdominal surgery, and to evaluate the predictive value of risk factors for PMV.Methods:A retrospective case-control study was conducted. The clinical data of patients with postoperative abdominal sepsis complicated with invasive mechanical ventilation who were admitted to the intensive care unit (ICU) of the Affiliated Hospital of Guizhou Medical University from January 1, 2018 to December 31, 2020 were collected. The patients were divided into PMV group (duration of mechanical ventilation longer than 48 hours) and non-PMV group (duration of mechanical ventilation shorter than 48 hours) according to the duration of mechanical ventilation in ICU. The patient's gender, age, body mass index (BMI), underlying diseases, mean arterial pressure (MAP), complete blood count, blood biochemistry, arterial blood gas, cardiac function indicators, procalcitonin (PCT) at admission to the ICU, the acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) and the sequential organ failure assessment (SOFA) scores in the first 24 hours of admission to the ICU, and other clinical information were recorded. Univariate and multivariate Logistic regression models were used to analyze the risk factors for PMV. Receiver operator characteristic curve (ROC curve) was drawn to evaluate the predictive value of related indicators for PMV.Results:A total of 195 patients with sepsis after abdominal surgery who received invasive mechanical ventilation were enrolled, including 127 males (65.1%) and 68 females (34.9%), with the median age of 65 (21, 93) years old. There were 91 patients (46.7%) in the non-PMV group and 104 patients (53.3%) in the PMV group. Univariate analysis showed that the APACHEⅡ score, SOFA score, cardiac troponin T (cTnT), N-terminal pro-B type natriuretic peptide (NT-proBNP) in the PMV group were significantly higher than those in the non-PMV group. Oxygenation index (PaO 2/FiO 2), total protein (TP) and prealbumin (PA) in the PMV group were all lower than those in the non-PMV group when admitted to ICU. In the PMV group, serum creatinine (SCr), blood urea nitrogen (BUN), cystatin C (Cys C) were significantly increased, prothrombin time (PT) was significantly prolonged, the proportion of patients with septic shock and hypertension were significantly increased as compared with those in the non-PMV group. Multivariate analysis showed that low PaO 2/FiO 2 at ICU admission [odds ratio ( OR) = 0.995, 95% confidence interval (95% CI) was 0.992-0.999, P = 0.010], high ln PCT ( OR = 1.301, 95% CI was 1.088-1.555, P = 0.004), high ln cTnT ( OR = 1.562, 95% CI was 1.079-2.261, P = 0.018) and septic shock ( OR = 4.967, 95% CI was 2.461-10.026, P = 0.000) were the independent risk factors for PMV in patients with sepsis after abdominal surgery. ROC curve analysis showed that the PaO 2/FiO 2, ln cTnT, ln PCT and septic shock had certain predictive value for PMV, the area under the ROC curve (AUC) of the four variables were 0.607, 0.638, 0.690 and 0.711, the sensitivity was 50.0%, 62.5%, 86.5% and 74.0%, and the specificity was 71.4%, 62.6%, 48.3% and 68.1%, respectively. The AUC for the joint prediction of the four variables was 0.803, with a sensitivity of 76.0% and a specificity of 78.0%. It suggested that the multivariate joint prediction of PMV was more accurate. Conclusions:Decreased PaO 2/FiO 2, increased PCT, increased cTnT and the occurrence of septic shock are independent risk factors for PMV in patients with sepsis complicated by abdominal surgery. The combination of above four indices was more accurate than one single variable in predicting PMV and had higher diagnostic value.
		                        		
		                        		
		                        		
		                        	
6.Comparative study of dexmedetomidine vs. midazolam on plasma catecholamine levels and hemodynamics in patients with septic shock
Ling YAO ; Bo LIU ; Tianhui HE ; Feng SHEN
Chinese Critical Care Medicine 2021;33(10):1193-1197
		                        		
