1.Median effective dose of remimazolam combined with sufentanil for inhibition of nasopharyngeal airway insertion reaction in elderly patients
Shuwen QIAN ; Xuan GUO ; Yun SHEN ; Mazhong ZHANG
The Journal of Clinical Anesthesiology 2024;40(1):61-64
Objective To calculate median effective dose(ED50)of remimazolam combined with sufentanil for inhibition of nasopharyngeal airway insertion reaction in elderly patients.Methods Thirty-eight patients,aged≥65 years,BMI 18-25 kg/m2,ASA physical statusⅠ-Ⅲ,undergoing elective cata-ract surgery under nasopharyngeal airway were selected.Patients received intravenous sufentanil injections of 0.1 μg/kg,followed by intravenous remimazolam injection 3 minutes later,then nasopharyngeal airway in-sertion after 2 minutes.The initial dose of remimazolam was 0.2 mg/kg and adjusted by 0.01 mg/kg,based on the modified Dixon sequential method.A positive reaction was defined as head shaking,choking,body movement,an increase in HR>20%of the basal value,or an increase in SBP or DBP>20%of the basal value at the time of nasopharyngeal airway placement.The procedure was stopped after seven crossovers.The ED50,95%effective dose(ED95)of remimazolam,and 95%confidence intervals(CI)were calculated by probabilistic unit(Probit)regression analysis.Results When combined with sufentanil,the ED50 and ED95 of remimazolam for inhibition of nasopharyngeal airway insertion in elderly patients was 0.193 mg/kg(95%CI 0.191-0.195 mg/kg)and 0.209 mg/kg(95%CI 0.205-0.213 mg/kg),respectively.Conclusion The ED50 of remimazolam combined with sufentanil for inhibition of nasopharyngeal airway in-sertion reaction in elderly patients were 0.193 mg/kg(95%CI 0.191-0.195 mg/kg).
2.Factors influencing pulmonary infection in elderly neurocritical patients and their predictive values: a data analysis for consecutive four-year
Jia YUAN ; Ying LIU ; Di LIU ; Difen WANG ; Feng SHEN ; Xu LIU ; Shuwen LI ; Dehua HE
Chinese Critical Care Medicine 2023;35(1):66-70
Objective:To analyze the factors influencing pulmonary infections in elderly neurocritical patients in the intensive care unit (ICU) and to explore the predictive value of risk factors for pulmonary infections.Methods:The clinical data of 713 elderly neurocritical patients [age ≥ 65 years, Glasgow coma score (GCS) ≤ 12 points] admitted to the department of critical care medicine of the Affiliated Hospital of Guizhou Medical University from 1 January 2016 to 31 December 2019 were retrospectively analyzed. According to whether or not they had HAP, the elderly neurocritical patients were divided into hospital-acquired pneumonia (HAP) group and non-HAP group. The differences in baseline data, medication and treatment, and outcome indicators between the two groups were compared. Logistic regression analysis was used to analyze the factors influencing the occurrence of pulmonary infection.The receiver operator characteristic curve (ROC curve) was plotted for risk factors and a predictive model was constructed to evaluate the predictive value for pulmonary infection.Results:A total of 341 patients were enrolled in the analysis, including 164 non-HAP patients and 177 HAP patients. The incidence of HAP was 51.91%. According to univariate analysis, compared with the non-HAP group, mechanical ventilation time, the length of ICU stay and total hospitalization in the HAP group were significantly longer [mechanical ventilation time (hours): 171.00 (95.00, 273.00) vs. 60.17 (24.50, 120.75), the length of ICU stay (hours): 263.50 (160.00, 409.00) vs. 114.00 (77.05, 187.50), total hospitalization (days): 29.00 (13.50, 39.50) vs. 27.00 (11.00, 29.50), all P < 0.01], the