ObjectiveTo explore the possible mechanism of Qishao Tongbi Capsule （芪芍通痹胶囊， QTC） in the treatment of intervertebral disc degeneration （IDD）. MethodsSeventy-five rats were randomly divided into control group， model group， low-dose QTC group， high-dose QTC group， high-dose QTC +agonist group， with 15 rats in each group. Except for the control group， all other groups were subjected to a fibrous ring puncture to prepare an IDD model. After modeling， rats in low-dose QTC group and high-dose QTC group were given QTC at doses of 0.2 and 0.8 g/（kg·d） by gavage， respectively. Rats in high-dose QTC+ agonist group was given QTC at 0.8 g/（kg·d） and SKL2001 solution at 10 mg/（kg·d） by gavage. The control group and model group were given 10 ml/（kg·d） distilled water by gavage. All treatments were given once a day for 4 consecutive weeks. After treatment， X-ray and magnetic resonance imaging （MRI） were used to detect IDD degree. Hematoxylin-eosin （HE） staining and Safranin O-Fast Green staining were used to observe the morphological changes of the intervertebral disc tissue. Immunohistochemical staining was performed to examine the levels of proteoglycan， type Ⅱ collagen （COL Ⅱ）， and matrix metalloproteinase-3 （MMP-3） in the intervertebral disc tissue. Western blotting was used to detect the extracellular matrix （ECM）-related proteins （proteoglycan， COL Ⅱ， MMP-3， MMP-9， MMP-13）， aging-related proteins （P53， P21， P16）， apoptosis related proteins， including B-cell lymphoma/leukemia 2 （BCL-2）， BCL-2 related X protein （BAX）， Cleaved Caspase-3， and Wnt/β-catenin pathway related proteins such as Wnt3a， glycogen synthase kinase-3β （GSK-3β） and β-catenin in the intervertebral disc nucleus pulposus （NP） tissue. Reverse Transcription Quantitative Polymerase Chain Reaction （RT-qPCR） was used to assess the mRNA expression of Wnt3a， GSK-3β， and β-catenin in intervertebral disc tissue. ResultsCompared with the model group， rats in the low-dose QTC group and high-dose QTC group exhibited improved DHI， decreased Pfirmann grading， and alleviated IDD. The structural integrity of the NP and annulus fibrosus increased， and the number of the NP increased. The levels of proteoglycan， COL Ⅱ， BCL-2 and GSK-3β increased， while the levels of MMP-3， MMP-9， MMP-13， P53， P21， P16， BAX， Cleaved Caspase-3， Wnt3a and β-catenin protein decreased. The mRNA expression of Wnt3a and β-catenin mRNA decreased， while GSK-3β mRNA expression increased （P＜0.05）. Compared with the low-dose QTC group， the high-dose QTC group showed further improvements in DHI， decrease in Pfirrmann grading （P＜0.05）， and greater alleviation of IDD. The structural integrity of NP and annulus fibrosus was further enhanced， and the number of NP cells further increased. The levels of proteoglycan， COL Ⅱ， BCL-2 and GSK-3β were higher， while the levels of MMP-3， MMP-9， MMP-13， P53， P21， P16， BAX， Cleaved Caspase-3， Wnt3a and β-catenin were lower. The mRNA expression of Wnt3a and β-catenin decreased， while GSK-3β mRNA expression increased （P＜0.05）. Compared with the high-dose QTC group， the high-dose QTC +agonist group showed a decrease of DHI， an increase of Pfirmann grading （P＜0.05）， significant aggravation of IDD， reduction in structural integrity of the NP and annulus fibrosus， a decrease of NP cell count， lower levels of proteoglycan， COL Ⅱ， BCL-2 and GSK-3β， and higher levels of MMP-3， MMP-9， MMP-13， P53， P21， P16， BAX and Cleaved Caspase-3. Additionally， GSK-3β mRNA expression decreased （P＜0.05）. ConclusionQTC can inhibit NP cell aging， apoptosis， and ECM degradation in IDD rats， and its therapeutic effect may be mediated through the inhibition of the Wnt/β-catenin pathway.

Cardiac troponin I (cTnI), a widely used biomarker for assessing cardiovascular risk, can provide a window for the evaluation of drug-induced myocardial injury. Label-free biosensors are promising candidates for detecting cell secretomes, since they do not require labor-intensive processes. In this work, a label-free electrochemical aptasensor is developed for in situ monitoring of cardiac cell secretomes in cell culture media based on target-induced strand displacement. The aptasensing system contains an aptamer-functionalized signal nanoprobe facing trimetallic metal-organic framework nanosheets and a gold nanoparticle-based detection working electrode modified with DNA nanotetrahedron-based complementary DNA for indirect target detection. The signal nanoprobes (termed CAHA) consisted of copper-based metal-organic frameworks, AuPt nanoparticles, horseradish peroxidase, and an aptamer. When the aptasensor is exposed to cardiac cell secretomes, cTnI competitively binds to the aptamer, resulting in the release of signal nanoprobes from the biorecognition interface and electrochemical signal changes. The aptasensor exhibited rapid response times, a low detection limit of 0.31 pg/mL, and a wide linear range of 0.001-100 ng/mL. We successfully used this aptasensor to measure cTnI concentrations among secreted cardiac markers during antitumor drug treatment. In general, aptasensors can be used to monitor a variety of cardiac biomarkers in the evaluation of cardiotoxicity.

