1.Correlation analysis of cytochrome P450c19 genes with osteoporotic fracture in a large sample of Shaanxi Guanzhong Population
Jintang WANG ; Shixun WU ; Yunhui JIA
Journal of Xi'an Jiaotong University(Medical Sciences) 2010;31(2):160-164
Objective To study the relationship between osteoporotic hip fracture (OHF) and polymorphism of cytochrome P450c19 genes. Methods By using a group correlation analysis, we made a case-control study of seven single nucleotide polymorphism (SNP) sites groupinge and haplotype in 400 OHF patients from Shaanxi Guanzhong population and 400 normal samples as controls. Results A total of 700 samples were successfully genotyped. Correlation analysis identified three blocks and selected seven tag SNPs. According to the variance analysis, rs7167343, rs8031463, and polymorphism of haplotype 3 were associated with hip fracture. Conclusion Cytochrome P450c19 genes have association with the incidence of osteoporotic fracture in the Han population we studied from Shaanxi Guanzhong region. Therefore, cytochrome P450c19 genes may affect the incidence of osteoporotic hip fracture in the Han population from Guanzhong region, Shaanxi Province.
2. High expression of death receptor membrane proteins in the cartilage of Kashin-Beck disease
Journal of Xi'an Jiaotong University(Medical Sciences) 2020;41(4):553-558
Objective To study the changes of apoptosis-related death receptor 4/5 (DR4/5) and FAS expressions in the cartilage from Kashin-Beck disease (KBD). Methods Cartilage specimens of KBD were collected according to the diagnostic criteria of KBD, primary osteoarthrosis (OA) was used as a case control and normal articular cartilage (NC) as a healthy control. The important molecule of DR signaling was detected by immunohistochemical staining. The results were counted under light microscope and the positive rate was calculated. The expression differences of the three groups of cartilage tissues were statistically analyzed. Results The FAS-positive cell rate in the superficial layer of KBD was significantly higher than that in the NC group, and it was higher in OA group than in NC group. The expressions of DR4 and DR5 in KBD group were not significant compared with those in NC group. In the middle layer, both FAS- and DR4-positive cell rates were higher in KBD and the OA cartilage than in the NC control. DR5 expression did not differ between NC group and KBD group. In the deep layer, the FAS positive cell rate in KBD was significantly higher than that in NC, and the DR4 positive cell rate in KBD and OA groups was significantly higher than that in NC group. DR5 did not differ between NC and KBD groups. Conclusion The expression of FAS/DR4 was observed in the deep and middle layers of KBD joints. It suggests that FAS/DR4-mediated cell death may be involved in the characteristic pathological changes of deep cartilage necrosis.
3.Expression changes of matrix metalloproteinases and tissue inhibitor of metalloproteinases in the articular cartilage of patients with Kashin-Beck disease
Jing MA ; Xiong GUO ; Xiaowei SHI ; Shixun WU ; Zengtie ZHANG ; Qiang ZHANG
Chinese Journal of Endemiology 2015;34(5):344-348
Objective To investigate the changes in the expression of matrix metalloproteinase-1 (MMP-1),matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-3 (TIMP-3) in the articular cartilage of patients with Kashin-Beck disease (KBD) and the role of these proteins in pathogenesis of KBD.Methods The postoperative cartilage samples were collected from patients with KBD,osteoarthritis and patients with non-bone disease (control).The expression of MMP-1,MMP-3 and TIMP-3 in the cartilage was detected by immuohistochemistry,and the positive chondrocytes were counted in different layers of the articular cartilage under microscope.Results The positive rates of MMP-1 in the upper [(67.00 ± 1.69)%] and deeper [(22.07 ± 29.66)%] layers of articular cartilage from patients with KBD,and in the deeper layer of articular cartilage from patients with osteoarthritis [(70.52 ± 37.84)%] were significantly higher than those in the control group [(51.73 ± 36.74)%,(3.75 ± 6.85)%,all P < 0.05].The positive rates of MMP-3 in the deeper layer of articular cartilage from patients with KBD [(28.84 ± 31.13)%] and in the middle and deeper layers of articular cartilage from patients with osteoarthritis [(55.69 ± 35.00)%,(45.96 ± 35.38%)] were significantly higher than those in normal cartilage [(34.09 ± 28.54)%,(14.46 ± 18.32)%,all P < 0.05].The positive rates of TIMP-3 in the middle layer of articular cartilage from patients with KBD [(21.25 ± 25.23)%] and in the middle and deeper layers of articular cartilage from patients with osteoarthritis [(20.40 ± 22.19)%,(18.10 ± 22.58)%] were significantly lower than those in normal cartilage [(36.74 ± 26.61)%,(7.81 ± 20.58)%,all P < 0.05].Conclusion MMP-1,MMP-3 and TIMP-3 play important roles in the articular cartilage damage of KBD.
