1.Association of physical activity and sugar sweetened beverage consumption with psychological sub health among middle school students in Bao an District, Shenzhen
Chinese Journal of School Health 2025;46(1):102-105
Objective:
To explore the association of physical activity and sugar sweetened beverage consumption with psychological sub health among middle school students in Bao an District, Shenzhen, so as to provide a reference for adolescent mental health promotion.
Methods:
A questionnaire survey was conducted in November 2024 by a stratified cluster random sampling method to select 6 926 junior and senior middle school students from 5 middle schools in Shenzhen. The questionnaire from Youth Risk Behavior Surveillance System was used to assess the consumption of sugar sweetened beverages, and physical activity Rating Scale was used to assess the level of physical activity, and Brief Instrument on Psychological Health of Youths was used to evaluate the psychological sub health status. The Chi -square test was used to analyze the differences in the detection rates of psychological sub health among different groups of middle school students, and a multivariate Logistic regression model was established to analyze the effects of physical activity and sugar sweetened beverage consumption and their combined effects on the psychological sub health of middle school students.
Results:
The detection rate of psychological sub health among middle school students in Bao an District, Shenzhen was 18.93%. Multivariate Logistic regression analysis showed that, after controlling for confounding factors such as gender, school stage, family residence, family economic status, parental literacy, academic stress and number of friends, lack of physical activity or excessive sugar sweetened beverage consumption were associated with increased risks of psychological sub health among middle school students ( OR =1.36, 1.45); and the highest risk of psychological sub health was found in middle school students who were lack of physical activity and excessive sugar sweetened beverage consumption ( OR =2.59) ( P <0.01). Further analysis by school stages showed that junior high school students with sufficient physical activity and excessive intake of sugary drinks ( ROR =2.10), lack of physical activity and excessive intake of sugary drinks ( ROR =2.31) were at higher risks of psychological sub health than senior high school students( P <0.05).
Conclusions
Insufficient physical activity and excessive sugar sweetened beverage consumption are closely associated with an increased risk of psychological sub health among middle school students. Effective interventions should be targeted to reduce the risk of psychological sub health problems among middle school students.
2.Meta analysis of physical activity on the improvement of health related quality of life among children and adolescents
LIU Shanjie, LI Gai, SONG Yunfeng, OUYANG Lingqing, XU Chi
Chinese Journal of School Health 2025;46(6):806-810
Objective:
To systematically review the effect of physical activity (PA) on health related quality of life (HRQoL) among children and adolescents and the magnitude of association, so as to provide an evidence based support for optimizing health promotion strategies for children and adolescents.
Methods:
Medline (PubMed), EMBASE, the Cochrane Library, SCIELO, PEDro, CNKI, Wanfang and VIP databases were searched to collect literature on PA and HRQoL in children and adolescents, which were published up to January 1, 2025. Cochrane ROB tool and specific evaluation system were used to assess literature quality, Stata 16.0 software was used for Meta analysis. For descriptive studies, correlation( r ) and 95% confidence interval( CI ) were used as effect indicators, and the inverse variance method was used to merge the data. Standardized mean difference( SMD ) and 95% CI were used to calculate the combined effect size of intervention studies.
Results:
A total of 22 studies involving 27 303 healthy children and adolescents were included, including 9 descriptive studies and 13 intervention studies. The results of descriptive analysis showed that PA level was positively correlated with HRQoL ( r =0.27, 95% CI =0.21-0.32, P <0.01). The analysis of intervention studies showed that PA had significant effects on overall HRQoL ( SMD =0.10, 95% CI =0.05-0.16), physical functioning ( SMD =0.16, 95% CI =0.08-0.23), and mental functioning ( SMD =0.17, 95% CI =0.10-0.25)( P <0.01). Subgroup analysis found that the effects of physical activity on overall HRQoL, physiological function, and psychological function in children and adolescents were maximized when the intervention duration was less than 6 months ( SMD =0.13, 0.18, 0.20), the frequency was≥5 times per week ( SMD =0.13, 0.19, 0.24) and there was a supplementary plan( SMD =0.10, 0.18, 0.23)( P <0.01).
