1.Establishment and Evaluation of New Mouse Model of Rheumatoid Arthritis Combined with Interstitial Lung Disease
Liting XU ; Qingyu ZHAO ; Chao YANG ; Lianhua HE ; Congcong SUN ; Shuangrong GAO ; Lili WANG ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2026;32(6):81-90
ObjectiveTo establish a mouse model of rheumatoid arthritis with interstitial lung disease (RA-ILD) in DBA/1 mice using Porphyromonas gingivalis (Pg) infection combined with collagen-induced arthritis (CIA), and to comprehensively evaluate pathological characteristics in joints, lungs, and serum. MethodsForty DBA/1 mice were randomly divided into four groups, i.e., Control, Pg infection (Pg), CIA, and Pg infection combined with CIA (Pg+CIA), with 10 mice in each group. Arthritis clinical symptoms were evaluated by recording arthritis incidence and clinical scores. Micro-CT scanning was used to assess knee joint pathology. Histopathological changes and collagen deposition in knee joints and lung tissues were analyzed using hematoxylin-eosin (HE) and Masson staining. Immunohistochemistry was performed to detect protein expression of α-smooth muscle actin (α-SMA), typeⅠ collagen (ColⅠ), and fibronectin (FN) in lung tissues. Real-time quantitative polymerase chain reaction(Real-time PCR)was used to measure mRNA expression levels of α-SMA, ColⅠ, FN, tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and IL-1β in lung tissues. Enzyme-linked immunosorbent assay (ELISA) was used to detect serum levels of Pg, cyclic citrullinated peptide (CCP), and immunoglobulin G (IgG). ResultsJoint lesions: The CIA and Pg+CIA groups showed 100% arthritis incidence, with evident joint redness, swelling, and deformity. The number of affected limbs was 27 and 28, and clinical scores were 68 and 70, respectively. No obvious clinical symptoms were observed in the Pg group. Histopathological and imaging analyses showed severe joint lesions in the CIA and Pg+CIA groups, with significantly increased histopathological scores, bone mineral density, bone volume fraction, trabecular thickness, and trabecular number compared to the Control group (P<0.01). No obvious joint pathology was observed in the Pg group. Lung lesions: The Pg+CIA group exhibited marked alveolar inflammation, interstitial inflammatory cell infiltration, and alveolar wall thickening, with pronounced blue staining of collagen fibers. Histopathological scores and collagen area ratios were significantly higher than those of the Control, Pg, and CIA groups (P<0.05). Lung protein and mRNA expression levels of α-SMA, ColⅠ, and FN were markedly increased, and mRNA levels of IL-6, TNF-α, and IL-1β were significantly elevated compared to the Control group (P<0.05). Serology: The Pg+CIA group showed significantly higher levels of CCP, Pg, and IgG compared with the Control, Pg, and CIA groups (P<0.05). ConclusionDBA/1 mice subjected to Pg infection combined with CIA exhibited pronounced symptoms and pathological features of RA-ILD, along with elevated serum anti-CCP antibody levels. This model represents a novel RA-ILD mouse model, providing a valuable experimental tool for investigating RA-ILD pathogenesis and developing new therapeutics, and serves as a basis for establishing anti-cyclic citrullinated peptide antibody (ACPA)-positive RA-ILD animal models.
