BACKGROUND: Oxidative stress is a crucial factor contributing to the occurrence and development of secondary damage in spinal cord injuries (SCI), ultimately impacting the recovery process. α-lipoic acid (ALA) exhibits potent antioxidant properties, effectively reducing secondary damage and providing neuroprotective benefits. However, the precise mechanism by which ALA plays its antioxidant role remains unknown. METHODS: We established a model of moderate spinal cord contusion in rats. Experimental rats were randomly divided into 3 distinct groups: the sham group, the model control group (SCI_Veh), and the ALA treatment group (SCI_ALA). The sham group rats were exposed only to the SC without contusion injury. Rats belonging to SCI_Veh group were not administered any treatment after SCI. Rats of SCI_ALA group were intraperitoneally injected with the corresponding volume of ALA according to body weight for three consecutive days after the surgery. Subsequently, three days after SCI, spinal cord samples were obtained from three groups of rats: the sham group, model control group, and administration group. Thereafter, total RNA was extracted from the samples and the expression of three sets of differential genes was analyzed by transcriptome sequencing technology. Real-time PCR was used to verify the sequencing results. The impact of ALA on oxidative stress in rats following SCI was assessed by measuring their total antioxidant capacity and hydrogen peroxide (H2O2) content. The effects of ALA on rat recovery following SCI was investigated through Beattie and Bresnahan (BBB) score and footprint analysis. RESULTS: The findings from the transcriptome sequencing analysis revealed that the model control group had 2975 genes with altered expression levels when compared to the ALA treatment group. Among these genes, 1583 were found to be upregulated while 1392 were down-regulated. Gene ontology (GO) displayed significant enrichment in terms of functionality, specifically in oxidative phosphorylation, oxidoreductase activity, and signaling receptor activity. The Kyoto encyclopedia of genes and genomes (KEGG) pathway was enriched in oxidative phosphorylation, glutathione metabolism and cell cycle. ALA was found to have multiple benefits for rats after SCI, including increasing their antioxidant capacity and reducing H2O2 levels. Additionally, it was effective in improving motor function (such as 7 days after SCI, the BBB score for SCI_ALA was 8.400 ± 0.937 compared to 7.050 ± 1.141 for SCI_Veh) and promoting histological recovery after SCI (The results of HE demonstrated that the percentage of damage area in was 44.002 ± 6.680 in the SCI_ALA and 57.215 ± 3.964 in the SCI_Veh at the center of injury.). The sequence data from this study has been deposited into Sequence Read Archive (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE242507). CONCLUSION: Overall, the findings of this study confirmed the beneficial effects of ALA on recovery in SCI rats through transcriptome sequencing, behavioral, as well histology analyses.
Inflammation is one of the key injury factors for spinal cord injury (SCI). Exosomes (Exos) derived from M2 macrophages have been shown to inhibit inflammation and be beneficial in SCI animal models. However, lacking targetability restricts their application prospects. Considering that chemokine receptors increase dramatically after SCI, viral macrophage inflammatory protein II (vMIP-II) is a broad-spectrum chemokine receptor binding peptide, and lysosomal associated membrane protein 2b (Lamp2b) is the key membrane component of Exos, we speculated that vMIP-II-Lamp2b gene-modified M2 macrophage-derived Exos (vMIP-II-Lamp2b-M2-Exo) not only have anti-inflammatory properties, but also can target the injured area by vMIP-II. In this study, using a murine contusive SCI model, we revealed that vMIP-II-Lamp2b-M2-Exo could target the chemokine receptors which highly expressed in the injured spinal cords, inhibit some key chemokine receptor signaling pathways (such as MAPK and Akt), further inhibit proinflammatory factors (such as IL-1ß, IL-6, IL-17, IL-18, TNF-α, and iNOS), and promote anti-inflammatory factors (such as IL-4 and Arg1) productions, and the transformation of microglia/macrophages from M1 into M2. Moreover, the improved histological and functional recoveries were also found. Collectively, our results suggest that vMIP-II-Lamp2b-M2-Exo may provide neuroprotection by targeting the injured spinal cord, inhibiting some chemokine signals, reducing proinflammatory factor production and modulating microglia/macrophage polarization.