Conclusion
PA is an important factor affecting the HRQoL of healthy children and adolescents, and the systematic PA intervention has a effect on improving the HRQoL.
3.Engineering of mesenchymal stem cell-derived exosomes and their application progress in the field of oral medicine
ZHAO Yunfeng ; LIU Qian ; LI Meng ; LI Luying ; ZHANG Wei ; HU Xiantong ; MA Chufan
Journal of Prevention and Treatment for Stomatological Diseases 2025;33(9):792-800
In recent years, mesenchymal stem cell-derived exosomes (MSC-EXO) have garnered increasing attention in the field of stomatology and have become an established research area in biomedical research. This article reviews the engineering of exosomes derived from mesenchymal stem cells and their application in the field of stomatology, in order to provide new ideas for the development of stomatology. Exosomes are nanoscale membrane vesicles secreted by cells and contain a variety of proteins, RNAs, lipids, and other biomolecules. They are transported through the circulatory system and can interact with other cells to regulate their biological behavior and participate in a variety of physiological and pathological processes. In the treatment of oral diseases, exosomes have shown great potential due to their natural biological activity and versatility. However, studies have found that relying solely on the function of natural exosomes may not fully meet the complex clinical requirements. Therefore, the concept of engineered exosomes has emerged. Engineered exosomes can be modified by bioengineering technology to enhance their targeting, allowing them to reach the lesion site more accurately. At the same time, engineered exosomes can also be surface modified or loaded internally to carry specific therapeutic molecules, such as drugs, gene editing tools or signaling molecules to improve the therapeutic effect. In addition, this engineered treatment can also confer greater stability to exosomes, making them better able to resist clearance by the immune system when circulating in the body, extending their half-life, and improving the effectiveness of treatment. Although engineered exosomes have attracted extensive attention in the fields of stomatology and other fields, their application is still mainly in the stage of basic research. To promote the clinical application of engineered exosomes, it is necessary to provide more sufficient evidence of biocompatibility and clarify their therapeutic effect and mechanism.
4.Progress and Evaluation of Animal Model of Heart Qi-Yin Deficiency Syndrome
Yayi LIU ; Yunfeng JIA ; Yiming ZUO ; Junping ZHANG ; Shichao LÜ
Laboratory Animal and Comparative Medicine 2025;45(4):411-421
Animal models combining disease and syndrome are important research tools to explore the nature of traditional Chinese medicine (TCM) syndromes. At present, the construction and evaluation methods of animal models have preliminarily established the foundation for standardized development. Qi-yin deficiency syndrome is a common type of TCM syndrome in cardiovascular diseases. It is an important pathogenic factor causing the onset, pathological damage, and chronic nature of cardiovascular diseases, as well as triggering other illnesses. The establishment of an animal model of cardiovascular disease with the characteristics of Qi-yin deficiency, along with an objective and standardized evaluation system, has become an important part of modern cardiovascular disease research. In recent years, research on the construction and evaluation of animal models of heart Qi-yin deficiency syndrome has increased, but the construction methods and evaluation criteria vary. Compared with other animal models, the literature is limited, lacking statistics and overall analysis. Therefore, based on the scientific connotation of heart Qi-yin deficiency syndrome, this article systematically reviews the evaluation system of its animal model, covering multidimensional methods such as macroscopic characterization assessment, physicochemical indicators and objective evaluation, and syndrome differentiation based on prescriptions. The specific model construction strategies are described, including single-factor induction methods (sleep deprivation, chronic intermittent hypoxia, arterial occlusion, high-salt feeding) and the compound-factor induction methods (sleep deprivation combined with drug method, chronic intermittent hypoxia combined with drug method, exhaustive swimming combined with drug method). Meanwhile, application examples of each model in the research are listed, the existing problems in the current model construction and evaluation are analyzed, and optimization directions are proposed, such as promoting the compound factor induction strategy and improving the objectivity of the evaluation criteria. This article aims to provide theoretical references for constructing an animal model of heart Qi-yin deficiency syndrome that conforms to TCM characteristics, and thereby laying a scientific foundation for the prevention and treatment of cardiovascular diseases with TCM.