		                        			
		                        			Objective:To compare the effects of dexmedetomidine (DEX) and midazolam on the endogenous plasma catecholamine levels and the dosage of exogenous norepinephrine (NE) to maintain blood pressure in patients with septic shock.Methods:From January 2018 to December 2019, patients admitted to the department of critical care medicine of the Affiliated Hospital of Guizhou Medical University who needed invasive mechanical ventilation and had a stay of more than 48 hours in the intensive care unit (ICU) for septic shock and received DEX or midazolam for sedation were enrolled in this study. The hemodynamic data, arterial blood lactic acid (Lac) level, arterial blood gas analysis and vasoactive drug dose at 0, 12, 24, 48, 72 hours after entering the ICU were dynamically recorded, and the plasma catecholamine levels at 0, 24, 48 hours after entering the ICU were recorded. The acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and sequential organ failure assessment (SOFA) score on ICU-admission were calculated. The parameters of prognosis were collected.Results:A total of 24 patients were enrolled, 12 in the DEX group and 12 in the midazolam group. There were similar dynamic trends in heart rate (HR), central venous pressure (CVP), venous-arterial carbon dioxide pressure difference (Pv-aCO 2), oxygenation index (PaO 2/FiO 2), and Lac level of patients between the two groups. Only the 12-hour CVP and 72-hour Pv-aCO 2 in the DEX group were significantly higher than those in the midazolam group [CVP (mmHg, 1 mmHg = 0.133 kPa): 13±3 vs. 10±3, Pv-aCO 2 (mmHg): 9.4±5.2 vs. 4.8±2.2, both P < 0.05], and the mean arterial pressure (MAP) of patients in the DEX group at 48 hours and 72 hours was significantly higher than that in the midazolam group (mmHg: 95±10 vs. 86±10, 96±9 vs. 88±7, both P < 0.05). There was no statistically significant difference in the duration of mechanical ventilation, ICU mortality or in-hospital mortality between the DEX group and the midazolam group [duration of mechanical ventilation (days): 5.6 (3.8, 9.5) vs. 10.5 (5.9, 15.0), ICU mortality: 16.7% vs. 33.3%, in-hospital mortality: 25.0% vs. 41.7%, all P > 0.05]. There was no significant difference in the dosage of propofol or sufentanil between the DEX group and the midazolam group [propofol (mg/kg): 0 (0, 9.35) vs. 4.07 (0, 13.75), sufentanil (μg/kg): 6.26 (4.90, 9.80) vs. 8.32 (3.52, 9.34), both P > 0.05]. The levels of plasma NE, dopamine and dobutamine in the DEX group at 48 hours were significantly lower than those in the midazolam group [NE (ng/L): 1 850.12 (342.16, 2 938.05) vs. 4 596.60 (3 310.56, 5 546.84), dopamine (ng/L): 119.10 (60.47, 200.71) vs. 275.40 (214.61, 418.88), dobutamine (ng/L): 51.20 (36.85, 75.59) vs. 98.97 (85.65, 107.10), all P < 0.05], but the amount of NE required to maintain MAP between 65 mmHg and 75 mmHg in the DEX group and the midazolam group was similar [μg/kg: 1 922 (1 170, 4 887) vs. 2 466 (2 043, 3 438), P > 0.05]. Conclusion:DEX can reduce plasma catecholamine levels in patients with septic shock more than midazolam, and does not increase the dose of exogenous NE, and has a smaller effect on hemodynamics in patients with septic shock than midazolam.
		                        		
		                        		
		                        		
		                        	
7.Meta-analysis and trials sequential analysis for the efficacy and safety of low- and medium-dose glucocorticoids in adult acute respiratory distress syndrome patients
Qing LI ; Chuan XIAO ; Feng SHEN ; Wei LI ; Shuwen LI ; Tianhui HE ; Jincheng QIN
Chinese Critical Care Medicine 2021;33(11):1302-1308
		                        		