proportion of open airway, diabetes, proton pump inhibitor (PPI), sedative, blood transfusion, glucocorticoids, and GCS ≤ 8 points were significantly increased than those in HAP group [open airway: 95.5% vs. 71.3%, diabetes: 42.9% vs. 21.3%, PPI: 76.3% vs. 63.4%, sedative: 93.8% vs. 78.7%, blood transfusion: 57.1% vs. 29.9%, glucocorticoids: 19.2% vs. 4.3%, GCS ≤ 8 points: 83.6% vs. 57.9%, all P < 0.05], prealbumin (PA) and lymphocyte count (LYM) decreased significantly [PA (g/L): 125.28±47.46 vs. 158.57±54.12, LYM (×10 9/L): 0.79 (0.52, 1.23) vs. 1.05 (0.66, 1.57), both P < 0.01]. Logistic regression analysis showed that open airway, diabetes, blood transfusion, glucocorticoids and GCS ≤ 8 points were independent risk factors for pulmonary infection in elderly neurocritical patients [open airway: odds ratio ( OR) = 6.522, 95% confidence interval (95% CI) was 2.369-17.961; diabetes: OR = 3.917, 95% CI was 2.099-7.309; blood transfusion: OR = 2.730, 95% CI was 1.526-4.883; glucocorticoids: OR = 6.609, 95% CI was 2.273-19.215; GCS ≤ 8 points: OR = 4.191, 95% CI was 2.198-7.991, all P < 0.01], and LYM, PA were the protective factors for pulmonary infection in elderly neurocritical patients (LYM: OR = 0.508, 95% CI was 0.345-0.748; PA: OR = 0.988, 95% CI was 0.982-0.994, both P < 0.01). ROC curve analysis showed that the area under the ROC curve (AUC) for predicting HAP using the above risk factors was 0.812 (95% CI was 0.767-0.857, P < 0.001), with a sensitivity of 72.3% and a specificity of 78.7%. Conclusions:Open airway, diabetes, glucocorticoids, blood transfusion, GCS ≤ 8 points are independent risk factors for pulmonary infection in elderly neurocritical patients. The prediction model constructed by the above mentioned risk factors has certain predictive value for the occurrence of pulmonary infection in elderly neurocritical patients.
3.Prognosis of patients planned and unplanned admission to the intensive care unit after surgery: a comparative study.
Wei LI ; Shuwen LI ; Feng SHEN ; Liang LI ; Daixiu GAO ; Bo LIU ; Lulu XIE ; Xian LIU ; Difen WANG ; Chunya WU
Chinese Critical Care Medicine 2023;35(7):746-751
OBJECTIVE:
To compare and analyze the effect of unplanned versus planned admission to the intensive care unit (ICU) on the prognosis of high-risk patients after surgery, so as to provide a clinical evidence for clinical medical staff to evaluate whether the postoperative patients should be transferred to ICU or not after surgery.
METHODS:
The clinical data of patients who were transferred to ICU after surgery admitted to the Affiliated Hospital of Guizhou Medical University from January to December in 2021 were retrospectively analyzed, including gender, age, body mass index, past history (whether combined with hypertension, diabetes, pulmonary disease, cardiac disease, renal failure, liver failure, hematologic disorders, tumor, etc.), acute physiology and chronic health evaluation II (APACHE II), elective surgery, pre-operative hospital consultation, length of surgery, worst value of laboratory parameters within 24 hours of ICU admission, need for invasive mechanical ventilation (IMV), duration of IMV, length of ICU stay, total length of hospital stay, ICU mortality, in-hospital mortality, and survival status at 30th day postoperative. The unplanned patients were further divided into the immediate transfer group and delayed transfer group according to the timing of their ICU entrance after surgery, and the prognosis was compared between the two groups. Cox regression analysis was used to find the independent risk factors of 30-day mortality in patients transferred to ICU after surgery.
