Objective To investigate the effect of MALAT1 expressions on cell proliferation and apoptosis in astrocytes by regulating mitogen-activated protein kinase(MAPK)/extracellular signal-regulated kinase(ERK1,2)pathway.Methods The MALAT1 gene was knocked down and over-expressed in C8-D1A cells by lentiviral and plasmid vectors,respectively.The expressions of MALAT1,cell proliferation-related markers(Ki67,MCM2,PCNA)and apoptosis-related proteins(Caspase-3,Bax,Bcl-2)were detected by quantitative real-time polymerase chain reaction(qPCR).CCK-8 assay and flow cytometry were used for cell proliferation and apoptosis in C8-D1A cells.Immunofluorescence was adopted to detect the protein expressions of Caspase-3 and Ki67.Western blotting was used to detect the protein expressions of Caspase-3,Bax,Bcl-2,ERK1/2,p-ERK1/2,p38MAPK and p-p38MAPK.Results Compared with the control group,over-expressed MALAT1 inhibited cell proliferation and induced cell apoptosis in C8-D1A cells while the knockdown of MALAT1 significantly enhanced cell proliferation and anti-apoptotic ability in C8-D1A cells.The proportion of C8-D1A cells in G0/G1-phase and G2/M-phase was higher than in the control group as evidenced by flow cytometry,but was lower in S-phase.Meanwhile,data showed that Caspase-3 was increased while p-ERK1/2 was decreased in terms of protein levels.The mRNA expressions of Ki67 and PCNA were decreased.After knockdown of MALAT1,the proportion of C8-D1A cells in S-phase was higher,but was lower in G2/M-phase.The protein expressions of Caspase-3 and Bax decreased while those of p-ERK1/2 and p-p38MAPK increased.The mRNA expressions of Ki67,MCM2 and PCNA were increased.The differences were all statistically significant(P＜0.05).Conclusion MALAT1 promotes astrocyte apoptosis and inhibits proliferation by regulating the MAPK/ERK1,2 signaling pathway.

Patients with severe trauma require an extremely timely treatment and transfusion plays an irreplaceable role in the emergency treatment of such patients. An increasing number of evidence-based medicinal evidences and clinical practices suggest that patients with severe traumatic bleeding benefit from early transfusion of low-titer group O whole blood or hemostatic resuscitation with red blood cells, plasma and platelet of a balanced ratio. However, the current domestic mode of blood supply cannot fully meet the requirements of timely and effective blood transfusion for emergency treatment of patients with severe trauma in clinical practice. In order to solve the key problems in blood supply and blood transfusion strategies for emergency treatment of severe trauma, Branch of Clinical Transfusion Medicine of Chinese Medical Association, Group for Trauma Emergency Care and Multiple Injuries of Trauma Branch of Chinese Medical Association, Young Scholar Group of Disaster Medicine Branch of Chinese Medical Association organized domestic experts of blood transfusion medicine and trauma treatment to jointly formulate Chinese expert consensus on blood support mode and blood transfusion strategies for emergency treatment of severe trauma patients ( version 2024). Based on the evidence-based medical evidence and Delphi method of expert consultation and voting, 10 recommendations were put forward from two aspects of blood support mode and transfusion strategies, aiming to provide a reference for transfusion resuscitation in the emergency treatment of severe trauma and further improve the success rate of treatment of patients with severe trauma.

