1.Effect of warm acupuncture on PINK1/Parkin pathway in the skeletal muscle of rats with chronic fatigue syndrome
Huayuan LI ; Chun LI ; Junwei LIU ; Ting WANG ; Long LI ; Yongli WU
Chinese Journal of Tissue Engineering Research 2025;29(8):1618-1625
BACKGROUND:It has been found that mitochondrial function is abnormal in patients with chronic fatigue syndrome,and the administration of coenzymes can improve the symptoms.Warm acupuncture is one of the most important treatments for this disease,but its mechanism of action is unclear. OBJECTIVE:To investigate the effects of warm acupuncture on the phosphatase and tensin inducible kinase 1(PINK1)/Parkin pathway in the skeletal muscle of rats with chronic fatigue syndrome. METHODS:After 3 days of adaptive feeding,32 male Sprague-Dawley rats were randomly divided into normal control,model,warm acupuncture,and coenzyme Q groups with 8 rats in each group.The chronic fatigue syndrome model was established by multiple factors,including swimming exhaustion,chronic immobilization and fasting.After successful modeling,the normal group and the model group were treated with the same fixation and gavage procedures,and the warm acupuncture group was treated with acupuncture at Guanyuan,Zhongwan and Zusanli(bilateral)points,once a day.After the needling was inserted,the moxa pillar was put on the needle handle and ignited,three sessions once.The coenzyme Q group was given 1 mL/kg coenzyme by gavage,once a day for 14 days.The body mass,exhaustive swimming time and food utilization rate during the treatment were recorded.After the treatment,the bilateral gastrocnemius muscles of rats in each group were collected.The pathological morphology of the gastrocnemius muscle was observed by hematoxylin-eosin staining,the mitochondrial morphology and autophagosome of the gastrocnemius muscle were observed by transmission electron microscope.The expression level of microtubule-associated protein light chain 3(LC3)Ⅱ protein in the skeletal muscle was detected by immunohistochemistry.Western blot was used to detect the expression of PINK1,Parkin,LC3 Ⅰ,and LC3 Ⅱ in the skeletal muscle. RESULTS AND CONCLUSION:Compared with the normal group,the gastrocnemius muscle nuclei of the model group were pyknotic,condensed,the number of cells was increased,the cells were arranged disorderly,and the fibers in the gastrocnemius muscle were tightly arranged in the model group.Compared with the model group,the intercellular space became smaller,the nuclei were reduced,and the cell arrangement was orderly in the warm acupuncture group and coenzyme Q group.Compared with the normal group,the skeletal muscle mitochondria in the model group were swollen,fused,and vacuolated seriously,the membrane was partially broken,the matrix was more dissolved,the cristae was broken and disappeared,and autophagy appeared.Compared with the model group,the number of mitochondria increased,the arrangement was relatively neat,mitochondrial vacuolization and rupture of cristae in the gastrocnemius muscle were improved,the membrane structure was relatively intact,and autophagy occurred.Compared with the normal group,the expression of PINK1 protein in the skeletal muscle of the model group was significantly increased(P<0.05),while the expression of Parkin,LC3 Ⅱ and LC3 Ⅱ/Ⅰ protein was slightly upregulated(P>0.05).Compared with the model group,the protein expressions of PINK1,Parkin,LC3 Ⅱ and LC3 Ⅱ/Ⅰ were significantly upregulated in the warm acupuncture and coenzyme Q groups(P<0.05),and the up-regulation was more significant in the warm acupuncture group.To conclude,warm acupuncture can play a role in the treatment of chronic fatigue syndrome by activating the PINK1/Parkin pathway,upregulating LC3 Ⅱ expression,forming mitochondrial autophagosomes,promoting the degradation of damaged mitochondria,and improving mitochondrial quality.
2.Ameliorative effect of patchouli alcohol on mice with lung-heat syndrome based on PI3K/Akt/NF-κB pathway
Linze LI ; Yi LI ; Haoyi QIAO ; Jiakang JIAO ; Qi ZHANG ; Xiaofang WU ; Xingyu ZHAO ; Yinming ZHAO ; Chun WANG ; Jianjun ZHANG ; Linyuan WANG
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):459-470
Objective:
To investigate the therapeutic effect of patchouli alcohol on mice with lung-heat syndrome based on the phosphoinositide 3-kinase(PI3K)/protein kinase B(Akt)/nuclear factor-kappa B(NF-κB) signaling pathway.
