BACKGROUND:Primary cortical neurons and microglial cells play a crucial role in exploring cell therapies for neurological disorders,and most of the current methods for obtaining the two types of cells are cumbersome and require separate extraction.It is therefore crucial to find a convenient and rapid method to extract both types of cells simultaneously. OBJECTIVE:To explore a novel method for simultaneous extraction of primary cortical neurons and microglial cells. METHODS:Newborn suckling SD rats were taken within 24 hours.The brain was removed and placed in a dish with DMEM,and the pia mater was removed for later use.Primary neurons were extracted from the same brain tissue,and then the remaining brain tissue was used to extract microglial cells.The whole process was performed on ice.Extraction and culture steps of primary cortical neurons:The cerebral cortex was taken 2.0-3.0 mm with forceps,and the tissue was digested with papain for 20 minutes.After aborting digestion,the blown tissue presented an adherent tissue suspension.The supernatant cell suspension was obtained,filtered,and dispensed into 15 mL centrifuge tubes.After centrifugation and re-suspension,the cells were inoculated onto 6-well plate crawls coated with L-polylysine.Neuronal morphology was observed at 1-day intervals,and staining could be performed for identification using immunofluorescence staining of MAP2 and β-Tubulin by day 7.Microglia extraction and culture steps:The remaining brain tissue at 8-10 mm thick was subjected to microglial cell extraction,digested by trypsin for 20 minutes.After digestion was stopped,the tissue was blown to a homogenate,and then the homogenate was transferred to the culture bottle for culture.On day 14,the culture flasks were sealed and subjected to constant temperature horizontal shaking for 2 hours.Microglial cells were shed in the supernatant.Purified microglial cells were taken and continued to be cultured for 3 days for identification by Iba1 immunofluorescence staining. RESULTS AND CONCLUSION:(1)After 24 hours of culture,the neurons were adherent to the wall,the cytosol was enlarged,and some neurons developed synapses.After 3 and 5 days of culture,the cytosol was further enlarged,and most of the neurons were in the form of synapses,and some neurons were growing in clusters.On day 7,neuronal synapses were prolonged and thickened,and they were connected with each other to form a network.The neurons were identified by β-Tubulin and MAP2 immunofluorescence staining.(2)The cells grew close to the wall on day 1 of culture.On days 3,5,and 7,the density of microglial cells was small,and the cell morphology was bright oval or round,but the cells basically grew in clumps on the upper layer of other cells.On day 10,the density of microglial cells increased significantly.On day 14,microglial cells grew in dense clumps on the upper layer of other cells,and then they could be isolated and purified.The isolated and purified cells were taken and re-cultured to day 3 and identified as microglial cells by Iba1 immunofluorescence;their purity was greater than 95%.(3)The results show that primary cortical neurons and microglial cells obtained by this method after extraction and culture are of high purity,good morphology,and high viability.

Objective:To investigate the acute gastrointestinal injury (AGI) in patients with extracorporeal membrane oxygenation (ECMO) at the early stage of operation and its influencing factors.Methods:A total of 70 patients with ECMO who were hospitalized in the Emergency Care Unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed, and a total of 70 patients with ECMO who were hospitalized in the emergency care unit of Guangxi Zhuang Autonomous Region People's Hospital from September 2020 to December 2021 were retrospectively analyzed. According to the 2012 guidelines of the European Society of Intensive Care Medicine on the classification of acute gastrointestinal injury in critically ill patients, the patients were divided into AGI group and non-AGI group. The incidence of acute gastrointestinal injury in the early stage was statistically analyzed, and the results of blood gas analysis during ECMO loading and ECMO parameters, hemodynamic indexes and biochemical indexes after ECMO transfer were statistically analyzed. To explore the influencing factors and independent risk factors of AGI in the early stage. In addition, 70 patients were divided into successful group and non-successful group according to whether they were successfully withdrawn. The occurrence of acute gastrointestinal injury between the two groups was compared, and the effect of acute gastrointestinal injury on ECMO patients was analyzed.Results:Among the 70 ECMO patients, the incidence of early AGI was 71.43% (50 cases), and the components of AGI Ⅰ, Ⅱ, Ⅲ and Ⅳ were 18.57% (13 cases), 41.43% (29 cases), 11.43% (8 cases) and 0% (0 cases), respectively. ① Univariate analysis showed that systolic blood pressure, diastolic blood pressure, mean arterial pressure (MAP), vasoactive drug index (VIS), pH, lactic acid and BMI were significantly different between AGI group and non-AGI group when ECMO was used ( P < 0.05). Logistic binary regression analysis showed that BMI was an independent risk factor for early AGI in ECMO patients (ROC area 0.657, 95% confidence interval 0.522-0.791 ( P < 0.05), and Yoden index 0.15). (3) The AGI composition ratio of the unsuccessful group was higher than that of the unsuccessful group ( P < 0.05). Conclusions:Patients with ECMO have a high incidence of AGI in the early stage, mainly occurring in grade I and Ⅱ. Systolic blood pressure, diastolic blood pressure, MAP, VIS, pH, lactic acid and BMI when ECMO is put on are influential factors for the early development of AGI in ECMO patients, among which BMI is an independent risk factor for the early development of AGI in ECMO patients. The occurrence of AGI reduces the probability of successful withdrawal in ECMO patients.

