BACKGROUND:Adipose-derived stem cels are regarded as the potential seed cels for tissue engineering. Colagenase digestion is used to isolate adipose-derived stem cels from fat pads currently. However, there are some problems, such as cumbersome operation and high cost.
OBJECTIVE: To study the basic biological characteristics of adipose-derived stem cels by tissue explants culture and to explore the differentiation potential into osteoblasts, adipocytes and endothelial progenitor cels in vitro.
METHODS:Adipose-derived stem cels were isolated by tissue explants technique from the bilateral groin fat pads of rats under aseptic conditions, and cultured in vitro. Cel counting kit-8 was used to detect the proliferative activity, and flow cytometry was employed to analyze the expression of cel surface markers. Passage 4 adipose-derived stem cels were cultured in osteogenic medium, adipogenic medium and endothelial progenitor cel medium for 2-3 weeks, and then the cels were identified.
RESULTS AND CONCLUSION:Adipose-derived stem cels that were isolated by tissue explants culture were easily cultured, and after subculture, cels were mainly spindle-shaped and grew in clone-like manner with swirling arrangement. Cels that experienced repeated subcultures stil kept stronger proliferative ability and the cel growth curve was shaped as a parabola. Immunochemical staining analysis revealed that adipose-derived stem cels were positive for CD44, CD90 and CD29, but negative for CD31, CD45. After adipogenic/osteogenic induction, the cels were respectively positive for oil red O staining and alizarin red staining. Induced endothelial progenitor cels were identified with CD34 and the ability to uptake Dil-ac-LDL and FITC-UEA. These findings indicate using the using tissue explants culture, high-purity adipose-derived stem cels easy to proliferate can be harvested, highly express stem cels-related antigens, and have the ability to differentiate into osteoblasts, adipocytes and endothelial progenitor cels, which meet the needs of seed cels in tissue engineering research.

ObjectiveTo evaluate the effect of high frequency oscillation ventilation (HFOV) vs. conventional mechanical ventilation (CV) on the treatment and prognosis of adult patients with acute respiratory distress syndrome (ARDS).Methods Published articles concerning randomized controlled trials (RCTs) about the effect of HFOV vs. CV on the prognosis of adult patients with ARDS published before May 2014 were retrieved from PubMed, EMBase, Cochrane central registry of controlled trials, CNKI and Wanfang Data. The mortality and data of physiological parameters were analyzed with STATA 12.0, and the mortality rate was also analyzed by trial sequential analysis with TSA 0.9, and the line chart was drawn with Microsoft Office Excel 2003.Results Seven trials with 1 731 patients met the criteria, all of them recorded the physiological parameters data, and mortality rate was mentioned in 6 trials (1 705 patients). Compared with CV, HFOV did not show any statistically significant beneficial effects on mortality [relative risk (RR) = 0.93, 95% confidence interval (95%CI) = 0.70-1.24,P = 0.63], and other clinical outcomes, including survival without mechanical ventilation (RR = 1.05, 95%CI = 0.72-1.54,P = 0.80), survival on mechanical ventilation (RR = 1.23, 95%CI = 0.65-2.35,P = 0.52), or treatment failure (RR = 0.89, 95%CI = 0.50-1.56,P = 0.67). The risk factors of adverse events including hypotension (RR = 0.89, 95%CI = 0.07-10.99,P =0.93), acidosis (RR = 1.05, 95%CI = 0.43-2.56,P = 0.91), and air leakage from ventilator (RR = 0.74, 95%CI = 0.31-1.80,P =0.51) were similar. But the physiologic parameters of patients and parameters of ventilator in HFOV group, including oxygenation index, positive end-expiratory pressure, tidal volume, mean airway pressure, arterial pH, partial pressure of arterial carbon dioxide, fraction of inspired oxygen, ratio of partial pressure of arterial oxygen to fraction of inspired oxygen, were better than those in the CV group. Methods adapted from formal interim monitoring boundaries applied to cumulative Meta-analysis showed that the evidence failed by a considerable degree to meet the standards for forgoing studies, and the necessary sample was 3 874 patients. Trial sequential analysis also showed that the accumulatedZ-score did not cross the traditional boundary (P = 0.05) and interim monitoring boundaries. This result indicated that there was no significant difference between CV and HFOV on mortality before the number of needed sample reached (3 874 cases). We could not get a definitive conclusion with current evidences.ConclusionsCompared with CV, the use of HFOV in ARDS was not associated with a significant reduction in mortality. But the physiologic parameters of patients in HFOV group were better than those in the CV group. More RCTs are needed to draw a definitive conclusion.