2.Relationship between psychological quality and non-suicidal self-injury behavior in middle school students: path analysis of coping style
Chunfang XIE ; Yang JIANG ; Xinlong TANG ; Jun ZHANG ; Lei CHEN ; Qinghao TONG
Sichuan Mental Health 2024;37(2):162-167
BackgroundNon-suicidal self-injury (NSSI) behavior is highly prevalent in middle school students, which poses a significant risk to the physical and mental health of middle school students, so finding ways to improve the NSSI behavior among middle school students is of great significance for promoting their healthy growth. ObjectiveTo investigate the relationship between psychological quality and NSSI behavior among middle school students in western Anhui, and to examine the pathway of coping style in the above relationship. MethodsFrom November to December 2020, 22 872 middle school students in western Anhui were selected using stratified cluster random sampling method, and were subjected to complete the assessment of Adolescent Non-suicidal Self-injury Assessment Questionnaire (ANSAQ), Adolescent's Psychological Suzhi Scale-Simplified Version (APSS-SV) and Coping Style Scale for Middle School Students (CSSMSS). Spearman correlation analysis was adopted to examine the correlation among scores of scales and the frequency of NSSI behavior in middle school students. Amos 23.0 was utilized to test the pathway of coping style in the relationship between psychological quality and NSSI behavior. ResultsA total of 21 718 (94.95%) middle school students completed the effective questionnaire survey. NSSI behavior was detected in 7 798 middle school students (35.91%). ANSAQ total score and CSSMSS positive coping style score of middle school students were negatively correlated with the frequency of NSSI behavior (r=-0.219, -0.179, P<0.01). CSSMSS negative coping style score was positively correlated with the frequency of NSSI behavior (r=0.093, P<0.01). The direct effect value of psychological quality on the frequency of NSSI behavior was -0.136, and the indirect effect value of coping style on the relationship between psychological quality and the frequency of NSSI behavior was -0.084. Among them, the effect value of positive coping style was -0.122, accounting for 55.45% of the total effect, while the effect value of negative coping style was 0.038, accounting for 17.27% of the total effect. ConclusionCoping style may mediate the relationship between psychological quality and NSSI behavior in middle school students, and the positive coping style and negative coping style play separate roles in the pathway of psychological quality on NSSI behavior.[Funded by 2020 Natural Science Research Project of Anhui Provincial Education Department (number, KJ2020B006)]
3.Network meta-analysis of the modeling effects of different factors on rabbit models of steroid-induced osteonecrosis of femoral head
Zhixing HU ; Qun LI ; Chao YANG ; Xiaoxiao WANG ; Luochangting FANG ; Wuqiong HOU ; Na LIN ; Weiheng CHEN ; Chunfang LIU ; Ya LIN
Chinese Journal of Tissue Engineering Research 2024;28(6):976-984
OBJECTIVE:The rabbit model of steroid-induced osteonecrosis of femoral head is the most commonly used animal model of femoral head necrosis.The pathological changes of the femoral head are close to clinical practice,however,the conditions,methods and evaluation standards of animal models reported in and outside China are not uniform,which leads to the low scientific value of animal models and is difficult to popularize.This study aimed to clarify the influence of different mold-making conditions on the establishment of steroid-induced osteonecrosis of femoral head rabbit model and analyze the appropriate conditions for the successful model establishment. METHODS:We searched the CNKI,WanFang,VIP,CBM,WoS,PubMed and EMbsae databases for the literature on the modeling of steroid-induced osteonecrosis of femoral head rabbits up to April 1,2022,completed the screening of the literature according to the inclusion and exclusion criteria and literature quality evaluation,and extracted the outcome index data in the literature.RevMan Stata and ADDIS statistical software were used to conduct a meta-analysis of the included data. RESULTS:(1)A total of 82 articles with 1 366 rabbits were included in the study.The steroid-induced osteonecrosis of femoral head modeling methods were divided into three types:steroid-alone method,steroid combined lipopolysaccharide method and steroid combined serum method.Among these,33 articles used steroid-alone method;20 articles used steroid combined lipopolysaccharide method;29 articles used steroid combined serum method.(2)Meta-analysis results showed that the three modeling methods significantly increased the rate of empty bone lacunae in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001),and significantly decreased the ratio of the trabecular bone area in the femoral head of steroid-induced osteonecrosis of femoral head rabbits(P<0.001).The order of empty bone lacunae rate of each modeling method was:steroid combined with lipopolysaccharide method>steroid-alone method>steroid combined with serum method>normal group,and the order of trabecular bone area rate of each modeling method was:normal group>steroid combined with serum method>steroid-alone method>steroid combined with lipopolysaccharide method.