RESULTS:
Finally, 377 patients were included in the post-operative admission to the ICU, including 232 in the planned transfer group and 145 in the unplanned transfer group (42 immediate transfers and 103 delayed transfers). Compared to the planned transfer group, patients in the unplanned transfer group had higher peripheral blood white blood cell count (WBC) at the time of transfer to the ICU [×109/L: 10.86 (7.09, 16.68) vs. 10.11 (6.56, 13.27)], longer total length of hospital stay [days: 23.00 (14.00, 34.00) vs. 19.00 (12.00, 29.00)], and 30-day post-operative mortality was higher [29.66% (43/145) vs. 17.24% (40/232)], but haemoglobin (Hb), arterial partial pressure of carbon dioxide (PaCO2), oxygenation index (PaO2/FiO2), and IMV requirement rate were lower [Hb (g/L): 95.00 (78.00, 113.50) vs. 98.00 (85.00, 123.00), PaCO2 (mmHg, 1 mmHg ≈ 0.133 kPa): 36.00 (29.00, 41.50) vs. 39.00 (33.00, 43.00), PaO2/FiO2 (mmHg): 197.00 (137.50, 283.50) vs. 238.00 (178.00, 350.25), IMV requirement rate: 82.76% (120/145) vs. 93.97% (218/232)], all differences were statistically significant (all P < 0.05). Kaplan-Meier survival curve showed that the 30-day cumulative survival rate after surgery was significantly lower in the unplanned transfer group than in the planned transfer group (Log-Rank test: χ2 = 7.659, P = 0.006). Univariate Cox regression analysis showed that unplanned transfer, APACHE II score, whether deeded IMV at transfer, total length of hospital stay, WBC, blood K+, and blood lactic acid (Lac) were associated with 30-day mortality after operation (all P < 0.05). Multifactorial Cox analysis showed that unplanned transfer [hazard ratio (HR) = 2.45, 95% confidence interval (95%CI) was 1.54-3.89, P < 0.001], APACHE II score (HR = 1.03, 95%CI was 1.00-1.07, P = 0.031), the total length of hospital stay (HR = 0.86, 95%CI was 0.83-0.89, P < 0.001), the need for IMV on admission (HR = 4.31, 95%CI was 1.27-14.63, P = 0.019), highest Lac value within 24 hours of transfer to the ICU (HR = 1.17, 95%CI was 1.10-1.24, P < 0.001), and tumor history (HR = 3.12, 95%CI was 1.36-7.13, P = 0.007) were independent risk factors for patient death at 30 days post-operative, and the risk of death was 2.45 times higher in patients unplanned transferred than in those planned transferred. Subgroup analysis showed that patients in the delayed transfer group had significantly longer IMV times than those in the immediate transfer group [hours: 43.00 (11.00, 121.00) vs. 17.50 (2.75, 73.00), P < 0.05].
CONCLUSIONS
The 30-day mortality, WBC and total length of hospital stay were higher in patients who were transferred to ICU after surgery, and PaO2/FiO2 was lower. Unplanned transfer, oncology history, use of IMV, APACHE II score, total length of hospital stay, and Lac were independent risk factors for patient death at 30 days postoperatively, and patients with delayed transfer to ICU had longer IMV time.
Humans
;
Retrospective Studies
;
Respiration, Artificial
;
Hospitalization
;
Prognosis
;
Intensive Care Units
4.Construction and practice of undergraduate training mode for clinical pharmacy specialty based on outcome-based education
Guoxiang HAO ; Yi ZHENG ; Xin HUANG ; Anchang LIU ; Chengwu SHEN ; Shuwen YU ; Rongmei WANG ; Lequn SU ; Wei ZHAO
China Pharmacy 2022;33(13):1635-1641
OBJECTIVE To introduce the construction of undergraduate specialty of clinical pharmacy based on the concept of outcome-based education (OBE),and to provide new idea and enlightenment for the construction of undergraduate specialty of clinical pharmacy in Chinese universities. METHODS Through the establishment and construction of training objectives and graduation requirements ,teaching reform was designed and implemented ,and the construction of teaching support system and teaching quality assurance system were completed. RESULTS The clinical pharmacy department of our university established the training direction of clinical pharmacy talents under the guidance of post competence ,including clarifying the training needs of undergraduate talents based on the overall requirements of national undergraduate education ;defining the social and industrial needs of clinical pharmacy talents based on the normative documents or concepts of clinical pharmacy ;clarifying the post and ability needs of clinical pharmacy talents based on the investigation of graduates and clinical pharmacists ;clarifying the development needs of clinical pharmacy based on the current situation and trends at home and abroad ;forming characteristic training objectives combined with the regional characteristics and school positioning , so as to construct training objectives and graduation requirements. The OBE concept was introduced into the undergraduate teaching reform of clinical pharmacy ;the pharmacy talent training direction were established under the guidance of post competence ;the training system was designed by reverse design method;a training mode of both innovation and practical ability was built so as to promote teaching reform ,strengthen the construction of grass-roots teaching organizations and teaching staff , and improve the construction of teaching quality assurance system. CONCLUSIONS The undergraduate training mode of clinical pharmacy specialty based on the concept of OBE is helpful to improve students ’personal comprehensive quality and professional knowledge and skills. The established undergraduate training model of clinical pharmacy specialty is in line with the modern educational concept and social needs ,and provides theoretical basis and practical experience for the training mode of clinical pharmacy professionals.
