1.Mechanism of Qingrun Prescription-containing Serum Improving Insulin Resistance in HepG2 Cells via Branched-chain α-keto Acid Dehydrogenase Regulation of Branched-chain Amino Acids (BCAAs)/mTOR Pathway
Xiangwei BU ; Xiaohui HAO ; Runyun ZHANG ; Meizhen ZHANG ; Ze WANG ; Haoshuo WANG ; Jie WANG ; Qing NI ; Lan LIN
Chinese Journal of Experimental Traditional Medical Formulae 2025;31(9):90-98
ObjectiveTo investigate the effect of Qingrun prescription(QRP)-containing serum on improving insulin resistance in HepG2 cells and its potential mechanisms. MethodsAn insulin resistance model was established in HepG2 cells with 1×10-6 mol·L-1 insulin. Branched-chain α-keto acid dehydrogenase (BCKDH) gene silencing was achieved using siRNA, and the cells were divided into 8 groups: normal group, model group (1×10-6 mol·L-1 insulin), metformin group (1 mmol·L-1 metformin), high-, medium-, and low-dose QRP groups (20%, 10%, and 5% QRP-containing serum, respectively), QRP + siRNA-silenced BCKDH (si-BCKDH) group (10% QRP-containing serum + si-BCKDH), and QRP + si-NC group (10% QRP-containing serum + si-NC). Glucose levels in the supernatant were measured with a glucose assay kit, while glycogen content was assessed using a glycogen assay kit. Levels of branched-chain amino acids (BCAAs) and branched-chain keto acids (BCKAs) were determined using ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). mRNA transcription and protein expression levels of BCKDH, dishevelled, Egl-10, and pleckstrin (DEP) domain-containing mammalian target of rapamycin (mTOR)-interacting protein (DEPTOR), mTOR, and ribosomal protein S6 kinase 1 (S6K1) were detected using real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot. ResultsCompared to the normal group, the model group exhibited significantly decreased glucose consumption and glycogen content, increased levels of BCAAs and BCKAs, downregulated expression of BCKDH and DEPTOR, and upregulated mTOR and S6K1 expression (P<0.01). In comparison to the model group, QRP treatment at all doses significantly enhanced glucose consumption and glycogen content while reducing BCAAs and BCKAs levels (P<0.01). The high- and medium-dose QRP groups demonstrated significant upregulation of BCKDH mRNA transcription and protein expression, as well as DEPTOR mRNA transcription. Moreover, the DEPTOR protein expression level was significantly increased in high-, medium-, and low-dose QRP groups, while mTOR and S6K1 mRNA and protein expression levels were markedly downregulated (P<0.05, P<0.01). Compared to the QRP + si-NC group, the QRP + si-BCKDH group exhibited increased BCAAs and BCKAs levels, significantly decreased BCKDH mRNA transcription and protein expression, downregulated DEPTOR mRNA and protein expression, and upregulated mTOR and S6K1 mRNA and protein expression (P<0.05, P<0.01). ConclusionQRP may improve insulin resistance by reprogramming BCAAs metabolism. This effect involves upregulating BCKDH, reducing BCAAs and BCKAs levels, and suppressing the mTOR pathway activation.
2.Mechanism of Qingrun Decoction in alleviating hepatic insulin resistance in type 2 diabetic rats based on amino acid metabolism reprogramming pathways.