(3)The results of subgroup analysis suggested that the rate of empty bone lacunae in the rabbit model induced by steroid alone might be related to the rabbit variety and the type of steroid used for modeling(difference between groups P<0.05),in which the combined effect amount of New Zealand white rabbits was higher than that of Chinese white rabbits(P<0.05)and Japanese white rabbits,and the combined effect amount of dexamethasone was higher than that of other steroids.The rate of empty bone lacunae induced by steroid combined with lipopolysaccharide was related to the administration mode of lipopolysaccharide and the type of steroid(P<0.05),among which the combined effect of methylprednisolone sodium succinate was significantly higher than that of other steroids(P<0.05),and the combined effect of prednisolone was significantly lower than that of other steroids(P<0.05).The combined effect of lipopolysaccharide 100 μg/kg×twice was significantly lower than 10 μg/kg×twice and 50 μg/kg×twice(P<0.05).The rate of empty bone lacunae in the model induced by steroid combined with serum was related to serum dose and steroid type(P<0.05),among which the combined effect amount of dexamethasone sodium phosphate was significantly higher than other steroid types(P<0.05),and the combined effect amount of dexamethasone was significantly lower than other steroid types(P<0.05);the combined effect amount of serum"10 mL/kg+6 mL/kg"combined dose was lower than other serum doses(P<0.05). CONCLUSION:(1)With the rate of empty bone lacunae and the ratio of trabecular bone area as the judgment standard for the successful establishment of the model,the three modeling methods can successfully construct the rabbit steroid-induced osteonecrosis of femoral head model,of which the steroid combined with lipopolysaccharide method is the best.(2)New Zealand white rabbits and dexamethasone are recommended when selecting the steroid-alone method.Methylprednisolone sodium succinate and low-dose lipopolysaccharide are recommended when selecting the steroid combined with lipopolysaccharide method.Dexamethasone sodium phosphate is recommended when selecting the steroid combined with serum modeling method.
4.Study on the effect of low-frequency rTMS combined with NJF technique in improving the enhancement of muscular tone after stroke
Liru CUI ; Hanming WANG ; Aoran YANG ; Jian TAN ; Dongliang YANG ; Chunfang LI
China Medical Equipment 2024;21(10):90-95
Objective:To analyze the clinical effect of low-frequency repetitive transcranial magnetic stimulation(rTMS)combined with neuromuscular joint facilitation(NJF)technique in improving the enhancement of muscle tone after stroke.Methods:From October 2020 to October 22,a total of 114 patients with increased muscle tone at lower limb during the recovery period of stroke who admitted to Beijing Rehabilitation hospital were selected,and they were randomly divided into the observation group(57 cases)and the control group(57 cases)according to the random number table method.The control group received routine rehabilitation treatment.The observation group added low-frequency rTMS and NJF training on the basis of the routine rehabilitation treatment.The differences of clinical efficacy and various indicators of muscle tone of lower limb before and after treatment between two groups were compared,which included modified Ashwonh score(MAS)scale,functional ambulation category(FAC)scale of the motor of lower limb,Fugl-Meyer motor function scale-Lower limbs(FMA-L),clinical spasmodic index(CSI),modified Barthel index(MBI)of activity ability of daily life,integrated electromyogram(IEMG)and root mean square(RMS)of tibialis anterior muscle of lower limb in electromyogram.Results:After intervention,29 cases appeared significant effect,and 23 cases appeared valid result,and 5 cases appeared invalid result in the observation group,and 20 cases appeared significant effect,and 20 cases appeared valid result,and 17 cases appeared invalid result in the control group.The total effective rate of the observation group was 91.23%,which was significantly higher than 70.18%of the control group,and the difference was statistically significant(x2=8.111,P<0.05).Before intervention,there was no statistically significant difference in each indicator between the two groups(P>0.05).The MAS score,CSI score,IEMG and RMS of two groups after intervention were significantly lower than those before intervention,while the FAC score,FMA-L score and MBI score were significantly higher than those before intervention,and the differences were significant(t=3.377,4.398,5.554,P<0.05),respectively.At the same time,the MAS score,CSI score,IEMG and RMS of the observation group were significantly higher than those of the control group,and the differences were statistically significant(t=11.160,9.625,16.993,8.095,P<0.05),respectively.Conclusion:The combined application of low-frequency rTMS and NJF training can significantly improve the enhancement of muscle tone after stroke,and effectively reduce the muscle tone of lower limb,and improve the motor function of lower limb and the degree of spasticity,and improve the activity ability of daily life.
