1.Morphological classification of CT reconstruction of the narrowest part of pediculoisthmic component
Shuai HAO ; Xun MA ; Yannan ZHANG ; Haoliang ZHAO ; Qingqing LIU
Chinese Journal of Tissue Engineering Research 2024;28(18):2876-2880
BACKGROUND:It has been suggested that CT multiplanar reconstruction should be performed prior to the placement of axial pedicle screws to determine the anatomy of the C2 pedicle in each patient,to design the appropriate screw locus and diameter,and to evaluate the feasibility of screw placement to reduce the incidence of surgery-related complications. OBJECTIVE:To evaluate the feasibility of axis pedicle screw placement by morphologic classification of pediculoisthmic component with CT multiplanar reconstruction. METHODS:The CT data of 200 patients(400 axial pedicle screws)with cervical spine were retrospectively studied by using Siemens Syngo.Via software.According to the direction of the axis of the pedicle,the CT multiplanar reconstruction positioning line was adjusted to reconstruct the sectional image of the narrowest part of the pediculoisthmic component.According to its morphological characteristics,the narrowest part of the pediculoisthmic component was divided into three types:type 1,"hook"type:Type 1a outer diameter width(a1)>0.4 cm,type 1b outer diameter width(a1)≤0.4 cm;type 2,"like circle/ellipse"type;type 3,"horizontal ellipse"type.The outer diameter width of the narrowest part of pediculoisthmic component(d1),medullary cavity width(d2),outer diameter height(a1),and medullary cavity height(a2)were compared among the three types,and the feasibility of pedicle screw placement of the three types was evaluated. RESULTS AND CONCLUSION:(1)A total of 400 axial pedicles included 269 cases of type 1,130 cases of type 2,and 1 case of type 3.(2)The mean external diameter height between types 1 and 2 was not significantly different(P>0.05).The mean medullary cavity height,mean outer diameter widths,and mean medullary cavity width were significantly different(P<0.001).There were 42 cases(15.6%)of type 1 and 0 cases(0.00%)of type 2 with mean external diameter width≤0.4 cm,and the difference was significant(P<0.001).There was only one case of type 3,whose external diameter height,medullary cavity height,outer diameter width and medullary cavity width were 1.20 cm,0.84 cm,0.64 cm and 0.31 cm,respectively.(3)These results confirm that axial pedicle screws can be safely inserted in patients with types 1a,2 and 3,which requires no further measurement and assessment.Pedicle screw insertion should be performed with caution in type 1b patients.Therefore,in type 1 patients,the width of the narrowest outer diameter of the pediculoisthmic component should be further measured to evaluate the feasibility of axial pedicle screw placement.
2.Clinical significance of cytokine levels in HBsAg sero-clearance in patients with severe hepatitis B
Xiaomei XIANG ; Xing WAN ; Juan ZHANG ; Jianmei XIAO ; Haoliang WANG ; Dongqing GU ; Guohong DENG
Journal of Army Medical University 2024;46(16):1913-1919
Objective To explore the clinical significance of serum cytokine expression in the hepatitis B surface antigen(HBsAg)sero-clearance in patients with severe hepatitis B.Methods A nested case-control trial was conducted on 14 inpatients with severe hepatitis B admitted in our hospital from 2006 to 2020.Of them,7 patients(aged 36.57±3.15 years)achieved HBsAg sero-clearance within 1 year after the onset of hepatitis B flares(with abrupt rise of ALT level to>5 times the upper limit of normal during HBV infection)and were assigned into HBsAg clearance group,while the other 7 patients(aged 34.14±2.97 years)only obtained HBsAg decreased less than 1 g within 1 year after the onset(HBsAg non-clearance group).Then,multiplex liquid-chip assay based on Luminex xMAP was used to detect the expression levels of 48 cytokines such as IFN-γ and IL-2 in serum samples of these 14 patients.Results The serum levels IFN-γ,IL-2Ra and SDF-1α were significantly lower in the HBsAg clearance group than the HBsAg non-clearance group(P<0.05),but no statistical differences were observed in other 39 cytokines between the 2 groups.And there were 5 cytokines having no mutual expression in both groups.The copy number of HBV DNA was positively correlated with serum HGF(r=0.675,P=0.008)and SDF-1α levels(r=0.587,P=0.027),while negatively with IP-10(r=-0.600,P=0.023)and MIG level(r=-0.640,P=0.014).Meanwhile,a positive correlation was found between HBsAg titer and IL12-p70 level(r=0.593,P=0.025),and a negative correlation between HBsAg titer and TNF-α level(r=-0.609,P=0.021).In addition,the serum total bilirubin level was positively correlated with the expression of SCGF-β(r=0.543,P=0.045).Conclusion Three differentially expressed cytokines and some cytokines related to HBV DNA level and HBsAg titer are found,which may provide new insights into the underlying immunological mechanism of HBV virus clearance caused by hepatitis flares.Meanwhile,it also provides potential biomarkers for HBsAg serological clearance in patients with severe hepatitis B.