Xiang-Wei BU ; Xiao-Hui HAO ; Run-Yun ZHANG ; Mei-Zhen ZHANG ; Ze WANG ; Hao-Shuo WANG ; Jie WANG ; Qing NI ; Lan LIN
China Journal of Chinese Materia Medica 2025;50(12):3377-3388
This study aims to investigate the mechanism of Qingrun Decoction in alleviating hepatic insulin resistance in type 2 diabetes mellitus(T2DM) rats through the reprogramming of amino acid metabolism. A T2DM rat model was established by inducing insulin resistance through a high-fat diet combined with intraperitoneal injection of streptozotocin. The model rats were randomly divided into five groups: model group, high-, medium-, and low-dose Qingrun Decoction groups, and metformin group. A normal control group was also established. The rats in the normal and model groups received 10 mL·kg~(-1) distilled water daily by gavage. The metformin group received 150 mg·kg~(-1) metformin suspension by gavage, and the Qingrun Decoction groups received 11.2, 5.6, and 2.8 g·kg~(-1) Qingrun Decoction by gavage for 8 weeks. Blood lipid levels were measured in different groups of rats. Pathological damage in rat liver tissue was assessed by hematoxylin-eosin(HE) staining and oil red O staining. Transcriptome sequencing and untargeted metabolomics were performed on rat liver and serum samples, integrated with bioinformatics analyses. Key metabolites(branched-chain amino acids, BCAAs), amino acid transporters, amino acid metabolites, critical enzymes for amino acid metabolism, resistin, adiponectin(ADPN), and mammalian target of rapamycin(mTOR) pathway-related molecules were quantified using quantitative real-time polymerase chain reaction(qRT-PCR), Western blot, and enzyme-linked immunosorbent assay(ELISA). The results showed that compared with the normal group, the model group had significantly increased serum levels of total cholesterol(TC), triglycerides(TG), low-density lipoprotein cholesterol(LDL-C), and resistin and significantly decreased ADPN levels. Hepatocytes in the model group exhibited loose arrangement, significant lipid accumulation, fatty degeneration, and pronounced inflammatory cell infiltration. In liver tissue, the mRNA transcriptional levels of solute carrier family 7 member 2(Slc7a2), solute carrier family 38 member 2(Slc38a2), solute carrier family 38 member 4(Slc38a4), and arginase(ARG) were significantly downregulated, while the mRNA transcriptional levels of solute carrier family 1 member 4(Slc1a4), solute carrier family 16 member 1(Slc16a1), and methionine adenosyltransferase(MAT) were upregulated. Furthermore, the mRNA transcription and protein expression levels of branched-chain α-keto acid dehydrogenase E1α(BCKDHA) and DEP domain-containing mTOR-interacting protein(DEPTOR) were downregulated, while mRNA transcription and protein expression levels of mTOR, as well as ribosomal protein S6 kinase 1(S6K1), were upregulated. The levels of BCAAs and S-adenosyl-L-methionine(SAM) were elevated. The serum level of 6-hydroxymelatonin was significantly reduced, while imidazole-4-one-5-propionic acid and N-(5-phospho-D-ribosyl)anthranilic acid levels were significantly increased. Compared with the model group, Qingrun Decoction significantly reduced blood lipid and resistin levels while increasing ADPN levels. Hepatocytes had improved morphology with reduced inflammatory cells, and fatty degeneration and lipid deposition were alleviated. Differentially expressed genes and differential metabolites were mainly enriched in amino acid metabolic pathways. The expression levels of Slc7a2, Slc38a2, Slc38a4, and ARG in the liver tissue were significantly upregulated, while Slc1a4, Slc16a1, and MAT expression levels were significantly downregulated. BCKDHA and DEPTOR expression levels were upregulated, while mTOR and S6K1 expression levels were downregulated. Additionally, the levels of BCAAs and SAM were significantly decreased. The serum level of 6-hydroxymelatonin was increased, while those of imidazole-4-one-5-propionic acid and N-(5-phospho-D-ribosyl)anthranilic acid were decreased. In summary, Qingrun Decoction may improve amino acid metabolism reprogramming, inhibit mTOR pathway activation, alleviate insulin resistance in the liver, and mitigate pathological damage of liver tissue in T2DM rats by downregulating hepatic BCAAs and SAM and regulating key enzymes involved in amino acid metabolism, such as BCKDHA, ARG, and MAT, as well as amino acid metabolites and transporters.
Animals
;
Drugs, Chinese Herbal/administration & dosage*
;
Rats
;
Insulin Resistance
;
Diabetes Mellitus, Type 2/genetics*
;
Male
;
Liver/drug effects*
;
Amino Acids/metabolism*
;
Rats, Sprague-Dawley
;
Humans
;
Metabolic Reprogramming
3.The role of 8-OxoG and its repair systems in liver diseases progression: responsible mechanisms and promising natural products.