5.Effect of transcranial direct current stimulation combined with robotic therapy on the rehabilitation of upper-limb motor function of hemiplegic stroke patients
Fei YANG ; Chunfang WANG ; Changcheng SUN ; Jingang DU
International Journal of Biomedical Engineering 2023;46(2):132-137
Objective:To investigate the effect of cathodic transcranial direct current stimulation (ctDCS) combined with upper limb robot therapy (RT) on the rehabilitation of upper limb motor function in stroke patients.Methods:Forty patients with stroke hemiplegia who met the enrollment criteria were randomly divided into a pseudo-stimulation group ( n = 20) and a stimulation group ( n = 20). In addition to conventional treatment in both groups, ctDCS + RT was used in the stimulation group, and sham stimulation + RT was used in the sham-stimulation group. Treatment was performed 10 times, 5 times per week, for 30 minutes each time. Patients in both groups were evaluated before, during, and after treatment using the Brunnstrom Staging Scale, the Modified Barthel Index (MBI) scale, and the Modified Ashworth Scale (MAS), respectively. Results:Compared with the same group before treatment, there were statistically significant differences in Brunnstrom’s rating grade Ⅳ for upper limbs and hands in both groups during and after treatment (all P < 0.05). Compared with the same group during treatment, there were statistically significant differences in Brunnstrom’s rating grade Ⅳ for upper limbs and hands in both groups after treatment (all P < 0.05). MBI scores were higher in two groups during and after treatment compared to the same group before treatment (all P < 0.05). MBI scores were higher in two groups after treatment compared to the same group during treatment (all P < 0.05). The MBI scores after treatment in the stimulation group were higher than those in the pseudo-stimulation group ( P < 0.05). The MAS scores of elbow flexion grade 2 and extension grade 4 and shoulder flexion grade 2, extension grade 2, adduction grade 2, and abduction grade 2 were lower in both groups after treatment compared with the same group before treatment (all P < 0.05). Conclusions:Cathodic transcranial direct current stimulation combined with robotic therapy can effectively promote upper limb motor function rehabilitation in stroke patients and is superior to upper limb robotic therapy alone.
6.Regulatory effects of the Nocardia rubra cell wall skeleton on the biological function of human neutrophils
Yunxi YANG ; Jiamin HUANG ; Lu LIU ; Linbin LI ; Chunfang ZHENG ; Yuying ZHOU ; Bingwei SUN
Chinese Journal of Burns 2023;39(7):625-632
Objective:To investigate the regulatory effects and mechanism of Nocardia rubra cell wall skeleton (Nr-CWS) on the biological function of human neutrophils. Methods:The experimental research method was used. Fifteen healthy adult volunteers (7 males and 8 females, aged 24 to 45 years) were recruited from Suzhou Physical Examination Center for physical examination from May to October 2022, the peripheral venous blood was collected, and neutrophils were extracted by immunomagnetic bead sorting. The cells were divided into normal control group without any treatment, Nr-CWS alone group treated with Nr-CWS of final mass concentration 60 ng/mL alone, endotoxin/lipopolysaccharide (LPS) alone group stimulated with LPS of final mass concentration 1 μg/mL alone, and LPS+Nr-CWS group stimulated with LPS first and then treated with Nr-CWS as before. After 1 h of culture, the chemotaxis distance, chemotactic cell percentage, chemotactic index, maximum chemotactic speed, and chemotactic function score of neutrophils were detected using the modified agarose chemotactic model; the proportion and fluorescence intensity