5.Perinatal characteristics and early postnatal circulatory function changes in the larger fetus in twin pregnancy with selective intrauterine growth restriction
Yahui ZHANG ; Dan WANG ; Yunfeng LIU ; Shi SHI ; Meihua PIAO
Chinese Journal of Perinatal Medicine 2024;27(3):188-195
Objective:To explore the perinatal characteristics and early postnatal circulatory function of the larger fetus in monochorionic diamniotic twin pregnancy complicated with selective intrauterine growth restriction (sIUGR).Methods:From February 2018 to August 2022, a total of 91 larger fetuses of the sIUGR pregnancies who were hospitalized in the neonatal intensive care unit of Peking University Third Hospital were retrospectively included. The perinatal factors, clinical monitoring indicators, and echocardiographic data of the larger twins in pregnancies with three types of sIUGR were compared using one-way analysis of variance and LSD test, Kruskal-Wallis H test, Chi-square test, and Bonferroni correction. Results:The gestational age, birth weight, and placental weight were (30.6±1.5) weeks, (1 503.9±286.4) g, and (548±120) g in the type Ⅱ sIUGR larger twins and were (30.5±2.3) weeks, (1 523.5±424.4) g, and (560±109) g in type Ⅲ, which were all smaller, lower, and lighter than those in the type Ⅰ[(33.0±1.7) weeks, (2 022.1±372.3) g, and (630±131) g, respectively] (LSD test, all P<0.05). Compared with type Ⅰ sIUGR larger twins, type Ⅱ and Ⅲ sIUGR larger twins had longer hospital stay [36.0 d (27.0-43.0 d) and 32.0 d (15.0-47.0 d) vs. 17.0 d (9.5-22.0 d)], higher proportion of preterm births due to fetal distress [63.6% (21/33) and 75.0% (15/20) vs. 31.6% (12/38), χ2=7.30 and 9.93] (Bonferroni correction, all P<0.017); Compared with type Ⅰ sIUGR larger twins, type Ⅱ sIUGR larger twins had higher proportion of postnatal use of vasoactive drugs [45.5% (15/33) vs. 18.4% (7/38), χ2=6.04, P=0.014]. The interventricular septum thickness was larger in the type Ⅲ sIUGR larger twins than those in the type Ⅰ and Ⅱ twins [(4.4±1.4) vs. (3.8±0.9) and (3.3±0.9) mm]; the thickness of left ventricular free wall was the largest in the type Ⅲ larger twins, followed by those in the type Ⅰ and type Ⅱ [(4.6±1.3) vs. (3.1±0.7) vs. (2.7±0.6) mm]; the left ventricular end-diastolic diameter, right ventricular outflow tract diameter, right ventricular anteroposterior diameter, and pulmonary artery diameter in type Ⅰ larger twins were increased comparing with those in type Ⅱ and Ⅲ [left ventricular end-diastolic diameter: (15.0±2.1) vs. (13.4±2.3) and (12.3±3.2) mm; right ventricular outflow tract diameter: (8.7±1.4) vs. (7.3±1.4) and (7.1±0.8) mm; right ventricular anteroposterior diameter: (7.1±1.5) vs. (6.5±0.9) and (6.4±1.0) mm; pulmonary artery diameter: (6.8±1.1) vs. (6.3±0.9) and (6.3±0.8) mm] (LSD test, all P<0.05). Conclusion:The larger fetuses of type Ⅱ and Ⅲ sIUGR pregnancies had smaller gestational age and lighter birth weight who are more prone to have fetal distress, so it is necessary to strengthen dynamic monitoring and circulatory support for such neonates during the perinatal period and early postnatal period. The thickening of the left ventricular wall and interventricular septum in the early postnatal period of type Ⅲ larger fetuses may lead to the decrease of ventricular diastolic function reserve, thereby the evaluation and monitoring of the myocardial diastolic function of these neonates in the early postnatal period are critical.
