Rheumatoid arthritis (RA) is an autoimmune disease with a complex etiology. Monocyte-derived macrophages (MDMs) infiltration are associated with RA severity. We have reported the deletion of G-protein-coupled receptor kinase 2 (GRK2) reprograms macrophages toward an anti-inflammatory phenotype by recovering G-protein-coupled receptor signaling. However, as more GRK2-interacting proteins were discovered, the GRK2 interactome mechanisms in RA have been understudied. Thus, in the collagen-induced arthritis mouse model, we performed genetic GRK2 deletion using GRK2^f/fLyz2-Cre^+/- mice. Synovial inflammation and M1 polarization were improved in GRK2^f/fLyz2-Cre^+/- mice. Supporting experiments with RNA-seq and dual-luciferase reporter assays identified peroxisome proliferator-activated receptor γ (PPARγ) as a new GRK2-interacting protein. We further confirmed that fms-related tyrosine kinase 1 (Flt-1), which promoted macrophage migration to induce angiogenesis, was inhibited by GRK2-PPARγ signaling. Mechanistically, excess GRK2 membrane recruitment in CIA MDMs reduced the activation of PPARγ ligand-binding domain and enhanced Flt-1 transcription. Furthermore, the treatment of mice with GRK2 activity inhibitor resulted in significantly diminished CIA pathology, Flt-1⁺ macrophages induced-synovial inflammation, and angiogenesis. Altogether, we anticipate to facilitate the elucidation of previously unappreciated details of GRK2-specific intracellular signaling. Targeting GRK2 activity is a viable strategy to inhibit MDMs infiltration, affording a distinct way to control joint inflammation and angiogenesis of RA.

OBJECTIVE: To analyze the predictors of successful weaning off extracorporeal membrane oxygenation (ECMO) after extracorporeal cardiopulmonary resuscitation (ECPR). METHODS: The clinical data of 56 patients with cardiac arrest who underwent ECPR in Hunan Provincial People's Hospital (the First Affiliated Hospital of Hunan Normal University) from July 2018 to September 2022 were retrospectively analyzed. According to whether ECMO was successfully weaning off, patients were divided into the successful weaning off group and the failed weaning off group. The basic data, duration of conventional cardiopulmonary resuscitation (CCPR, the time from cardiopulmonary resuscitation to ECMO), duration of ECMO, pulse pressure loss, complications, and the use of distal perfusion tube and intra-aortic balloon pump (IABP) were compared between the two groups. Univariate and multivariate Logistic regression analyses were performed to identify the risk factors for weaning failure of ECMO. RESULTS: Twenty-three patients (41.07%) were successfully weaned from ECMO. Compared with the successful weaning off group, patients in the failed weaning off group were older (years old: 46.7±15.6 vs. 37.8±16.8, P < 0.05), higher incidence of pulse pressure loss and ECMO complications [81.8% (27/33) vs. 21.7% (5/23), 84.8% (28/33) vs. 39.1% (9/23), both P < 0.01], and longer CCPR time (minutes: 72.3±19.5 vs. 54.4±24.6, P < 0.01), shorter duration of ECMO support (hours: 87.3±81.1 vs. 147.7±50.8, P < 0.01), and worse improvement in arterial blood pH and lactic acid (Lac) levels after ECPR support [pH: 7.1±0.1 vs. 7.3±0.1, Lac (mmol/L): 12.6±2.4 vs. 8.9±2.1, both P < 0.01]. There were no significant differences in the utilization rate of distal perfusion tube and IABP between the two groups. Univariate Logistic regression analysis showed that the factors affecting the weaning off ECMO of ECPR patients were pulse pressure loss, ECMO complications, arterial blood pH and Lac after installation [pulse pressure loss: odds ratio (OR) = 3.37, 95% confidence interval (95%CI) was 1.39-8.17, P = 0.007; ECMO complications: OR = 2.88, 95%CI was 1.11-7.45, P = 0.030; pH after installation: OR = 0.01, 95%CI was 0.00-0.16, P = 0.002; Lac after installation: OR = 1.21, 95%CI was 1.06-1.37, P = 0.003]. After adjusting for the effects of age, gender, ECMO complications, arterial blood pH and Lac after installation, and CCPR time, showed that pulse pressure loss was an independent predictor of weaning failure in ECPR patients (OR = 1.27, 95%CI was 1.01-1.61, P = 0.049). CONCLUSIONS Early loss of pulse pressure after ECPR is an independent predictor of failed weaning off ECMO in ECPR patients. Strengthening hemodynamic monitoring and management after ECPR is very important for the successful weaning off ECMO in ECPR.
Humans ; Extracorporeal Membrane Oxygenation ; Blood Pressure ; Retrospective Studies ; Perfusion ; Cardiopulmonary Resuscitation

