Objective To investigate how culture duration affects the expression of immune membrane proteins in human monocyte-derived dendritic cells (DCs) and their exosomes (DEXs). Methods Human monocytes were induced with recombinant granulocyte-macrophage colony-stimulating factor (GM-CSF) and interleukin 4 (IL-4) to differentiate into DCs and were subsequently matured with tumor necrosis factor α(TNF-α). Exosomes were isolated by ultracentrifugation, and DEXs were identified by transmission electron microscopy and Amnis imaging flow cytometry, which were also used to quantify the expression of immune membrane proteins on DCs and DEXs. Results On the 10th day of culture, DCs displayed high surface expression of CD11c, CD80, CD86, major histocompatibility complex class I (MHC-I), and MHC-II. Expression peaked at day 18(CD11c: 78.66%±20.33%, CD80: 76.41%±10.02%, CD86: 96.43%±0.43%, MHC-I: 84.71%±2.96%, MHC-II: 80.01%±7.03%). After day 24, the overall expression showed a declining trend, with statistically significant differences observed for all markers except CD80 and MHC-II. By day 30, 80% of the DCs still expressed CD80, CD86, and MHC-II. The expression of immune membrane proteins on DEX surfaces also reached its peak on day 18, followed by an overall decline with prolonged culture time, with statistically significant differences observed for all markers except CD80. Correlation analysis revealed a significant positive relationship between the expression levels of immune membrane proteins on DC and DEX surfaces (CD11c: r=0.98; CD80: r=0.65; CD86: r=0.82; MHC-I: r=0.86; MHC-II: r=0.93). Conclusion Human monocyte-derived DCs in vitro express high expression of immune membrane proteins and maintain stable expression over a specific period. The exosomes secreted by these cells similarly demonstrate high surface expression of immune membrane proteins, with temporal trends aligned with those of the parent DCs.
Humans ; Dendritic Cells/immunology* ; Exosomes/immunology* ; Monocytes/metabolism* ; Cells, Cultured ; Time Factors ; B7-1 Antigen/metabolism* ; Membrane Proteins/immunology* ; Cell Culture Techniques/methods* ; B7-2 Antigen/metabolism* ; Cell Differentiation ; CD11c Antigen/metabolism* ; Granulocyte-Macrophage Colony-Stimulating Factor/pharmacology*

OBJECTIVES: To investigate the therapeutic mechanism of Danzhi Jiangtang Capsule (DZJTC) for repairing renal vascular endothelial injury in rats with diabetic nephropathy (DN). METHODS: Fifty male SD rat models of DN, established by left nephrectomy, high-sugar and high-fat diet and streptozotocin injection, were randomized into DN model group, low-, medium-, and high-dose DZJTC treatment groups, and DAPT (a γ-secretase inhibitor) treatment group, with 10 rats with normal feeding as the control group. DZJTC was administered by daily gavage at 0.315, 0.63, or 1.26 g/kg, and DAPT (20 mg/kg, dissolved in 50% CMC-Na solution) was given by gavage every other day for 4 weeks; normal saline was given in the control and model groups. After treatment, the levels of creatinine (CRE), blood urea nitrogen (BUN), and microalbuminuria (mALB) were detected with ELISA, and renal pathologies were observed by transmission electron microscopy. Renal expressions of vascular endothelial growth factor (VEGF) and endothelin-1 (ET-1) were measured by immunohistochemistry, and the protein expressions of CD31 and Notch signaling pathway components were detected using Western blotting. RESULTS: The rat models of DN showed significantly increased CRE, BUN, and mALB levels, obvious renal pathologies under electron microscopy, increased renal VEGF, ET-1 and CD31 expressions, and upregulated Notch1, NICD, and MAML1 protein levels. Treatment with DZJTC at the 3 doses and DAPT significantly reduced CRE, BUN, and mALB levels, improved renal pathology, decreased VEGF, ET-1 and CD31 expressions, and lowered Notch1, NICD and MAML1 levels, and the effects were the most pronounced with high-dose DZJTC. CONCLUSIONS DZJTC ameliorates hyperproliferation and dysfunction of renal vascular endothelium in DN rats possibly by regulating renal VEGF and ET-1 levels via inhibiting NICD- and MAML1-mediated Notch signaling pathway.
Animals ; Male ; Drugs, Chinese Herbal/therapeutic use* ; Rats ; Rats, Sprague-Dawley ; Signal Transduction/drug effects* ; Diabetic Nephropathies/drug therapy* ; Receptor, Notch1/metabolism* ; Kidney/blood supply* ; Diabetes Mellitus, Experimental ; Down-Regulation ; Endothelium, Vascular/metabolism* ; Nuclear Proteins/metabolism*

