Aim To screen and study the effects of Tao Hong Si Wu decoction(THSWD)-mediated treat-ment on circular RNA(circRNA)expression profiles in rats with middle cerebral artery occlusion(MCAO),and investigate the possible roles and molecular mecha-nisms of THSWD.Methods Next-generation RNA sequencing was conducted to identify circRNA expres-sion profiles in MCAO rats after treatment with THSWD and compared with the MCAO model group and control group.Bioinformatics analysis was performed to predict the potential target microRNAs and mRNAs.Gene On-tology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway analyses for the potential target mRNAs were applied to explore the potential roles of differentially expressed circRNAs.RT-qPCR was performed to verify circRNAs with significant differences in expression.Results We identified 87 significantly differentially expressed circRNAs between the MCAO group versus the control group,and 86 sig-nificantly differentially expressed circRNAs between the MCAO group versus the THSWD group.respective-ly.Among them,17 circRNAs induced by the MCAO model were reversed via treatment with THSWD.To demonstrate the roles of mRNAs targeted by DECs,the GO and KEGG databases were used.Further analysis revealed that five circRNAs may play important roles in the development of MCAO.Conclusions The com-prehensive expression profile of circRNAs in rats with middle cerebral artery occlusion after THSWD treat-ment is determined for the first time,suggesting that the therapeutic effect of THSWD on MCAO may be a-chieved by regulating the expression of circRNAs.

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.

Aim To screen and study the expression of long non-coding RNA (IncRNA) in rats with middle cerebral artery occlusion (MCAO) with MCAO treated with Tao Hong Si Wu decoction (THSWD) and determine the possible molecular mechanism of THSWD in treating MCAO rats. Methods Three cerebral hemisphere tissue were obtained from the control group, MCAO group and MCAO + THSWD group. RNA sequencing technology was used to identify IncRNA gene expression in the three groups. THSWD-regulated IncRNA genes were identified, and then a THSWD-regu-lated IncRNA-mRNA network was constructed. MCODE plug-in units were used to identify the modules of IncRNA-mRNA networks. Gene ontology (GO) and kyoto encyclopedia of genes and genomes (KEGG) were used to analyze the enriched biological functions and signaling pathways. Cis- and trans-regulatory genes for THSWD-regulated IncRNAs were identified. Reverse transcription real-time quantitative pol-ymerase chain reaction (RT-qPCR) was used to verify IncRNAs. Molecular docking was used to identify IncRNA-mRNA network targets and pathway-associated proteins. Results In MCAO rats, THSWD regulated a total of 302 IncRNAs. Bioinformatics analysis suggested that some core IncRNAs might play an important role in the treatment of MCAO rats with THSWD, and we further found that THSWD might also treat MCAO rats through multiple pathways such as IncRNA-mRNA network and network-enriched complement and coagulation cascades. The results of molecular docking showed that the active compounds gallic acid and a-mygdalin of THSWD had a certain binding ability to protein targets. Conclusions THSWD can protect the brain injury of MCAO rats through IncRNA, which may provide new insights for the treatment of ischemic stroke with THSWD.