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.Association of metabolic syndrome with cardiovascular disease mortality and all-cause mortality in peritoneal dialysis patients
Jichao GUAN ; Haiying XIE ; Shuwen GONG ; Hongya WANG ; Shimin WANG ; Shuijuan SHEN
Chinese Journal of Endocrinology and Metabolism 2022;38(10):873-879
Objective:To investigate the association of metabolic syndrome(MS) with cardiovascular disease(CVD) mortality and all-cause mortality in peritoneal dialysis patients.Methods:A retrospective analysis was performed on patients who underwent peritoneal dialysis from January 1, 2013 to July 31, 2021 in the Shaoxing People′s Hospital. Patients were divided into MS group and non-MS group. The differences in baseline biochemical variables, comorbidities, and clinical outcomes between the two groups were compared. Kaplan-Meier method was used to obtain survival curves, the Cox regression model was used to evaluate the influence of MS for survival rates, and the inverse probability of treatment weighting(IPTW) was used to eliminate influence of the confounders in the groups.Results:A total of 494 peritoneal dialysis patients were enrolled in this study, which were divided into MS group( n=266) and non-MS group( n=228). The total median follow-up time was(31±22) months. At baseline, the standard mean difference( SMD) in smoking history, drinking history, CVD history, prevalence of chronic glomerulonephritis, left ventricular ejection fraction, B-type natriuretic peptides, hemoglobin, blood calcium, hypersensitive C-reactive-protein, intact parathyroid hormone, ultrafiltration and 4 h dialysate/plasma creatinine in the two groups were greater than 0.1. Their SMD decreased to under 0.1 after IPTW, showing a good balance between the two groups. The analysis of the survival curve of Kaplan Meier showed that the cumulative survival rate and cumulative CVD survival rate in MS group were significantly lower than those in non-MS group before and after IPTW( P<0.05). After IPTW was used to eliminate the effect of confounders, multivariate Cox regression analysis still displayed that MS was an independent risk factor for all-cause mortality( HR=1.824, 95% CI 1.121-2.968, P=0.015) and CVD mortality( HR=2.470, 95% CI 1.324-4.609, P=0.004)in peritoneal dialysis patients. Conclusion:The prevalence of metabolic syndrome is high in peritoneal dialysis patients. MS is an independent risk factor for all-cause mortality and CVD mortality in peritoneal dialysis patients.