Methods:
First, network pharmacology was used to predict the potential targets of patchouli alcohol in the treatment of lung-heat syndrome, and a "component-disease-key target" network was constructed for pathway analysis. Then, 40 BALB/c mice were assigned to the normal, lung-heat model, honeysuckle, and low-dose and high-dose patchouli alcohol groups. All groups, except the blank group, were intranasally infected with 50 μL (103 TCID50) of influenza virus solution. After two hours of infection, mice were treated once a day for seven consecutive days. The therapeutic mechanism of patchouli alcohol was explored by measuring pulmonary inflammatory factors, the PI3K/Akt/NF-κB pathway, hypothalamic fever markers (PGE2, cAMP, cGMP levels), rectal temperature, and tissue energy metabolism.
Results:
Network pharmacology identified 135 target genes related to patchouli alcohol and lung-heat syndrome, with the key targets being STAT3, H1F1A, and NF-κB1. In animal experiments, patchouli alcohol significantly alleviated influenza virus-induced lung inflammatory damage in mice with lung-heat syndrome, inhibited the expression of TNF-α and IL-6 in lung tissues(P<0.01), and suppressed the activation of the PI3K/Akt/NF-κB pathway. It also reduced hypothalamic levels of PGE2 and cAMP(P<0.01), suppressed the increase in rectal temperature, significantly decreased liver glycogen and pyruvate levels(P<0.01), and increased the activities of SDH, LDH, and Na+ -K+ -ATPase in the liver(P<0.01)
Conclusion
Patchouli alcohol improves the symptoms of lung-heat syndrome in mice by inhibiting the activation of the PI3K/Akt/NF-κB pathway, reducing proinflammatory cytokines and inflammatory damage, and regulating hypothalamic fever markers and energy metabolism.
3.Study on the effects of carvacrol on stomach-heat and stomach-cold rats and its mechanism of cooling and clearing based on energy metabolism and gastrointestinal function
Qi ZHANG ; Yi LI ; Hongye LI ; Fengwei ZHANG ; Minghui JIANG ; Xingyu ZHAO ; Linze LI ; Xiaofang WU ; Yinming ZHAO ; Songrui DI ; Jianjun ZHANG ; Chun WANG ; Linyuan WANG
Journal of Beijing University of Traditional Chinese Medicine 2025;48(4):471-482
Objective:
To investigate the biological effects of carvacrol on rats with stomach-heat and stomach-cold and its regulation on transient receptor potential(TRP) channels in rats with stomach-heat, and to study the cold and heat properties of carvacrol and its possible mechanism.
Methods:
According to the random number method, 100 SD rats were divided into stomach-heat blank group, stomach-heat model group, Coptidis Rhizoma group, stomach-heat low-dose and high-dose carvacrol group, stomach-cold blank group, stomach-cold model group, Baked ginger group, stomach-cold low-dose group and high-dose carvacrol group, 10 rats in each group. The rat model of stomach-heat was established by intragastric administration of pepper aqueous solution (0.80 g/kg) and anhydrous ethanol, and the rat model of stomach-cold was established by intragastric administration of water extract of Anemarrhena asphodeloides and sodium hydroxide (10.40 g/kg). On the day of modeling, the rats in the Baked ginger group were given Baked ginger decoction (0.78 g/kg), and the rats in the Coptidis Rhizoma group were given Coptidis Rhizoma decoction (0.43 g/kg).The stomach-cold and stomach-heat low-dose group of carvacrol was given carvacrol emulsion (40 mg/kg), high-dose group was given carvacrol emulsion (80 mg/kg).All rats of the blank and model groups were given the equal volume of emulsion prepared by 5% dimethyl sulfoxide, 1% Tween 80, 1% polyethylene glycol 400, and 93% normal saline, once a day, for 7 days. The general condition of rats was observed and the body mass was recorded. The pathological morphology of gastric tissue was observed by hematoxylin-eosin staining. The changes of material and energy metabolism, cyclic nucleotide (cAMP), thyroid hormone and gastrointestinal hormone in each group were determined by enzyme-linked immunosorbent assay. The expression levels of transient receptor potential vanilloid subtype 1 (TRPV1), transient receptor potential channel M8 (TRPM8) and uncoupling protein-1 (UCP1) in rats with gastric fever were detected by Western blotting.