of phagocytosis cells, the level of reactive oxygen species (ROS), the protein expression levels of granular protein CD35, CD66b, and CD63, and the concentrations of inflammatory cytokines of interleukin 2 (IL-2), IL-4, IL-6, IL-10, IL-17A, tumor necrosis factor alpha (TNF-α), and interferon-γ in cell culture supernatant were detected by flow cytometry. The number of samples in each group in the above experiments was 15. Data were statistically analyzed with analysis of variance for factorial design and independent sample t test. Results:After 1 h of culture, the chemotactic function score of cells in normal control group, Nr-CWS alone group, LPS alone group, and LPS+Nr-CWS group were 15.0, 14.5±0.5, 1.5±0.5, 12.0±1.5, respectively. Compared with those in normal control group, the chemotaxis distance, chemotactic cell percentage, chemotactic index, maximum chemotactic speed, and chemotactic function score of cells were significantly decreased in LPS alone group and LPS+Nr-CWS group (with t values of 18.36, 18.88, 54.28, 18.36, 46.77, 10.58, 14.74, 6.84, 10.58, and 4.24, respectively, P<0.05); compared with those in LPS alone group, the five chemotactic function indexes as above in LPS+Nr-CWS group were significantly increased (with t values of 11.47, 14.65, 11.62, 11.47, and 13.75, respectively, P<0.05). After 1 h of culture, compared with those in normal control group, the proportion and fluorescence intensity of phagocytosis cells were significantly increased in Nr-CWS alone group (with t values of 6.86 and 6.73, respectively, P<0.05), and the above two indexes were significantly decreased in LPS alone group (with t values of 7.35 and 22.72, respectively, P<0.05) and LPS+Nr-CWS group (with t values of 21.37 and 13.10, respectively, P<0.05). After 1 h of culture, compared with that in normal control group, the level of ROS of cells in LPS alone group was significantly increased ( t=6.64, P<0.05); compared with that in LPS alone group, the level of ROS of cells in LPS+Nr-CWS group was significantly decreased ( t=5.46, P<0.05). After 1 h of culture, compared with those in normal control group, the protein expressions of CD35, CD66b, and CD63 of cells were significantly increased in LPS alone group and LPS+Nr-CWS group (with t values of 16.75, 17.45, 10.82, 5.70, 19.35, and 15.37, respectively, P<0.05); compared with those in LPS alone group, the protein expressions of CD35, CD66b, and CD63 of cells were significantly decreased in LPS+Nr-CWS group (with t values of 4.92, 5.72, and 3.18, respectively, P<0.05). After 1 h of culture, compared with those in normal control group, the concentrations of IL-2, IL-4, IL-6, IL-10, IL-17A, TNF-α, and interferon-γ in cell culture supernatant were significantly increased in LPS alone group (with t values of 22.10, 9.50, 7.21, 10.22, 24.88, 8.43, and 47.48, respectively, P<0.05), and the concentrations of IL-6, IL-10, IL-17A, TNF-α, and interferon-γ in cell culture supernatant were significantly increased in LPS+Nr-CWS group (with t values of 4.68, 5.12, 8.02, 5.58, and 7.13, respectively, P<0.05); compared with those in LPS alone group, the concentrations of IL-2, IL-4, IL-6, IL-10, IL-17A, TNF-α, and interferon-γ in cell culture supernatant were significantly decreased in LPS+Nr-CWS group (with t values of 5.39, 2.83, 5.79, 2.90, 5.87, 4.88, and 39.64, respectively, P<0.05). Conclusions:Nr-CWS can enhance the phagocytosis ability of neutrophils in normal condition and improve the chemotactic function, ROS level, degranulation protein level, and inflammatory factor level of human neutrophils in infectious condition. Nr-CWS can enhance the anti-infection ability of human neutrophils by regulating its biological behavior in innate immunity.