3.circDDX17 targets miR-223-3p / RIP3 to regulate the proliferation and apoptosis of non-small cell lung cancer cells
Chengzhi DING ; Guolei WANG ; Gongqian JIANG ; Hongtao WANG ; Yuanyuan LIU ; Haoliang ZHANG ; Fang SUN ; Li WEI
Chinese Journal of Oncology 2024;46(3):239-248
Objective:To explore the molecular mechanism of circDDX17 regulating the proliferation and apoptosis of non-small cell lung cancer cells by targeting the miR-223-3p/RIP3 molecular axis.Methods:The expression levels of circDDX17, miR-223-3p, and RIP3 in human normal lung epithelial cell lines BEAS-2B and non-small cell lung cancer cells H1299, A549, and H446 were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). The plasmids of pcDNA, pcDNA-circDDX17, anti-miR-con, anti-miR-223-3p, pcDNA-circDDX17 and miR-con, pcDNA-circDDX17 and miR-223-3p mimics were transfected into H1299 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay was used to detect the cell proliferation. Flow cytometry was used to detect the cell cycle and cell apoptosis. Plate cloning experiment was used to detect cell proliferation ability. The dual luciferase report experiment was applied to verify the targeting relationship between miR-223-3p with circDDX17 and RIP3. Western blot was used to detect the protein expression of cyclinD1, CDK2, cleaved caspase-3 and Bax.Results:The expression levels of circDDX17 and RIP3 mRNA in H1299, A549, and H446 cells were significantly reduced ( P<0.05), the expression level of miR-223-3p mRNA was significantly increased ( P<0.05) compared with BEAS-2B. The cell viability [(69.46±4.68)%], the number of cell clones (83.49±7.86), the proportion of cells in S phase [(22.52±1.41) %], the protein expression levels of cyclinD1 and CDK2 in PCDNa-CircDDX17 group were lower than those in pcDNA group [(97.54±7.72)%, 205.03±13.37, (28.69±1.49)%, respectively, P<0.05], while the percentage of G 0/G 1 phase cells [(64.45±3.56)%], apoptosis rate [(18.36±1.63)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17 group were higher than those of pcDNA group [(51.33±2.76) % and (5.21±0.54) %, respectively, P<0.05]. The viability [(72.64±5.44)%], the number of cell clones (78.16±8.23), the proportion of S-stage cells [(21.34±1.59) %], the protein expression levels of CyclinD1 and CDK2 in anti-miR-223-3p group were lower than those in anti-miR-con group [(103.47±6.25)%, 169.32±14.53, (28.43±1.26)%, respectively, P<0.05]. Percentage of G 0/G 1 phase cells [(62.86±3.28)%], apoptosis rate [(14.64±1.67)%], the protein expression levels of cleaved caspase-3 and Bax in the anti-miR-223-3p group were higher than those of anti-miR-con group [(51.33±2.71)% and (4.83±0.39)%, respectively, P<0.05]. MiR-223-3p has complementary sites with circDDX17 or RIP3. The viability [(135.45±9.28)%], the number of cell clones (174.64±10.68), the proportion of S-phase cells [(26.39±2.25)%], the protein expression levels of cyclinD1 and CDK2 in pcDNA-circDDX17+miR-223-3p group were higher than those in pcDNA-circDDX17+miR-con group [(101.56±6.68)%, 107.65±7.62, (21.64±1.72)%, P<0.05]. Percentage of G 0/G 1 phase cells [(56.64±2.76)%], apoptosis rate [(8.34±0.76)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17+miR-223-3p group were lower than those of pcDNA-circDDX17+miR-con group [(64.03±3.48)% and (15.21±1.18)%, respectively, P<0.05]. Conclusion:circDDX17 could inhibit the proliferation and induce apoptosis of non-small cell lung cancer cells via targeting the miR-223-3p / RIP3 molecular axis.