Objective To investigate the protective effect of dexmedetomidine(Dex)pretreatment on myocardial ischemia-reperfusion mice and the effect of Nod-like receptor protein 3(NLRP3)inflammatory signaling pathway.Methods C57BL/6J mice were randomly divided into sham group(only threading without ligation),model group(recovery after ligation of left anterior descending coronary artery),positive group(modeling after intraperitoneal injection of 1 mg·kg-1 trimetazidine),Dex group(modeling after intraperitoneal injection of 20 μg·kg-1 dexmedetomidine),MCC950 group(modeling after intraperitoneal injection of 10 mg·kg-1 NLRP3 inhibitor MCC950),12 mice in each group.Cardiac function indexes were detected at 24 h after reperfusion,the expression level of related proteins in myocardial tissue was detected by Western blot,enzyme-linked immunosorbent assay(ELISA)was used to detect the expression level of serum factor,myocardial antioxidant index was detected by kit method,and apoptosis was detected by Tunel method.Results The NLRP3 protein expression levels of sham group,model group,positive group,Dex group and MCC950 group were 0.31±0.05,1.06±0.07,0.52±0.05,0.65±0.07 and 0.39±0.04,respectively;the expression levels of apoptosis-associated granuloid protein(ASC)were 0.27±0.08,0.88±0.09,0.46±0.05,0.59±0.07 and 0.34±0.04,respectively;CK-MB levels were(25.64±2.94),(102.08±7.04),(49.61±7.70),(60.86±5.24)and(63.24±5.38)U·L-1,respectively;IL-6 levels were(104.78±10.73),(231.54±15.56),(158.20±16.54),(165.10±14.77)and(141.17±14.08)pg·mL-1,respectively;Tunel positive cell rates were(4.34±0.16)％,(25.98±1.58)％,(8.74±0.93)％,(11.06±1.07)％and(9.19±0.88)％,respectively.Sham group were compared with model group;Dex group and MCC950 group were compared with model group,the above indexes were statistically significant(all P＜0.05).Conclusion Dexmedetomidine preconditioning may prevent ischemia-reperfusion myocardial injury by inhibiting NLRP3/ASC/caspase-1 inflammatory pathway,inflammatory response and myocardial cell apoptosis.

COVID-19 has been prevalent for three years. The virulence of SARS-CoV-2 is weaken as it mutates continuously. However, elderly patients, especially those with underlying diseases, are still at high risk of developing severe infections. With the continuous study of the molecular structure and pathogenic mechanism of SARS-CoV-2, antiviral drugs for COVID-19 have been successively marketed, and these anti-SARS-CoV-2 drugs can effectively reduce the severe rate and mortality of elderly patients. This article reviews the mechanism, clinical medication regimens, drug interactions and adverse reactions of five small molecule antiviral drugs currently approved for marketing in China, so as to provide advice for the clinical rational use of anti-SARS-CoV-2 in the elderly.

Objective:To establish an isotope dilution liquid chromatography-tandem mass spectrometry (ID-LC-MS/MS) method for the determination of L-tryptophan and its metabolites in serum.Methods:The methodology was established and evaluated using serum samples collected from 166 healthy subjects undergoing physical examinations at West China Hospital from November 2022 to January 2023 were collected. Isotope-labeled markers of L-tryptophan (Trp), L-kynurenine (Kyn), and kynurenic acid (KA) were used as internal standards. After protein precipitation treatment of serum samples, LC-MS/MS was used to determine Trp, Kyn, and KA simultaneously. The selectivity, specificity, linearity, detection limit (LOD), quantification limit (LOQ), carry-over, precision, recovery rate, matrix effect, and dilution integrity of the method were evaluated.Results:The linearity of Trp, Kyn, and KA was demonstrated to be 0.999. The LODs were 0.10 μmol/L, 0.01 μmol/L and 1.00 nmol/L, respectively. The LOQs were 0.20 μmol/L, 0.04 μmol/L and 2.00 nmol/L, respectively. The intra-batch precision and inter-batch precision were below<10%. The average recovery rate and the relative matrix effect were all about 100%. The samples over the upper limit of quantitation can be diluted up to 16 times. The Trp concentration, Kyn concentration, KA concentration, Kyn/Trp ratio, and KA/Kyn ratio in serum of healthy subjects were 59.55±10.92 μmol/L, 1.85±0.43 μmol/L, 39.89±17.93 nmol/L, (31.64±8.19)×10 -3 and 21.51±6.72, respectively. Conclusion:An ID-LC-MS/MS method was successfully established for the quantitative determination of Trp, Kyn, and KA in serum. The method proved to be simple, rapid, sensitive, accurate, and reliable, providing robust support for clinical research related to these analytes.