Ying ZHENG ; Junxin CHEN ; Ze LIU ; Kaibo WANG ; Hao ZHANG
Chinese Journal of Natural Medicines (English Ed.) 2025;23(7):815-823
The accumulation of deoxyribonucleic acid (DNA) oxidative damage mediated by reactive oxygen species (ROS) is closely associated with liver diseases. 8-Oxoguanine (8-OxoG), a prevalent DNA oxidation product, plays a significant role in liver disease progression. The base excision repair (BER) pathway, comprising over 30 proteins including 8-OxoG DNA glycosylase1 (OGG1), MutY homolog (MUTYH), and MutT homolog protein 1 (MTH1), is responsible for the clearance and mismatch repair of 8-OxoG. Abnormally high levels of 8-OxoG and dysregulated expression and function of 8-OxoG repair enzymes contribute to the onset and development of liver diseases. Consequently, targeting the 8-OxoG production and repair system with agonists or inhibitors may offer a promising approach to liver disease treatment. This review summarizes the impact of 8-OxoG accumulation and dysregulated repair enzymes on various liver diseases, including viral liver disease, alcoholic liver disease (ALD), metabolic dysfunction-associated steatotic liver disease (MASLD), cholestatic liver disease (CLD), liver fibrosis, cirrhosis, and liver cancer. Additionally, we review natural constituents as potential therapeutic agents that regulate 8-OxoG production, repair enzymes, and repair system-related signal pathways in oxidative damage-induced liver diseases.
Humans
;
Liver Diseases/genetics*
;
Biological Products/pharmacology*
;
DNA Repair/drug effects*
;
Guanine/metabolism*
;
Animals
;
Disease Progression
;
DNA Damage
;
Oxidative Stress
4.Research on Automatic Microalgae Detection System Based on Deep Learning
Rui-Jie XIANG ; Hao LIU ; Zhen LU ; Ze-Yu XIAO ; Hai-Peng LIU ; Yin-Chu WANG ; Xiao PENG ; Wei YAN
Progress in Biochemistry and Biophysics 2024;51(1):177-189
ObjectiveThe scale of microalgae farming industry is huge. During farming, it is easy for microalgae to be affected by miscellaneous bacteria and other contaminants. Because of that, periodic test is necessary to ensure the growth of microalgae. Present microscopy imaging and spectral analysis methods have higher requirements for experiment personnel, equipment and sites, for which it is unable to achieve real-time portable detection. For the purpose of real-time portable microalgae detection, a real-time microalgae detection system of low detection requirement and fast detection speed is needed. MethodsThis study has developed a microalgae detection system based on deep learning. A microscopy imaging device based on bright field was constructed. With imaged captured from the device, a neural network based on YOLOv3 was trained and deployed on microcomputer, thus realizing real-time portable microalgae detection. This study has also improved the feature extraction network by introducing cross-region residual connection and attention mechanism and replacing optimizer with Adam optimizer using multistage and multimethod strategy. ResultsWith cross-region residual connection, the mAP value reached 0.92. Compared with manual result, the detection error was 2.47%. ConclusionThe system could achieve real-time portable microalgae detection and provide relatively accurate detection result, so it can be applied to periodic test in microalgae farming.
5.Development and application of portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma
Ze WANG ; Hao CUI ; Runkai CHEN ; Zhen YUAN ; Bo WEI ; Jianxin CUI
China Medical Equipment 2024;21(10):29-35
Objective:To design and develop a portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma,which aimed at a scene of rescue environment at frontline,and its feasibility was evaluated preliminarily through animal experiment.Methods:Based on the actual demands of the rescue environment at frontline,a set of portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma(abbreviation:minimally invasive diagnostic and therapeutic device)was researched,developed and assembled,which included portably integrated host,disposable flexible lens of endoscope,disposable apparatus of minimally invasive surgery,extendable channel device of avoiding pneumoperitoneum and so on.A male Bama miniature pig was selected,and it received two different surgeries included portably minimally invasive diagnostic and therapeutic device,and conventionally laparoscopic surgery after it received general anesthesia.The damage controls included hemostasis of intraoperative parenchyma organ,sealing and repairing of gastrointestinal perforation and drainage of indwelling catheter in abdominal and pelvic cavity between two groups were compared,and the difference of the mobility performance between them also was compared.The operational evaluation of minimally invasive surgery of damage control surgery and the potential of its clinical conversion were conducted.Results:Compared to conventional laparoscopy,this minimally invasive diagnostic and therapeutic device had better mobility,and the transfer time of this device was(3.3±1.0)min,which was significantly shorter than(14.5±3.2)min of conventional laparoscopy,and the difference of that between two device was significant(t=-5.786,P<0.05).The minimally invasive diagnostic and therapeutic device could successfully realize a series of operation of damage control surgery included exploration,flushing,suction,hemostasis,repair and drainage under the pneumoperitoneum or without pneumoperitoneum,which operation was safety and feasibility.Conclusion:The portably minimally invasive diagnostic and therapeutic device for abdominally warfare trauma can realize integration and optimization,and mobility and portability on the basis of the current laparoscopic platform,which can successfully realize the operation of damage control surgery.It has favorable application prospects and capabilities of clinical conversion.