Exosomes , Macrophages , Mice, Inbred C57BL , Microglia , Spinal Cord Injuries , Animals , Spinal Cord Injuries/pathology , Spinal Cord Injuries/metabolism , Spinal Cord Injuries/genetics , Exosomes/metabolism , Exosomes/transplantation , Mice , Macrophages/metabolism , Microglia/metabolism , Microglia/drug effects , Microglia/pathology , Lysosomal-Associated Membrane Protein 2/metabolism , Lysosomal-Associated Membrane Protein 2/genetics , Cell Polarity/drug effects , Cell Polarity/physiology , Female , Neuroprotection/physiology , Signal Transduction/drug effects , Chemokines/metabolism
<p><b>OBJECTIVE</b>To study the molecular mechanism of Yangjing Zhongyu Decoction (YZD) n-butanol extracts (ZDC) and ethyl acetate extracts (YSYZ) in reducing androgen in porcine granulose cells by mitogen-activated protein kinase (MAPK) pathway.</p><p><b>METHODS</b>Porcine granulose cells were isolated and cultured. They were inoculated by MAPK inhibitor PD98059 at different concentrations, and then they were divided into the blank control group (0), 1, 3, 10, and 25 micromol/L groups. After 24-h culture the cytochrome P450c17a (CYP17) mRNA expression level was detected using Real-time fluorescent quantitative PCR. Contents of androgen (testosterone) in the supernate were detected using RIA and optimal PD98059 concentration screened. After intervened by 10 micromol/L PD98059 for 24 h, the culture solution was intervened by effective ingredients of with or without YZD or YSYZ at various concentrations (0, 1 , 5, 25, 50 mg/mL) at various time points (3, 6, 18, 24 h). Expression levels of p-ERK1/2, c-Fos and CYP17 were detected by Western blot. Testosterone content in the supernate was determined by radioimmunoassay (RIA).</p><p><b>RESULTS</b>Ten pLmol/L PD98059 could obviously decrease p-ERK1/2 protein expression and increase CYP17 mRMA expression, and elevate testosterone content in the supernate (P < 0.05). ZDC and YSYZ at 25 ng/mL could increase p-ERK1/2 protein expression and c-Fos levels, and reduce CYP17 protein expression, and lower testosterone content in the supernate after 6-h intervention (P < 0.01).</p><p><b>CONCLUSION</b>Effective ingredients of YZD could reduce androgen production in porcine granulose cells through increasing activities of MAPK.</p>
Animals , Female , Androgens , Drugs, Chinese Herbal , Pharmacology , Flavonoids , Granulosa Cells , Metabolism , Mitogen-Activated Protein Kinase 3 , Metabolism , Mitogen-Activated Protein Kinases , Metabolism , RNA, Messenger , Swine
<p><b>OBJECTIVE</b>To observe the effect of Yangjing Zhongyu Decoction (YZD) on mRNA and protein expression of PCNA, StAR, and FSHR in ovarian granulose cells (GCs) cultured by excess androgen.</p><p><b>METHODS</b>Ovarian GCs from porcine follicles were isolated and cultured in vitro. Follicular stimulating hormone (FSH) or YZD was added in the GCs treated by excess testosterone propionate. Totally 48 h later mRNA and protein expression of PCNA, StAR, and FSHR were detected by RT-PCR and Western blot.</p><p><b>RESULTS</b>Excess androgen inhibited mRNA and protein expression of PCNA, StAR, and FSHR of GCs. FSH and YZD could antagonize inhibition of excess androgens, and promote mRNA and protein expression of PCNA, StAR, and FSHR in GCs.</p><p><b>CONCLUSION</b>YZD could antagonize the inhibition of excess androgen on mRNA and protein expression of PCNA, StAR and FSHR in GCs. Thus, we inferred that YZD could improve the follicle dysplasia by promoting mRNA and protein expression of PCNA, StAR and FSHR in GCs.</p>
Animals , Female , Androgens , Pharmacology , Cells, Cultured , Drugs, Chinese Herbal , Pharmacology , Follicle Stimulating Hormone , Pharmacology , Granulosa Cells , Cell Biology , Metabolism , Membrane Transport Proteins , Genetics , Metabolism , Ovarian Follicle , Cell Biology , Proliferating Cell Nuclear Antigen , Genetics , Metabolism , RNA, Messenger , Genetics , Receptors, FSH , Genetics , Metabolism , Swine
<p><b>OBJECTIVE</b>To observe the therapeutic effect of acupoint sticking of Wuzhuyu (Evodia Rutaecarpa) for treatment of bradycardia.</p><p><b>METHODS</b>Sixty cases were randomly divided into an observation group and a control group, 30 cases in each group. The observation group was treated with acupoint sticking of Wuzhugu (Evodia Rutaecarpa) at Neiguan (PC 6) and Xinshu (BL 15) once each day. The control group was treated with oral administration of Xinbao pills, 3 pills each time, thrice each day. The therapeutic effects and changes of 24 hours' holter were observed after 4 weeks.</p><p><b>RESULTS</b>After treatment, 24 hours' average heart rate was significantly improved in the two groups, with significant differences as compared with those before treatment (both P<0.01) and with no significant difference between the two groups (P>0.05). The total effective rate was 86.7% (26/30) in the observation group and 83.3% (25/30) in the control group, their therapeutic effect being similar.</p><p><b>CONCLUSION</b>Acupoint sticking of Wuzhugu (Evodia Rutaecarpa) can significantly raise the levels of 24 hours' average heart rate in the patients of bradycardia. This therapy and Xinbao pills have similar therapeutic effect on the improvement of clinical symptom and heart rate levels.</p>