The community structure and diversity of the gut microbiota are associated with human diseases. However, the analysis of different community structure might be influenced by experimental approaches such as the quality of DNA extraction. Therefore, evaluating the efficiency of different DNA extraction methods for specific intestinal species is a guideline for obtaining a comprehensive human gut microbial profile, which may assist the in-depth investigation into the structure of the gut microbial community. The aim of this study was to perform a comparative analysis of five different DNA extraction methods. With the aid of qPCR, the efficiency of five DNA extraction kits was evaluated in terms of the purity of the extracted DNA, the DNA concentration, and the abundance of genomic DNA extracted from specific intestinal species. The results showed that the kit Q gave the best extraction results, especially for Gram-positive bacteria such as Lactobacillus and Bifidobacterium. The average DNA concentration of the N kit was lower than that of the Q kit, but there was no significant difference between the two in terms of the purity. Compared to the other three commercial kits (M, PSP, TG), the efficiency of the N kit in extracting the genomic DNA of the specified microorganisms were the least different from those of the Q kit. In contrast, the DNA extracted by the M kit was of higher quality but of lower concentration, and was not very efficient for Gram-positive bacteria. The DNA extracted by the TG and PSP kits was inferior to the other validated kits in terms of the concentration, quality and bacterial abundance. These results provide a basis for the selection of genomic DNA extraction methods in microecological research experiments.
DNA/genetics* ; DNA, Bacterial/genetics* ; Feces/microbiology* ; Humans ; Microbiota/genetics* ; RNA, Ribosomal, 16S/genetics*

OBJECTIVE: To establish quantitative surface plasmon resonance (SPR) assay for antibodies against human platelet antigen-1a (HPA-1a). METHODS: Recombinant protein was fixed on the chip surface by amino coupling method. SPR assay was used to detect the standard antibodies against HPA-1a at different conceatration. The optimal experimental parameters were determined, and standard curves were constructed with linear regression. Moreover, the sensitivity, specificity, accuracy and precision of the assay were evaluated. RESULTS: The quantitative SPR assay for HPA-1a antibodies was established. The determination ranges were 0-20 IU, with accuracy (recovery rate) was 97.75%-103.08%. The intra-assay precision [coefficients of variation (CV)] was 3.53%-4.29%, and the inter-assay precision (CV) was 2.08%-4.40%. For specificity test, several kinds of monoclonal and human antibodies against platelet membrane protein were tested and no positive result was observed. CONCLUSION The established quantitative SPR assay for HPA-1a antibodies shows good sensitivity, specificity, accuracy and precision, and this rapid and simple method provides a new reference method for scientific research and clinical antibody detection.
Antigens, Human Platelet ; Blood Platelets ; Humans ; Isoantibodies ; Surface Plasmon Resonance