4.circDDX17 targets miR-223-3p / RIP3 to regulate the proliferation and apoptosis of non-small cell lung cancer cells
Chengzhi DING ; Guolei WANG ; Gongqian JIANG ; Hongtao WANG ; Yuanyuan LIU ; Haoliang ZHANG ; Fang SUN ; Li WEI
Chinese Journal of Oncology 2024;46(3):239-248
Objective:To explore the molecular mechanism of circDDX17 regulating the proliferation and apoptosis of non-small cell lung cancer cells by targeting the miR-223-3p/RIP3 molecular axis.Methods:The expression levels of circDDX17, miR-223-3p, and RIP3 in human normal lung epithelial cell lines BEAS-2B and non-small cell lung cancer cells H1299, A549, and H446 were detected by reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR). The plasmids of pcDNA, pcDNA-circDDX17, anti-miR-con, anti-miR-223-3p, pcDNA-circDDX17 and miR-con, pcDNA-circDDX17 and miR-223-3p mimics were transfected into H1299 cells. 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide (MTT) assay was used to detect the cell proliferation. Flow cytometry was used to detect the cell cycle and cell apoptosis. Plate cloning experiment was used to detect cell proliferation ability. The dual luciferase report experiment was applied to verify the targeting relationship between miR-223-3p with circDDX17 and RIP3. Western blot was used to detect the protein expression of cyclinD1, CDK2, cleaved caspase-3 and Bax.Results:The expression levels of circDDX17 and RIP3 mRNA in H1299, A549, and H446 cells were significantly reduced ( P<0.05), the expression level of miR-223-3p mRNA was significantly increased ( P<0.05) compared with BEAS-2B. The cell viability [(69.46±4.68)%], the number of cell clones (83.49±7.86), the proportion of cells in S phase [(22.52±1.41) %], the protein expression levels of cyclinD1 and CDK2 in PCDNa-CircDDX17 group were lower than those in pcDNA group [(97.54±7.72)%, 205.03±13.37, (28.69±1.49)%, respectively, P<0.05], while the percentage of G 0/G 1 phase cells [(64.45±3.56)%], apoptosis rate [(18.36±1.63)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17 group were higher than those of pcDNA group [(51.33±2.76) % and (5.21±0.54) %, respectively, P<0.05]. The viability [(72.64±5.44)%], the number of cell clones (78.16±8.23), the proportion of S-stage cells [(21.34±1.59) %], the protein expression levels of CyclinD1 and CDK2 in anti-miR-223-3p group were lower than those in anti-miR-con group [(103.47±6.25)%, 169.32±14.53, (28.43±1.26)%, respectively, P<0.05]. Percentage of G 0/G 1 phase cells [(62.86±3.28)%], apoptosis rate [(14.64±1.67)%], the protein expression levels of cleaved caspase-3 and Bax in the anti-miR-223-3p group were higher than those of anti-miR-con group [(51.33±2.71)% and (4.83±0.39)%, respectively, P<0.05]. MiR-223-3p has complementary sites with circDDX17 or RIP3. The viability [(135.45±9.28)%], the number of cell clones (174.64±10.68), the proportion of S-phase cells [(26.39±2.25)%], the protein expression levels of cyclinD1 and CDK2 in pcDNA-circDDX17+miR-223-3p group were higher than those in pcDNA-circDDX17+miR-con group [(101.56±6.68)%, 107.65±7.62, (21.64±1.72)%, P<0.05]. Percentage of G 0/G 1 phase cells [(56.64±2.76)%], apoptosis rate [(8.34±0.76)%], the protein expression levels of cleaved caspase-3 and Bax in pcDNA-circDDX17+miR-223-3p group were lower than those of pcDNA-circDDX17+miR-con group [(64.03±3.48)% and (15.21±1.18)%, respectively, P<0.05]. Conclusion:circDDX17 could inhibit the proliferation and induce apoptosis of non-small cell lung cancer cells via targeting the miR-223-3p / RIP3 molecular axis.