		                        			
		                        			Objective:To explore the efficacy and safety of low- and medium-dose of glucocorticoids in adult patients with acute respiratory distress syndrome (ARDS) through Meta-analysis and trials sequential analysis (TSA).Methods:Databases associated with adult ARDS treatment with low- and medium-dose glucocorticoids both in English and in Chinese were searched from PubMed, Medline, China Biology Medicine (CBM), Cochrane Library, CNKI, Wanfang Data and VIP, of which the search duration was from the establishment of the database to December 2020. Low-dose glucocorticoids were defined as methylprednisolone ≤ 1 mg·kg -1·d -1, and medium dose glucocorticoids were defined as methylprednisolone ≤ 2 mg·kg -1·d -1. According to the Cochrane Collaboration bias risk assessment tool, the quality of the included literature was evaluated, and the data were extracted. Meta-analysis and TSA were used to evaluate the effects of low- and medium-dose glucocorticoids on the hospital mortality, intensive care unit (ICU) mortality, and mechanical ventilation free time in ICU for 28 days, PaO 2/FiO 2, and the occurrence of nosocomial infections and hyperglycemia. Results:A total of 996 patients in 7 literatures were finally included, including 515 patients in the low- and medium-dose glucocorticoid group (hormone group) and 481 patients in the conventional treatment group (control group). The research quality of 7 literatures was relatively high. The results of Meta-analysis and TSA showed that, compared with the control group, the hospital mortality in the hormone group was significantly decreased [relative risk ( RR) = 0.77, 95% confidence interval (95% CI) was 0.66-0.89, P = 0.000 6], and mechanical ventilation free time in ICU for 28 days was significantly prolonged [standardized mean difference ( SMD) = 0.50, 95% CI was 0.36-0.65, P < 0.000 1]. Although Meta-analysis showed that the ICU mortality of the hormone group was significantly lower than that of the control group ( RR = 0.61, 95% CI was 0.38-0.99, P = 0.04), the TSA results showed that the cumulative Z value crossed the traditional threshold, but did not cross the TSA cut-off value, and the sample size did not reach required information size (RIS, n = 3 252), needed more research to confirm. Although Meta-analysis showed that PaO 2/FiO 2 in the hormone group was significantly higher than that in the control group ( SMD = 0.78, 95% CI was 0.13-1.43, P = 0.02), TSA showed that the cumulative Z value did not pass the traditional and TSA cut-off values. More research was needed for verification. Meta-analysis also showed that there was no significant difference in the incidence of new infection ( RR = 0.93, 95% CI was 0.74-1.17, P = 0.54) and the incidence of hyperglycemia ( RR = 1.11, 95% CI was 1.00-1.23, P = 0.05) between the hormone group and the control group. Conclusion:low- and medium-dose of glucocorticoids therapy can reduce the hospital mortality of adult ARDS patients and shorten the mechanical ventilation duration in ICU for 28 days, and low- and medium-dose of glucocorticoids therapy does not increase the risk of infection and hyperglycemia.
		                        		
		                        		
		                        		
		                        	
8.Risk factors for death in elderly patients admitted to intensive care unit after elective abdominal surgery: a consecutive 5-year retrospective study
Shuwen LI ; Tianhui HE ; Feng SHEN ; Difen WANG ; Xu LIU ; Jingcheng QIN ; Chuan XIAO ; Wei LI ; Qing LI ; Daixiu GAO
Chinese Critical Care Medicine 2021;33(12):1453-1458
		                        		