Objective To establish an HPLC method for the simultaneous determination of harpagide and harpagoside content in Scrophularia ningpoensis (SN). Methods An Eclipse C₁₈ column was used for determination of methanol extract of S. ningpoensis with a HPLC-PDA method and mobile phase of acetonitrile-0.03% phosphate solution in a gradient elution manner. The flow rate of mobile phase was 1.0 ml/min, and the detection wavelengths were 210 nm and 280 nm. Results Harpagide and harpagoside contents in SN showed good linear relationships within 0.1020-0.5100 mg/ml (r=0.9999) and 0.0340-0.1700 mg/ml (r=0.9999). Their average recovery rates were 97.44% and 97.08%.The RSDs were 0.93% and 1.24%.. There were significant differences in the contents of harpagide and harpagoside in SN from 15 origins (P<0.01). The content of harpagoside in Sichuan Long-dong, Zhejiang Lin-an, Zhejiang Pan-an and Henan Nan-feng is higher. Conclusion This method is stable, accurate and reproducible and can be used for the quality control of SN.

Objective:To analyze the influence of factors before initiation of extracorporeal cardiopulmonary resuscitation (ECPR) on the prognosis of patients, so as to explore the intervention timing and improvement strategy of ECPR.Methods:A retrospective analysis was performed on 29 patients who underwent ECPR in the First Affiliated Hospital of Hunan Normal University (Hunan people's Hospital)from July 2018 to April 2021. Patients were divided into the survival group ( n = 13) and death group ( n = 16) according to whether they survived at discharge. The duration of conventional cardiopulmonary resuscitation (CCPR), initial heart rate before ECPR, the ratio of out-of-hospital cardiac arrest (OHCA) and in-hospital cardiac arrest (IHCA), and the ratio of transported cases outside the hospital were compared between the two groups. According to different CCPR time, the patients were divided into the ≤45 min group, 45-60 min group and >60 min group to compare the hospital survival and sustained return of spontaneous circulation (ROSC) rate . According to the location of cardiac arrest, the patients from emergency department and other department were divided to compare the survival of IHCA. Results:The total survival rate was 44.83%, the average duration of extracorporeal membrane oxygenation (ECMO) was 114 (33.5, 142.5) h, and the average duration of CCPR time was 60 (44.5, 80) min. The duration of ECMO was longer in the survival group than in the death group ( P = 0.001). The duration of CCPR (the time from CPR to ECMO) in the survival group was significantly shorter than that in the death group ( P = 0.010). Patients with defibrillatory rhythm had higher hospital survival rate ( P = 0.010). OHCA patients had higher mortality than IHCA patients ( P = 0.020). Mortality of patients transferred from other hospitals was higher ( P = 0.025). Hospital survival and ROSC decreased in turn by CCPR duration ≤ 45 min, 45-60 min, and > 60 min ( P = 0.001). The location of CA occurrence had no impact on the hospital survival rate of IHCA patients ( P=0.54). Conclusions:Hospital survival of ECPR is higher than that of CCPR. ECPR is effective for refractory cardiac arrest. The prognosis of ECPR is significantly related to the duration of CCPR, initial heart rate, and location of CA. Education and team training should be strengthened to improve the survival rate of ECPR.