7.Intervention of Osteoking in Rats with Myofascial Pain Syndrome
Xiaoxiao WANG ; Qun LI ; Zhixing HU ; Changting FANGLUO ; Ruirui MING ; Tengteng XU ; Chao YANG ; Wuqiong HOU ; Lili WANG ; Yini JIANG ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):19-29
ObjectiveTo clarify the intervention effect of Osteoking (OK) in rats with myofascial pain syndrome (MPS) and preliminarily explore the pharmacological mechanism of OK in relieving chronic pain from the perspective of anti-inflammatory disease. MethodThe 60 SD rats were divided into normal group, model group, low, medium, and high dose OK groups (0.66, 1.31, 2.63 mL·kg-1), and positive celecoxib group (21 mg·kg-1). The MPS rat model was established by beating combined with the centrifugal exercise method, and the OK and celecoxib were given at the same time. SMALGO paw pressure pain manometer detected the shock pain point tenderness threshold of rats, and the Von-Frey needle and acetone stimulation method detected the mechanical hyperalgesia threshold and cold hyperalgesia stimulation response respectively. Eight weeks and 10 weeks after modeling, the spontaneous discharge state and convulsion response of MPS rats were determined by electromyograph (EMG) instrument. The gait changes of MPS rats were detected using a CatWalk gait analyzer. The expression levels of interleukin-1 β (IL-1β), tumor necrosis factor-α (TNF-α), substance P (SP), and bradykinin (BK) were measured by enzyme-linked immunosorbent assay (ELISA). The protein expression levels of nuclear transcription factor-κB (NF-κB) inhibiting protein α (IκBα), phosphorylates (p)- IκBα, NF-κB p65, and p-NF-κB p65 were detected in MPS rats by Western blot. The positive expression of p-NF-κB p65 was detected by immunofluorescence. ResultCompared with the normal group, the model group shows 100% positive rates for EMG signal and local convulsions response at both the 8th and 10th weeks. The tenderness threshold and mechanical hyperalgesia threshold are significantly reduced. Cold hyperalgesia score is significantly increased, and gait is abnormal. The expression levels of serum and trigger points IL-1β, TNF-α, SP, BK, p-IκBα, and p-NF-κB p65, as well as the positive expression intensity of p-NF-κB p65 are significantly increased (P<0.01). Compared with the model group, the positive rate of EMG detection and local convulsion response is significantly reduced in the medium and high dose OK groups (P<0.05). The tenderness threshold and mechanical hyperalgesia threshold increase significantly in the medium and high dose OK groups, and the cold hyperalgesia score is significantly reduced in the high dose OK group (P<0.01). The standing time, swing time, and walking period are significantly increased. The swing speed, maximum contact area, and maximum contact intensity are significantly decreased in the high dose OK group (P<0.05). Moreover, the protein expression levels of p-IκBα/IκBα and p-NF-κB p65/NF-κB p65 are significantly reduced in the medium and high dose OK groups (P<0.05,P<0.01). The positive expression intensity of p-NF-κB p65 is significantly decreased in the high dose OK group (P<0.01). ConclusionThe mechanism of OK in relieving the pain in trigger points of MPS and improving gait abnormalities is related to the downregulation of the NF-κB p65 inflammatory signaling pathway to reduce the expression of inflammatory factors and pain mediators in blood and trigger point tissue.
8.Mechanism of Osteoking in Improving Energy Metabolism in Rats with Myofascial Pain Syndrome Based on cAMP/PKA/PGC1α Signaling Pathway
Xiaoxiao WANG ; Qun LI ; Chao YANG ; Changting FANGLUO ; Zhixing HU ; Lili WANG ; Suya ZAHNG ; Xueting LIU ; Yanqiong ZHANG ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(24):54-62
ObjectiveFrom the perspective of energy metabolism, the mechanism of Osteoking (OK) in the treatment of myofascial pain syndrome (MPS) was revealed through systems biology prediction combined with holistic animal experimental validation methods. MethodFirstly, the key targets of MPS and their related molecular mechanisms were predicted by the systems biology method, and the core network targets were screened. Then, the network-predicted targets were verified by animal experiments. Specifically, 60 SD rats were randomly divided into normal group, model group, low, medium, and high dose OK groups (0.66, 1.31, 2.63 mL·kg-1), and positive celecoxib group (21 mg·kg-1). The MPS model was established by beating combined with a centrifugal exercise method for eight weeks. Except for two days after modeling, the intervention of OK or celecoxib was performed. After the completion of the model, the drug was administered for two weeks. The histopathological changes of trigger point muscle tissue were observed by hematoxylin-eosin staining. The content/activity of Na-K-ATP enzyme (Na+-K+-ATPase), Ca2+ pump (Ca2+ATPase), Ca2+, lactate dehydrogenase (LDH), glutathione (GSH), malondialal (MDA), superoxide dismutase (SOD), cyclic adenosine phosphate (cAMP), and protein kinase A (PKA) in serum and/or trigger point muscle tissue in MPS rats was