Objective:To investigate the recovery of renal function and its influencing factors in patients receiving extracorporeal membrane oxygenation (ECMO) support and complicated with acute kidney injury(AKI).Methods:This was a retrospective observational study. The clinical data of patients with ECMO support and AKI admitted to the Emergency intensive care unit of the People's Hospital of Guangxi Zhuang Autonomous Region from October 2019 to December 2021 were collected. The patients were divided into renal function recovery group and renal function non-recovery group according to the recovery of renal function after 28 days of ECMO. With renal function non-recovery at 28 days as the end point of the study, and the variables with significant differences in baseline were selected for stepwise backward regression to determine the independent risk factors. The receiver operator characteristic (ROC) curve was drawn, and the area under the curve (AUC) was used to evaluate the diagnostic value of independent risk factors.Results:A total of 40 patients were enrolled, of which 28 patients (70%) had recovery of renal function, and 12 patients (30%) did not have recovery of renal function. Stepwise backward multivariate logistic regression analysis showed that lactate level at ECMO initiation was an independent risk factor for non-recovery of renal function ( OR = 1.380, 95% CI: 1.096-1.738, P = 0.006). The ROC curve showed that the AUC and 95% CI were 0.863 (0.751-0.975), the sensitivity was 100%, and the specificity was 75%. Conclusion:Lactate level at ECMO initiation was an independent risk factor for non-recovery of renal function on 28 days after ECMO initiation among patients undergoing ECMO support complicated with AKI. Lactate has a high predictive value for the non-recovery of renal function.

