Based on copper death, this study investigates the effect and mechanism of Shenmai Injection on isoproterenol(ISO)-induced myocardial fibrosis(MF) in rats. SPF-grade male SD rats were randomly divided into a normal group, model group, captopril(5 mg·kg~(-1)) positive control group, and Shenmai Injection low(6 mL·kg~(-1)), medium(9 mL·kg~(-1)), and high(12 mL·kg~(-1)) dose groups. Except for the normal group, the rats in the other groups were subcutaneously injected with ISO(5 mg·kg~(-1)) once a day for 10 consecutive days to establish an MF model. Starting from the second day after successful modeling, intraperitoneal injections of the respective treatments were administered for 28 consecutive days. Hematoxylin-eosin(HE) and Masson staining were used to observe pathological changes and fibrosis levels in the myocardial tissue. Colorimetry was employed to detect serum Cu~(2+) concentration in rats. The levels of inflammatory cytokines interleukin-6(IL-6), interleukin-1β(IL-1β), interleukin-18(IL-18), tumor necrosis factor-α(TNF-α), as well as mitochondrial energy metabolites adenosine triphosphate(ATP), adenosine diphosphate(ADP), and adenosine monophosphate(AMP) in serum were measured using enzyme-linked immunosorbent assay(ELISA). Western blot was performed to detect the expression of collagen Ⅰ(Col-Ⅰ), collagen Ⅲ(Col-Ⅲ), and copper death-related proteins dihydrolipoamide acetyltransferase(DLAT), ferredoxin 1(FDX1), lipoic acid synthetase(LIAS), and heat shock protein 70(HSP70) in myocardial tissue. Immunofluorescence was used to detect the expression of DLAT, FDX1, and HSP70, while immunohistochemistry was conducted to examine the expressions of DLAT, FDX1, LIAS, and HSP70. The results showed that, compared to the model group, the myocardial structure disorder and collagen fiber deposition in the drug treatment groups were significantly improved, the cardiac index level was reduced, serum Cu~(2+), IL-6, IL-1β, IL-18, TNF-α, ADP, and AMP levels were significantly decreased, ATP levels were significantly increased, and the expressions of Col-Ⅰ, Col-Ⅲ, and HSP70 proteins in myocardial tissue were significantly reduced, while the expressions of DLAT, FDX1, and LIAS proteins were significantly elevated. In conclusion, Shenmai Injection effectively alleviates myocardial structure disorder and interstitial collagen fiber deposition in ISO-induced MF rats, promotes copper excretion, and reduces copper death in the ISO-induced rat MF model.
Animals ; Male ; Drugs, Chinese Herbal/administration & dosage* ; Rats, Sprague-Dawley ; Rats ; Myocardium/metabolism* ; Drug Combinations ; Fibrosis/metabolism* ; Copper/blood* ; Cardiomyopathies/genetics* ; Humans

OBJECTIVE: To study the efficacy of lumbar oblique manipulation in the treatment of lumbar disc herniation with different herniation locations based on MSU classification. METHODS: A total of 272 patients with lumbar disc herniation who were treated from June 2023 to December 2023 were divided into central type group, paracentral type group, and far lateral type group. Among them, there were 73 cases in the central type group, including 41 males and 32 females, with an age of (46.39±6.89) years;161 cases in the paracentral type group, including 88 males and 73 females, with an age of (37.14±5.89) years;and 38 cases in the far lateral type group, including 22 males and 16 females, with an age of (28.56±4.89) years. The visual analogue scale (VAS) and straight leg raising angle of the three groups of patients before treatment, after treatment, and at 1 and 3 months after treatment were recorded, and inter-group, intra-group, and correlation comparisons were made. RESULTS: A total of 272 patients were followed up, with a follow-up time of (3.0±0.2) months. The VAS score of central type patients after treatment was 2(2, 3) points, which was lower than 4(3, 5) points before treatment, and the difference was statistically significant (P<0.05). There was no statistically significant difference between 1 month and 3 months after treatment and before treatment (P>0.05). The VAS score of paracentral type patients after treatment 2(2, 3) points and 1 month after treatment 3(2, 4) points were lower than that before treatment 5(4, 6) points, and the differences were statistically significant (P<0.05). There was no statistically significant difference in VAS between 3 months after treatment and before treatment (P>0.05). There were no statistically significant differences in VAS scores of far lateral type patients before treatment, after treatment, and at 1 and 3 months after treatment (P>0.05). The straight leg raising angle of central type patients after treatment 64(58, 69) and 1 month after treatment 58(52, 65) were significantly different from that before treatment 44(40, 51) (P<0.05);there was no statistically significant difference between 3 months after treatment and before treatment (P>0.05). The straight leg raising angle of paracentral type patients after treatment 61(55, 67)°, 1 month after treatment 61(53, 66)°, and 3 months after treatment 47(41, 56)° were significantly different from that before treatment 44(36, 52)° (P<0.05). There were no statistically significant differences in the straight leg raising angle of far lateral type patients before treatment, after treatment, and at 1 and 3 months after treatment (P>0.05). There was a correlation between VAS and straight leg raising angle in the three groups of patients, but there was no linear relationship. CONCLUSION Lumbar oblique manipulation has a better effect in treating patients with central and paracentral lumbar disc herniation, but a poor effect in treating far lateral type;after treatment, the curative effect of paracentral type patients lasts longer than that of central type patients.
Humans ; Male ; Female ; Intervertebral Disc Displacement/physiopathology* ; Adult ; Lumbar Vertebrae/physiopathology* ; Middle Aged ; Manipulation, Spinal