Objective:To investigate the safety, feasibility and accuracy of esophageal sponge cytology in esophageal carcinoma screening in high-incidence districts.Methods:Opportunistic screening for esophageal carcinoma was conducted on individuals aged 40-75 years with high-risk factors for esophageal carcinoma and visited out-patient clinic in Lianshui People's Hospital from May 2021 to June 2022. A new esophageal cell collector independently developed in China was used for esophageal sponge cytology sampling followed by cytopathological analysis. Atypical squamous cells or more severe lesions were defined as positive esophageal sponge cytology. Then gastroscopy was performed, and all suspicious areas under the endoscopy were biopsied for histopathological examination. Gastroscopy, biopsy histopathology and esophageal sponge cytology were conducted blindly in pairs. Outcome measures included adverse reactions during sampling, subject tolerability (using a visual simulation score), sampling quality, and diagnostic efficacy of esophageal sponge cytology using gastroscopy plus biopsy histopathology as the gold standard.Results:A total of 1 590 patients completed the screening program. During esophageal sponge cytology sampling, no serious adverse events were observed, and the adverse reactions were mainly manifested as vomiting during sampling [0.31% (5/1 590)] and sore throat after sampling [2.45% (39/1 590)], all of which resolved spontaneously without further medical intervention. The majority of subjects [98.62% (1 568/1 590)] reported good tolerance during the procedure. After sampling, 1 526 (95.97%) subjects had completely expanded sponge material, meeting the standard of good sampling quality. The scanning analysis of the digital pathology system showed that the number of sampled cells in 1 590 subjects ranged (2.01-4.00)×10 6, with a median of 3.48×10 6 cells, which could meet the requirements for interpreting cytological results. Using the positive esophageal sponge cytology for the diagnosis of esophageal carcinoma including high-grade intraepithelial neoplasia, esophageal squamous cell carcinoma and adenocarcinoma of esophagogastric junction, the sensitivity, specificity, positive predictive value, negative predictive value, and accuracy were 98.57% (69/70), 91.51% (1 391/1 520), 34.85% (69/198), 99.93% (1 391/1 392), and 91.82% (1 460/1 590), respectively. Conclusion:Esophageal sponge cytology presents promising diagnostic efficacy for esophageal carcinoma screening, offering a simple, safe, convenient, and effective approach in high-incidence esophageal carcinoma regions.

Objective To investigate the value of depth of invasion (DOI) and pattern of invasion (POI) in predicting recurrence and evaluating prognosis of early oral squamous cell carcinoma (OSCC). Methods Data of 109 patients with primary OSCC were retrospectively collected. The chi-square test was used to compare categorical variables of clinical data. Univariate and multivariate Logistic regression analyses were used to analyze the correlation between clinical data and tumor recurrence. Cox regression analysis was used to analyze the correlation between clinical data and overall survival of patients. Results Univariate analysis showed that POI, DOI and pathological grade were correlated with recurrence and poor prognosis of early OSCC. Multivariate Logistic regression analysis indicated that DOI was an influencing factor for OSCC recurrence (OR=4.515, 95%CI, 1.283 to 15.894, P<0.05) and prognosis (HR=2.993, 95%CI, 1.225 to 7.317, P<0.05). There was a high correlation between POI and DOI of OSCC. Conclusion DOI ≥5 mm is considered as a relevant factor for recurrence and poor prognosis of early OSCC.

Objective:To explore the efficacy of balloon dilation percutaneous kyphoplasty (PKP) combined with prying reduction bone grafting in the treatment of thoracolumbar fractures, and its impact on patients′neurological function.Methods:The clinical data of 184 patients with thoracolumbar fractures from March 2017 to May 2020 in Cangzhou Integrated Traditional Chinese and Western Medicine Hospital of Hebei Province were retrospectively analyzed. Among them, 86 patients were treated with balloon dilation PKP combined with prying reduction and bone grafting (combined group), and 98 patients were treated with open reduction decompression and internal fixation (internal fixation group). The intraoperative bleeding volume, surgical time, intraoperative fluoroscopy frequency, efficacy and complications were recorded. One year after surgery, X-ray films were taken to measure the height of the anterior border and posterior border of vertebral body and the Cobb angle, and the pain visual analogue score (VAS), Barthel index and neurological Frankel grade were evaluated.Results:The intraoperative bleeding volume and surgical time in combined group were significantly higher than those in internal fixation group: (205.64 ± 45.63) ml vs. (180.37 ± 30.08) ml and (110.22 ± 28.91) min vs. (81.66 ± 20.77) min, and there were statistical differences ( P<0.01); there was no statistical difference in intraoperative fluoroscopy frequency between the two groups ( P>0.05). The total effective rate in combined group was significantly higher than that in internal fixation group: 91.86% (79/86) vs. 79.59% (78/98), the total incidence of complications was significantly lower than that in internal fixation group: 13.95% (12/86) vs. 38.78% (38/98), and there were statistical differences ( P<0.05 or <0.01). The height of the anterior border and posterior border of vertebral body 1 year after surgery in combined group were significantly higher than those in internal fixation group: (95.78 ± 1.59) mm vs. (62.74 ± 1.80) mm and (98.53 ± 3.80) mm vs. (95.06 ± 3.28) mm, the Cobb angle was significantly smaller than that in internal fixation group: (6.53 ± 2.80)° vs. (18.06 ± 2.68)°, and there were statistical differences ( P<0.01). The VAS and Barthel index 1 year after surgery in both groups improved significantly, and there were statistical differences ( P<0.01), but there were no statistical differences between the two groups ( P>0.05). The rate of neurological Frankel grade E 1 year after surgery in combined group was significantly higher than that in internal fixation group: 56.98% (49/86) vs. 23.47% (23/98), and there was statistical difference ( P<0.01). Conclusions:Compared with open reduction decompression and internal fixation, the balloon dilation PKP combined with prying reduction and bone grafting patients with thoracolumbar fractures is better for vertebral reduction and deformity repair, with higher safety, more significant pain relief, and better recovery of daily life and neurological function.