Objective:To explore the mechanism of dexmedetomidine (DEX) regulating microglial (MG) polarization and neuroinflammation after traumatic brain injury (TBI) in rats.Methods:Forty-two adult male SD rats were randomly (random number) divided into the sham group, TBI group, TBI+DEX group (further divided into 1 d, 3 d and 7 d subgroups), TBI+NF-κB inhibitor (pyrrolidine dithiocarbamate, PDTC) group and TBI+DEX+PDTC group, with 6 animals in each group. The rat TBI model was established according to the modified Feeney free fall method. PDTC was intraperitoneally injected 1 h after modeling with a dose of 100 mg/kg, and DEX was intraperitoneally injected 2 h after modeling with a dose of 100 μg/kg. Modified neurological severity score (mNSS) was used to evaluate rat neurological function, ELISA was used to detect serum inflammatory factors, and rats’ damaged cortex was collected to detect the phenotype markers of MG and protein expressions of MyD88 and NF-κB p65, and immunofluorescence staining was used to observe the expression and nuclear entry of NF-κB p65 in MG in injured cortex. One-way and two-way ANOVA were used to compare the measurement data among multiple groups.Results:Compared with the sham group, the mNSS score was significantly higher in the TBI group, and DEX treatment significantly decreased the mNSS score of TBI rats ( P<0.05). ELISA and Western blot results showed that in the TBI group, the tumor necrosis factor-α (TNF-α), interleukin (IL)-1β in serum and M1 phenotype marker (TNF-α, IL-1β) in brain were increased, the expression of anti-inflammatory factor IL-10 in serum and M2 phenotype markers (arginase-1 and IL-10) in brain were decreased ( P<0.05), and DEX downregulated the expression of TNF-α, IL-1β in serum and M1 phenotype markers in brain, while upregulated the level of L-10 in serum and the M2 phenotype marker in brain ( P<0.05). In addition, the expression of MyD88 and the nuclear translocation of NF-κB p65 were inhibited in the DEX group, and this effect could be enhanced by PDTC. Conclusions:DEX modulates MG activation in TBI rats by inhibiting NF-κB nuclear translocation and reduces neuroinflammation.

Background: Cardiopulmonary resuscitation (CPR) strategies in COVID-19 patients differ from those in patients suffering from cardiogenic cardiac arrest. During CPR, both healthcare and non-healthcare workers who provide resuscitation are at risk of infection. The Working Group for Expert Consensus on Prevention and Cardiopulmonary Resuscitation for Cardiac Arrest in COVID-19 has developed this Chinese Expert Consensus to guide clinical practice of CPR in COVID-19 patients. Main recommendations: 1) A medical team should be assigned to evaluate severe and critical COVID-19 for early monitoring of cardiac-arrest warning signs. 2) Psychological counseling and treatment are highly recommended, since sympathetic and vagal abnormalities induced by psychological stress from the COVID-19 pandemic can induce cardiac arrest. 3) Healthcare workers should wear personal protective equipment (PPE). 4) Mouth-to-mouth ventilation should be avoided on patients suspected of having or diagnosed with COVID-19. 5) Hands-only chest compression and mechanical chest compression are recommended. 6) Tracheal-intubation procedures should be optimized and tracheal-intubation strategies should be implemented early. 7) CPR should be provided for 20-30 min. 8) Various factors should be taken into consideration such as the interests of patients and family members, ethics, transmission risks, and laws and regulations governing infectious disease control. Changes in management: The following changes or modifications to CPR strategy in COVID-19 patients are proposed: 1) Healthcare workers should wear PPE. 2) Hands-only chest compression and mechanical chest compression can be implemented to reduce or avoid the spread of viruses by aerosols. 3) Both the benefits to patients and the risk of infection should be considered. 4) Hhealthcare workers should be fully aware of and trained in CPR strategies and procedures specifically for patients with COVID-19.