6.Effects of Chinese herbal formula Er-miao-san on Gnαq expression and improvement of inflammatory reaction in CIA model rats
Wenjing LU ; Zhen LI ; Huiqin HAO ; Yuting GAO ; Ze WANG ; Jin LIU
Chinese Journal of Immunology 2024;40(6):1222-1227
Objective:To explore the effect of different concentrations of Chinese herbal formula,Er-miao-san(EMS)immune regulatory gene on Gnαq expression in rats with collagen-induced arthritis(CIA).Methods:In the study,we established a CIA model,and randomly divided into model groups,EMS group with 1.5,3.0 and 4.5 g/kg(L-EMS,M-EMS,H-EMS)were adminis-tered to CIA rats by gavage,methotrexate was used as a positive control group(MTX).The rats in each group were given correspond-ing drugs for treatment,and the normal control group and model group were given the same amount of normal saline gavage.ELISA detects the expression of IL-1β,IL-6 and TNF-α in rats'serum.The mRNA and protein expression of Gnαq were analyzed using qRT-PCR and Western blot.The localization of Gnαq was performed by IHC.Results:The joint of rats in CIA group showed obvious damage,tissue hyperplasia and inflammatory infiltration;the joint tissue destruction and proliferation of MTX and EMS groups were reduced;the expressions of IL-1β,IL-6 and TNF-α in CIA group were extremely increased(P<0.01).The expressions of IL-1β(except H-EMS group),IL-6 and TNF-α in MTX and EMS groups were significantly lower than those in CIA group(P<0.05).The expressions of IL-1β,IL-6 and TNF-α in L-EMS group were lower than those in the M-EMS and H-EMS groups(P<0.05,P<0.01).The expression of Gnαq mRNA and protein in spleen and joints of CIA group were higher than those of Control group(P<0.01).The expression of Gnαq mRNA and protein in spleen and joints of EMS and MTX groups were lower than those of CIA group(P<0.05).The expression of Gnαq mRNA and protein in the dose of 1.5 g/kg of EMS group was lower than those of the other groups(P<0.01,P<0.05).Different degree of posi-tive signal was detected in spleen and joints in different groups.Conclusion:Gnαq is expressed in spleen and joints of CIA rats,may participate in the formation of inflammatory response in CIA rats,and then mediate the formation and development of RA by down-reg-ulating the expression of Gnαq.
7.Maintenance and improvement program for WYD2000 field surgical lamp
Ming-Ming ZHU ; Ji-Su CAO ; Ze-Rui ZHANG ; Jiang-Hui HAO ; Rui-Zhe WANG ; Feng ZHOU ; Lu-Chuan XUN
Chinese Medical Equipment Journal 2024;45(4):116-118
The steps for installation and withdrawal of WYD2000 field surgical lamp were introduced.The failures and causes of broken cross-arm connector of WYD2000 field surgical lamp were analyzed.The problems of WYD2000 field surgical lamp in vulnerability to breaking and difficulty in maintenance were solved by designing and manufacturing a special maintenance tool and optimizing the materials and fixing mode of cross-arm connection.References were provided for main-tenance and improvement of WYD2000 field surgical lamp.[Chinese Medical Equipment Journal,2024,45(4):116-118]
8.Comparative study of two fixation methods in single segment interbody fusion and fixation with unilateral dual channel spinal endoscopic technique.