		                        			
		                        			Objective:To investigate the risk factors that were associated with the death of elderly patients who were admitted to the intensive care unit (ICU) after elective abdominal surgery, and to find reliable and sensitive predictive indicators for early interventions and reducing the mortality.Methods:A retrospective case-control study was conducted. The clinical data of elderly (age≥65 years old) patients after elective abdominal surgery admitted to the ICU of the Affiliated Hospital of Guizhou Medical University from January 1st 2016 to December 31st 2020 were collected, including the patient's gender, age, body mass index (BMI), medical history, American Society of Anesthesiologists (ASA) grades, surgical classification, intraoperative blood loss, duration of operation, interval time between end of operation and admission to the ICU, acute physiology and chronic health evaluationⅡ(APACHEⅡ) score and the worst laboratory examination results within 24 hours of ICU admission, the first blood gas analysis in ICU, the duration of invasive mechanical ventilation, and the length of ICU stay. Postoperative abdominal infection was evaluated by the pathogenic culture of peritoneal drainage fluid and clinical symptoms and signs. The patients were divided into death group and survival group based on clinical outcomes, and clinical data were compared between the two groups. Binary multivariate Logistic regression analysis was used to screen the risk factors of death, and the receiver operator characteristic curve (ROC curve) was plotted to analyze the predictive values of these risk factors.Results:A total of 226 elderly patients with elective abdominal surgery were admitted to the ICU of our hospital during the past 5 years, of whom, two patients who did not undergo laboratory examinations within 24 hours of admission to the ICU were excluded. Finally, 224 patients met the criteria, with 158 survivors and 66 deaths. Univariate analysis showed that: compared with survival group, APACHEⅡscore, blood lactate acid (Lac) and the proportion of postoperative abdominal infection were higher in death group [APACHEⅡ score: 27.5 (25.0, 31.3) vs. 23.0 (18.0, 27.0), Lac (mmol/L): 2.9 (1.8, 6.6) vs. 1.8 (1.1, 2.8), the proportion of postoperative abdominal infection: 65.2% (43/66) vs. 35.4% (56/158), all P < 0.01], prothrombin time (PT), activated partial thromboplastin time (APTT) and interval time between end of surgery and admission to ICU were longer [PT (s): 17.20 (14.50, 18.63) vs. 14.65 (13.90, 16.23), APTT (s): 45.15 (38.68, 55.15) vs. 39.45 (36.40, 45.70), interval time between end of surgery and admission to ICU (hours): 39.2 (0.7, 128.9) vs. 0.7 (0.3, 2.0), all P <0.01], postoperative hemoglobin (Hb), platelet count (PLT), prealbumin (PA), mean arterial pressure (MAP) and oxygenation index (PaO 2/FiO 2) were lower in death group [Hb (g/L): 95.79±23.64 vs. 105.58±19.82, PLT (×10 9/L): 138.5 (101.0, 177.5) vs. 160.5 (118.5, 232.3), PA (g/L): 80.88±43.63 vs. 116.54±50.80, MAP (mmHg, 1 mmHg = 0.133 kPa): 76.8±19.1 vs. 91.6±19.8, PaO 2/FiO 2 (mmHg): 180.0 (123.5, 242.5) vs. 223.5 (174.8, 310.0), all P < 0.05]. Binary multivariate Logistic regression analysis showed that APACHEⅡscore [odds ratio ( OR) = 1.187, 95% confidence interval (95% CI) =1.008-1.294, P < 0.001], interval time between end of operation and admission to ICU ( OR = 1.005, 95% CI = 1.001-1.009, P = 0.016) and postoperative abdominal infection ( OR = 2.630, 95% CI = 1.148-6.024, P = 0.022) were independent risk factors for prognosis in these patients. MAP ( OR = 0.978, 95% CI = 0.957-0.999, P = 0.041) and PaO 2/FiO 2 ( OR = 0.994, 95% CI = 0.990-0.998, P = 0.003) were protective factors for the patients' prognosis. Lac, Hb, PLT, PA, PT and APTT had no predictive value for the prognosis of elderly patients admitted to ICU after elective abdominal surgery [ OR value and 95% CI were 1.075 (0.945-1.223), 1.011 (0.99-1.032), 1.000 (0.995-1.005), 0.998 (0.989-1.007), 1.051 (0.927-1.192) and 1.003 (0.991-1.016), respectively, all P > 0.05. ROC curve analysis showed that APACHEⅡscore, interval time between end of operation and admission to the ICU and the postoperative abdominal infection had certain predictive values for the prognosis of elderly patients, the area under ROC curve (AUC) were 0.755, 0.732 and 0.649 respectively, all P < 0.001; When the cut-off of APACHEⅡscore and interval time between end of operation and admission to the ICU were 24.5 scores and 2.15 hours, the sensitivity were 78.8% and 66.7%, respectively, and the specificity were 62.0% and 76.6%, respectively. The combined predictive value of the three variables was the highest, which AUC was 0.846, the joint prediction probability was 0.27, the sensitivity was 83.3%, and the specificity was 75.3%. Conclusion:APACHEⅡscore, interval time between end of surgery and admission to ICU, and postoperative abdominal infection may be independent risk factors for the death of elderly patients who were admitted to the ICU after elective abdominal surgery, there would be far greater predictive values when the three variables were combined.
		                        		