Integrating multisensory inputs to generate accurate perception and guide behavior is among the most critical functions of the brain. Subcortical regions such as the amygdala are involved in sensory processing including vision and audition, yet their roles in multisensory integration remain unclear. In this study, we systematically investigated the function of neurons in the amygdala and adjacent regions in integrating audiovisual sensory inputs using a semi-chronic multi-electrode array and multiple combinations of audiovisual stimuli. From a sample of 332 neurons, we showed the diverse response patterns to audiovisual stimuli and the neural characteristics of bimodal over unimodal modulation, which could be classified into four types with differentiated regional origins. Using the hierarchical clustering method, neurons were further clustered into five groups and associated with different integrating functions and sub-regions. Finally, regions distinguishing congruent and incongruent bimodal sensory inputs were identified. Overall, visual processing dominates audiovisual integration in the amygdala and adjacent regions. Our findings shed new light on the neural mechanisms of multisensory integration in the primate brain.
Animals ; Macaca ; Acoustic Stimulation ; Auditory Perception/physiology* ; Visual Perception/physiology* ; Amygdala/physiology* ; Photic Stimulation

In this study, the transmittance of tanshinone Ⅱ_A(Tan Ⅱ_A) and cryptotanshinone(CTS) through the blood-prostate barrier and their distributions in the prostate tissue were compared between tanshinone extract(Tan E) treatment group and the corresponding monomer composition group under the equivalent dose conversion in vitro and in vivo. First, the human prostate epithelial cell line RWPE-1 was cultured in vitro for 21 days for the establishment of a blood-prostate barrier model, and the transmission of Tan Ⅱ_A and CTS through the barrier model was investigated after administration of Tan E and corresponding single active components. Second, SD rats were administrated with 700 mg·kg~(-1) Tan E, 29 mg·kg~(-1) CTS, and 50 mg·kg~(-1) Tan Ⅱ_A by gavage, and plasma and prostate tissue samples were collected at the time points of 2, 4, 8, 12, and 24 h. The Tan Ⅱ_A and CTS concentrations in the samples were determined. The results showed that in the cell model, the cumulative transmission amounts of CTS and Tan Ⅱ_A in the extract at each time point were higher than those of the corresponding single active components(P<0.01). In rats, after the administration of Tan E, the concentrations of Tan Ⅱ_A and CTS in rat plasma and prostate were higher than those of the corresponding single active components. This study demonstrated that the coexisting components in Tan E promoted the penetration of its main pharmacological components Tan Ⅱ_A and CTS through the blood-prostate barrier. The findings provide a theoretical and experimental basis for the application of Tan E in the clinical treatment of prostate-related diseases.
Male ; Rats ; Humans ; Animals ; Prostate ; Rats, Sprague-Dawley ; Abietanes/pharmacology* ; Permeability