5.Circular RNA-Encoded Proteins in Gastrointestinal Cancer:A Review
Jie JIANG ; Zai LUO ; Haoliang ZHANG ; Zhengjun QIU ; Chen HUANG
Acta Academiae Medicinae Sinicae 2024;46(1):72-81
Circular RNAs(CircRNAs)are a class of non-coding RNAs with a covalently closed-loop structure,high stability,and tissue specificity,with the production mechanisms different from linear RNAs.Recent studies have discovered that some CircRNAs can encode proteins via cap-independent translation mechanisms such as internal ribosome entry site,N6-methyladenosine,and rolling loop translation.The encoded proteins regulate homologous linear proteins or downstream signaling pathways via protein bait or other mecha-nisms,thereby exerting biological functions.Studies have shown that CircRNAs play a role in various diseases,especially in tumor progression,proliferation,invasion,and metastasis and immune regulation.Therefore,by elucidating the expression and roles of proteins encoded by CircRNAs in tumorigenesis and development,this pa-per is expected to provide new tumor markers and potential targets for tumor diagnosis and treatment.
6.Finite element analysis of the influence of acetabular cup position on the acetabular side stress in hip dysplasia after joint replacement
Sijia XIA ; Heping WANG ; Haoliang ZHANG ; Guochun ZHA
Chinese Journal of Orthopaedics 2024;44(18):1215-1223
Objective:To investigate the effects of acetabular cup positions on the acetabular side stress in hip dysplasia after total hip arthroplasty (THA) using finite element analysis.Methods:Data were obtained from a 36-year-old female patient with developmental dysplasia of the hip. Three-dimensional finite element models were established for different acetabular cup positions using finite element analysis. Each model was categorized based on the center of rotation into four groups: anatomical rotation center, high rotation center, lateralized rotation center, superior-lateral rotation center. ANSYS software applied loads to the model to simulate the stress around the acetabulum in standing (588 N vertical stress) and walking conditions ( X=325 N, Y=-195 N, Z=1 462.5 N). Quantitative analyses of the relative displacement and stress at the acetabular-bone interface were conducted for each region under the two different loading conditions in all eight models. Results:In the standing position with a cup coverage of 90%, the relative displacement at the acetabular-bone interface was: 45.16 μm for the anatomical rotation center group, 47.57 μm for the high rotation center group, 77.27 μm for the lateralized rotation center group, and 96.13 μm for the superior-lateral rotation center group. Acetabular stress values were 9.07 MPa for the anatomical rotation center group, 11.23 MPa for the high rotation center group, 10.88 MPa for the lateralized rotation center group, and 17.75 MPa for the superior-lateral rotation center group. With a cup coverage of 70%, the relative displacements were 64.15, 65.71, 104.10, and 144.53 μm for the respective groups. The corresponding stresses were 9.30, 11.31, 13.98, and 21.45 MPa. In the walking state with a cup coverage of 90%, the relative displacements at the acetabular-bone interface were 189.67 μm for the anatomical rotation center group, 173.55 μm for the high rotation center group 311.03 μm for the lateralized rotation center group, and 572.93 μm for the superior-lateral rotation center group. The stresses were 39.92, 37.33, 47.92, and 71.94 MPa, respectively. With a cup coverage of 70%, the relative displacements were 239.09 μm for the anatomical rotation center group, 248.83 μm for the high rotation center group, 381.84 μm for the lateralized rotation center group, and 1105.90 μm for the superior-lateral rotation center group. The corresponding stresses were 40.62, 58.42, 56.26, and 3,606.30 MPa.Conclusion:With cup coverage at 70% and 90%, the high rotation center and anatomical rotation center exhibited lower and less frequent relative displacements at the acetabular-bone contact surface.