OBJECTIVES: To investigate the role of long noncoding RNA (lncRNA) HClnc1 in regulating proliferation, invasion, and migration of hepatocellular carcinoma (HCC) cells and the regulatory mechanism. METHODS: HClnc1 expression levels in liver cancer tissues were analyzed using data from the TCGA database. BrdU incorporation, plate cloning, and transwell assays were employed to examine the effects of HClnc1 silencing/overexpression and/or ODC1 silencing on proliferation, invasion, and migration of liver cancer cells. The effects of HClnc1 silencing on ODC1 protein and mRNA expression in the liver cancer cells were analyzed using qRT-PCR and Western blotting. The activity of ODC1 promoter was analyzed using a dual luciferase reporter gene assay. Pull-down experiment, mass spectrometry analysis, and chromatin immunoprecipitation (ChIP) assay were used for identification of HClnc1-binding proteins and their interactions. Protein interactions with the ODC1 promoter region and their binding efficiencies were investigated using RNA interference and ChIP analysis. RESULTS: HClnc1 was significantly overexpressed in HCC tissues. In liver cancer cells, HClnc1 silencing significantly inhibited cell proliferation, invasion, and migration, while HClnc1 overexpression promoted these behaviors. ODC1 silencing also suppressed malignant behaviors of liver cancer cells, and counteracted the effects of HClnc1 overexpression. Interference of HClnc1 obviously inhibited ODC1 promoter activity. RBBP5 and KAT2B proteins were identified to bind simultaneously with HClnc1. HClnc1 overexpression upregulated ODC1 protein expression, while interference of RBBP5 or KAT2B downregulated ODC1 protein expression and blocked HClnc1-induced upregulation of ODC1 protein. Both RBBP5 and KAT2B could directly bind to ODC1 promoter region; knocking out KAT2B or RBBP5 reduced the binding efficiency, while knocking out HClnc1 reduced the binding of both RBBP5 and KAT2B to ODC1 promoter region. CONCLUSIONS By targeting the RBBP5/KAT2B epigenetic modification complex, HClnc1 increases ODC1 promoter activity to enhance ODC1 transcription and promote the proliferation and migration of liver cancer cells.
Humans ; Cell Proliferation ; RNA, Long Noncoding/genetics* ; Cell Movement ; Liver Neoplasms/metabolism* ; Cell Line, Tumor ; Carcinoma, Hepatocellular/genetics* ; Promoter Regions, Genetic ; Gene Expression Regulation, Neoplastic