6.Study on the Effect of Intestinal Flora on Intestinal Motility in Rats with Slow Transit Constipation of Qi Stagnation Pattern Based on Pseudo-Sterile Experiment and Fecal Microbiota Transplantation
Qihong LIU ; Xiao KE ; Yunfeng LUO ; Lunan HU ; Yan REN ; Wenyi FANG ; Peilin ZHAO ; Jinxian YAN
Journal of Traditional Chinese Medicine 2024;65(9):943-948
ObjectiveTo clarify the relationship between intestinal flora and intestinal motility in rats with slow transit constipation (STC) and qi stagnation syndrome by conducting a pseudo-sterile experiment and fecal microbiota transplantation (FMT) technology. MethodsTwenty-four Wistar rats were randomly divided into normal group (n=6), STC with qi stagnation pattern group (n=6) and pseudo-sterile group (n=12). In the STC group with qi stagnation pattern, 3 mg/kg of loperamide suspension by intragastric administration combined with tail clamping stimulation were performed to establish the rat model of STC with qi stagnation pattern. After successful modeling, fresh feces from the rats in the STC with qi stagnation pattern group and the normal group were collected to prepare 100 mg/ml of fecal bacterial suspension. In the pseudo-sterile group, the antibiotic cocktail method was used (a mixed antibiotic suspension containing bacitracin, streptomycin sulfate, and neomycin sulfate at 20 mg/ml each was administered intragastrically) to establish pseudo-sterile rats model. After successful modeling, the rats were randomly divided into normal fecal bacterial liquid group and STC with qi stagnation pattern fecal bacterial liquid group, with six rats in each group, and then were given 10 ml/kg of the prepared corresponding rat fecal bacterial suspension by gavage. Rats in STC with qi stagnation pattern group were given an equal volume of sterile water by gavage. All groups were administered once a day for 7 consecutive days. The small intestinal propulsion rate of the STC with qi stagnation pattern group, the normal fecal bacterial liquid group, and STC with qi stagnation pattern fecal bacterial liquid group were compared. ELISA method was used to detect serum 5-hydroxytryptamine (5-HT) levels. Immunohistochemistry was used to detect the positive expression levels of 5-hydroxytryptamine 3 receptor (5-HT3R) and 5-hydroxytryptamine 4 receptor (5-HT4R) in colon tissue. Western blot method was used to detect the protein expression levels of tryptophan hydroxylase 1 (TPH1), tryptophan hydroxylase 2 (TPH2), serotonin transporter (SERT), and monoamine oxidase A (MAO-A) in colon tissue. ResultsCompared to those in the normal fecal bacterial liquid group, the small intestinal propulsion rate, serum 5-HT level, positive expression of 5-HT3R and 5-HT4R in colon tissue, and protein expression of TPH1, TPH2, SERT and MAO-A significantly decreased in the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group (P<0.05). There was no statistically significant difference in the indicators between the STC with qi stagnation pattern group and STC with qi stagnation pattern fecal bacterial liquid group (P>0.05). ConclusionThe intestinal flora in STC rats with qi stagnation pattern can lead to a slowdown in intestinal transmission function, whose mechanism may be related to intestinal motility disorders affected by the synthesis, transport, metabolism and other pathways of 5-HT.