7.Effects of electroacupuncture at Zusanli on duodenal Caspase-1 and GSDMD of rats with functional dyspepsia
Jiabao LIU ; Shuwen JIN ; Zhaoxia KANG ; Yiming LIU ; Li ZHOU ; Feng SHEN ; Paidi XU ; Hongxing ZHANG ; Xiaoli PAN
Chinese Journal of Physical Medicine and Rehabilitation 2022;44(12):1057-1063
Objective:To explore any effect of electroacupuncture (EA) at the Zusanli point on the scorch death of duodenal cells in rats with functional dyspepsia (FD) and possible mechanisms.Methods:Twenty-four 7-day-old Sprague-Dawley rats were randomly divided into a blank group, a model group and an EA group, each of 8. FD was induced in both the model and EA group rats using iodoacetamide gavage with tail-clip stimulation. After successful modeling the EA group was given acupuncture at the Zusanli point and then connected with a Korean acupuncture point nerve stimulator for 2 weeks. The other 2 groups were not given any intervention. The rats′ body weight was recorded before and after the modeling, as well as 7 and 14 days later. The gastric emptying rate and the small intestine propulsion rate of the three groups were detected right after the EA intervention, and the serum expression levels of interleukin-1β (IL-1β) and interleukin-6 (IL-6) were measured using enzyme-linked immunoassays. Real-time fluorescence quantitative polymerase chain reactions were used to detect the transcription levels of IL-1β and IL-6 in the rats′ duodenums, while western blotting was employed to assess the expression of caspase-1 P20 and dermatin D (GSDMD) in their duodenums.Results:After successful modeling, the average body weight of the rats in the model and EA groups was significantly different from that in blank group, and after 7 and 14 days the average body weight of the former groups was significantly different from that of the blank group, with significant differences between the two groups as well. After the EA intervention significant differences were observed in gastric reside and small intestine propulsion rate between the EA group and the model group, as well as between the model and the blank group. After the intervention, there were significant differences between the blank group and the other two groups in the average expression of IL-1β and IL-6 in serum, IL-1β and IL-6 mRNA in the duodenum, as well as the GSDMD and caspase-1 p20 proteins in the duodenum. There were significant differences between the model and EA groups in all of the above measurements.Conclusions:EA at the Zusanli point can significantly reduce the level of scorch death in the duodenum of FD rats, as well as relieve low-grade duodenal inflammation and the clinical symptoms of FD. Its mechanism may be related to the down-regulation of the expression of caspase-1 P20 and GSDMD-N protein, and of inflammatory factors such as IL-1β and IL-6, relieving low-grade duodenal inflammation.
8.Comparison between transoral radiofrequency coblation surgery and open partial laryngectomy for the treatment of supraglottic laryngeal carcinoma
Shuwen GUAN ; Feng WEN ; Hong SHEN ; Enmin ZHAO ; Yong QIN ; Shuifang XIAO
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2022;57(12):1457-1462
Objective:To explore the feasibility and efficacy of radiofrequency coblation assisted transoral surgery for the treatment of supraglottic laryngeal carcinoma by comparing with concurrent patients treated with conventional transcervical approach. To clarify the advantages of different surgical methods and to summarize the experience of supraglottic carcinoma radiofrequency ablation.Methods:Forty-six patients with supraglottic laryngeal carcinoma treated in department of otorhinolaryngology head and neck surgery, Peking University First Hospital from March 2014 to January 2021 were analyzed retrospectively. Among them(43 males, 3 females, aged from 45 to 79 years old), 23 patients were treated with radiofrequency coblation and 23 patients with partial laryngectomy with conventional transcervical approach. The operation time, intra-operative blood loss volume, recovery time, inpatient total medical cost and follow-up information of the two groups were analyzed. SPSS 26.0 software was used for statistical analysis.Results:There were no significant differences in age, gender, TNM staging,tumor staging and postoperative radiotherapy between the two groups (all P>0.05).The operation time, intra-operative blood loss volume, recovery time, inpatient total medical cost of the RFC-TOS group were110.0(60.0,150.0)min,5.0(5.0,30.0)ml,3.0(2.0,5.0)days,6.0(4.0,14.0)days and 26 100.7(16 145.5,47 044.4)yuan. The data of conventional transcervical approach group were 205.0(156.5,272.3)min, 150.0(50,200) ml, 18.0(16.3,22.8)days and 56520.1(440 992.5,67 109.9)yuan, ( Z=-4.03, -4.94, -4.97, -4.98 and -4.13;all P<0.001).The 5-year local control rate, disease-specific survival rate and overall survival rate of the two groups were 86.96%,95.65%,91.30% and 86.96%,91.30%,73.90% renspectively, which had no significant difference between the two groups(all P>0.05). Conclusions:Compared with conventional transcervical surgeries, RFC-TOS could be a reliable new surgical option for organ-function preservation strategy in the treatment of supraglottic laryngeal carcinoma.The RFC is a suitable new technique and deserving more multi-center clinical trials for its clinical promotion.
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.

Result Analysis
Print
Save
E-mail