Ze-Xuan WU ; Tao WANG ; Zhi-Gang LEI ; Xiao-Lin LI ; Hao-Zhe LIU ; Xiao-Yang LI ; Xin-Xin BAI ; Hong-Hai XU
China Journal of Orthopaedics and Traumatology 2024;37(12):1158-1163
OBJECTIVE:
To explore clinical efficacy of unilateral and bilateral nail-rod system fixation in single-level intervertebral fusion fixation of lumbar disc herniation (LDH) and instability with unilateral dual-channel spinal endoscopy.
METHODS:
The clinical data of 63 patients with LDH complicated with instability treated by unilateral double-channel spinal endoscopy from March 2021 to June 2022 were retrospectively analyzed. According to intraoperative fixation methods, the patients were divided into two groups, included unilateral nail rod system fixation group(unilateral group) and bilateral nail rod system fixation group(bilateral group). There were 31 patients in unilateral group, including 15 males and 16 females; aged from 40 to 67 years old with an average of (54.65±7.32) years old; 3 patients with L3,4, 15 patients with L4,5, 13 patients with L5S1. There were 32 patients in bilateral group, including 18 males and 14 females, aged from 43 to 68 years old with an average of (56.19±6.63) years old;4 patients with L3,4, 17 patients with L4,5, 11 patients with L5S1. The time of operation, length of incision, operation time, length of hospital stay, cost of treatment, occurrence of complications and adverse events were recorded and compared between two groups. Visual analogue scale (VAS) of lumbar and leg pain, Japanese Orthopaedic Association (JOA) and Oswestry disability index (ODI) were compared before operation, 3 and 12 months after operation. MacNab standard was used to evaluate clinical effect.
RESULTS:
Both of two groups were successfully completed operation and followed up. In unilateral fixation group, follow-up time, operation time, incision length, postoperative ambulation time, hospital stay and treatment cost were (11.81±1.08) months, (122.48±13.86) min, (3.15±0.45) cm, (3.06±0.73) d, (6.87±1.09) d, and (3.49±0.09) ten thousand yuan;while (11.47±1.16) months, (134.94±10.94) min, (6.45±0.83) cm, (3.19±0.86) d, (7.03±1.03) d, (3.77±0.12) ten thousand yuan in bilateral fixation group, respectively;there were significant differences in operation time, incision length and treatment cost between two groups (P<0.05), no significant difference in follow-up time, postoperative ambulation time and hospital stay (P>0.05). There were no significant difference in VAS of low back pain, JOA score, ODI and MacNab grade between two groups before and after operation (P>0.05). No significant complications and adverse events were observed between two groups after operation.
CONCLUSION
Both of unilateral and bilateral nail rod system for the treatment of LDH with instability could achieve the expected results. Unilateral fixation has more advantageous than bilateral fixation in terms of operation time, incision length and treatment cost. For patients with osteoporosis before operation or insufficient fixation strength of unilateral pedicle nails during surgery, bilateral fixation is feasible to enhance reliability of fixation.
Humans
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Male
;
Female
;
Middle Aged
;
Spinal Fusion/methods*
;
Endoscopy/methods*
;
Aged
;
Adult
;
Intervertebral Disc Displacement/surgery*
;
Lumbar Vertebrae/surgery*
;
Retrospective Studies
9.Current treatment for male infertility: an umbrella review of systematic reviews and meta-analyses.