7.Finite element model establishment and stress analysis of lumbar-sacral intervertebral disc in ankylosing spondylitis
Zhijie KANG ; Zhenhua CAO ; Yangyang XU ; Yunfeng ZHANG ; Feng JIN ; Baoke SU ; Lidong WANG ; Ling TONG ; Qinghua LIU ; Yuan FANG ; Lirong SHA ; Liang LIANG ; Mengmeng LI ; Yifei DU ; Lin LIN ; Haiyan WANG ; Xiaohe LI ; Zhijun LI
Chinese Journal of Tissue Engineering Research 2024;28(6):840-846
BACKGROUND:Ankylosing spondylitis is a chronic inflammatory disease with chronic rheumatic immunity.Soft tissue ossification and fusion and spinal stiffness can cause biomechanical changes. OBJECTIVE:To reconstruct the lumbar-sacral intervertebral disc in ankylosing spondylitis patients with lumbar kyphosis by finite element analysis,and to study the range of motion of each segment of T11-S1 and the biomechanical characteristics of annulus fibrosus and nucleus pulposus. METHODS:The imaging data were obtained from an ankylosing spondylitis patient with lumbar kyphosis.The original CT image data of continuously scanned spine were imported into Mimics 21.0 in DICOM format,and T11-S1 was reconstructed respectively.The established model was imported into 3-Matic software in the format of"Stl"to reconstruct the intervertebral disc,and the fibrous intervertebral disc model was obtained.The improved model was further imported into Hypermesh software,and the vertebra,nucleus pulposus,annulus fibrosus and ligament were mesh-divided.After the material properties were given,the model was imported into ABAQUS software to observe the range of motion of each vertebral body in seven different working conditions of T11-S1,and analyze the biomechanical characteristics of each segment of annulus fibrosus and nucleus pulposus. RESULTS AND CONCLUSION:(1)The range of motion of L1 vertebrae was higher than that of other vertebrae under six different working conditions:extension,forward flexion,rotation(left and right),and lateral flexion(left and right).The maximum range of motion was 2.18° during L1 vertebral flexion,and the minimum range of motion was 0.12° during L5 vertebral extension.(2)The annular fiber flexion at L2-L3 segments was greater than the extension(P<0.05),and the annular fiber flexion at L3-L4 and L4-L5 segments was less than the extension(P<0.05).The left rotation of L1-L2 annular fibers was greater than the right rotation(P<0.05).The left flexion of the annulus was greater than the right flexion in L1-L2,L2-L3,L3-L4,L4-L5 and L5-S1 segments(P<0.05).(3)The nucleus pulposus stresses of T11-L12,L1-L2,L2-L3,L3-L4 and L4-L5 segments in forward flexion were greater than in extension(P<0.05).The left rotation of T12-L1 and L3-L4 segments was smaller than the right rotation(P<0.05),and that of T11-T12,L1-L2,and L2-L3 segments was larger than the right rotation(P<0.05).The left flexion was larger than the right flexion in the T11-S1 segment.(4)It is concluded that in ankylosing spondylitis patients with lumbar kyphosis,the minimum range of motion of the vertebral body is located at the L5 vertebral body in extension.To prevent fractures,it is recommended to avoid exercise in the extension position.During the onset of lumbar kyphosis in patients with ankylosing spondylitis,the maximum stress of the annulus fibrosus and nucleus pulposus is located in the L1-L2 segment,which is fixed and will not alter with the change of body position.The late surgical treatment and correction of deformity should focus on releasing the pressure of the annulus fibrosus and nucleus pulposus in this segment to avoid the rupture of the annulus fibrosus and the injury of the nucleus pulposus.