Objective:To detect the expression of interleukin 2 (IL-2)/Janus kinase 3/signal transduction and transcriptional activator 5 (JAK3/STAT5) signaling pathway in peripheral blood of patients with ankylosing spondylitis (AS) and explore its mechanism in the development and progression of AS.Methods:Clinical data, peripheral blood and laboratory tests of 30 patients with active AS (ASA), 30 patients with stable AS (ASS) and 50 healthy subjects (HC) were collected. The mRNA expression levels of JAK3, signal transduction and transcription activator 5a (STAT5a) and signal transduction and transcription activator 5b (STAT5b) were detected by quantitative real-time-polymerase chain reaction (RT-qPCR). The expression levels of JAK3, STAT5a and STAT5b proteins and phosphorylated proteins were detected by Western-blot. Plasma IL-2 concentration was determined by enzyme-linked immunosorbent assay (ELISA). Two independent samples t-test or one-way analysis of variance were used for measurement data consistent with normal distribution, LSD- t test was used for pairwise comparison between the three groups, Mann-Whitney U test or Kruskal-Wallis H test was used for non-normal distribution, χ2 test was used for correlation analysis of categorical variables. Spearman correlation analysis was used for correlation analysis between variables, and receiver operating characteristic (ROC) curve was used to evaluate the value of JAK3, STAT5a and STAT5b mRNA expression levels in monitoring AS activity. Results:① The mRNA expression levels of JAK3, STAT5a and STAT5b were significantly different among the three groups ( F=65.98, P<0.001; F=21.15, P<0.001; F=13.67, P<0.001). JAK3 mRNA expression in ASA group (2.5±0.9) was significantly higher than that in ASS group (1.1±0.4) and healthy subjects (1.0±0.5), the difference was statistically significant (both P<0.001). The mRNA expression level of STAT5a in ASA group (1.4±0.3) was significantly higher than that in ASS group (0.9±0.3) and healthy subjects group (1.0±0.3), the difference was statistically significant (both P<0.001). STAT5b mRNA expression level in ASA group (1.5±0.6) was significantly higher than that in ASS group (1.0±0.4) and healthy subjects (1.0±0.4), the difference was statistically significant (both P<0.001). The expression level of JAK3 mRNA in HLA-B27 positive group (1.9±1.0) was higher than that in HLA-B27 negative group (1.4±0.6), and the difference was statistically significant ( t=-2.22, P=0.032). The phosphorylation levels of JAK3, STAT5a and STAT5b showed statistically significant differences among the three groups ( F=91.56, P<0.001; F=25.15, P< 0.001; F=178.59, P<0.001). The phosphorylation level of JAK3 protein in ASA group (1.04±0.08) was significantly higher than that in ASS group (0.568±0.019) and healthy subjects (0.536±0.064), the difference was statistically significant (both P<0.001). The phosphorylation level of STAT5a protein in ASA group (1.166±0.096) was significantly higher than that in ASS group (0.923±0.018) and healthy subjects (0.911±0.017), the difference was statistically significant (both P<0.001). The phosphorylation level of STAT5b protein in ASA group (0.81±0.05) was significantly higher than that in ASS group (0.21±0.03) and healthy subjects (0.24± 0.07), the difference was statistically significant (both P<0.001). The difference of plasma IL-2 concentration among the three groups was statistically significant ( F=3.32, P=0.040). The IL-2 concentration in the ASA group [(110±40) pg/ml] was significantly higher than that in the ASS group [(89±40) pg/ml] and the healthy group [(88±39) pg/ml], the difference was statistically significant ( P=0.044, P=0.016). ② Spearman correlation analysis showed that STAT5a mRNA expression level was positively correlated with platelets in AS patients ( r=0.353, P=0.006). JAK3 mRNA expression level in ASA group was positively correlated with IL-2 concentration ( r=0.766, P<0.001), and negatively correlated with estimated glomerular filtration rate ( r=-0.485, P=0.007). STAT5a mRNA expression level was positively correlated with erythrocyte sedimentation rate ( r= 0.680, P<0.001), and STAT5b mRNA expression level was positively correlated with hypersensitive C-reactive protein (CRP) ( r=0.823, P<0.001). ③ The ROC curve showed that JAK3 mRNA expression level predicted the area under ROC curve (AUC) of ASA with a 95% CI of 0.920 (0.853, 0.987), sensitivity and specificity of 86.7% and 90.0%, respectively. STAT5a mRNA expression level predicted the AUC 95% CI of ASA was 0.874 (0.787, 0.961), and the sensitivity and specificity were 96.7% and 66.7%, respectively. STAT5b mRNA expression level predicted the AUC 95% CI of ASA was 0.749 (0.617, 0.881), and the sensitivity and specificity were 73.3% and 80.0%, respectively. Conclusion:This study suggests that IL-2/JAK3/STAT5 may be involved in the pathogenesis of AS, and JAK3 mRNA can be used as a biological indicator to monitor the activity of AS disease.

Unlike healthy, non-transformed cells, the proteostasis network of cancer cells is taxed to produce proteins involved in tumor development. Cancer cells have a higher dependency on molecular chaperones to maintain proteostasis. The chaperonin T-complex protein ring complex (TRiC) contains eight paralogous subunits (CCT1-8), and assists the folding of as many as 10% of cytosolic proteome. TRiC is essential for the progression of some cancers, but the roles of TRiC subunits in osteosarcoma remain to be explored. Here, we show that CCT4/TRiC is significantly correlated in human osteosarcoma, and plays a critical role in osteosarcoma cell survival. We identify a compound anticarin-β that can specifically bind to and inhibit CCT4. Anticarin-β shows higher selectivity in cancer cells than in normal cells. Mechanistically, anticarin-β potently impedes CCT4-mediated STAT3 maturation. Anticarin-β displays remarkable antitumor efficacy in orthotopic and patient-derived xenograft models of osteosarcoma. Collectively, our data uncover a key role of CCT4 in osteosarcoma, and propose a promising treatment strategy for osteosarcoma by disrupting CCT4 and proteostasis.