Objective To explore the role of miR-429 in synovial mesenchymal stem cell-derived exosomes(SMSC-Exos)in repairing cartilage damage in temporomandibular joint osteoarthritis(TMJ OA)by extracting SMSC-Exos from human synovial tissue and screening differentially expressed microRNA(miRNA)through transcriptome sequencing.Methods Human synovial tissues were obtained from 6 patients who underwent surgery at the First Medical Center of the Chinese PLA General Hospital from June to December 2023,including 3 patients with osteoarthritis(OA group)and 3 control patients(control group),all of whom were male.SMSC-Exos were extracted from the synovial tissues for miRNA sequencing and differential expression analysis.Further,Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed on the target genes of differentially expressed miRNA to identify key functional miRNA and construct miRNA-target gene regulatory networks and protein-protein interaction(PPI)networks of target genes.An in vitro model of rabbit condylar cartilage cell inflammatory microenvironment induced by interleukin-1β(IL-1β)was established,with the control group cultured in DMEM/F12 basic medium and the inflammation-induced group cultured in DMEM/F12 basic medium containing 10 ng/ml IL-1β.RT-qPCR was used to detect the effects of overexpressed target miRNA on the mRNA expression levels of cartilage phenotype factors such as type Ⅱ collagen α1 chain(Col2a1),aggrecan(Acan),as well as inflammatory factors including a disintegrin and metalloproteinase with thrombospondin motifs 5(Adamts5)and cyclooxygenase-2(Cox-2).Results(1)SMSC-Exos were successfully isolated,cultured,and identified.(2)miRNA sequencing of SMSC-Exos from OA and control groups revealed 16 differentially expressed miRNAs(|log2FC|>2,P<0.05).Compared with control group,7 miRNAs were up-regulated and 9 were down-regulated in OA group.GO and KEGG analysis indicated that the target genes of miR-429 were mainly involved in development process,anatomical structure development,system development,cell development and differentiation,and were enriched in inflammation-related pathways such as mitogen-activated protein kinase(MAPK)and phosphatidylinositol 3-kinase-protein kinase B(PI3K-Akt).(3)Functional validation of miR-429 in the rabbit condylar cartilage cell inflammatory model showed that overexpression of miR-429 increased the mRNA expression levels of Col2a1 and Acan(P<0.05)and decreased the mRNA expression levels of Adamts5 and Cox-2(P<0.05)in the inflammation-induced group.Conclusions miRNA sequencing of SMSC-Exos isolated and identified from human synovial tissues reveals a specific miRNA expression profile in OA patients,with miR-429 significantly down-regulated.Functional validation demonstrates that overexpression of miR-429 has reparative and anti-inflammatory effects on condylar cartilage cells in an inflammatory microenvironment.

Objective To explore the molecular mechanism of miR-125b promoting invasion,metastasis and epithelial-mesenchymal transition(EMT)of gastric cancer cells.Methods The expression of miR-125b in gastric cancer and its adjacent tissues was studied by qRT-PCR.After upregulating or downregulating miR-125b in gastric cancer cells,the protein expressions of DKK3 and SERPINA4 were detected by Western blotting.Dual luciferase reporting assay was used to verify whether miR-125b can target DKK3 and SERPINA4.MKN45 cells were co-transfected with miR-125b inhibitor and target gene siRNA.Migration and invasion experiments were conducted to explore whether miR-125b can regulate the biological function of MKN45 cells through DKK3 and SERPINA4.Then,the regulatory mechanism of SRF on miR-125b was investigated.Finally,by in vivo experiments,the expression of SRF in gastric cancer cells was upregulated or downregulated by lentivirus transfection;the number of lung metastases in nude mice was detected to explore the effect of SRF on gastric cancer cell metastasis.Results In this study,the expression of miR-125b increased in gastric cancer tissues,which was correlated with clinical stage and lymph node metastasis.Dual luciferase reporting experiments showed that DKK3 and SERPINA4 were the direct targets of miR-125b in gastric cancer cells,and could activate the Wnt/β-catenin signaling pathway,thereby promoting the transcription process of EMT-related transcription factors Twist1 and Slug,inducing the occurrence of EMT,and promoting the metastasis of gastric cancer.In vitro and in vivo experiments confirmed that SRF promoted the invasion and metastasis of gastric cancer cells by positively regulating the expression of miR-125b.Conclusion Taken together,SRF/miR-125b axis promotes the EMT and metastasis of gastric cancer cells,and these regulators represent new potential therapeutic targets or biomarkers for gastric cancer.