8.Repetitive magnetic stimulation of S3 nerve root and M1 area for treating urinary retention after spinal cord injury
Zihan XU ; Yunfeng BI ; Jiang LI ; Zongliang ZHANG ; Chen SONG ; Jie DONG ; Dong LIU
Chinese Journal of Tissue Engineering Research 2024;28(11):1719-1723
BACKGROUND:Repetitive magnetic stimulation of either S3 nerve root or M1 area can improve the urination function of patients with urinary retention after spinal cord injury,but there are few reports on the repetitive magnetic stimulation of both sites in patients with urinary retention after spinal cord injury. OBJECTIVE:To observe the effect of repetitive magnetic stimulation of both S3 nerve root and M1 area on urinary retention after spinal cord injury. METHODS:Forty patients with urinary retention after spinal cord injury were enrolled and were randomly divided into two groups(n=20 per group):group A(repetitive magnetic stimulation in both S3 nerve root and M1 area)and group B(repetitive magnetic stimulation in the S3 nerve root and sham stimulation in the M1 area).Patients in both groups were given 4-week repetitive magnetic stimulation based on conventional bladder function intervention.The stimulation time and duration of treatment were same in both groups,with a treatment time of 21 minutes daily,5 days per week,for 4 weeks in total.The urination diary and urodynamics were compared between two groups. RESULTS AND CONCLUSION:Before treatment,there were no statistically significant differences in the average daily catheterization times,average daily catheterization volume,average single urinary volume,urinary storage period(maximum bladder volume,bladder pressure),and urinary voiding period(detrusor pressure,residual urine volume)between the two groups(P>0.05).After 4 weeks of treatment,the average daily catheterization times in group A were lower than before treatment(P<0.05),while the average single urination volume in group A was higher than that before treatment(P<0.05);and the average daily catheterization times in group B were lower than before treatment(P<0.05).After 4 weeks of treatment,the average daily catheterization times in group A were lower than those in group B,and the average single urination volume was higher than that in group B(P<0.05).After 4 weeks of treatment,the maximum bladder volume and detrusor pressure during urination were increased in both groups compared with before treatment(P<0.05),while the bladder pressure and residual urine volume at the maximum volume of the two groups were decreased compared with those before treatment(P<0.05).Compared with group B,the maximum bladder volume and detrusor pressure during urination were higher in group A,while the bladder pressure and residual urine volume at maximum volume were lower in group A(P<0.05).To conclude,two treatments can both improve the urination function of patients with urinary retention after spinal cord injury,and repetitive magnetic stimulation of both S3 nerve root and M1 area is superior to repetitive magnetic stimulation of S3 nerve root alone.Repetitive magnetic stimulation of both S3 nerve root and M1 area can effectively improve the urination function of patients with urinary retention after spinal cord injury.
9.Biomechanical features of posterior"Y"osteotomy and fixation in treatment of ankylosing spondylitis based on finite element simulation analysis
Le ZHANG ; Zhenhua CAO ; Yunfeng ZHANG ; Yangyang XU ; Feng JIN ; Baoke SU ; Lidong WANG ; Xing WANG ; Ling TONG ; Qinghua LIU ; Yuan FANG ; Lirong SHA ; Haiyan WANG ; Xiaohe LI ; Zhijun LI
Chinese Journal of Tissue Engineering Research 2024;28(12):1842-1848
BACKGROUND:Ankylosing spondylitis is a progressive inflammation of spinal stiffness deformity caused by tissue ossification and fibrosis.The posture of ankylosing spondylitis patients is abnormal and their activities are limited that minor injuries can lead to thoracolumbar fractures.Traditional medical image observation limits doctors'preoperative decision planning and postoperative disease prevention for ankylosing spondylitis treatment. OBJECTIVE:Based on the spinal model of ankylosing spondylitis patients before and after posterior spinal cancellous ossification osteotomy("Y"osteotomy for short),to explore the biomechanical changes of"Y"osteotomy and fixation in the treatment of ankylosing spondylitis. METHODS:Based on the preoperative and postoperative CT images of an ankylosing spondylitis patient who went to the Second Affiliated Hospital of Inner Mongolia Medical University,a three-dimensional spine model(T11-S1)before and after"Y"osteotomy(L3 osteotomy)was reconstructed in Mimics 19.0 software.A 7.5 Nm torque was applied to the top of T11 vertebral body to simulate the movement of the spine under six conditions:flexion,extension,left bending,right bending,left rotation and right rotation.Finally,the range of motion of each vertebral body,the stress of each intervertebral disc,and the stress of the screw rod system were simulated. RESULTS AND CONCLUSION:(1)After"Y"type osteotomy and posterior fixation,the range of motion of all vertebrae in the spine decreased,and the loss rate of upper vertebrae was large(L1:77.95%).(2)The maximum stress of the spinal intervertebral disc before operation occurred at the L1-L2 segment(0.55 MPa),and the maximum stress of the spinal intervertebral disc after operation occurred at the T11-T12 segment(0.50 MPa),and the stress of intervertebral disc below T12 was far less than that before operation.(3)The maximum stress of the screw rod system(166.67 MPa)occurred in the upper and middle segments of the rod body and the root of the pedicle screw.(4)In conclusion,the"Y"type posterior fixation operation enhances the stability of the spine and reduces the range of motion of the spine.The vertebral body decompression of the fixed segment is great and the stress-shielding phenomenon of the lower vertebral body is significant.The stiffness of the rod body and the stress concentration area of the pedicle screw should be strengthened to avoid the fracture of the rod caused by stress fatigue.