Objective To conduct a systematic review to evaluate efficacy and safety of bladder neck preservation techniques during radical prostatectomy.Methods We searched PubMed,Embase,Medline,Central (the Cochrane Library,Issue 1,2013),CNKI,CBM from the inception to July 2015 for randomized or observational studies assessing the influence of preservation of bladder neck on the continence recovery and cancer control outcomes after radical prostatectomy.Two researchers evaluated the quality of included studies.A meta-analysis was conducted using Review Manager 5.0 software.Results Twelve observational and two randomized studies were analyzed.BNP group had a faster recovery of early continence than non-BNP group at 1 month[OR =2.88,95％ CI 1.80-4.60,P ＜ 0.05],3-4 months [OR =2.32,95％ CI 1.42-3.80,P ＜ 0.05].However there was no statistically significant difference in terms of continence rate at 6 months [OR =1.49,95％ CI 0.97-2.29,P =0.07] and 12 months [OR =0.99,95％ CI 0.70-1.39,P =0.93] among two groups.There was no significant difference in terms of PSM rate [OR =0.92,95 ％ CI 0.71-1.19,P =0.52] between BNP and non-BNP groups.Conclusions We found that preservation of the bladder neck result in earlier return of urinary continence without adversely affecting cancer control.

Objective:To investigate the effect of ultrasound-guided midline catheter placement on the incidence of catheter-related bloodstream infection (CRBSI) in severe emergency patients.Methods:Five hundred and twenty-nine patients were chosen as the research objects from March 2018 to December 2019 at Emergency Intensive Care Unit, which was divided into the experimental group ( n=278) and the control group ( n=251). In the experimental group, ultrasound-guided midline catheter was used as central venous catheter (CVC) removal method of sequential, and in the control group, peripheral venous indwelling needle was used as sequential method after removal of CVC. CVC, midline catheter and the indwelling time of indwelling needle were counted. The utilization rate of CVC was compared between the two groups. Kaplan-Meier survival curve was plotted to describe the CVC indwelling time of the two groups and log-rank test was performed. Cox regression analysis was performed to analyze the influencing factors of CVC indwelling time and compare the incidence of CRBSI and other catheter-related complications. Results:The CVC indwelling time of the experimental group was significantly shorter than that of the control group (8 d vs. 13 d, P=0.000). The CVC utilization rate of the experimental group was significantly lower than that of the control group (49.83% vs. 80.45%, P=0.000). Multivariate Cox regression analysis showed that difficult intravenous access, length of ICU stay, the site of catheter placement, and midline catheter implantation without ultrasound-guidance were independent risk factors for prolonged CVC indwelling time ( P=0.000). The CRBSI rate of the experimental group was significantly lower than that of the control group (0.571‰ vs. 3.802‰, P=0.038). There was no significant difference in the incidence of other catheter-related complications between the two groups ( P=0.403). Conclusions:Ultrasound-guided midline catheter implantation can shorten the indwelling time of CVC, reduce the utilization rate of CVC, and reduce the incidence of CRBSI, which is worthy of clinical promotion.