7.Perioperative management strategy and efficacy of the left ventricular assist device HeartCon in the treatment of adult patients with end-stage heart failure
Wei ZHANG ; Yongfeng SHAO ; Buqing NI ; Linfei ZHANG ; Yuanyuan SONG ; Sheng ZHAO ; Haoliang SUN
Chinese Journal of Thoracic and Cardiovascular Surgery 2024;40(5):262-268
Objective:To discuss the perioperative management strategy of the HeartCon left ventricular assist device in the treatment of adult patients with end-stage heart failure and evaluate the effectiveness of blood pump.Methods:Ten consecutive patients with end-stage heart failure treated with the LVAD HeartCon at the Department of Cardiovascular Surgery, Jiangsu Province Hospital from July 2021 to July 2023 were enrolled in this study. The clinical data and follow-up results were retrospectively analyzed. Blood pump parameter, cardiac function classification, liver and kidney function, coagulation, myocardial markers, N-terminal pro-B-type natriuretic peptide, von Willebrand factor antigen, echocardiography, cardiothoracic ratio, and 6-minute walking distance test (6MWT) were evaluated before and after implantation of LVAD in 30, 60, 90, 180, 360, 540, and 720 days. EQ-5D-5L questionnaire was used to evaluate the quality of life.Results:There were 9 males and 1 female with a mean age of (53.7±9.7) years. All patients survived. Renal insufficiency occurred in 1 patient and recurrent aseptic granuloma occurred in 1 patient. There were no significant differences in aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), serum creatinine (Scr), blood urea nitrogen (BUN), right ventricular fractional area change (FAC) and tricuspid annular plane systolic excursion (TAPSE) between pre-operation and 30, 60 and 90 days post-operation( P>0.05). The levels of N-terminal pro-B-type natriuretic peptide (NT-proBNP), von Willbrand factor (vWF) antigen, left ventricular end-diastolic diameter (LVDd) and cardiothoracic ratio decreased significantly on the 30th, 60th and 90th day after operation ( P<0.05). The left ventricular ejection fraction (LVEF), 6MWT and EQ-5D-5L scores were significantly increased at 30, 60 and 90 days after operation compared with those before operation ( P<0.05). Conclusion:With the left ventricular assist device HeartCon, the cardiac function and quality of life of patients were significantly improved within 3 months after operation, and no serious complications were observed, proving that the device is safe and effective.
8.Clinical analysis of 15 cases of traumatic duodenal injury
Jing ZHANG ; Jie PEI ; Ji LI ; Yuejiao XU ; Ji GUO ; Haoliang ZHAO ; Ruochong HE
Chinese Journal of General Practitioners 2020;19(10):938-941
The clinical data of 15 patients with duodenal trauma who were admitted to Shanxi Bethune Hospital from January 2012 to June 2019 were retrospectively analyzed. There were 13 patients with blunt injury and 2 with penetrating injury. The surgical procedure was selected by patient′s condition and extent of injury combined with the clinical symptoms, imaging examination and the Organ Injury Scale grading system of the American Association for the Surgery of Trauma (AAST-OIS). All patients were followed up through outpatient examination and telephone interview till February 2020. Ten patients were diagnosed as duodenal trauma by CT scan before operation, and 5 patients were diagnosed during the operation. According to the AAST-OIS, 1 patient was with grade Ⅰ injury, 6 in grade Ⅱ, 5 in grade Ⅲ, 2 in grade Ⅳ and 1 in grade Ⅴ. All 15 patients received surgical treatment, including 1 with simple suture, 5 with break suture and duodenal diverticularization, 6 with break suture and biliary drainage (3 with hepatocystic duct drainage and 3 with cholecystostomy), 2 with pancreaticoduodenectomy. Postoperative complications occurred in 3 patients with Clavien system classification of Ⅲ b, Ⅱ and Ⅱ. One patient with duodenal stricture and severe abdominal infection was cured after gastrectomy and Billroth Ⅱ gastrojejunostomy 6 months after operation, and 2 cases with duodenal fistula were cured after conservative treatment. One patient who underwent pancreaticoduodenectomy was followed up for 6 months in the outpatient department, and 14 patients were followed up for 6-24 months. For emergency abdominal trauma patients with suspected duodenal injury, surgical exploration should be carried out actively. The site and range of intestinal wall injury should be considered in order to select a reasonable operation. Effective duodenal decompression and complete peritoneal drainage are important for the success of surgery. Early postoperative enteral nutrition support is one of the key measures for successful wound healing.