Background and Objectives: Osteoarthritis (OA) is a degenerative disease that leads to the progressive destruction ofarticular cartilage. Current clinical therapeutic strategies are moderately effective at relieving OA-associated pain but cannot induce chondrocyte differentiation or achieve cartilage regeneration. We investigated the ability of wedelolactone, a biologically active natural product that occurs in Eclipta alba (false daisy), to promote chondrogenic differentiation. Methods: and Results: Real-time reverse transcription–polymerase chain reaction, immunohistochemical staining, and immunofluorescence staining assays were used to evaluate the effects of wedelolactone on the chondrogenic differentiation of mesenchymal stem cells (MSCs). RNA sequencing, microRNA (miRNA) sequencing, and isobaric tags for relative and absolute quantitation analyses were performed to explore the mechanism by which wedelolactone promotes the chondrogenic differentiation of MSCs. We found that wedelolactone facilitates the chondrogenic differentiation of human induced pluripotent stem cell-derived MSCs and rat bone-marrow MSCs. Moreover, the forkhead box O (FOXO) signaling pathway was upregulated by wedelolactone during chondrogenic differentiation, and a FOXO1 inhibitor attenuated the effect of wedelolactone on chondrocyte differentiation. We determined that wedelolactone reduces enhancer of zeste homolog 2 (EZH2)-mediated histone H3 lysine 27 trimethylation of the promoter region of FOXO1 to upregulate its transcription. Additionally, we found that wedelolactone represses miR-1271-5p expression, and that miR-1271-5p post-transcriptionally suppresses the expression of FOXO1 that is dependent on the binding of miR-1271-5p to the FOXO1 3’-untranscribed region. Conclusions These results indicate that wedelolactone suppresses the activity of EZH2 to facilitate the chondrogenic differentiation of MSCs by activating the FOXO1 signaling pathway. Wedelolactone may therefore improve cartilage regeneration in diseases characterized by inflammatory tissue destruction, such as OA.

Objective:To isolate and culture WU polyomavirus (WUPyV), and to analyze the genome-wide evolutionary patterns, homology and population dynamics.Methods:Real-time quantitative PCR was used to detect the nasopharyngeal aspirate samples of hospitalized children with respiratory tract infection in Beijing Friendship Hospital during 2020 to 2022. Primary human airway epithelial cells cultured at the air-liquid interface were used to isolate and culture WUPyV. Whole genome sequence of the isolated strain was obtained by Sanger sequencing. For phylogenetic and evolutionary dynamics analysis, the whole genome was compared with the published whole genome sequences in GenBank database.Results:The detection rate of WUPyV was 4.7% (31/659) during 2020 to 2022, and a clinical strain BJ0593 of WUPyV type Ⅲc was successfully isolated. The homology of the whole genome and gene fragments of WUPyV was high. The average evolutionary rate of VP2 gene was about 1.256×10 -4 substitution/site every year, and the population dynamics of WUPyV tended to be flat in the last decade. Conclusions:This study successfully isolated a clinical WUPyV type Ⅲ strain for the first time, which provided the basis for further investigation on the molecular evolution and pathogenicity of WUPyV.