Objective:To investigate the function of gasdermin D (GSDMD) in intestinal damage of mice with severe acute pancreatitis (SAP).Methods:The healthy C57BL/6 mice were divided into four groups randomly, including normal saline (NS) group, small interfering RNA (siRNA)-NS group, SAP model group and siRNA-SAP group, with 6 mice in each group. The SAP mouse model was reproduced by intraperitoneal injection of caerulein 50 μg/kg combined with lipopolysaccharide (LPS) 10 mg/kg; the NS group was given the same amount of NS; in the siRNA-SAP group and siRNA-NS group, siRNA 50 mg/kg was injected through the tail vein three times before modeling or injection of NS. The blood of mice eyeball in each group was taken 12 hours after modeling, and serum interleukins (IL-1β, IL-18) levels were detected by enzyme linked immunosorbent assay (ELISA). The mice were sacrificed to observe the general changes in abdominal cavity, the pancreas and ileum tissues were taken to observe the pathological changes under a light microscope. The expression of long-chain non-coding RNA uc.173 (lnc uc.173) was detected by reverse transcription-polymerase chain reaction (RT-PCR). Immunohistochemical method was used to detect the expression of tight junction proteins zonula occluden-1 (ZO-1) and Occludin in intestinal mucosal epithelial cells. Western blotting was used to detect the GSDMD protein expression level in the intestinal tissue.Results:The serum levels of IL-1β and IL-18 in the SAP model group were significantly higher than those in the NS group and the siRNA-NS group [IL-1β (ng/L): 146.66±1.40 vs. 44.48±5.76, 81.49±10.75, IL-18 (ng/L): 950.47±177.09 vs. 115.43±16.40, 84.84±21.90, all P < 0.05]; and the levels of IL-1β and IL-18 in the siRNA-SAP group were significantly lower than those in the SAP model group [IL-1β (ng/L): 116.26±15.54 vs. 146.66±1.40, IL-18 (ng/L): 689.96±126.08 vs. 950.47±177.09, both P < 0.05]. General observation showed that there were no obvious abnormalities in the abdominal cavity of the mice in the NS and siRNA-NS groups; the mice in the SAP model group and the siRNA-SAP group had different degrees of edema and congestion in the intestine; compared with the SAP model group, the abnormalities in the siRNA-SAP group was significantly reduced. Under light microscope, there were no obvious changes in the pancreas and intestinal mucosa in the NS group and the siRNA-NS group; the pancreatic tissue of the SAP model group and the siRNA-SAP group had different degrees of edema, inflammatory cell infiltration, and lobular structure damage, and the intestinal mucosa was damaged to a certain degree, and the villi were broken to varying degrees, but the damage in the siRNA-SAP group was lighter. The results of RT-PCR showed that the expression of lnc uc.173 in the intestinal tissues of the model SAP group was significantly lower than that of the NS group and the siRNA-NS group (2 -ΔΔCt: 0.26±0.12 vs. 1.01±0.37, 0.67±0.32, both P < 0.05), while the expression of lnc uc.173 in the siRNA-SAP group was significantly higher than that in the SAP model group (2 -ΔΔCt: 0.60±0.39 vs. 0.26±0.12, P < 0.05). Immunohistochemistry showed that ZO-1 and Occludin proteins in the NS group were distributed along the epithelial cells of the intestinal mucosa, showing a strong expression; ZO-1 and Occludin expressions were significantly reduced in the SAP model group and siRNA-SAP group, but the expressions in the siRNA-SAP group was higher than that in the SAP model group. Western blotting showed that the expression level of GSDMD protein in the intestinal tissues of the SAP model group was significantly higher than that of the NS group and the siRNA-NS group [GSDMD protein (GSDMD-N/β-actin): 1.99±0.46 vs. 1, 1.00±0.78, both P < 0.05]. Compared with the SAP model group, the expression of GSDMD protein in the siRNA-SAP group was significantly decreased [GSDMD protein (GSDMD-N/β-actin): 1.42±0.42 vs. 1.99±0.46, P < 0.05]. Conclusions:The systemic inflammatory response and intestinal mucosal barrier damage of SAP mice may be related to the increase of GSDMD expression in intestinal tissues. GSDMD mediates cell pyrolysis to promote the release of inflammatory factors, cause intestinal injury, and down-regulate the expression of intestinal epithelial cell tight junction proteins such as ZO-1 and Occludin, resulting in intestinal mucosal damage.