Objective:To investigate the people's cardiopulmonary resuscitation (CPR) ability in Hunan Province and whether there are differences in the skill level of CPR among respondents of different ages and education levels.Methods:A self-made questionnaire was conducted to survey people in Hunan Province by means of WeChat from May 2018 to April 2019. The questionnaire referred to 2016 National consensus on cardiopulmonary resuscitation in China issued by Cardiopulmonary Resuscitation Specialized Committee of Chinese Research Hospital Association and 2017 version of American Heart Association (AHA) CPR operational guide, and combined with the CPR assessment scale of Hunan Provincial People's Hospital. The contents of the survey included the basic information of the subjects, the common knowledge and skills of first aid, the willingness to learn CPR skills and implement CPR, and the operational requirements of high quality CPR, etc. Results:A total of 6 563 people received the on-site first aid knowledge questionnaire, and 4 355 people completed and submitted the questionnaire. The recovery rate was 66.36%, of which 3 602 valid questionnaires were from IP in Hunan Province, and the qualified rate was 82.71%. Among the subjects, 1 532 were men (42.53%) and 2 070 were women (57.47%). The majority aged group from 19 to 30 (59.41%) and from 31 to 50 (36.70%). 307 were rural (8.52%) and 3 295 were urban (91.48%); and the majority levels of education were senior or technical secondary school (38.26%) and undergraduate or junior college (44.50%). Of the 3 602 respondents, 39.09% indicated that they had been exposed to CPR knowledge and only 0.36% indicated that they did not wish to participate in CPR training. 69.93% of the respondents said they would actively to help if they wet cardic arrest, and 97.92% of those were willing to do so if the patients were relatives or friends. The survey results showed that only 8.91% of respondents chose relatively high-quality CPR options (chest compressions were performed first, the pressing point was located in the sternum of the midpoint of bilateral nipple line, the pressing frequency was 100-120 times/min, pressing depth was 5-6 cm, the ratio of chest compression to artificial ventilation was 30∶2). Among these people, the correct rate was higher in the ages of 31-50 years old and ≤ 18, 19-30 than those ≥51 years old (12.71% vs. 0%, 6.87%, 8.70%, χ2 = 41.420, P < 0.01). The correct rate of education at graduate level and above was higher than those in junior middle school and below, high school or technical secondary school, undergraduate or junior college (19.57% vs. 2.07%, 3.41%, 13.72%, χ2 = 152.262, P < 0.01). Conclusions:The public in Hunan Province have a strong sense of first aid, and some of the theoretical knowledge of CPR. People between 31 years old and 50 years old of age and with graduate education and above know more about CPR, but the overall mastery of CPR skills is poor. It is necessary to further improve the ability of the public as the first witness by teaching various skills of CPR in various forms.

Objective:To Analyze the risk factors and prognosis of patients with hyperfibrinolysis after severe trauma.Methods:A prospective case-control study was conducted to analyze the clinical data of severe trauma patients whose injury severity score (ISS)≥16 points admitted to First People's Hospital of Taicang from October 2017 to December 2018. Blood clot dissolution rate parameter (LY30) ≥3% was diagnosed as hyperfibrinolysis. The patients were divided into hyperfibrinolysis group ( n=27) and non-hyperfibrinolysis group ( n=27). Data were collected including patients' general information, hemoglobin (Hb), hematocrit (Hct), platelet count (PLT), fibrinogen (Fg), lactic acid value and thrombelastograghy (TEG). Univariate and multivariate Logistic regression analysis were used to identify risk factors of hyperfibrinolysis after severe trauma. Prognostic indicators and K-M survival curve of the patients were analyzed. Results:(1) A total of 142 patients with severe trauma were enrolled. There were 101 males and 41 females, aged 45-65 years (mean, 56 years). The incidence of hyperfibrinolysis was 14.8%. (2) Univariate analysis showed that ISS, abbreviated injury scale (AIS) ratio of pelvic limbs ≥3 points, shock index (SI) ratio >1, temperature, PLT, Fg and lactic acid value were correlated with the occurrence of hyperfibrinolysis after severe trauma ( P<0.05). (3) Multivariate Logistic regression analysis showed that PLT ( OR=1.035), ISS ( OR=0.898) and lactate acid value ( OR=0.735) were independent risk factors for hyperfibrinolysis after severe trauma ( P<0.05 or 0.01). (4) Hyperfibrinolysis group required more massive transfusion (38.1% vs. 2.5%), were more prone to multiple organ dysfunction syndrome (MODS) (90.5% vs. 48.8%), and had a higher 24-hour mortality (47.6% vs. 8.3%), when compared with non-hyperfibrinolysis group ( P<0.01). K-M survival curve indicated that the median survival time in hyperfibrinolysis group after trauma was 1.33 days ( IQR, 0.40-17.20 days). Conclusions:The incidence of hyperfibrinolysis after severe trauma is low. ISS, PLT and lactic acid value are the independent risk factors for hyperfibrinolysis after severe trauma. Patients with severe post-traumatic hyperfibrinolysis are characterized by high probabilities of massive transfusion and MODS, short survival time and high early mortality.