4.Expressions and significance of erythropoietin producing hepatocellular cell line receptor A2 in pediatric brain glioma
Qiao SHAN ; Xinjun WANG ; Shixun WANG ; Peidong LI ; Jianheng WU ; Xiaohui LI
Chinese Journal of Applied Clinical Pediatrics 2014;29(5):357-360
Objective To investigate the expressions and significance of erythropoietin producing hepatocellular cell line receptor A2 (EphA2) in pediatric brain glioma.Methods Seventy-eight fresh pediatric glioma specimens with a defined histological grade were collected in the Fifth Affiliated Hospital of Zhengzhou University from Jan.2009to Mar.2013,which included 36 of low grade glioma(Ⅰ-Ⅱ grade),42 of high grade glioma(Ⅲ-grade),another 33 cases with brain trauma line pressure to remove children brain tissues were collected as control group.The expressions of EphA2 mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR),Western blot and immunohistochemistry.Results 1.RT-PCR and Western blot showed that EphA2 did not express in control brain tissue,but the expression levels of EphA2 mRNA were over-expressed in pediatric brain glioma,and the difference was statistically significant(F =36.271,P < 0.05) ;the expression levels of EphA2 protein were significantly higher in high-grade pediatric glioma group than in low-grade pediatric glioma group,and the difference was statistically significant(F =42.839,P < 0.05).2.Immunohistochemistry showed that EphA2 expression was negative in control group,the positive expression in low-grade glioma group was 88.57%,and the positive expression in high-grade glioma group was 100.00%.Moreover,the higher the grade glioma,distribution of EphA2 expression was stronger,and the difference was statistically significant(Z =4.962,P < 0.05).Conclusions The mRNA and protein expression levels of EphA2 were significantly high in pediatric brain glioma which were associated with the grade of glioma.Therefore,EphA2 may participate in the development and progression of pediatric brain glioma.
5.Comparison of the expression profiles of cell death factors in articular cartilage between Kashin-Beck disease and osteoarthritis.
Shixun WU ; Xiong GUO ; Feng ZHANG ; Jingjing ZHENG ; Zengtie ZHANG
Journal of Southern Medical University 2014;34(12):1785-1789
OBJECTIVETo compare the expressions of programmed cell death 5 (PDCD5) and early growth response protein-1 (EGR-1) in the articular cartilage between Kashin-Beck disease (KBD) and primary osteoarthritis and the roles of these factors in KBD cartilage.
METHODSCartilage specimens were collected from 10 confirmed KBD patients, 15 osteoarthritic patients and 6 healthy subjects. The expression levels of PDCD5 and EGR-1 in the cartilage were detected by immunohistochemistry staining, and the positive chondrocyte counts were recorded in the different layers of KBD and OA cartilages.