detected by enzyme-linked immunosorbent assay. Protein expression levels of PKA and the peroxisome proliferator-activated receptor γ coactivator 1α (PGC1α) in MPS rats were detected by immunohistochemistry. The protein expression levels of PKA, PGC1α, and mitochondrial transcription factor A (TFAM) in MPS rats were detected by Western blot. ResultThe network prediction results suggest that OK acts on the key target of energy metabolism related to the occurrence and development of MPS and may participate in the activation of the cAMP/PKA/PGC1α signaling pathway. The experimental validation results show that compared with the normal group, contracture nodules and disordered arrangement of muscle fibers appear in the trigger point muscle tissue of MPS rats. Na+-K+-ATPase, Ca2+ATPase, SOD activity, Ca2+, and GSH contents in serum and/or trigger point muscle tissue are significantly decreased (P<0.01). Both LDH activity and MDA contents are significantly increased (P<0.01), and the protein expression levels of cAMP, PKA, PGC1α, and TFAM are significantly decreased (P<0.01). Compared with the model group, OK improves the histopathological morphology of trigger point muscle fibers in MPS rats, and after the intervention of OK, Na+-K+-ATPase, Ca2+ATPase, SOD activity, Ca2+, and GSH contents in serum and/or trigger point muscle tissue in MPS rats are significantly increased (P<0.05, P<0.01). LDH activity and MDA contents are significantly reduced (P<0.05, P<0.01). The protein expression levels of cAMP, PKA, PGC1α, and TFAM are significantly increased (P<0.05, P<0.01). ConclusionThe mechanism of OK's intervention in MPS rats may be related to its effective activation of the cAMP/PKA/PGC1α signaling pathway, thus promoting mitochondrial energy metabolism and trigger point muscle fiber damage repair in muscle cells.
9.Anti-swelling and Analgesic Mechanism of Jianpi Tongluo Prescription from MAPKs Signaling Pathway
Ruirui MING ; Changting FANG-LUO ; Xiaoxiao WANG ; Zhixing HU ; Chao YANG ; Tengteng XU ; Hongjie WANG ; Chunfang LIU ; Na LIN
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(12):85-93
ObjectiveTo observe the anti-swelling and analgesic effects of Jianpi Tongluo prescription (JPTL) and to explore its mechanism initially. MethodA total of 120 ICR mice were divided into normal group, model group, JPTL low-, medium- and high-dose groups (5, 10, 20 g·kg-1) and positive drug (celecoxib, 0.03 g·kg-1) group, with 10 in each group (po,once a day). Complete freund's adjuvant (CFA) was used to induce the model of chronic inflammatory pain, and xylene-induced ear swelling test, hot plate test and acetic acid writhing test were performed to observe the anti-swelling and analgesic effects of different doses of JPTL in these four acute and chronic models. Further, enzyme-linked immunosorbent assay (ELISA) was used to detect the expressions of prostaglandin E2 (PGE2), interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum and inflammatory paw of mice with chronic inflammatory pain, and the expressions of aquaporin 1 (AQP1), aquaporin 3 (AQP3), cyclooxygenase 1 (COX1), cyclooxygenase 2 (COX2) and mitogen-activated protein kinases (MAPKs) in inflammatory paw were detected by Western blot, to explore the preliminary mechanism of JPTL. ResultCompared with the conditions in the normal group, there was a significant increase in the ear swelling of xylene-induced model mice, a shortened paw withdrawal latency in the hot plate test (P<0.01). Compared with the model group, JPTL remarkably increased the inhibition rate of xylene-induced ear swelling (P<0.05, P<0.01), prolonged the latency period of writhing caused by acetic acid and reduced the number of writhing (P<0.05, P<0.01). Compared with normal group, the degree of feet swelling in chronic inflammatory pain mice was significantly increased, the threshold of mechanical pain was decreased and the threshold of cold pain was increased (P<0.05, P<0.01), the protein contents of AQP1 and AQP3 in inflammatory feet were increased, and the contents of IL-1β, IL-6, TNF-α, PGE2 and COX2 in inflammatory feet were increased in serum and/or inflammatory feet. The protein expression levels of p-p38 MAPK, p-JNK and p-ERK in inflammatory feet were increased (P<0.01). Compared with the model group, JPTL relieved paw swelling of mice with chronic inflammatory pain, elevated mechanical withdrawal threshold while decreased cold withdrawal threshold, with analgesia lasting for 4 h and the optimal time point for analgesia being 2 h after administration (P<0.05, P<0.01). Moreover, JPTL down-regulated AQP1, AQP3, COX2, p-p38 MAPK, p-JNK and p-ERK in inflammatory paw of mice with chronic inflammatory pain and reduced IL-1β, IL-6, TNF-α, and PGE2 in serum and/or inflammatory paw, but it had no significant effect on COX1 (P<0.05, P<0.01). ConclusionJPTL has anti-swelling and analgesic effects, and its mechanism is related to inhibiting the production of cytokines and inflammatory mediators via the down-regulation of MAPKs signaling pathway, which provides an experimental basis for the clinical application of JPTL.