BACKGROUND:Studies have found that activation of nuclear factor-erythroid 2-related factor 2/heme oxidase-1(Nrf2/HO-1)pathway can alleviate oxidative stress caused by cerebral ischemia-reperfusion injury,but whether human bone marrow mesenchymal stem cells(hBMSC)can activate Nrf2/HO-1 pathway to alleviate cerebral ischemia-reperfusion injury is still lacking relevant studies. OBJECTIVE:To investigate whether intracranial transplantation of hBMSC alleviates oxidative stress injury in cerebral ischemia-reperfusion animal models by activating Nrf2/HO-1 pathway. METHODS:Totally 40 male SPF SD rats were randomly divided into sham operation group,model group,hBMSC transplantation group,hBMSC+solvent group and hBMSC+Nrf2 inhibitor group.Each group consisted of eight animals.In the model group and the hBMSC transplantation group,middle cerebral artery occlusion model was prepared by thread embolization method.The thread embolization was removed 1 hour later,and 30 μL PBS or hBMSC cultured to at least passage 5 was injected into the right cortex and striatum of rats.In the hBMSC+Nrf2 inhibitor group and hBMSC+solvent group,the left ventricle was injected with Nrf2 inhibitor Brusatol and its solvent dimethyl sulfoxide respectively 24 hours before model establishment,then the middle cerebral artery occlusion model was prepared,and hBMSC was injected.Relevant indexes were detected 3 days after transplantation. RESULTS AND CONCLUSION:(1)CT and TTC staining showed the same area and volume of cerebral infarction:model group>hBMSC+Nrf2 inhibitor group>hBMSC+solvent group>hBMSC transplantation group>sham operation group.(2)Hematoxylin-eosin staining and Nissl's staining showed that the ischemic brain tissue was intact and the neurons were normal in the sham operation group.Compared with the model group,the pathological morphology and neuronal injury of the hBMSC transplantation group and the hBMSC+solvent group were significantly improved.Compared with the hBMSC+solvent group,the hBMSC+Nrf2 inhibitor group had more serious pathological morphology and neuronal damage.(3)Western blot assay and oxidative stress index detection results showed that compared with the sham operation group,Nrf2 and HO-1 proteins were decreased(all P<0.05),malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the model group.Compared with the model group,the expression levels of Nrf2 and HO-1 proteins were increased(all P<0.05),malondialdehyde was decreased and superoxide dismutase was increased(all P<0.05)in the hBMSC transplantation group and the hBMSC+solvent group.Compared with the hBMSC+solvent group,the expression levels of Nrf2 and HO-1 proteins were simultaneously decreased(all P<0.05),and malondialdehyde was increased and superoxide dismutase was decreased(all P<0.05)in the hBMSC+Nrf2 inhibitor group.(4)These results indicate that hBMSC can alleviate cerebral ischemia-reperfusion injury possibly by activating Nrf2/HO-1 pathway.

Objective: To examine the mediating effects of mindfulness level on resilience and symptoms of anxiety and depression among healthcare workers, so as to provide the reference for developing effective psychological intervention. Methods: The clinical doctors, nurses and public health professionals were selected using the stratified random cluster sampling method from hospitals, disease prevention and control centers, and health departments in five cities in Shandong Province, including Qingdao, Jinan, Rizhao, Jining and Liaocheng in January 2023. Basic information, mindfulness level, resilience and symptoms of anxiety and depression among healthcare workers were collected using general demographic questionnaires, the 5-item Mindful Attention Awareness Scale, 10-item Connor-Davidson Resilience Scale, Generalized Anxiety Disorder Scale and Patient Health Questionnaire, respectively. The Process program was used to analyze the mediating effects of mindfulness level on resilience and symptoms of anxiety and depression. Results: A total of 1 836 healthcare workers were investigated, including 472 males (25.71%) and 1 364 females (74.29%), and the median age was 39 (interquartile range, 12) years. There were 629 clinical doctors (34.26%), 963 nurses (52.45%) and 244 public health professionals (13.29%). The median scores of mindfulness level and resilience were 22 (interquartile range, 7) and 20 (interquartile range, 4) points, respectively. The detection rates of anxiety and depression symptoms were 49.78% and 72.28%, respectively. The mediation analysis showed that mindfulness level exerted a partial mediating effect between resilience and anxiety symptoms (β=-0.510, P<0.001), with a direct effect value of -0.130 and a mediating effect value of -0.046, and the mediating effect accounted for 26.14% of the total effect; mindfulness level also exerted a partial mediating effect between resilience and depression symptoms (β=-0.575, P<0.001), with a direct effect value of -0.120 and a mediating effect value of -0.052, and the mediating effect accounted for 30.23% of the total effect. Conclusion Mindfulness level plays a mediating effect between resilience and symptoms of anxiety and depression among healthcare workers.

  Objective  To understand the current status of surgical treatment for hemophilia osteoarthropathy (HO) in China.  Methods  Using an online questionnaire, select domestic hospitals that partici-pated in the compilation of the 'Guideline for perioperative management of hemophilia patients undergoing orthopaedic surgery in China ', in addition to members of the National Joint Surgery Group, and the Orthopedic Branch of the Chinese Medical Association for targeted investigation and analysis.  Results  A total of 17 domestic hospitals were included, all of which were general hospitals. Hospitals that started HO surgery treatment before 2000 accounted for 35.29%. A total of 3057 surgical cases of HO were reported by those hospitals. The most commonly performed surgical procedures were hip and knee joint replacement. The most commonly used coagulation factor replacement regimen was recombinant coagulation factor preparation. Ten hospitals reported finding patients with transfusion-related infectious diseases. Bleeding and hematoma formation were the most frequently reported surgical complications. Excessive length of hospital stay and high economic costs were the most frequently reported problems.  Conclusions  Surgical treatment for HO in 17 hospitals is mainly carried out in some large comprehensive medical centers in the eastern region. Compared with the patient base, the popularity and number of surgeries are still relatively insufficient. It is necessary to further standardize the treatment system by standardizing factor replacement and strengthening rehabilitation to improve surgical treatment outcomes.