9. The application of turbocharging technique in repairing large defect with free perforator flap
Haoliang HU ; Xueyuan LI ; Mintao TIAN ; Weisheng MAO ; Xin WANG ; Hong CHEN ; Weiwen ZHANG
Chinese Journal of Plastic Surgery 2019;35(9):862-867
Objective:
To investigate the feasibility and clinic outcome of the turbocharging technique in repairing large defect with free perforator flap.
Methods:
From January 2017 to December 2018, 6 patients with defect of over length or large size were repaired with free perforator flaps, anterolateral thigh(ALT) flap in 3 cases and deep inferior epigastric artery perforator(DIEP) flap in 3 cases in Department of Hand Surgery, Ningbo No.6 Hospital. There were 4 males and 2 females, aged from 29 to 54 years old, with an average age of 41 years old.It was found that the size of the flap beyond the perforasome, while multiple perforators in the flaps were not from the same pedicle. The turbocharging technique was applied in the operation. The distal perforator of the flap was anastomosed with the proximal pedicle branch. The flap size ranged from 20 cm×8 cm to 25 cm×12 cm. The donor sites were closed directly for 3 cases and skin grafting for 3 cases.
Results:
All the flaps survived successfully, no distal necrosis occurred. The patients were followed up for 3 to 12 months, with an average of 9 months. All flaps survived well with satisfactory appearance and pliable texture. The healing of skin graft was satisfactory in 3 cases in donor site. No graft skin contracture occurred. The donor sites closed directly in 3 cases had linear scar in donor site, no obvious contracture occurred. The flap sensation returned to S2-S3.
Conclusions
If the size of the flap is super long or large, beyond the perforasome, while multiple perforators in the flaps were not from the same pedicle, the application of turbocharging technique can avoid partial necrosis of the flap and improve the survival rate of the flap, which is an ideal alternative.
10.Changes in cytoskeleton and glycosaminoglycan synthesis in passaged articular chondrocytes
Liangliang LI ; Zhiqiang ZHANG ; Na GUO ; Haoliang ZHAO ; Xiaohu WANG ; Xiaochun WEI
Chinese Journal of Orthopaedic Trauma 2018;20(9):792-797
Objective To observe the changes in cytoskeleton (CSK) and glycosaminoglycan (GAG) synthesis following passage culture of articular chondrocytes and the correlation between CSK and GAG.Methods Eight male New Zealand White rabbits (8-month-old) were sacrificed by air embolism.After the chondrocytes from their knee joints were isolated by enzymolysis method,monolayer culture was performed.The chondrocytes of primary passage (P0) and passages 1 & 2 (P1,P2) were inoculated into 24-well plates with round cover slips put at the bottoms.Cell climbing slices were fixed after attachment of chondrocytes.The CSK proteins,actin,vimentin,tubulin and vinculin were stained by immuuofluorescence antibody on P0,P1 and P2 cell climbing slices,respectively.The CSK morphology was observed by laser confocal scanning microscopy and the fluorescence intensities of CSK proteins were detected by the fluorescence intensity software.The medium was changed for each generation after cell fusion and the GAG concentrations in the supernatants were measured at 24,36,48,60 h after medium change by alcian blue method.Results The intermediate filament networks became loosen and the dense distributions surrounding the nucleus decreased;more microtubule processes formed at the cell periphery with passage.The fluorescence intensity of actin of P1 chondrocytes was significantly increased than that of P0 (P < 0.05),but there were no such significant differences between P0 and P2 or between P1 and P2 (P > 0.05).The fluorescence intensities of vimentin and tubulin were significantly decreased with passage respectively,and there were such significant differences between any two of P0,P1 and P2 (P < 0.05).The GAG concentrations in the supernatants were significantly decreased with passage at each time point,and there were such significant differences between any two of P0,P1 and P2 (P < 0.05).Conclusions Passage culture of articular chondrocytes may lead to changes in morphology and protein expression intensity of the main components of CSK,and accordingly to decreased synthesis amount of GAG,one of the extracellular matrix of chondrocytes,indicating the changed characteristics of chondrocytes after passage and a certain correlation between CSK and GAG.

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