Results:
Compared with the stomach-heat blank group, the body mass of rats in the stomach-heat model group decreased at the fifth and seventh day (P<0.05). The contents (or ratio) of hepatic glycogen (HGlyc), total cholesterol (TC), triglyceride (TG), and vasoactive intestinal peptide (VIP) were decreased (P<0.05), and Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, cytochrome C oxidase (COX), NADH dehydrogenase (ND), cyclic adenosine phosphate (cAMP), cAMP/cyclic guanosine phosphate (cGMP), triiodothyronine (T3), thyroxine (T4), gastrin (GAS), motilin (MTL), and α-amylase (α-AMS) all increased (P<0.05). Compared with the stomach-heat model group, the body mass of rats in the Coptidis Rhizoma group decreased at the third, fifth, and seventh day, the contents (or ratio) of HGlyc, TC, TG, VIP and α-AMS were increased, and Na+ -K+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, and GAS all decreased (P<0.05). The body mass of rats in the stomach-heat low-dose carvacrol group decreased at the seventh day. The contents (or ratio) of HGlyc, TC, and VIP were increased, Na+ -K+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, and MTL all decreased, the expression of TRPV1 and UCP1 in gastric tissue decreased, while TRPM8 increased (P<0.05) in rats of the stomach-heat low-dose and high-dose carvacrol groups. Compared with the stomach-cold blank group, the body mass of rats in the stomach-cold model group decreased at the third, fifth, and seventh day, the contents (or ratio) of HGlyc, TC, TG, α-AMS, and VIP all increased, while Na+ -K+ -ATPase, Ca2+ -Mg2+ -ATPase, COX, ND, cAMP, cAMP/cGMP, T3, T4, GAS, and MTL all decreased (P<0.05). Compared with the stomach-cold model group, the body mass of rats in the Baked ginger group was increased at the seventh day, and the contents (or ratio) of HGlyc, VIP, and α-AMS all decreased, while Na+ -K+ -ATPase, COX, ND, cAMP/cGMP, T3, T4, GAS, and MTL all increased (P<0.05). The contents of HGlyc, cAMP, α-AMS, and VIP of rats in the stomach-cold low and high-dose carvacrol group all decreased (P<0.05). TG in the stomach-cold low-dose carvacrol group was increased. TC, Ca2+ -Mg2+ -ATPase, and cGMP all increased, while cAMP/cGMP decreased (P<0.05) in the high-dose carvacrol group.
Conclusion
In this study, the rat model of stomach-cold and stomach-heat were successfully established by using cold and heat factors. The result showed that carvacrol had a certain inhibitory effect on body mass, material energy metabolism, cyclic nucleotide level, thyroid hormone and gastrointestinal function in rats with stomach-heat, indicating that the drug was cold. Carvacrol′s cold medicinal property could be biologically explained by TRPV1 activation, UCP1 induction, and TRPM8 suppression.
4.Non-linear association between long-term air pollution exposure and risk of metabolic dysfunction-associated steatotic liver disease.
Wei-Chun CHENG ; Pei-Yi WONG ; Chih-Da WU ; Pin-Nan CHENG ; Pei-Chen LEE ; Chung-Yi LI
Environmental Health and Preventive Medicine 2024;29():7-7
BACKGROUND:
Metabolic Dysfunction-associated Steatotic Liver Disease (MASLD) has become a global epidemic, and air pollution has been identified as a potential risk factor. This study aims to investigate the non-linear relationship between ambient air pollution and MASLD prevalence.
METHOD:
In this cross-sectional study, participants undergoing health checkups were assessed for three-year average air pollution exposure. MASLD diagnosis required hepatic steatosis with at least 1 out of 5 cardiometabolic criteria. A stepwise approach combining data visualization and regression modeling was used to determine the most appropriate link function between each of the six air pollutants and MASLD. A covariate-adjusted six-pollutant model was constructed accordingly.