Allergic rhinitis (AR) is a global health problem that causes major illnesses and disabilities worldwide. Epidemiologic studies have demonstrated that the prevalence of AR has increased progressively over the last few decades in more developed countries and currently affects up to 40% of the population worldwide. Likewise, a rising trend of AR has also been observed over the last 2–3 decades in developing countries including China, with the prevalence of AR varying widely in these countries. A survey of self-reported AR over a 6-year period in the general Chinese adult population reported that the standardized prevalence of adult AR increased from 11.1% in 2005 to 17.6% in 2011. An increasing number of original articles and imporclinical trials on the epidemiology, pathophysiologic mechanisms, diagnosis, management and comorbidities of AR in Chinese subjects have been published in international peer-reviewed journals over the past 2 decades, and substantially added to our understanding of this disease as a global problem. Although guidelines for the diagnosis and treatment of AR in Chinese subjects have also been published, they have not been translated into English and therefore not generally accessible for reference to non-Chinese speaking international medical communities. Moreover, methods for the diagnosis and treatment of AR in China have not been standardized entirely and some patients are still treated according to regional preferences. Thus, the present guidelines have been developed by the Chinese Society of Allergy to be accessible to both national and international medical communities involved in the management of AR patients. These guidelines have been prepared in line with existing international guidelines to provide evidence-based recommendations for the diagnosis and management of AR in China.
Adult ; Asian Continental Ancestry Group* ; China ; Comorbidity ; Developed Countries ; Developing Countries ; Diagnosis* ; Epidemiologic Studies ; Epidemiology ; Global Health ; Humans ; Hypersensitivity* ; Prevalence ; Rhinitis, Allergic*

Objective To study the changes of immune function and liver function in patients with primary biliary cirrhosis.Methods From March 2013 to March 2016 in our hospital patients with primary biliary cirrhosis 30 cases as the observation group.There were 17 patients with mild liver injury and 13 with severe hepatic dysfunction.And at the same time for the admission of 30 patients with other liver diseases in patients with liver disease as a group,30 cases underwent physical examination as healthy group were detected by flow cytometry in three groups of subjects of immunity comparison of three groups of cells,dendritic cell subsets in peripheral blood and liver function index.Results The CD4+ cells and CD8+ cells in the observation group were significantly higher than those in the healthy group(P＜0.05),while the CD11c+ cells,CD123+ cells and NK cells were significantly lower than those in the healthy group(P＜0.05).In the observation group,CD4+ cells,CD8+ cells,CD11c+ cells and CD123+ cells were significantly higher than those of other liver diseases but the NK cells were significantly lower than those of the other groups (P＜0.05).In healthy group,CD4+ cells,CD8+ cells,CD11c+ cells,CD123+ cells and NK cells were significantly higher than those of other liver disease group(P＜0.05) The CD4+,CD8+ and CD123+ cells in the liver injury group were significantly higher than those in the liver function damage group,while the CD11c+ and NK cells were significantly lower than those in the liver function group,and the difference was statistically significant(P＜0.05).The CD4+ and CD8+ cells in the AMA-M2 negative group were significantly lower than that in the AMA-M2 positive group,while those in CD123+,CD1 1c+ and NK cells were significantly higher than those in the AMA-M2 positive group,and the difference was statistically significant (P＜ 0.05).Conclusion The immune function of patients with primary biliary cirrhosis and liver function injury are associated with production of serum AMA-M2 antibodies,including CD11c+ and CD123+ or as the main factors influence the development of the disease and the AMA-M2 antibody.