Objective To explore the effect of adipose-derived stem cells (ADSCs) on inflammatory factors in rats with lipopolysaccharide (LPS)-induced acute lung injury (ALI) and the possible mechanism of anti-inflammatory. Methods Seventy male Sprague-Dawley (SD) rats were randomly divided into normal control group (n = 10), LPS model group (n = 30), and ADSCs intervention group (n = 30) by random number table. ALI model was reproduced by intraperitoneal injection of 8 mg/kg LPS, and the rats in ADSCs intervention group received tail vein injection of 300 μL ADSCs 30 minutes after the model reproduction, the samples of normal control group were harvested immediately without any intervention, and the specimens in remained two groups were taken at 6, 24, 72 hours respectively. Arterial partial pressure of oxygen (PaO2) and lactate level in femoral artery were determined. Enzyme-linked immunosorbent assay (ELISA) was used to detect the serum myeloperoxidase (MPO) and interleukin-10 (IL-10) in the blood of left ventricle. Lung wet/dry weight (W/D) ratio was detected by thoracotomy, and the pathological changes of lung tissue were observed under an optical microscope. Western Blot was used to detect the protein expression of nuclear factor-κB (NF-κB) in lung tissue of rats. Results Compared with the normal control group, the damage degree of lung tissue of LPS model group was significantly heavier from 6 hours, and lung W/D ratio, blood lactate, MPO, IL-10 and expression level of NF-κB in lung tissue were significantly increased respectively, while PaO2 was decreased significantly. Compared with LPS model group, the damage degree of lung tissue of ADSCs intervention group was significantly reduced from 6 hours, and lung W/D ratio, blood lactate, MPO, and NF-κB expression in lung tissue were significantly decreased, while PaO2 was increased significantly, and it became normal at 72 hours [lung W/D ratio: 5.33±0.29 vs. 5.77±0.42 at 6 hours, 5.14±0.46 vs. 5.43±0.38 at 72 hours; blood lactate (mmol/L): 3.6±1.0 vs. 5.7±1.1 at 6 hours, 3.1±1.0 vs. 3.8±1.2 at 72 hours; blood MPO (μg/L): 1.50±0.90 vs. 2.70±1.85 at 6 hours, 0.46±0.30 vs. 0.71±0.22 at 72 hours; NF-κB (gray value): 0.40±0.11 vs. 0.50±0.09 at 6 hours, 0.24±0.03 vs. 0.33±0.06; PaO2 (mmHg, 1 mmHg = 0.133 kPa): 78.0±4.1 vs. 74.5±3.2 at 6 hours, 89.3±9.4 vs. 81.9±3.4 at 72 hours; all P < 0.05]. The IL-10 level was significantly higher than that of LPS model group only at 24 hours (ng/L: 27.75±15.49 vs. 17.52±6.56, P < 0.05). Conclusion ADSCs can effectively relieve the inflammatory response of ALI induced by LPS, probably by inhibiting the expressions of NF-κB and blocking the release of inflammatory cytokines.

Objective To compare the intestinal microbiota of septic shock patients and healthy subjects,and study the composition of the intestinal microbiota and its effect on septic shock patients in the intensive care unit (ICU).Methods A total of 15 stool samples were prospectively collected from septic shock patients admitted to the ICU in the First Affiliated Hospital of Zhengzhou University between June 2015 and February 2016,while 15 samples from healthy subjects served as controls.Bacterial DNA was submitted for 16S rDNA gene sequencing.The association between gut microbiota composition and clinical parameters was evaluated.Shannon index was used to assess the bacterial diversity.Results Compared with the healthy subjects,the composition of intestinal microbiota in septic shock patients changed significantly.The abundance of Proteobacteria and Fusobacteria were significantly higher in septic shock patients than in healthy subjects (23.71％ vs 3.53％,P=0.000 6;1.27％ vs 0.12％,P=0.059,respectively).In this study,29 species were identified,and the composition of intestinal microbiota in each patient was highly individualized.There was no significant difference in Shannon index between septic shock patients and healthy subjects (P=0.12).Conclusions The composition of intestinal microbiota in septic shock patients was characterized by high diversity and individualization,but there was the phenomenon of overproduction of single bacteria genus.The relationship between the composition of intestinal microbiota and clinical outcomes requires further exploration by large sample studies.