RESULTS:
A total of 131,592 participants were included, with 40.6% met the criteria of MASLD. "Threshold link function," "interaction link function," and "restricted cubic spline (RCS) link functions" best-fitted associations between MASLD and PM2.5, PM10/CO, and O3 /SO2/NO2, respectively. In the six-pollutant model, significant positive associations were observed when pollutant concentrations were over: 34.64 µg/m3 for PM2.5, 57.93 µg/m3 for PM10, 56 µg/m3 for O3, below 643.6 µg/m3 for CO, and within 33 and 48 µg/m3 for NO2. The six-pollutant model using these best-fitted link functions demonstrated superior model fitting compared to exposure-categorized model or linear link function model assuming proportionality of odds.
CONCLUSION
Non-linear associations were found between air pollutants and MASLD prevalence. PM2.5, PM10, O3, CO, and NO2 exhibited positive associations with MASLD in specific concentration ranges, highlighting the need to consider non-linear relationships in assessing the impact of air pollution on MASLD.
Humans
;
Nitrogen Dioxide
;
Cross-Sectional Studies
;
Air Pollution/analysis*
;
Air Pollutants/analysis*
;
Particulate Matter/analysis*
;
Liver Diseases
;
Environmental Exposure/analysis*
5.Improvement on Quality Standard of Yuanhu Zhitong Oral Liquid
Lu FU ; Chengyu CHEN ; Jin GAO ; Dan WU ; Chun LI ; Zhiming CAO ; Jianli GUAN ; Ping WANG ; Haiyu XU
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(9):125-131
ObjectiveTo improve the quality standard of Yuanhu Zhitong oral liquid in order to strengthen the quality control of this oral liquid. MethodThin layer chromatography(TLC) was used for the qualitative identification of Corydalis Rhizoma and Angelicae Dahuricae Radix in Yuanhu Zhitong oral liquid by taking tetrahydropalmatine, corydaline reference substances and Corydalis Rhizoma reference medicinal materials as reference, and cyclohexane-trichloromethane-methanol(5∶3∶0.5) as developing solvent, Corydalis Rhizoma was identified using GF254 glass thin layer plate under ultraviolet light(365 nm). And taking petroleum ether(60-90 ℃) -ether-formic acid(10∶10∶1) as developing solvent, Angelicae Dahuricae Radix was identified using a silica gel G TLC plate under ultraviolet light(305 nm). High performance liquid chromatography(HPLC) was performed on a Waters XSelect HSS T3 column(4.6 mm×250 mm, 5 μm) with acetonitrile(A)-0.1% glacial acetic acid solution(adjusted pH to 6.1 by triethylamine)(B) as the mobile phase for gradient elution(0-10 min, 20%-30%A; 10-25 min, 30%-40%A; 25-40 min, 40%-50%A; 40-60 min, 50%-60%A), the detection wavelength was set at 280 nm, then the fingerprint of Yuanhu Zhitong oral liquid was established, and the contents of tetrahydropalmatine and corydaline were determined. ResultIn the thin layer chromatograms, the corresponding spots of Yuanhu Zhitong oral liquid, the reference substances and reference medicinal materials were clear, with good separation and strong specificity. A total of 12 common peaks were identified in 10 batches of Yuanhu Zhitong oral liquid samples, and the peaks of berberine hydrochloride, dehydrocorydaline, glaucine, tetrahydropalmatine and corydaline. The similarities between the 10 batches of samples and the control fingerprint were all >0.90. The results of determination showed that the concentrations of corydaline and tetrahydropalmatine had good linearity with paek area in the range of 0.038 6-0.193 0, 0.034 0-0.170 0 g·L-1, respectively. The methodological investigation was qualified, and the contents of corydaline and tetrahydropalmatine in 10 batches of Yuanhu Zhitong oral liquid samples were 0.077 5-0.142 9、0.126 1-0.178 2 g·L-1, respectively. ConclusionThe established TLC, fingerprint and determination are simple, specific and reproducible, which can be used to improve the quality control standard of Yuanhu Zhitong oral liquid.