Aged , Female , Humans , Male , Middle Aged , Acupuncture Points , Acupuncture Therapy , Bradycardia , Therapeutics , Treatment Outcome
<p><b>OBJECTIVE</b>To screen serum biomarkers in patients with mycosis fungoides (MF) using surface-enhanced laser desorption and ionization with time-of-flight detection mass spectrometry (SELDI-TOF-MS) technique.</p><p><b>METHODS</b>Serum was analyzed from 14 patients with MF and 17 controls using CM10 Protein-chip to capture serum proteins, followed by Biomarker Wizard software analysis.</p><p><b>RESULTS</b>In all specimens, about 131 protein peaks could be detected when the relative molecular weight ranged from 0 to 50 000. When comparing the protein fingerprint between these two groups, 14 differentially expressed protein peaks were found. By searching SWISS-PRO database, we found 7 670Da peaks accord with C-C motif chemokine 22.</p><p><b>CONCLUSION</b>SELDI-TOF-MS technique can be used for screening serum protein biomarkers in patients with MF.</p>
Humans , Biomarkers, Tumor , Blood , Blood Proteins , Mycosis Fungoides , Blood , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods
Serum samples from endometrial cancer (EC) patients and healthy females were analyzed using surface-enhanced laser desorption-ionization time-of-flight mass spectrometry (SELDI-TOF-MS) to discover the potential diagnostic biomarker for detection of EC. A preliminary training set of spectra derived from 40 EC patients and 30 healthy women were used to develop a proteomic model that effectively discriminated cancer patients from healthy women. The training set had a specificity of 100% and sensitivity of 92.5% in the EC detection. A blind test set, including 20 new cancer cases and 10 healthy women, was used to validate the sensitivity and specificity of this multivariate model, which had a corresponding results of 60% in specificity and 75% in sensitivity, respectively. The combination of SELDI-TOF-MS with bioinformatics tools could help find new biomarkers and establish the detection of EC with high sensitivity and specificity.
Female , Humans , Biomarkers, Tumor , Metabolism , Case-Control Studies , Endometrial Neoplasms , Diagnosis , Metabolism , Gene Expression Regulation, Neoplastic , Medical Oncology , Methods , Models, Biological , Neoplasm Proteins , Neoplasm Staging , Protein Array Analysis , Proteomics , Methods , Sensitivity and Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Methods
<p><b>OBJECTIVE</b>Human selenoprotein P (HSelP) is unique protein that contains 10 selenocysteines encoded by 10 inframe UGA, which typically function as stop codon. The function of HSelP remains unclear, in part due to the inability to express it by gene recombinant technique. This study is to investigate expression and purification of recombinant HSelP in prokaryotic expression system, and its activity to induce apoptosis in vitro.</p><p><b>METHODS</b>The shorter HSelP isoform was cloned. After the selenocysteine (SeCys) at 40th position from N terminus of the HSelP shorter isoform was mutated into cysteine by PCR, it was expressed in E. coli. The expressed product was purified with DEAE column and identified by Western blot. Subsequently, its function on induction of mitochondrial apoptotic activity was studied.</p><p><b>RESULTS</b>The mutant HSelP shorter isoform expressed in prokaryotic system was purified by DEAE column to 90% homogeneity. The purified product, HSelP280m, induced the opening of mitochondrial permeability transition pore (PTP) and decreased the transmembrane potential in a dose-dependent manner. These events could be abolished by PTP specific inhibitors.</p><p><b>CONCLUSION</b>HSelP280m can induce the opening of mitochondrial PTP, which provides a basis for investigating the structure and function of recombinant HSelP.</p>
Animals , Humans , Male , Mice , Apoptosis , Cloning, Molecular , Cysteine , Genetics , Escherichia coli , Metabolism , Ion Channels , Membrane Potentials , Mice, Inbred BALB C , Mitochondria, Liver , Physiology , Mitochondrial Membrane Transport Proteins , Mutation , Protein Isoforms , Proteins , Genetics , Metabolism , Pharmacology , Selenium , Selenocysteine , Genetics , Selenoprotein P , Selenoproteins
Objective To analyze the alterations of serum protein fingerprint in patients with hypertensive disorder complicating pregnancy(HDCP),screen serum biomarker and establish a diagnostic model of HDCP.Methods Surface-enhanced laser desorption lionization-time of flight-mass spectrometry (SELDI-TOF-MS)technology was used to analyze serum including 25 cases of HDCP patients and 30 cases of age-,gravity-and parity-matched healthy term pregnant women on IMAC3-Cu proteinchip before delivery. Biomarker Wizard and Biomarker Pattern software was used to detect protein peaks significantly different between HDCP and controls,and establish a primary diagnostic model of HDCP.This model was further evaluated by blind test using other 16 parts of serum protein fingerprint.Results Ten protein peaks were significantly different at the molecular range of 2000-50 000(P