10.Sampling data of fresh frozen plasma, cryoprecipitates and leukocyte-free platelet quality monitoring in Hebei Province from 2015 to 2021: a retrospective analysis
Ying CHANG ; Zixuan ZHANG ; Xiaotong ZHANG ; Hongwei NIU ; Jianpeng ZHAO ; Chunfang LI ; Lexia LIU ; Wei ZHANG ; Fengxia YANG ; Xiaohong SUN
Chinese Journal of Blood Transfusion 2023;36(6):519-528
【Objective】 To form the sampling data interval by retrospectively analyzing the sampling data of quality monitoring of fresh frozen plasma, cryoprecipitates and leukocyte-free platelets in all blood stations in Hebei Province during the past 7 years. 【Methods】 The data of blood component sampling from 12 blood station quality control laboratories in Hebei from 2015 to 2021 were collected. The FⅧ content and plasma protein content of fresh frozen plasma, the FⅧ content and fibrinogen content of cryoprecipitates, and the leukocyte residuals, red blood cell mixed and platelet content of leukocyte-free platelets were taken as the objects for discrete point and fitted curve analysis. 【Results】 The FⅧ level of fresh frozen plasma: (1.36±1.1) IU/mL, 5 blood stations showed a representative overall high or low or fluctuated characteristics; Fresh frozen plasm-plasma protein items: overall mean ±SD: (61.13±16.7) g/L, four blood stations showed scattered distribution or continuous high value scattered points; Cryoprecipitates FⅧ: the overall mean ±SD: (134.25±58.7) IU/mL, four blood stations showed the differentiation characteristics of continuous high, low or stable in the middle; Cryoprecipitates-fibrinogen items: the overall mean ±SD: (215.27±83.5) mg, five blood stations showed the overall high or low and fluctuated. Leukocyte-free apheresis platelet-to-leukocyte residual items: overall mean ±SD: 0.37±0.96 (×106/bag), two blood stations showed a relatively high representative overall characteristics, and the rest were concentrated between 0 and 1; The total mean ±SD of platelet-to-red blood cell mixture without leukocyte was 2.45±2.82 (×109/bag), with obvious segmented concentrated distribution, and scattered distribution in 3 blood centers. Platelet content: the overall mean ±SD was 3.14±1.55 (×1011/bag), many deviations were noticed in 3 blood stations, and 1 blood station showed representative overall high characteristics. 【Conclusion】 This analysis shows that the distribution status of each blood station in different items is similar. The distribution status of discrete point groups and the change trend of the concentrated part of the fitting curve show that there are some differences in the monitoring level between the quality control laboratories of each blood station, and the update of detection instruments and reagents and the selection of detection methods greatly affect the test results. The summary data presented the index interval framework formed in the past 7 years, which helped to understand the difference between the results of each laboratory, correct the accuracy of the test results, better play the guiding role of quality monitoring in the blood preparation process, and continue to enhance the standardization of the whole process of blood collection and supply in the province.

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