AIM:To investigate the impact of homocysteine(Hcy)on the inflammatory response mediated by BV2 mouse microglia,and to explore the mechanism of Ras-related protein 1a(Rap1a)in the Hcy-induced inflammatory response in BV2 cells.METHODS:Mouse microglial cell line BV2 was cultured in vitro,and Hcy intervention was used to establish a hyperhomocysteinemia cellular model.The cells were divided into 4 groups:blank control group,and 50 μmol/L,100 μmol/L and 150 μmol/L Hcy groups.The mRNA expression levels of M1 polarization markers,inflammatory factors IL-6,IL-1β and TNF-α,and Rap1a in BV2 cells were detected by RT-qPCR.Additionally,the levels of inflamma-tory factors IL-6,IL-1β and TNF-α in BV2 cells were measured using an ELISA kit.The protein expression level of Rap1a was detected by Western blot assay.To verify the function of Rap1a,viral transfection was employed for both over-expression and knockdown experiments.RESULTS:Under the intervention of Hcy concentration above 100 μmol/L,BV2 cells exhibited inflammatory polarization,as indicated by the increased mRNA expression of M1 polarization markers CD80 and CD86(P＜0.05).The mRNA and protein expression levels of inflammatory factors IL-6,IL-1β and TNF-α were significantly up-regulated(P＜0.05).Additionally,the mRNA and protein expression levels of Rap1a also showed a significant increase(P＜0.05).Moreover,Rap1a mRNA level was positively correlated with CD80 mRNA,IL-1β content and TNF-α content(P＜0.05).The multiplicities of infection of the viruses with Rap1a overexpression and Rap1a knock-down were both 80,and the effective transfection was observed through fluorescence microscopy.Overexpression of Rap1a exacerbated the inflammatory polarization of BV2 cells induced by Hcy(P＜0.05),while knockdown of Rap1a attenuated this polarization(P＜0.05).CONCLUSION:Hcy can promote M1 polarization of BV2 mouse microglia,leading to in-flammatory response,which indicates that Rap1a could potentially serves as a critical regulatory factor in the Hcy-induced inflammatory response of BV2 cells.

Tocilizumab has been reported to attenuate the "cytokine storm" in COVID-19 patients. We attempted to verify the effectiveness and safety of tocilizumab therapy in COVID-19 and identify patients most likely to benefit from this treatment. We conducted a randomized, controlled, open-label multicenter trial among COVID-19 patients. The patients were randomly assigned in a 1:1 ratio to receive either tocilizumab in addition to standard care or standard care alone. The cure rate, changes of oxygen saturation and interference, and inflammation biomarkers were observed. Thirty-three patients were randomized to the tocilizumab group, and 32 patients to the control group. The cure rate in the tocilizumab group was higher than that in the control group, but the difference was not statistically significant (94.12% vs. 87.10%, rate difference 95% CI-7.19%-21.23%, P = 0.4133). The improvement in hypoxia for the tocilizumab group was higher from day 4 onward and statistically significant from day 12 (P = 0.0359). In moderate disease patients with bilateral pulmonary lesions, the hypoxia ameliorated earlier after tocilizumab treatment, and less patients (1/12, 8.33%) needed an increase of inhaled oxygen concentration compared with the controls (4/6, 66.67%; rate difference 95% CI-99.17% to-17.50%, P = 0.0217). No severe adverse events occurred. More mild temporary adverse events were recorded in tocilizumab recipients (20/34, 58.82%) than the controls (4/31, 12.90%). Tocilizumab can improve hypoxia without unacceptable side effect profile and significant influences on the time virus load becomes negative. For patients with bilateral pulmonary lesions and elevated IL-6 levels, tocilizumab could be recommended to improve outcome.
Antibodies, Monoclonal, Humanized ; COVID-19/drug therapy* ; Humans ; SARS-CoV-2 ; Treatment Outcome