RESULTSThe KBD cartilages contained a significantly higher percentage of PDCD5-positive chondrocytes in the middle layer [(41.35 ± 2.97)%] than OA cartilages [(26.48 ± 2.04)%, P=0.001] and normal cartilages [(19.02 ± 1.88)%, P=0.000] with also obvious PDCD5 over-expression in the deeper layer compared to OA (P=0.000) and normal cartilages (P=0.029), but PDCD5 expression in the superficial layer of the cartilages showed no significant difference among the 3 groups(P>0.05). The average EGR-1 positivity rate in the superficial layer of the cartilage was significantly higher in KBD patients than in OA patients (P=0.000) and healthy controls (P=0.000), but in the middle layer, its positivity rate in KBD patients was higher than that in the normal control (P=0.017) but lower than that of OA cartilage (P=0.002); EGR-1 expression in the deeper layer was comparable in KBD and OA cartilages but both was higher than that in normal cartilages. PDCD5 and EGR-1 expressions were not correlated in either KBD or normal cartilages, but were positively correlated in the superficial layer of OA cartilages.
CONCLUSIONSKBD cartilages show a significantly increased PDCD5 expression in the deeper layer and enhanced EGR-1 expression in both superficial and deeper layers, suggesting the involvement of PDCD5 and EGR-1 in the pathogenesis of KBD.
Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Cartilage, Articular ; metabolism ; pathology ; Chondrocytes ; metabolism ; Early Growth Response Protein 1 ; metabolism ; Humans ; Immunohistochemistry ; Kashin-Beck Disease ; metabolism ; Neoplasm Proteins ; metabolism ; Osteoarthritis ; metabolism ; Transcriptome
6.The effects of HBx on the proliferation and invasion of hepatocellular carcinoma and sorafenib resistance
Luzheng LIU ; Jiacheng CHEN ; Liang CHEN ; Cheng CHEN ; Dafeng XU ; Shixun LIN ; Xiangxiang LUO ; Jincai WU
Chinese Journal of Hepatobiliary Surgery 2021;27(11):842-846
Objective:To investigate the effects of hepatitis B virus X (HBx) on hepatocellular carcinoma (HCC) proliferation, invasion, and sorafenib resistance.Methods:HepG2 cell line infected with HBx ORF lentivirus was set as the HBx high expression group and infected with empty vector was set as the negative control group. The interference group was infected with the HBx siRNA virus based on the HBx high expression group to reduce HBx expression. Interference control group as interference group but with infected empty vector virus. Western blotting was used to measure the protein level of HBx. Cell proliferation, invasion ability, and sorafenib semi-inhibitory concentration (IC50) of HCC cells under different HBx expression levels were respectively detected by cell proliferation assay kit, Transwell invasion assay, and cell titer-glo kit.Results:Western blotting showed that the stable cell lines were successfully established. Cell proliferation of the HBx high expression group was better than that of the blank control and negative control groups, and the cell proliferation of the interference group was lower than that of the interference control and HBx high expression groups, and the differences were all statistically significant ( P<0.05). The number of cells crossing Matrigel gel was (46.2±4.1), (50.7±5.1) and (48.2±5.2) in the blank control group, negative control group, and interference group, respectively. The number of cells crossing Matrigel gel in the HBx high expression group (124.2±8.3) and the interference control group (117.2±7.5) were higher than the above three groups, respectively, and the differences were all statistically significant ( P<0.05). The IC50 of cells in the HBx high expression group and the interference control group were (5.36±0.31) μmol/L and (5.48±0.20) μmol/L, respectively, which were higher than those in the blank control group, the negative control group, and the interference group (4.75±0.22) μmol/L, (4.60±0.14) μmol/L and (3.98±0.03) μmol/L. The differences were all statistically significant ( P<0.05). Conclusion:HBx promoted the tumor proliferation and invasion of HepG2 HCC cells, enhanced the ability to sorafenib resistance, and inhibited apoptosis.