Although it is widely believed that abnormal energy metabolism exists in cancer cells and affects the biological behavior of cancers, the exact mechanism of energy metabolic reprogramming and specific mechanism of its effect on proliferation, invasion and metastasis of cancer cells have not been clarified. In recent years, studies have shown that long non-coding RNA (lncRNA) can affect energy metabolism, development and progression of cancer cells through binding to specific nucleic acids and proteins at the transcriptional and post-transcriptional stages, and specifically through transcriptional interference, epigenetic regulation of genes, changes in protein activity, competitive binding to microRNA (miRNA) and other related mechanisms. The further study on the mechanism of lncRNA regulating energy metabolism reprogramming of cancer cells is expected to find new markers and targets for diagnosis and treatment of cancer. This paper reviews the current research progress of the mechanism of lncRNA regulating metabolic reprogramming of glucose, fatty acid, protein and nucleotide in cancer, and provides a new idea of lncRNA's regulation of energy metabolism pathways for targeted anticancer therapy.

Due to its advantages of low wear, high stability and flexibility, the new generation metal-on-metal hip prosthesis is favored by many patients with hip diseases, especially young patients. However, in recent years, several studies have indicated that adverse reactions to metal debris (ARMD) caused the formation of inflammatory pseudotumor, which ultimately led to a higher revision rate after metal-on-metal hip arthroplasty. This aroused the widespread concern from doctors and patients. Moreover, revision surgery for metal-on-metal hip arthroplasty in the setting of inflammatory pseudotumor is faced with a great risk of failure because of the large defects of bone and surrounding soft tissue and difficulty in removing the original prosthesis and the fixing of the modified prosthesis. Therefore, the use of such products is restricted with caution in their choice. We summarized the recent developments in the research in the risk factors, diagnosis and treatment of inflammatory pseudotumor after metal-on-metal hip arthroplasty. The risk factors for the formation of inflammatory pseudotumor around the hip prosthesis mainly include the increase of metal ion concentration, the position of prosthesis implantation and the patient's own factors. The diagnosis mainly depends on physical examination, imaging examination, laboratory examination, arthroscopy and histological examination. The treatment strategies for clinical symptomatic and asymptomatic patients are also varies. Through the detailed analysis, evaluation and summary of the above contents, we may provide guidance for the selection of hip prosthesis, and lay the foundation for further exploration of the mechanism of inflammatory pseudotumor caused by ARMD.