Jian-Jun YE ; Ze-Yu CHEN ; Qi-Hao WANG ; Xin-Yang LIAO ; Xing-Yuan WANG ; Chi-Chen ZHANG ; Liang-Ren LIU ; Qiang WEI ; Yi-Ge BAO
Asian Journal of Andrology 2024;26(6):645-652
This umbrella review aimed to summarize and provide a general evaluation of the effectiveness of current treatments for male infertility and assess the quality of evidence and possible biases. An umbrella review of systematic reviews and meta-analyses available in PubMed, Web of Science, and Scopus, covering studies published up to October 2023, was conducted. Sperm concentration, morphology, and motility were used as endpoints to evaluate the effectiveness of the treatments. Of 2998 studies, 18 published meta-analyses were extracted, yielding 90 summary effects on sperm concentration ( n = 36), sperm morphology ( n = 26), and sperm motility ( n = 28) on 28 interventions. None of the meta-analyses were classified as having low methodological quality, whereas 12 (66.7%) and 6 (33.3%) had high and moderate quality, respectively. Of the 90 summary effects, none were rated high-evidence quality, whereas 53.3% ( n = 48), 25.6% ( n = 23), and 21.1% ( n = 19) were rated moderate, low, and very low, respectively. Significant improvements in sperm concentration, morphology, and motility were observed with pharmacological interventions (N-acetyl-cysteine, antioxidant therapy, aromatase inhibitors, selective estrogen receptor modulators, hormones, supplements, and alpha-lipoic acid) and nonpharmacological interventions (varicocele repair and redo varicocelectomy). In addition, vitamin supplementation had no significant positive effects on sperm concentration, motility, or morphology. Treatments for male infertility are increasingly diverse; however, the current evidence is poor because of the limited number of patients. Further well-designed studies on single treatment and high-quality meta-analysis of intertreatment comparisons are recommended.
Humans
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Male
;
Antioxidants/therapeutic use*
;
Infertility, Male/therapy*
;
Meta-Analysis as Topic
;
Sperm Count
;
Sperm Motility
;
Systematic Reviews as Topic
10.Effects of Anterior Cruciate Ligament Reconstruction Timing on Post-Operative Motor Performance and Proprioception.
Xiang-Yi WANG ; Jing-Yi SUN ; Chen HE ; Yi QIAN ; Sen GUO ; Xiao-Han ZHANG ; Hao XU ; Zhuang LIU ; Ya-Wei GONG ; Lei LI ; Ming-Ze LIU ; Feng GAO ; Jing-Bin ZHOU
Acta Academiae Medicinae Sinicae 2024;46(6):797-804
Objective To evaluate the effects of anterior cruciate ligament (ACL) reconstruction timing on the motor performance and proprioception by clinical evaluation as well as proprioception and motor performance tests on the patients more than 2 years after ACL reconstruction. Methods The patients who underwent ACL reconstruction in the National Institute of Sports Medicine,General Administration of Sport of China from January 2015 to January 2021 and met the inclusion criteria were followed up,and the postoperative data were collected retrospectively.Fifty-six patients who met the inclusion criteria were included in this study and categorized into two groups:early surgery (n=28,who underwent ACL reconstruction ≤3 weeks after injury) and delayed surgery (n=28,who underwent ACL reconstruction >3 weeks after injury).The basic information,clinical evaluation results,proprioception,and motor performance were compared between the two groups. Results The ACL return to sport after injury scale (ACL-RSI) score in the early surgery group was higher than that in the delayed surgery group [(68.68±22.04)scores vs. (55.82±24.87)scores,P=0.045].There was no difference in the range of motion of the knee joint,the positive rate of pivot shift test,or the scores of Tegner,Marx,Lysholm,knee injury and osteoarthritis outcome score (KOOS),and international knee documentation committee (IKDC) between the two groups (all P>0.05).Although there was no significant difference in range of motion of the knee joint between the two groups,the proportion of knee flexion and extension affected in the early surgery group was smaller than that in the delayed surgery group.Neither motor performance (isokinetic strength test,Y-balance test,and single-leg jump test) nor proprioception had difference between the two groups (all P>0.05). Conclusions Early ACL reconstruction outperformed delayed ACL reconstruction in improving the psychological health,emotions,and confidence in returning to sport,accelerating functional recovery of the patients.The timing of ACL reconstruction has no significant effect on the short-term postoperative knee stability,knee function,motor performance,or proprioceptive recovery of the patients.Early ACL reconstruction is recommended for improving the clinical outcomes.
Humans
;
Anterior Cruciate Ligament Reconstruction/methods*
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Proprioception/physiology*
;
Retrospective Studies
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Male
;
Female
;
Range of Motion, Articular
;
Anterior Cruciate Ligament Injuries/physiopathology*
;
Adult
;
Postoperative Period
;
Time Factors
;
Return to Sport
;
Recovery of Function
;
Knee Joint/physiopathology*
;
Young Adult

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