7.Determining the accuracy of lumbar 4/5 pedicle screw entry point based on digital three-dimensional technology
Shixun WU ; Shizhang LIU ; Ming LING ; Xianglong DUAN ; Zhengming SUN ; Jiyuan SHI
Journal of Xi'an Jiaotong University(Medical Sciences) 2022;43(2):263-268
【Objective】 To establish a digital model of lumbar 4 and lumbar 5 vertebral bodies through three-dimensional imaging technology so as to explore the precise placement of pedicle screws during the lumbar posterior internal fixation operation. 【Methods】 CT scan image data set of lumbar spine included six specimens. Then lumbar modeling was produced using Mimics software, implanting pedicle screws was simulated with the computer to determine the reliability of pedicle screw for herringbone crest method, Weinstein method, and Magerl method. 【Results】 This study included six specimens (4 males and 2 females), with an average age of 42.83 years. The distance from the right Magerl entry point to the actual entry point of the lumbar 4 vertebrae was significantly greater than that of the left side. The distance from the left herringbone crest entry point of lumbar 4 vertebrae to the actual entry point was significantly greater than Weinstein method and Magerl method (P<0.001 and P<0.001), and the distance from the right herringbone crest entry point of lumbar 4 vertebrae to the actual entry point was significantly greater than Weinstein method (P=0.003). Both left and right abduction angles of lumbar 4 vertebrae were true abduction angle >Magerl abduction angle > Weinstein abduction angle > herringbone abduction angle. The distances of true-Weinstein and true-herringbone ridges on the left side of lumbar 5 vertebrae were significantly greater than those on the right side (P=0.002 and P=0.004), and the Weinstein abduction angle on the right side of lumbar 5 vertebrae was greater than that on the left side (P=0.003). For the left and right sides of lumbar 5 vertebrae, the distance from herringbone crest entry point to the actual entry point was significantly greater than that of Weinstein method and Magerl method (P<0.001 and P<0.001), and the distance from the Magerl entry point to the real entry point was significantly greater than that of the Weinstein method. The abduction angle of left and right sides of lumbar 5 vertebrae was as follows: true abduction angle > Magerl abduction angle > Weinstein abduction angle > herringbone abduction angle. 【Conclusion】 Both Weinstein entry point and Magerl entry point for lumbar 4 and lumbar 5 vertebra are close to the real entry point. The Weinstein abduction angle and Magerl abduction angle of lumbar 4 and lumbar 5 vertebrae have minor differences compared with real abduction angle. Therefore, it is recommended that Weinstein method is the preferred choice for lumbar 4 pedicle screw placement, while Weinstein method or Magerl method is the preferred choice for lumbar 5 pedicle screw placement.
8.Difference in extracellular matrix loss between KBD and osteoarthritis
Shixun WU ; Cuiyan WU ; Xiong GUO
Journal of Xi'an Jiaotong University(Medical Sciences) 2021;42(2):306-310,316
【Objective】 To study the major missing components of extracellular matrix in cartilage from Kashin-Beck disease (KBD) and osteoarthritis (OA). 【Methods】 The articular cartilage specimens discarded in clinical surgery were collected; paraffin sections were prepared; and HE staining, toluidine blue, and crocusin staining were used to semi-quantitatively analyze the differences in the content of extracellular matrix in cartilage from KBD and OA. 【Results】 The percentage of HE staining area in the normal group (NC group) (83.65±8.38)% was significantly higher than that in the primary osteoarthropathy group (OA group) (57.90±21.88)% and the KBD group (KBD group) (43.67±23.91)%. The stained area percentage of the OA group was higher than that of the KBD group (P=0.034). In the NC group (75.66±12.54)% of the saffron essence O stained area percentage was significantly higher than that of the OA group (53.81±10.48)% and the KBD group (62.07±14.66)%; the KBD group was higher than the OA group (P=0.011). No statistically significant difference in area percentage was found among the NC group, OA group and KBD group stained with toluidine blue (P>0.05). 【Conclusion】 The total loss of extracellular matrix of KBD cartilage tissue is more than that of OA cartilage. The loss of proteoglycan in OA articular cartilage is more serious than that in KBD cartilage, suggesting that the lost extracellular matrix of KBD cartilage tissue is mainly type 2 collagen.