Objective To investigate the mid-and long-term effects of metal-on-metal hip resurfacing arthroplasty (HRA),and to analyze its related factors.Methods A total of 64 patients (81 hips) who underwent metal-on-metal HRA from June 2005 to January 2013 were recruited in the present study.There were 35 males (44 hips) and 29 females (37 hips) with the mean age of 48.26±10.45 years (20-65 years),including 47 unilateral and 17 bilateral HRAs.The cohort consisted of osteoarthritis secondary to the developmental dysplasia of the hip (23 cases,29 hips),necrosis of the femoral head (19 cases,22 hips),osteoarthritis (8 cases,10 hips),rheumatoid arthritis (5 cases,9 hips),ankylosing spondylitis (6 cases,8 hips),pigmented villonodular synovitis (2 cases,2 hips) and Otto's disease (1 case,1 hip).During the follow-up duration,radiographic features,including acetabular inclination angle,stem-femoral shaft angle,component loosening,osteolysis,femoral neck narrowing and heterotopic ossification,were evaluated by hip X-rays in straight and froglike position.The size and type of pseudotumor were assessed by MRI and ultrasonography.Clinical efficacy was evaluated by Harris hip score and the University of California at Los Angeles (UCLA) hip score.Considering revision surgery as the end point,the component survivorship was calculated.Results The mean follow-up was 7.98±2.21 years,ranging from 5.0 to 12.8 years.The mean postoperative Harris hip score (92.01 ±5.69) was higher than the preoperative score (41.93 ±9.09).The mean postoperative UCLA pain,walking,function,activity scores (9.37±0.86,9.14± 1.01,8.77± 1.09,6.47± 1.27,respectively) were improved when compared with the preoperative UCLA scores (3.57± 1.23,5.99± 1.30,5.00± 1.01,3.84± 1.41,respectively).The postoperative flexion,abduction and adduction,medial and lateral rotation of the hip was larger than the preoperative ones.Complications occurred in 10 hips (12.3％,10/81).Seven patients (8 hips) experienced early and intermediate complications,including one intraoperative femoral nerve injury,one deep femoral artery and saphenous nerve injuries during the same surgery,one unexplained pain of hip,one femoral neck fracture,three hips of heterotopic ossification,and one pseudotumour.There were mid-and long-term complications in two hips,including one narrowing of the neck and one pseudotumour which was occurred at 9 years.There was one patient (2 hips) underwent revision surgery twice at 5 months and 9 years.The former cause of revision was femoral neck fracture and the latter one was pseudotumour.The Kaplan-Meier survivorship was 98.8％ at five years,and 95.0％ at ten years.Conclusion Patients who underwent metal-on-metal HRA could obtain good mid and long-term results.Pseudotumour and unexplained pain of the hip are critical factors which can affect the mid-and long-term results and survivorship of metal-on-metal HRA.

Objective To explore the appropriate target ranges of blood glucose in intensive insulin therapy (ⅡT) for acute hyperglycemia following traumatic brain injury (TBI).Methods A randomized,open-label and controlled clinical trial was performed on 208 patients,admitted to our hospitals from Junuary 2014 to Sepember 2016.They were divided into ⅡT group (n=156),who were subdivided into slight (10.1-13.0 mmol/L),moderate (7.1-10.0 mmol/L),and strict (4.4-7.0 mmol/L) control blood glucose groups (n=52),and non-ⅡT group (n=52).Survival analysis 6 months after treatment was performed by Kaplan-Meier method.Modified Rankin scale (mRS) scores and Barthel index (BI),Glasgow Outcome scale (GOS) scores,concentrations of lactic acid in cerebrospinal fluid (CSF) and glycosylated hemoglobin,Glasgow coma scale (GCS) scores,Acute Physiology and Chronic Health Evaluation (APACHE Ⅱ) scores,Length of staying in intensive care unit (ICU) and incidence of adverse events were compared between the patients from different groups at different treatment times.Results Blood glucose level within 7 d of admission in patients ofⅡT group was in target ranges.The survival rate of patients from slight and moderate control blood glucose groups was significantly higher than that in the non-ⅡT group and strict control blood glucose group 6 months after treatment (x2=4.237,P=0.040;x2=5.621,P=0.018).As compared with those in the non-ⅡT group and strict control blood glucose group,the mRS scores 3 months after treatment were significantly decreased,and GOS scores and BI one,3 and 6 months after treatment were significantly increased in patients from slight and moderate control blood glucose groups (P＜0.05).As compared with that in the non-ⅡT group,and slight and moderate control blood glucose groups,the glycosylated hemoglobin level 7 d after treatment was significantly decreased in strict control blood glucose group (P＜0.05).As compared with those in the non-ⅡT group and strict control blood glucose group,the concentration of lactic acid in CSF 7 d after treatment,APACHE Ⅱ scores 7 and 14 d after treatment,length of staying in ICU and incidence of adverse events were significantly decreased in patients from slight and moderate control blood glucose groups (P＜0.05).The mean value of blood glucose in slight and moderate control blood glucose groups was (8.40±0.39) mmol/L.Conclusion Proper ⅡT improves the outcomes of TBI patients and (8.40±0.39) mmol/L are established as the target ranges in ⅡT for TBI.