Objective:To investigate the different outcomes of two types of acute kidney injury (AKI) according to standard of Kidney Disease: Improving Global Outcomes-AKI (KDIGO-AKI), and to analyze the risk factors that affect the prognosis of intensive care unit (ICU) patients in China.Methods:A secondary analysis was performed on the database of a previous study conducted by China Critical Care Clinical Trial Group (CCCCTG), which was a multicenter prospective study involving 3 063 patients in 22 tertiary ICUs in 19 provinces and autonomous regions of China. The demographic data, scores reflecting severity of illness, laboratory findings, intervention during ICU stay were extracted. All patients were divided into pure AKI (PAKI) and acute on chronic kidney disease (AoCKD). PAKI was defined as meeting the serum creatinine (SCr) standard of KDIGO-AKI (KDIGO-AKI SCr) and the estimated glomerular filtration rate (eGFR) at baseline was ≥ 60 mL·min -1·1.73 m -2, and AoCKD was defined as meeting the KDIGO-AKI SCr standard and baseline eGFR was 15-59 mL·min -1·1.73 m -2. All-cause mortality in ICU within 28 days was the primary outcome, while the length of ICU stay and renal replacement therapy (RRT) were the secondary outcome. The differences in baseline data and outcomes between the two groups were compared. The cumulative survival rate of ICU within 28 days was analyzed by Kaplan-Meier survival curve, and the risk factors of ICU death within 28 days were screened by Cox multivariate analysis. Results:Of the 3 063 patients, 1 042 were enrolled, 345 with AKI, 697 without AKI. The AKI incidence was 33.11%, while ICU mortality within 28 days of AKI patients was 13.91% (48/345). Compared with PAKI patients ( n = 322), AoCKD patients ( n = 23) were older [years old: 74 (59, 77) vs. 58 (41, 72)] and more critical when entering ICU [acute physiology and chronic health evaluation Ⅱ (APACHEⅡ) score: 23 (19, 27) vs. 15 (11, 22)], had worse basic renal function [eGFR (mL·min -1·1.73 m -2): 49 (38, 54) vs. 115 (94, 136)], more basic complications [Charlson comorbidity index (CCI): 3 (2, 4) vs. 0 (0, 1)] and higher SCr during ICU stay [peak SCr for diagnosis of AKI (μmol/L): 412 (280, 515) vs. 176 (124, 340), all P < 0.01]. The mortality and RRT incidence within 28 days in ICU of AoCKD patients were significantly higher than those of PAKI patients [39.13% (9/23) vs. 12.11% (39/322), 26.09% (6/23) vs. 4.04% (13/322), both P < 0.01], while no significant difference was found in the length of ICU stay. Kaplan-Meier survival curve analysis showed that the 28-day cumulative survival rate in ICU in AoCKD patients was significantly lower than PAKI patients (Log-Rank: χ2 = 5.939, P = 0.015). Multivariate Cox regression analysis showed that admission to ICU due to respiratory failure [hazard ratio ( HR) = 4.458, 95% confidence interval (95% CI) was 1.141-17.413, P = 0.032], vasoactive agents treatment in ICU ( HR = 5.181, 95% CI was 2.033-13.199, P = 0.001), and AoCKD ( HR = 5.377, 95% CI was 1.303-22.186, P = 0.020) were independent risk factors for ICU death within 28 days. Conclusion:Further detailed classification (PAKI, AoCKD) based on KDIGO-AKI SCr standard combined with eGFR is related to ICU mortality in critical patients within 28 days.