Objective To explore the effect of endoplasmic reticulum stress activating transcription factor 6(ATF6)on the expression of reproduction related gene heat shock protein family A member 1 like(HSPA1L)and preliminari-ly clarify its regulatory molecular mechanism.Methods The ATF6 over-expression plasmid was transfected into HEK-293T cells and the over-expression efficiency was verified by RT-qPCR and Western blot.The transcriptome sequen-cing information of testis tissue of male ATF6 knockout mice was used to screen five reproduction related genes down-stream of ATF6.The dual luciferase reporter assay selected the downstream genes with high promoter activity and de-tected the effect of over-expression of ATF6 on the promoter activity of downstream genes.The possible binding sites of ATF6 and downstream gene promoters were predicted by gene-regulation.RT-qPCR and Western blot were used to detect the effect of over-expression of ATF6 on downstream gene expression in HEK-293T cells.Whether ATF6 binds to downstream gene promoters was determined by electrophoretic mobility shift assay(EMSA).Results The expres-sion of ATF6 mRNA(P＜0.001)and protein(P＜0.001 and P＜0.05)in HEK-293T cells was significantly increased after transfection.HSPA1L(P＜0.001 and P＜0.05),a reproductive related gene downstream of ATF6 was screened by transcriptome sequencing and dual luciferase reporter assay.ATF6 promoted the truncated promoter activity of HSPA1L(P＜0.001).After over-expression of ATF6,the expression of HSPA1L was significantly increased(P＜0.001).The differences were statistically significant.ATF6 protein could bind to the aagtcgtcac DNA sequence of HSPA1L promoter.Conclusions ATF6,a key molecule of endoplasmic reticulum stress,regulates the expression of reproduction related gene HSPA1L by binding to the promoter of HSPA1L.This result will lay a foundation for further research on the prevention or treatment of endoplasmic reticulum stress(ERS)related male infertility.

BACKGROUND:Minimally invasive surgery is developing rapidly.Robot-assisted minimally invasive transforaminal lumbar interbody fusion and robot-assisted unilateral biportal endoscopic transforaminal/posterior lumbar interbody fusion are important posterior minimally invasive surgical approaches to treat lumbar degenerative diseases.However,it is worth discussing which operation method is more advantageous. OBJECTIVE:To compare the clinical efficacy and imaging examination between different operation groups,and discuss the clinical application value of robot-assisted minimally invasive lumbar posterior fusion technology to treat lumbar degenerative diseases. METHODS:Clinical data of 83 patients with lumbar degenerative diseases from January 2018 to June 2022 at the Department of Orthopedics,Sichuan Academy of Medical Sciences&Sichuan Provincial People's Hospital were retrospectively analyzed.Of them,27 patients received robot-assisted minimally invasive transforaminal lumbar interbody fusion treatment(group A);30 patients received robot-assisted unilateral biportal endoscopic transforaminal/posterior lumbar interbody fusion treatment(group B),and 26 traditional minimally invasive transforaminal lumbar interbody fusion patients were selected as the control group(group C).There were no significant differences in gender,age,body mass index,surgical segment,preoperative visual analog scale score and Oswestry Disability Index among the three groups(P＞0.05).The operation time,intraoperative blood loss,complications,fluoroscopic dose,fluoroscopic time,and fluoroscopic frequency were compared among the three groups.Gertzbein-Robbins'classification was used to evaluate the accuracy of percutaneous pedicle screw.Visual analog scale and Oswestry Disability Index scores were evaluated after surgery.The excellent and good rate of the three surgical options was evaluated using Macnab's criteria. RESULTS AND CONCLUSION:(1)The operation time of group A was significantly shorter than that of groups B and C(P＜0.05),but there was no significant difference between group B and group C(P＞0.05).The intraoperative blood loss in group B was significantly less than that in group A,and that in group A was significantly less than that in group C(P＜0.05).(2)The fluoroscopic dose,fluoroscopic time,and fluoroscopic frequency of group C were significantly higher than those of groups A and B(P＜0.05).(3)Visual analog scale score and Oswestry Disability Index in the three groups significantly improved after operation when compared with that before operation(P＜0.05),but there was no significant difference among the three groups 1 day and 6 months after surgery(P＞0.05).(4)Postoperative imaging showed that the accuracy of percutaneous pedicle screw placement in groups A and B was better than that in group C(P＜0.05).(5)There was no significant difference in the excellent and good rate of MacNab criteria among the three groups(P＞0.05).(6)There was no significant difference in complications among the three groups(P＞0.05).(7)The results indicated that robot-assisted minimally invasive transforaminal lumbar interbody fusion and robot-assisted unilateral biportal endoscopic transforaminal/posterior lumbar interbody fusion are effective surgery methods for lumbar degenerative diseases.Compared with traditional minimally invasive transforaminal lumbar interbody fusion,robot-assisted minimally invasive transforaminal lumbar interbody fusion surgery has higher efficiency,less intraoperative radiation and higher internal fixation accuracy,which has a good clinical application value.