10.Human osteoarthritic chondrocytes up-regulate the expression of osteoprotegerin in osteoblasts via the Indian hedgehog signaling pathway
Jiale LI ; Dasheng LUO ; Liujie ZHENG ; Wei LIU ; Yunfeng YAO
Chinese Journal of Tissue Engineering Research 2024;28(26):4194-4201
BACKGROUND:Upregulation of hedgehog protein signaling can increase the expression of osteoarthritis markers,Runx2,a disintegrin and metalloproteinase with thrombospondin motifs,collagen type X alpha 1,and matrix metalloproteinase 13,while inhibition of hedgehog proteins attenuates the severity of osteoarthritis.It is speculated that osteoarthritic chondrocytes can influence bone formation by affecting osteoblasts through the Indian hedgehog protein(IHH)signaling pathway. OBJECTIVE:To investigate the effect of human osteoarthritic chondrocytes on subchondral osteoblasts. METHODS:Tibial plateau specimens from patients with osteoarthritis were collected.Chondrocytes were extracted using enzymatic digestion,and osteoblasts were extracted using enzymatic pre-digestion + bone block method.Chondrocytes were identified by toluidine blue staining and immunofluorescence and osteoblasts were identified by alkaline phosphatase staining and immunofluorescence.Chondrocytes were cultured in sodium alginate beads to maintain chondrocyte phenotype and co-cultured with osteoblasts.The co-culture system was added with IHH signaling pathway inhibitor(cyclopamine,10 nmol/L)and activator(purmorphamine,10 nmol/L)separately.After 48 hours of co-culture,osteoblasts from each group were collected,mRNA expressions of Gli1,osteoprotegerin,Runx2,parathyroid hormone-related peptide,alkaline phosphatase,receptor activator of nuclear factor-kB ligand(RANKL)and osteocalcin were detected by qRT-PCR,and protein expressions of GLi1,oseoprotegerin and RANKL in osteoblasts were detected by western blot. RESULTS AND CONCLUSION:The mRNA expression levels of GLi1,osteoprotegerin and RUNX2 in osteoblasts were significantly increased,while the mRNA expression levels of parathyroid hormone-related peptide were decreased(P<0.05)when co-cultured with human osteoarthritic chondrocytes.The mRNA and protein levels of Gli1 were significantly decreased after the addition of IHH signaling pathway inhibitor(cyclopamine)(P<0.05),and the mRNA and protein levels of Gli1 were significantly increased after the addition of IHH signaling pathway activator(purmorphamine)(P<0.05).Osteoprotegerin showed the same trend as Gli1 in the experiment.The osteoprotegerin/RANKL ratio followed the same trend as osteoprotegerin.To conclude,human osteoarthritic chondrocytes can promote the expression of Gli1,osteoprotegerin,Runx2 and other proteins in osteoblasts.The upregulation of osteoprotegerin is related to the IHH signaling pathway.Osteoarthritic chondrocytes can up-regulate the expression of osteoprotegerin in osteoblasts through the IHH signaling pathway and thus up-regulate the osteoprotegerin/RANKL ratio,which will contribute to bone formation in subchondral bone.


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