6. Analysis of cerebral gray matter structure in multiple sclerosis and neuromyelitis optica
Xiao-Li LIU ; Ai-Xue WU ; Ru-Hua LI ; An-Ting WU ; Cheng-Chun CHEN ; Lin XU ; Cai-Yun WEN ; Dai-Qian CHEN
Acta Anatomica Sinica 2024;55(1):17-24
Objective The volume and cortical thickness of gray matter in patients with multiple sclerosis (MS) and neuromyelitis optica (NMO) were compared and analyzed by voxel⁃based morphometry (VBM) and surface⁃based morphometry (SBM), and the differences in the structural changes of gray matter in the two diseases were discussed. Methods A total of 21 MS patients, 16 NMO patients and 19 healthy controls were scanned by routine MRI sequence. The data were processed and analyzed by VBM and SBM method based on the statistical parameter tool SPM12 of Matlab2014a platform and the small tool CAT12 under SPM12. Results Compared with the normal control group (NC), after Gaussian random field (GRF) correction, the gray matter volume in MS group was significantly reduced in left superior occipital, left cuneus, left calcarine, left precuneus, left postcentral, left central paracentral lobule, right cuneus, left middle frontal, left superior frontal and left superior medial frontal (P<0. 05). After family wise error (FWE) correction, the thickness of left paracentral, left superiorfrontal and left precuneus cortex in MS group was significantly reduced (P<0. 05). Compared with the NC group, after GRF correction, the gray matter volume in the left postcentral, left precentral, left inferior parietal, right precentral and right middle frontal in NMO group was significantly increased (P<0. 05). In NMO group, the volume of gray matter in left middle occipital, left superior occipital, left inferior temporal, right middle occipital, left superior frontal orbital, right middle cingulum, left anterior cingulum, right angular and left precuneus were significantly decreased (P<0. 05). Brain regions showed no significant differences in cortical thickness between NMO groups after FWE correction. Compared with the NMO group, after GRF correction, the gray matter volume in the right fusiform and right middle frontal in MS group was increased significantly(P<0. 05). In MS group, the gray matter volume of left thalamus, left pallidum, left precentral, left middle frontal, left middle temporal, right pallidum, left inferior parietal and right superior parietal were significantly decreased (P<0. 05). After FWE correction, the thickness of left inferiorparietal, left superiorparietal, left supramarginal, left paracentral, left superiorfrontal and left precuneus cortex in MS group decreased significantly (P<0. 05). Conclusion The atrophy of brain gray matter structure in MS patients mainly involves the left parietal region, while NMO patients are not sensitive to the change of brain gray matter structure. The significant difference in brain gray matter volume between MS patients and NMO patients is mainly located in the deep cerebral nucleus mass.
7.Effect of ANAs on hormone response in patients with AIH-PBC overlap syndrome and AIH-only
Qiong LI ; Ai-Ping TIAN ; Yong-Wu MAO ; Fu-Chun WANG ; Xiao-Rong MAO
Medical Journal of Chinese People's Liberation Army 2024;49(1):64-69
Objective To investigate the effect of antinuclear antibodies(ANAs)on hormone response in patients with autoimmune hepatitis(AIH)-primary biliary cholangitis(PBC)overlap syndrome(AIH-PBC OS)and AIH-only within half a year.Methods A retrospective analysis of 77 patients with autoimmune liver disease(AILD)admitted to First Clinical Medical College of Lanzhou University from January 2018 to December 2021,all of whom were confirmed by liver biopsy and receiving glucocorticoid treatment.Among them,46 patients were in AIH-PBC OS group and 31 were in AIH-only group.The general clinical characteristics,liver puncture-related indexes,autoantibodies and immunoglobulin indexes of patients in each group at the time of diagnosis were collected and compared,and the biochemical and immunoglobulin indexes of patients at the time of hormone use and at the time of review within 6 months were also collected,and the hormone response within 6 months was evaluated according to the levels of glutamic transaminase(AST),glutamic alanine transaminase(ALT)and immunoglobulin G(IgG),and the effect of ANAs on hormone response outcomes in both groups over a six-month period was analyzed.Multifactorial ordered logistic analysis was performed to evaluate the effect of ANAs on hormone response between two groups.Results There was no statistically significant difference in the percentage of AIH-PBC OS and AIH-only patients among both ANAs-positive and-negative AILD patients(55.6%vs.44.4%and 65.6%vs.34.4%,P>0.05).Among 46 patients with AIH-PBC OS,there were 25 in ANAs-positive group and 21 in ANAs-negative group.The rate of complete hormone response within 6 months was lower than that of ANAs-negative group(44.0%vs.76.2%),while the rate of hormone non-response was higher than that of ANAs-negative group(20.0%vs.0),and the difference was statistically significant(P<0.05).There were 20 cases of ANAs-positive and 11 cases of ANAs-negative in the 31 AIH-only patients.There was no statistically significant difference in the results of hormone response within 6 months between the two groups(P>0.05).Multifactorial ordered logistic analysis showed that AIH-PBC OS patients were more likely to have a higher likelihood of 6-month hormone non-response rate in ANAs-positive patients,and the difference was statistically different(P<0.05).And there was no significant effect of ANAs type on hormone response outcome in AIH-only patients(P>0.05).Conclusion AIH-PBC OS ANAs-positive patients have a poor hormone response within half a year.In AIH-only patients,ANAs have no significant effect on hormone response results.