		                        		
		                        		
		                        	
9.Berberine dose-dependently inhibits the expression of procoagulant and fibrinolytic inhibitory factors in lipopolysaccharide-induced rat type Ⅱ alveolar epithelial cells
Xiang LI ; Guixia YANG ; Feng SHEN ; Xinghao ZHENG ; Tianhui HE ; Shuwen LI ; Yumei CHENG ; Qing LI ; Wei LI ; Jincheng QIN
Chinese Critical Care Medicine 2021;33(1):53-58
		                        		
		                        			
		                        			Objective:To observe the effects of berberine on procoagulant and fibrinolytic inhibitory factors produced by rat type Ⅱ alveolar epithelial cell (AECⅡ) induced by lipopolysaccharide (LPS).Methods:AECⅡ cells (RLE-6TN cells) were cultured in vitro, and the cells in logarithmic growth phase were collected. The cytotoxicity text of berberine was detected by cell counting kit-8 (CCK-8) to determine the drug concentration range according to inhibition concentration of half cells (IC 50). The RLE-6TN cells were divided into five groups, the cells in blank control group were cultured in DMEM; the cells in LPS group were stimulated with 5 mg/L LPS; and the cells in berberine pretreatment groups were pretreated with 20, 50 and 80 μmol/L berberine for 1 hour, and then were co-cultured with 5 mg/L LPS. The cells were collected after LPS induced for 24 hours. The protein and mRNA expression levels of tissue factor (TF), tissue factor pathway inhibitor (TFPI) and plasminogen activator inhibitor-1 (PAI-1) in the cells were detected by Western blotting and real-time fluorescence quantification reverse transcription-polymerase chain reaction (RT-qPCR). The levels of activated protein C (APC), precollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT) and antithrombin Ⅲ (ATⅢ) in the cell supernatant were measured by enzyme linked immunosorbent assay (ELISA). Results:According to the inhibition rate curve, the IC 50 of berberine on RLE-6TN cells was 81.16 μmol/L. Therefore, 20, 50 and 80 μmol/L were selected as the intervention concentration of berberine. Compared with the blank control group, the expression and secretion of procoagulant and fibrinolytic inhibitory factors were abnormal in RLE-6TN cells after LPS induced for 24 hours. The protein and mRNA expression levels of TF and PAI-1 in the LPS group were significantly increased, but the protein and mRNA expression levels of TFPI were significantly decreased. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly decreased, while the levels of PⅢP and TAT were significantly increased. After pretreatment with berberine, the abnormal expression and secretion of procoagulant and fibrinolytic inhibitory factors induced by LPS were corrected in a dose-dependent manner, especially in 80 μmol/L. Compared with the LPS group, the protein and mRNA expression levels of TF and PAI-1 in the berberine 80 μmol/L group were significantly decreased [TF protein (TF/GAPDH): 0.45±0.02 vs. 0.55±0.03, TF mRNA (2 -ΔΔCt): 0.39±0.08 vs. 1.48±0.11, PAI-1 protein (PAI-1/GAPDH): 0.37±0.02 vs. 0.64±0.04, PAI-1 mRNA (2 -ΔΔCt): 1.14±0.29 vs. 4.18±0.44, all P < 0.01] and those of TFPI were significantly increased [TFPI protein (TFPI/GAPDH): 0.53±0.02 vs. 0.45±0.02, TFPI mRNA (2 -ΔΔCt): 0.94±0.08 vs. 0.40±0.05, both P < 0.01]. Meanwhile, the levels of APC and ATⅢ in the cell supernatant were significantly increased [APC (μg/L): 1 358.5±26.0 vs. 994.2±23.1, ATⅢ (μg/L): 118.0±7.4 vs. 84.4±2.7, both P < 0.01], while those of PⅢP and TAT were significantly decreased [PⅢP (μg/L): 11.2±0.4 vs. 18.6±0.9, TAT (ng/L): 222.1±2.8 vs. 287.6±7.0, both P < 0.01]. Conclusions:Berberine could inhibit the LPS-induced expressions of procoagulant and fibrinolytic inhibitory factors in rat AECⅡ cells and promote the expressions of anticoagulant factors in a dose-dependent manner. Berberine may be a new therapeutic target for alveolar hypercoagulability and fibrinolysis inhibition in acute respiratory distress syndrome (ARDS).
		                        		