[Objective] To investigate the correlation between human leukocyte antigen (HLA) gene polymorphism and hepatitis B virus (HBV) infection. [Methods] Venous blood samples were collected from 501 healthy individuals undergoing physical examinations at Yan’an Hospital in Kunming, Yunnan Province. Enzyme linked immunosorbent assay (ELISA) was used to detect HBV halves. Based on the results of HBV half detection, the patients were divided into three groups: HBV carrier group, previous infection group, and healthy control group. The HLA-A antigen genotype was detected using polymerase chain reaction with sequence specific primers (PCR-SSP) genotyping technology, and the distribution frequency of HLA-A gene polymorphism was compared between HBV carrier group and healthy control group, as well as between previous infection group and healthy control group. SPSS17.0 software was used for data statistical analysis. [Results] In the healthy control group, the HLA-A2 positivity rate was 47.49%, and the allele frequency was 31.29%.The overall frequency of gene distribution in the healthy control group was consistent with the HLA-A allele table commonly and confirmed in China published by the Chinese Bone Marrow Bank. The HLA-A2 positivity rate and allele frequency in the HBV carrier group were 63.04% and 42.23%, respectively; The difference in HLA-A2 positivity rate and allele frequency among carriers was statistically significant (P<0.05). the HLA-A2 positivity rate and allele frequency in the HBV previous infection group were 56.14% and 35.97%, respectively, which did not significantly differ from those in the healthy control group (P>0.05). [Conclusion] HLA-A2 gene may be a susceptibility gene for chronic hepatitis B HBV carriers.

Objective:To explore the effect of psychodrama therapy in adolescent inpatients with mental disor-ders.Methods:Totally 22 patients with mental disorders(aged 12-18 years)were recruited from the inpatient children's ward of a certain hospital.According to the interview outline,a semi-structured interview was conducted in adolescent inpatients with mental disorders who participated in 4 sessions of psychodrama therapy.These patients took psychotropic drugs regularly.The interview data were analyzed in depth by using the grounded theory meth-od.The self-compiled Psychodrama Therapy Participation Feeling and Evaluation Questionnaire was used to investi-gate the feelings,gains or changes of patients after psychodrama therapy.Results:The interview analysis found that the effects of psychodrama therapy included reducing symptoms,improving self-esteem,promoting functional recov-ery and improving social adaptability.The results of the self-compiled Psychodrama Therapy Participation Feeling and Evaluation Questionnaire showed that the top 4 aspects were promoting interpersonal relationship,being inter-esting,willing to continue to participate and improving self-confidence.Conclusion:Psychodrama therapy has an auxiliary therapeutic effect in adolescent inpatients with mental disorders,and can promote the improvement of self-esteem and social skills.

Objective To establish the UPLC fingerprint chromatogram combined with chemometric analysis for the quality evaluation of classical formula Linggui Zhugan Decoction.Methods SHIMADZU Shim-Pack GIST C18 column(100 mm×2.1 mm,2.0 μm)was used with acetonitrile-0.1%phosphoric acid aqueous solution as mobile phase,gradient elution;flow rate was 0.2 mL/min;the detection wavelength was 266 nm for the first 30 minutes and 235 nm for the last 36 minutes;the column temperature was 30℃.The UPLC fingerprint of Linggui Zhugan Decoction was established by Similarity Evaluation System for Chromatographic Fingerprint of TCM(2012.130723 version),and the common peak was determined and the similarity evaluation was carried out.Based on the peak area determination results of the common peak of the fingerprint,the quality of different batches of Linggui Zhugan Decoction was evaluated by chemometrics such as clustering analysis and principal component analysis.Results A total of 24 common peaks were confirmed and 14 components were identified by using reference substances.The similarity of 10 batches of Linggui Zhugan Decoction samples was greater than 0.950,which could be divided into two categories by chemometrics,and the principal component 1-4 were the main factors affecting its quality evaluation.OPLS-DA identified 6 differential markers.Conclusion The fingerprint research method established in the study is simple,reliable and reproducible.Through the method of fingerprint combined with chemometrics analysis,the differences between Linggui Zhugan Decoction from different origins of medicinal materials are identified,which provides a reference for the internal quality evaluation of Linggui Zhugan Decoction.