ObjectiveTo assess the quality of randomized controlled trials （RCTs） of compound traditional Chinese medicine （TCM） prescriptions in the treatment of nonalcoholic steatohepatitis （NASH）， and to provide recommendations for standardizing the design and reporting of RCTs in this field. MethodsDatabases such as PubMed， Web of Science， Embase， the Cochrane Library， CNKI， VIP， and Wanfang Data were searched for RCTs of compound TCM prescriptions in the treatment of NASH published from January 1， 2018 to December 31， 2023， and the articles were screened and assessed based on the Cochrane risk-of-bias assessment tool （RoB 2）， the unified standard for clinical trial reporting （CONSORT 2010）， and CONSORT-CHM Formulas 2017 for compound TCM prescriptions. ResultsA total of 45 articles were finally included， and most of these studies were rated as high-risk bias by RoB 2.0. The analysis based on the CONSORT control checklist showed a relatively low reporting rate for most of the key items regarding the quality of RCT studies. ConclusionA relatively large risk of bias is observed in the clinical studies on compound TCM prescriptions in the treatment of NASH published in the past six years， which may lead to the poor quality of reporting and evidence. It is suggested that the top-level design of clinical studies should be taken seriously in addition to investigating the advantages of TCM， so as to improve the quality of clinical studies.

Objective To observe the safety and effectiveness of single dose intravenous infusion of tranexamic acid(TX-A)in dual level posterior lumbar interbody fusion(PLIF),and to explore the changes and trends in perioperative white blood cell(WBC),erythrocyte sedimentation rate(ESR),and C-reactive protein(CRP).Methods Between October 2020 and September 2022,46 patients with lumbar degenerative disease were treated with dual level PLIF,including 18 males and 28 females,with an average age of(60.24±10.68)years old,from 34 to 80 years old.They were divided into observation group and control group according to different treatment methods.There were 28 patients in the observation group,including 12 males and 16 females,with an average age of(61.04±9.03)years old.There were 3 cases with lumbar disc herniation(LDH),lumbar spinal stenosis(LSS)18 cases,lumbar spondylolisthesis(LS)7 cases.TXA(1 g/100 ml)was administered intravenously 15 min before skin incision after general anesthesia.The control group consisted of 18 patients,including 6 males and 12 females,with an average age of(59.00±13.04)years old.There were 5 cases with LDH,LSS 9 cases,LS 4 cases,and TXA was not used.The operation time,intraoperative bleeding volume,postoperative drainage volume,postoperative deep vein thrombosis(DVT),postoperative hospital stay,postoperative activated partial thromboplastin time(APTT),prothrombin time(PT),thrombin time(TT),fibrinogen(FIB),platelet(PLT),red blood cell(RBC),hemoglobin(HB),hematocrit(HCT),the first day,the fourth day,the seventh day and the last tested after operation WBC,ESR and CRP were recorded.Results The postop-erative wounds of the patients healed well and there was no DVT.46 patients were followed up from 3 to 6 months.The intraop-erative blood loss was 400.0(300.0,500.0)ml and the postoperative drainage was 260.0(220.0,450.0)ml in the observation group,which were lower than the control group[600.0(400.0,1000.0)ml,395.0(300.0,450.0)ml],P＜0.05.There was no significant difference between the two groups in operation time,postoperative hospital stay,postoperative APTT,PT,TT,FIB,PLT,RBC,HB,HCT,and postoperative WBC,ESR and CRP at different times(P＞0.05).Conclusion Single dose intravenous infusion of TXA can reduce the blood loss of bi-segmental PLIF,and has no significant effect on WBC,ESR and CRP after op-eration.

Objective To investigate the in vitro anti-inflammatory effects of Sophora davidii Hance leaves total alkaloids(SDLTAs)and possible molecular mechanisms.Methods The lipopolysaccharide(LPS)-induced inflammation model of RAW264.7 cells was used,and different concentrations of SDLTAs(50,100 and 200 μg/mL)were administered,and the effect of SDLTAs on cellular NO expression was detected by the Griess method;ELISA method was used to detect the effect of SDLTAs on the expression of IL-6,TNF-α and IL-1β;The expression of iNOS,NF-κB p65 and IκBα mRNA was detected by RT-qPCR;Western blotting was used to detecte the expres-sion of p-p38,p-p65 and p-JNK in the cells and NF-κB p65 in the nucleus.Results SDLTAs could significantly inhibit the LPS-induced inflammatory response in RAW264.7 cells.SDLTAs significantly decreased the secretion of NO,IL-6,TNF-α and IL-1β in cells(P<0.01),and significantly decreased the mRNA expressions of iNOS,NF-κB p65 and IκBα in cells(P<0.01).Significantly decreased the protein expression of p-p38,p-p65 and p-JNK in cells and NF-κB p65 in nucleus(P<0.01).Conclusion SDLTAs can exert anti-inflammatory effects by regulating the MAPK/NF-κB signalling pathway.