OBJECTIVE: To observe the effects of a traditional Chinese medicine (TCM) capsule for replenishing qi, nourishing yin and activating blood on Notch/Hes1 signaling pathway in the renal tissue and vascular endothelial CD34 and CD144 expressions in a rat model of diabetic nephropathy. METHODS: Rat models of early-stage diabetic nephropathy were established by left nephrectomy and high- fat and high- sugar feeding combined with intraperitoneal injection of STZ. The rats were randomized into model group, benazepril group, and high-, moderate-, and low-dose TCM capsule groups for corresponding treatments, with 6 normal rats as the control group. After 8 weeks of drug treatment, blood glucose and 24-h urinary albumin of the rats were measured, and the renal histopathology was observed with HE staining; Hes1 expression in the renal tissue was detected with immunohistochemical staining, and the renal expressions of CD34 and CD144 were detected using Western blotting. RESULTS: Compared with the normal control group, the rat models of diabetic nephropathy showed obvious abnormalities in 24- h urinary albumin and expressions of Hes1, CD34 and CD144d. The TCM capsule at both the high and moderate doses significantly reduced 24-h urinary albumin in the rats; the renal expressions of Hes1 and CD34 was significantly reduced in all the dose groups, and the expression of CD144 was significantly reduced in the high- dose group. Compared with benazepril group, the TCM capsule obviously reduced CD34 expression at all the 3 doses and lowered CD144 expression at the low dose. Histopathologically, the rats in the model group showed glomerular hypertrophy, increased mesenteric matrix, thickening and widening of the mesenteric membrane, and nodular hyperplasia. These pathologies were obviously alleviated by treatment with the TCM capsule at the high and moderate doses. CONCLUSIONS The Traditional Chinese medicine (TCM) capsule for replenishing qi, nourishing yin and activating blood can reduce Hes1, CD34 and CD144 in kidney tissue of model rats, play a protective role on kidney function and delay the development of DN.
Animals ; Diabetic Nephropathies ; Drugs, Chinese Herbal ; Medicine, Chinese Traditional ; Qi ; Rats ; Signal Transduction ; Transcription Factor HES-1

Objective To evaluate the effect of high flow nasal catheter oxygen (HFNCO) therapy on extubation failure rate,reintubation rate,and incidence of related complications within 48 h after extubation in mechanically ventilation patients.Methods A prospective,single-center,randomized controlled trial was conducted in the ICU of a teaching hospital affiliated to a medical university.A total of 77 patients with mechanical ventilation duration of ≥ 48 h and met the condition of spontaneous breathing test (SBT) were selected.The patients whose LUS ≥ 14 at 30 min of SBT were enrolled,and were randomly (random number) divided into 3 groups according to different oxygen therapies:the traditional oxygen therapy group,the noninvasive ventilation (NIV) group,and the HFNCO group.The effect of oxygen therapy and outcomes after extubation were compared among the three groups.The measurement data were presented as the mean±standard deviation (SD),and the numeration data were expressed as ratio or constituent ratio.The independent sample t test and LSD-t test were used for the comparisons between the two groups and the one-way ANOVA for differences between multiple groups.The differences between enumeration data were assessed by chi-square test.A P＜0.05 was considered statistically significant.Results There was no significant difference in gender,age and other general conditions between the two groups (P＞0.05).The NIV group and HFNCO group had lower extubation failure rate (14.29％,15.38％ vs 34.87％) and reintubation rate (10.7％,11.54％ vs 21.74％) than the convertional oxygen therapy group (P＜0.05).In addition,the traditional oxygen therapy group had longer mechanical ventilation duration [(24.33±4.42) d vs (8.58±1.09) d,(8.37±2.43) d],antibiotic use time [(19.21±4.37) d vs (8.34±2.54) d,(7.41±1.06) d],and ICU hospitalization time [(27.27±4.24) d vs (10.38±2.07) d,(9.44±0.79) d],all P＜0.05.Conclusions Treatment with HFNCO or NIV after extubation can effectively reduce the rates of extubation failure and reintubation,and improve the outcome of the mechanical ventilation.There is no difference in clinical efficiency between the NIV group and HFNCO group.However,compared with NIV,HFNCO can effectively reduce respiratory rate and avoid the retention of CO2,which has a wider application prospect in clinical practice.