8.Scutellarin inhibitting BV-2 microglia-mediated neuroinflammation via the cyclic GMP-AMP synthase-stimulator of interferon gene pathway
Zhao-Da DUAN ; Li YANG ; Hao-Lun CHEN ; Teng-Teng LIU ; Li-Yang ZHENG ; Dong-Yao XU ; Chun-Yun WU
Acta Anatomica Sinica 2024;55(2):133-142
Objective To explore the effect of scutellarin on lipopolysaccharide(LPS)induced neuroinflammation in BV-2 microglia cells.Methods BV-2 microglia were cultured and randomly divided into 6 groups:control group(Ctrl),cyclic GMP-AMP synthetase(cGAS)inhibitor RU320521 group(RU.521 group),LPS group,LPS+RU.521 group,LPS+scutellarin pretreatment group(LPS+S)and LPS+S+RU.521 group.The expressions of cGAS,stimulator of interferon gene(STING),nuclear factor kappa B(NF-κB),phosphorylated NF-κB(p-NF-κB),neuroinflammatory factors PYD domains-containing protein 3(NLRP3)and tumor necrosis factor α(TNF-α)in BV-2 microglia were detected by Western blotting and immunofluorescent double staining(n= 3).Results Western blotting and immunofluorescent double staining showed that compared with the control group,the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in BV-2 microglia increased significantly after LPS induction(P<0.05),while the expression of cGAS,STING,p-NF-κB,NLRP3 and TNF-α in LPS+S group were significantly lower than those in LPS group(P<0.05).Treatment with cGAS pathway inhibitor RU.521 showed similar effects as the pre-treatment group with scutellarin.In addition,the change of NF-κB in each group was not statistically significant(P>0.05).Conclusion Scutellarin inhibits the neuroinflammation mediated by BV-2 microglia cells,which may be related to cGAS-STING signaling pathway.
9.Effect of warm-needling moxibustion on anterior cruciate ligament injury repair and related growth factors in rabbits with knee osteoarthritis
Chun LI ; Yanlin ZHANG ; Di LIU ; Minglei WANG ; Duo WANG ; Junwei LIU ; Yongli WU
Chinese Journal of Tissue Engineering Research 2024;28(23):3621-3626
BACKGROUND:Warm-needling moxibustion can effectively treat knee osteoarthritis.Degeneration,injury and fracture of the anterior cruciate ligament can affect the local stability of the knee joint,and then induce the formation of knee osteoarthritis.Whether warm-needling moxibustion can repair the injured cruciate ligament and the mechanism of action are still unclear. OBJECTIVE:To observe the effects of warm-needling moxibustion on the morphology of the anterior cruciate ligament and the expression of insulin growth factor-1 and transforming growth factor-β in rabbits with knee osteoarthritis and to clarify the mechanism of anterior cruciate ligament repair by warm-needling moxibustion. METHODS:Thirty New Zealand rabbits were randomly divided into blank group,model group and warm-needling moxibustion group,with 10 rabbits in each group.Knee osteoarthritis model was established by plaster cast immobilization.The blank group was not intervened.Rabbits in the model group rabbits were fixed in a rabbit holder for 15 minutes every day.The warm-needling moxibustion group was treated with warm acupuncture,once a day,7 days as a course of treatment,a total of two courses.After treatment,the imaging changes of the anterior cruciate ligament were observed by MRI and MRI grading statistics were performed.Morphological changes of the anterior cruciate ligament were observed by transmission electron microscope and hematoxylin-eosin staining.mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were detected by RT-PCR and western blot,respectively. RESULTS AND CONCLUSION:MRI examination:Compared with the blank control group,the anterior cruciate ligament in the model group was thickened,edematous,and partially torn,and the difference in grading statistics was statistically significant(P<0.05).Compared with the model group,the anterior cruciate ligament in the warm-needling moxibustion group was slightly thickened,with mild edema and no tearing,and