Background and Aims:Accurate early pathogen diagnosis is a breakthrough for improving the prognosis of infectious pancreatic necrosis(IPN)patients.However,there is currently a lack of efficient methods for early identification of IPN in clinical settings.This study was performed to assess the application value of next-generation sequencing technology based on metagenomic capture(MetaCAP)in the pathogen diagnosis of IPN. Methods:A prospective study was conducted on 29 patients suspected of having acute necrotizing pancreatitis at Xiangya Hospital of Central South University between January and July 2024.Blood samples were tested using MetaCAP and conventional pathogen culture.The results of peritoneal fluid pathogen culture were used as the gold standard to compare the diagnostic efficacy of the two methods. Results:Due to three cases lacking peritoneal fluid culture results,a total of 26 cases were included in the final analysis.The overall mortality rate was 23.1％(6/26).During hospitalization,9 cases(34.6％)were diagnosed with IPN.The sensitivity and negative predictive value of MetaCAP for diagnosing IPN were significantly higher than those of conventional pathogen culture(77.8％vs.11.1％,P=0.031;86.7％vs.65.2％,P=0.032),while the differences in specificity(76.5％vs.88.2％,P=0.689)and positive predictive value(63.6％vs.33.3％,P=0.347)between the two methods were not statistically significant.The average detection time for MetaCAP was 33(20-49)h,while microbial culture took 125(45-142)h,with a significant difference(P＜0.001).The average cost for blood MetaCAP testing was 2 500 yuan per case,but it accounted for only 1.19％of the average hospitalization cost. Conclusion:MetaCAP has significant value in the early pathogen diagnosis of IPN,with a shorter detection time,good testing efficacy,and health-economic value,demonstrating a promising clinical application prospect.

Objective:To investigate the mechanism of DNA damage and repair in MLL-rearranged acute myeloid leukemia(MLL-r AML)cells by the combination of Chidamide and the BRD4 inhibitor(+)-JQ-1.Methods:MLL-r AML cell lines Molm-13,MV4-11 and non-MLL-r AML cell line Kasumi were divided into control group(contr),Chidamide group(chida),(+)-JQ-1 group and Combination group(combi),respectively.Cell viability of Molm-13 was measured by CCK-8 to determine optimal the concentrations of Chidamide and(+)-JQ-1.The cell cycle was detected by flow cytometry,and apoptosis-related factors Bcl-2,Bax and caspase-3 were detected by Western blot.DNA damage marker γH2AX was detected by immunofluorescence.The protein expressions of DNA damage factor γH2AX,DNA damage checkpoint kinases p-ATR,p-CHK1,p-ATM,p-CHK2 and DNA damage repair factors Rad51 and 53BP1 were detected by Western blot.The expression of DNA damage repair factors Rad51 and 53BP1 mRNA was detected by qRT-PCR.Results:Under the treatment of Chidamide(300 nmol/L)and(+)-JQ-1(400 nmol/L),the proportion of G1 phase cells in MLL-r AML cell lines Molm-13 and MV4-11 was increased in combination group compared with control group.In non-MLL-r AML cell line Kasumi,compared with control group,the proportion of G1 phase cells in combination group was increased(P＜0.05).In Molm-13 and MV4-11 cell lines,compared with control group,the expression level of DNA damage marker γH2AX in combination group was increased(P＜0.05).The expression levels of DNA damage checkpoint and damage repair factors p-ATR,p-CHK1,p-ATM,p-CHK2,Rad51,53BP1 were decreased(P＜0.05).In Kasumi cell line,compared with control group,there was no significant change in the expression of some of the above factors in combination group(P＞0.05),but the expression trend of some factors was opposite.In MLL-r AML cell lines Molm-13 and MV4-11,compared with control group,the expression levels of Bax and caspase-3 protein were increased in combination group,while the expression levels of Bcl-2 protein were decreased(P＜0.05).In non-MLL-r AML cell line Kasumi,there was no significant change in apoptotic factor protein expression in combination group compared with control group(P＞0.05).Conclusion:Chidamide combined with(+)-JQ-1 can inhibit the proliferation of MLL-r AML cells,inhibit the initiation of protective self-repair of these leukemia cells by inhibiting the DNA damage response pathway,and ultimately increase the apoptosis of these cells,but non-MLL-r AML cells have no similar results.