Objective? To explore the effect of enteral nutrition(EN) combined with high quality nursing in the treatment of severe acute pancreatitis(SAP). Methods? By convenience sampling, a total of 100 non-surgical SAP patients in the 901 Hospital of the Chinese People＇s Liberation Army Joint Logistics Support Force from September of 2012 to September of 2015 were selected as subjects and divided into the experimental group(n=50) and control group (n=50) according to random number table. The control group received the parenteral nutrition (PN) support combined with high quality nursing while the experimental group received the EN support combined with high quality nursing. After treatment, the two groups were compared in terms of the improving condition of the nutritional index, change of scores in Acute Physical and Chronic Health EvaluationⅡ (APACHEⅡ) , overall nursing satisfaction and incidence of adverse events. Results? 14 days after treatment, the experimental group was better than the control group in the serum amylase level, serum albumin, fasting blood glucose recovery and APACHE Ⅱ score with statistical difference (P＜0.05); the overall nursing satisfaction rate was 92% in the experimental group, higher than the control group with 66% and the difference was statistically significant (P＜ 0.05); the incidence of complications after intervention in the experimental group was 14%, lower than the control group with 36% and the difference was statistically significant (P＜0.05). Conclusions? EN support combined with high quality nursing has important clinical values for SAP patients. It can better alleviate patients＇ anxiety and stress, improve their diet structure, reduce the incidence of complications to the maximum extent, increase the clinical nursing satisfaction rate, and is worthy of clinical popularization.

Objective To implement a two?level community?based health education pattern of schistosomiasis in residents of endemic areas in marshland and lake regions,so as to explore the suitable pattern of health education under hypo?endemic situa?tion. Methods Two schistosomiasis endemic villages in Jiangling County,Hubei Province were collected as study areas,and among which,one village was treated as an intervention group,where the two?level community?based health education pattern as well as regular control measures was implemented;the other village was a control group,where only regular control measures were implemented. The awareness rates on schistosomiasis control,the rates of correct behavior and the compliance rates of ex?amination,treatment and chemotherapy of the two groups before and after the intervention were compared. Results According to the results of the baseline survey in 2014,the awareness rates of schistosomiasis control of the intervention and control groups were 84.00%and 77.45%,respectively,the correct rates of behavior of the two groups were 72.00%and 63.73%,respectively, and the compliance rates of the treatment were 80.36%and 82.28%,respectively,there were no statistically significant differ? ences between all the above rates of the two groups(all P>0.05). After the intervention of the two?level community?based health education,the correct rates of behavior,and the compliance rates of examination and chemotherapy of the two groups were 92.31%and 80.37%,95.11%and 82.55%,84.13%and 63.64%,respectively,and the differences between all the rates above of the two groups were statistically significant(all P<0.05). When compared to those before intervention,the growing rates of the compliance rates of examination,treatment and chemotherapy of the intervention group were 20.97%,15.33%and 23.29%, respectively,while those of control group were 14.27%,4.17%,-3.77%,respectively,the growing rates of the intervention groups were higher than those of the control groups. Conclusions Through the two?level community?based pattern of health edu?cation,the compliance rates of examination and treatment of the residents have improved,and therefore,the pattern is suitable for popularization and application in marshland and lake regions.

Objective To discuss the current status of application of the four kinds of clinical decision‐making method (mode recognition method ,hypothesis deductive method ,event‐driven method and applying regulation method ) in the clinical practice of the emergency resident doctors and to understand their mastery situation of the clinical decision‐making methods and the influencing factors .Methods One thousand and thirty‐five patients collected by 207 resident doctors (5 cases were randomly collected from the patients diagnosed and treated by each resident doctor ) were divided into 3 groups according to different year systems .The clinical decision‐making methods ,diagnosis accuracy of different clinical decision‐making methods and the influencing factors of decision‐making methods were compared among different grades .Results The difference in the decision‐making methods among different year systems had statistical significance (P<0 .05);furthermore ,the difference in the diagnostic accuracy among different decision‐making methods had statistical significance (P< 0 .05) ,the differences in the defensive behaviors and different decision‐making methods were statistically significant (P<0 .05) ,the multi‐classification Logistic regression of different decision‐making methods showed that the differences in the different residency year system ,residency education ,residency clinical contacting time ,inpatient symptoms ,defensive behaviors and interrogation reliability degree had statistical significance (P< 0 .05) .Conclusion The most commonly used decision‐making method by the resident doctors is the applying regulation method .The residency year system ,resi‐dency education ,residency clinical contacting time ,inpatient symptoms ,defensive behaviors and interrogation reliability degree are the influencing factors of application of the clinical decision‐making methods in the clinical practice of the emergency medicine spe‐cialty .