Objective:To investigate the effects of poly (ADP-ribose) polymerase-1(PARP-1) in intestinal mucosal barrier injury in rat model with severe acute pancreatitis (SAP).Methods:Twenty healthy male Wistar rats were divided into four groups ( n=5 each group) using a random table method: control, SAP, 3-aminobenzamide (3-AB), and 3-AB control groups. The SAP model was induced by intraperitoneal injection of cerulean with lipopolysaccharide. At 30 min, the rats were treated with the PARP-1 inhibitor, 3-AB, or normal saline,separately. After 12 h, all rats were sacrificed to harvest pancreas tissues, intestines tissues, and blood from the hearts for index detection. Serum amylase (AMY) and interleukin (IL)-6 levels were measured using an automatic biochemical instrument and enzyme-linked immunosorbent assay (ELISA), respectively.The protein expression of PARP-1 and nuclear factor (NF-κB) were measured using Western blot and that of occludin was measured using an immunohistochemical test. One-way analysis of variance was used for comparison of multiple groups of variables. Non-parametric tests of rank conversion were used when variances were not uniform. A P <0.05 was considered statistically significant. Results:Compared to the control group, the following indexes in the SAP group were significantly increased: ascites (with serious hemorrhage and necrosis in the pancreas and disordered intestinal villi),serum AMY and IL-6 levels, and the expression of PARP-1 and NF-κB. However, Occludin expression was significantly decreased. There was no significant difference between 3-AB group and 3-AB control group. Compared to the SAP group, the severity of SAP and pancreatitis-associated intestinal injury was significantly attenuated with the administration of 3-AB. Serum AMY and IL-6 levels were significantly decreased (serum AMY: 1 879.25 ± 736.6 U/L vs 5 569.33 ± 1993.48 U/L; IL-6: 77.98 ± 20.65 pg/mL vs 209.14 ± 79.08 pg/mL, both P<0.05), but the expression of PARP-1 and NF-κB were significantly increased (PARP-1: 1.44 ± 0.09 vs 1.49 ± 0.13; NF-κB: 0.63 ± 0.09 vs 0.96±0.08, both P<0.05). Similarly, Occludin expression was significantly decreased (6.7±1.5 vs 3.2±1.1, P<0.05). Conclusions:Inhibition of PARP-1 has protective effects on SAP associated intestinal mucosal barrier damage. The mechanism may be related to the inhibition of NF-κB signaling pathway and increase intestinal mucosal Occludin protein expression.

Objective:To analyze the risk factors of renal function recovery in patients with severe acute pancreatitis (SAP) combined with acute kidney injury (AKI).Methods:A retrospective study was conducted in 105 SAP patients with AKI who were admitted to ICU or EICU of the Affiliated Hospital of Qingdao University from January 2013 to October 2019. According to the recovery of renal function at 28 days, the patients were divided into the renal function recovery group and the poor recovery group. Multivariate logistic regression analysis was used to analyze the clinical data of the two groups and to determine the risk factors related to renal function recovery.Results:According to the recovery of renal function, 105 patients were divided into the renal function recovery group ( n=73) and the poor recovery group ( n=32). Compared with the renal function recovery group, patients in the poor recovery group were older, had a higher prevalence of diabetes and coronary heart disease and a higher score on the acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ); More patients had abdominal necrosis infection and abdominal hemorrhage. The proportion of patients who applied mechanical ventilation was higher in the poor recovery group. Multivariate logistic regression analysis showed that abdominal necrosis infection ( OR=5.088, 95% CI:1.041-24.871, P=0.044) and mechanical ventilation ( OR=4.615, 95% CI:1.126-18.904, P=0.034) were the independent risk factors of renal function recovery in SAP patients with AKI. Conclusions:Abdominal necrosis infection and mechanical ventilation are the independent risk factors for renal function recovery in patients with SAP and AKI.

Objective To compare the impacts of percutaneous coronary intervention (PCI) and medical therapy on mortality in patients with stable coronary artery disease.Methods We searched PubMed,Embase,Cochrane central register of controlled trials,Wanfang data and CNKI to find relevant randomized controlled trials on PCI versus medical therapy for treating patients with stable coronary artery disease,which were reported before December 2013.Publications were selected according to inclusion and exclusion standard.Meta-analyses was performed with the software of STATA 12.0.Results Five randomized controlled trials and 5 567 patients were enrolled for this analysis.Compared with medical therapy,PCI could not significantly decrease the long-term all-cause mortality (RR =0.96,95％ CI 0.80-1.15),the cardiac death rate (RR =1.02,95％ CI 0.77-1.36),the myocardial infarction rate (RR =1.05,95％ CI 0.89-1.23),the acute coronary syndrome (RR =0.70,95％ CI 0.27-1.82),the rate of freedom from angina (RR =1.09,95％ CI0.98-1.21),and the rate of stroke (RR =1.27,95％ CI 0.75-2.15).However,the revascularization rate was significantly lower for patients in PCI group (RR =0.60,95％ CI 0.42-0.86).Conclusions Long-term mortality is similar for patients with stable coronary artery disease underwent PCI or medical therapy.