Objective To investigate the mechanism of Dayuan Decoction in the treatment of Helicobacter pylori(Hp)-positive perioral acne based on network pharmacology and animal experiments.Methods The effective active components and their corresponding targets of Dayuan Decoction were obtained from TCMSP.Targets related to Hp and perioral acne were retrieved from GeneCards,OMIM and TTD databases,and the intersection of drug targets and disease targets was considered as the target of Dayuan Decoction in the treatment of Hp-positive perioral acne.A protein-protein interaction network for the drug-disease intersection targets was constructed by using Cytoscape 3.7.2 and the STRING database,and the core targets were screened.GO and KEGG pathway enrichment analyses of the core target genes were performed using the DAVID database and the bioinformatics platform.By establishing an Hp-positive perioral acne rat model through gastric gavage with Hp bacterial liquid and intradermal injection of Propionibacterium acnes suspension,the intervention effect of Dayuan Decoction was observed and the core targets and related pathways were preliminary validated.Morphology in the perioral skin and gastric tissue were observed through HE staining.The expressions of TLR4,MyD88,NF-κB in perioral skin tissue,the contents of TNF-α and IL-6 in serum were measured using ELISA.Results A total of 145 active components were screened from Dayuan Decoction,corresponding to 253 targets,with 57 targets overlapping between drugs and diseases.The core targets for the treatment of Hp-positive perioral acne with Dayuan Decoction were identified as IL6,TNF,IL1B,AKT1,TP53,etc.KEGG pathway enrichment analysis revealed 153 signaling pathways,primarily involved in pathways in cancer,lipids and atherosclerosis,AGE-RAGE signaling pathway in diabetic complications,TLR signaling pathway and TNF signaling pathway.Dayuan Decoction attenuated the expression of TLR4,MyD88 and NF-κB in the perioral skin of the Hp-positive perioral acne rat model,as well as reduced the contents of TNF-α and IL-6 in serum(P<0.01).Conclusion Dayuan Decoction may exert therapeutic effects on Hp-positive perioral acne by modulating the TLR4/NF-κB signaling pathway,reducing the levels of inflammatory factors TNF-α and IL-6,and alleviating the inflammatory damage in the perioral skin.

Objective To explore the mechanism of Dayuan Decoction in treating Helicobacter pylori(Hp)positive oral ulcer based on network pharmacology and experimental validation.Methods Active components of Dayuan Decoction and its targets were collected using the TCMSP database;relevant targets for Hp positive oral ulcer were screened through the GeneCards database;the Venny 2.1.0 online platform was used to screen the intersection targets of drug active components and diseases;a protein-protein interaction network was built using STRING 11.0 database and core key targets were screened using Cytoscape 3.7.1 software;GO and KEGG enrichment analysis was conducted on intersection targets using the DAVID database.Oral administration of standard Hp bacterial solution and burning of oral mucosa with glacial acetic acid were used to construct an animal model of Hp positive oral ulcer,and the model rats were intervented with Dayuan Decoction by gavage.The morphology of oral and gastric mucosal tissues was observed,and the expression of core targets in oral and gastric mucosal tissues were detected.Results A total of 165 targets were obtained for the treatment of Hp positive oral ulcer with Dayuan Decoction.The main active components were quercetin,kaempferol,5,8,2'-trihydroxy-7-methoxyflavone,etc.,the core targets were AKT1,IL6,TP53,TNF,etc.GO and KEGG enrichment analysis suggested that Dayuan Decoction in the treatment of Hp positive oral ulcer was mainly related to the inflammatory pathway.Animal experiments showed that compared with the blank group,the structure of the oral and gastric tissues in the model group rats were significantly disrupted,with significant infiltration of inflammatory cells;the contents of IL-6 and TNF-α in the serum significantly increased(P<0.05),the contents of TLR4 and NLRP3 in oral and gastric mucosal tissues significantly increased(P<0.05),and the protein expression of TLR4 and NF-κB in oral mucosal tissues significantly increased(P<0.05).Compared with the model group,the histopathological manifestations of rats in all doseage of Dayuan Decoction groups significantly improved,the contents of IL-6 and TNF-α in the serum,and the contents of TLR4 and NLRP3 in the oral and gastric mucosal tissues significantly decreased,the protein expression of TLR4 and NF-κB in the oral mucosal tissue significantly decreased(P<0.05).Conclusion Dayuan Decoction is possible to regulate the inflammatory development of Hp positive oral ulcer and improve their symptoms by downregulating TLR4 and NF-κB levels.

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.