the difference in grading statistics was statistically significant(P<0.05).General observation:In the model group,the surface of the anterior cruciate ligament was glossy and faded,with the edge being covered with flocculent periosteum and obvious tissue necrosis;in the warm-needling moxibustion group,the surface of the ligament was glossy,and the ligament was in a normal helical shape.Hematoxylin-eosin staining:In the model group,there was obvious tissue necrosis in the anterior cruciate ligament,a large number of new capillaries,loosely arranged fibroblasts and collagen fibers.In the warm-needling moxibustion group,there was a small amount of tissue necrosis and few new vessels in the anterior cruciate ligament,and the cells and collagen fibers were loosely and irregularly arranged.Transmission electron microscopy:In the model group,the fibers in the anterior cruciate ligament were arranged in a disordered way with uneven thickness and distribution,and there are more fibroblasts that were irregular in morphology;in the warm acupuncture group,the fibers were basically arranged longitudinally,with uneven thickness and distribution,and a small number of oval-shaped fibroblasts were observed.RT-PCR and western blot assay:mRNA and protein expressions of insulin growth factor-1 and transforming growth factor-β were significantly decreased in the model group compared with the blank control group(P<0.05),but significant increased after treatment with warm-needling moxibustion(P<0.05).To conclude,warm-needling moxibustion can alleviate anterior cruciate ligament injury and regulate the expression of insulin growth factor-1 and transforming growth factor-β to treat knee osteoarthritis.
10.Development and Application of Detection Methods for Capture and Transcription Elongation Rate of Bacterial Nascent RNA
Yuan-Yuan LI ; Yu-Ting WANG ; Zi-Chun WU ; Hao-Xuan LI ; Ming-Yue FEI ; Dong-Chang SUN ; O. Claudio GUALERZI ; Attilio FABBRETTI ; Anna Maria GIULIODORI ; Hong-Xia MA ; Cheng-Guang HE
Progress in Biochemistry and Biophysics 2024;51(9):2249-2260
ObjectiveDetection and quantification of RNA synthesis in cells is a widely used technique for monitoring cell viability, health, and metabolic rate.After exposure to environmental stimuli, both the internal reference gene and target gene would be degraded. As a result, it is imperative to consider the accurate capture of nascent RNA and the detection of transcriptional levels of RNA following environmental stimulation. This study aims to create a Click Chemistry method that utilizes its property to capture nascent RNA from total RNA that was stimulated by the environment. MethodsThe new RNA was labeled with 5-ethyluridine (5-EU) instead of uracil, and the azido-biotin medium ligand was connected to the magnetic sphere using a combination of “Click Chemistry” and magnetic bead screening. Then the new RNA was captured and the transcription rate of 16S rRNA was detected by fluorescence molecular beacon (M.B.) and quantitative reverse transcription PCR (qRT-PCR). ResultsThe bacterial nascent RNA captured by “Click Chemistry” screening can be used as a reverse transcription template to form cDNA. Combined with the fluorescent molecular beacon M.B.1, the synthesis rate of rRNA at 37℃ is 1.2 times higher than that at 15℃. The 16S rRNA gene and cspI gene can be detected by fluorescent quantitative PCR,it was found that the measured relative gene expression changes were significantly enhanced at 25℃ and 16℃ when analyzed with nascent RNA rather than total RNA, enabling accurate detection of RNA transcription rates. ConclusionCompared to other article reported experimental methods that utilize screening magnetic columns, the technical scheme employed in this study is more suitable for bacteria, and the operation steps are simple and easy to implement, making it an effective RNA capture method for researchers.


Result Analysis
Print
Save
E-mail