		                        		
		                        		
		                        	
10.Effects of andrographolide on the expression of procoagulant and fibrinolytic inhibition factors in rat type Ⅱ alveolar epithelial cells stimulated by lipopolysaccharide
Guixia YANG ; Xiang LI ; Feng SHEN ; Xinghao ZHENG ; Tianhui HE ; Shuwen LI ; Yumei CHENG ; Qing LI ; Wei LI ; Jincheng QIN
Chinese Critical Care Medicine 2021;33(2):155-160
		                        		
		                        			
		                        			Objective:To determine the effect of andrographolide (AD) on the expression of procoagulant and fibrinolytic inhibitory factors in rat type Ⅱ alveolar epithelial cells (AECⅡ) stimulated by lipopolysaccharide (LPS).Methods:The AECⅡ cells RLE-6TN in the logarithmic growth phase were divided into 5 groups: the normal control (NC) group, the LPS group, and the 6.25, 12.5, and 25 mg/L AD groups (AD 6.25 group, AD 12.5 group, AD 25 group). The NC group was cultured with RPMI 1640 conventional medium. In the LPS group, 5 mg/L LPS was added to the RPMI 1640 conventional medium for stimulation. Cells in the AD groups were treated with 6.25, 12.5, and 25 mg/L AD in advance for 1 hour and then given LPS to stimulate the culture. The cells and cell culture supernatant were collected 24 hours after LPS stimulation. The protein and mRNA expressions of tissue factor (TF), tissue factor pathway inhibitor (TFPI), and plasminogen activator inhibition-1 (PAI-1) in cells were detected by Western blotting and real-time fluorescent quantitative polymerase chain reaction (RT-qPCR). The levels of procollagen Ⅲ peptide (PⅢP), thrombin-antithrombin complex (TAT), antithrombin Ⅲ (AT-Ⅲ) and activated protein C (APC) in the cell supernatant were detected by enzyme linked immunosorbent assay (ELISA).Results:Compared with the NC group, the protein and mRNA expressions of TF and PAI-1 in the LPS group were significantly increased, and the protein and mRNA expressions of TFPI were significantly reduced. At the same time, the levels of PⅢP and TAT in the cell supernatant were significantly increased, the levels of AT-Ⅲ, APC were significantly reduced. Compared with the LPS group, the protein and mRNA expressions of TF and PAI-1 in AD 6.25 group, AD 12.5 group, AD 25 group were significantly reduced [TF/GAPDH: 0.86±0.08, 0.45±0.04, 0.44±0.04 vs. 1.32±0.10, TF mRNA (2 -ΔΔCt): 2.59±0.25, 2.27±0.05, 1.95±0.04 vs. 4.60±0.26, PAI-1/GAPDH: 2.11±0.07, 1.45±0.04, 0.86±0.09 vs. 2.56±0.09, PAI-1 mRNA (2 -ΔΔCt): 3.50±0.22, 2.23±0.29, 1.84±0.09 vs. 6.60±0.27, all P < 0.05], while the protein and mRNA expressions of TFPI were significantly increased [TFPI/GAPDH: 0.78±0.05, 0.81±0.03, 0.84±0.07 vs. 0.36±0.02, TFPI mRNA (2 -ΔΔCt): 0.46±0.09, 0.69±0.07, 0.91±0.08 vs. 0.44±0.06, all P < 0.05]. Also the levels of PⅢP and TAT in the cell supernatant were significantly reduced, and the levels of AT-Ⅲ and APC were significantly increased [PⅢP (μg/L): 13.59±0.23, 12.66±0.23, 10.59±0.30 vs. 15.82±0.29, TAT (ng/L): 211.57±6.41, 205.69±4.04, 200.56±9.85 vs. 288.67±9.84, AT-Ⅲ (μg/L): 102.95±3.86, 123.92±2.63, 128.67±1.67 vs. 92.93±3.36, APC (μg/L): 1 188.95±14.99, 1 366.12±39.93, 1 451.15±29.69 vs. 1 145.55±21.07, all P < 0.05]. With the increase of the dose of AD, the above-mentioned promotion and inhibition effects became more obvious. In the AD 25 group, TF, PAI-1 protein and mRNA expressions decreased, TFPI mRNA expression increased, PⅢP level in the supernatant decreased and AT-Ⅲ, APC levels increased compared with AD 6.25 group, the difference was statistically significant, and the decrease of PAI-1 protein expression and PⅢP level in the supernatant were also statistically significant compared with AD 12.5 group. Conclusions:Andrographolide in the dose range of 6.25-25 mg/L can dose-dependently inhibit the expression and secretion of procoagulant and fibrinolytic inhibitor-related factors in AECⅡ cells RLE-6TN stimulated by LPS, and promote the secretion of anticoagulant factors. 25 mg/L has the most obvious effect.
		                        		
		                        		
		                        		
		                        	
            

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