1.Clinical Study on Treating Lumbar Intervertebral Disc Protrusion Accompanied with Secondary Lumbar Spinal Stenosis by Acupuncture Combined with Medication Plus Joint Mobilization Treatment
Xun WANG ; Yuxin ZHENG ; Yujie CAO ; Yuelong CAO ; Daofang DING
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(4):16-19
Objective To study the clinical efficacy of acupuncture combined with medication plus joint mobilization in the treatment of lumbar intervertebral disc protrusion accompanied with secondary lumbar spinal stenosis.MethodsTotally 66 cases of lumbar intervertebral disc protrusion accompanied with secondary lumbar spinal stenosis were collected randomly and divided into treatment group (34 cases) and control group (32 cases). The control group was given treatment of simple acupuncture and TCM medication, while the treatment group was given joint mobilization treatment besides acupuncture and TCM medication. Functions of lumbar vertebra were evaluated by ODI scale and the degrees of pain were evaluated by VAS. The clinical efficacy in the two groups was compared. Results ODI in both groups were significantly improved after treatment compared with that before treatment. However, the changing range of the ODI of the treatment group was more significant than that in control group (P<0.01). After treatment, VAS scores were relieved (P<0.05) in both groups, and treatment group was more significant than that in control group (P<0.05). The total clinical efficacy was 97.06% (33/34) in the treatment group, and 84.38% (27/32) in the control group, with statistical significance (P<0.05).Conclusion Acupuncture combined with medication plus joint mobilization in the treatment of lumbar intervertebral disc protrusion accompanied with secondary lumbar spinal stenosis has good efficacy.
2.Perinatal management and outcomes of triplet and multiple pregnancy
Chun YANG ; Yong WANG ; Daofang WANG ; Qimei WANG ; Hong CHANG ; Mingfeng ZHUANG
Chinese Journal of General Practitioners 2008;7(6):381-383
Objective To analyze clinical management and outcomes of perinatal fetuses born in triplet and multiple pregnancy.Methods Data of clinical management and outcomes of 55 perinatal fetuses born during 1996 to 2005 by women with triplet and multiple pregnancy were analyzed retrospectively.Results Antenatal check-up was performed regularly for 1/6 of those pregnant women during 1996 to 2000,as compared to that of 8/11 during 2001 to 2005(P<0.05).No significant difference in incidence of varied obstetric complications was found between those during 1996 to 2000 and during 2001 to 2005.Average gestational week at birth was(32.7±2.8)and(35.1±1.9)weeks(P<0.05),and averagebirth weight was(1561±471)grams and(1987±453)grams(P<0.01),respectively for them during 1996 to 2000 and during 2001 to 2005.Caesarean section was performed more for the pregnant women in the earlier first five years than in the later five years,but not reaching statistical significance.Incidence of neonatal complications significantly decreased in the later five years than that in the earlier five yeats,including hyaline membrane disease,neonatal asphyxia,infectious diseases,intracranial hemorrhage and pulmonary hemorrhage(P<0.05),but difference in incidence of apnea and low body temperature between the earlier and later five years did not reach statistical significance.Perinantal mortality was 8 in 35 births in the later five years,as compared to that of 12 in 20 births in earlier five years(P<0.01).Conclusions Outcomes of perinatal fetuses in triplet and multiple pregnancy can be improved by combined action of obstetricians and pediatricians,including regular antenatal examination,active prevention and treatment for pregnant complications and preterm infant diseases,earlier admission waiting for delivery to prolong gestational weeks and increase birth weight,and applying antenatal dexamethasone therapy.
3.Mechanism of resveratrol on the promotion of induced pluripotent stem cells.
Daofang DING ; Xiaofeng LI ; Hao XU ; Zhen WANG ; Qianqian LIANG ; Chenguang LI ; Yongjun WANG
Journal of Integrative Medicine 2013;11(6):389-96
To investigate the effects of resveratrol (RV) in reprogramming mouse embryonic fibroblasts (MEFs) into induced pluripotent stem cells (iPSCs) and the related mechanism.
4.Embryonic Stem Cell Transplantation for Spinal Cord Injury (review)
Leqin XU ; Daofang DING ; Xiaofeng LI ; Yongjun WANG ; Qi SHI ; Chongjian ZHOU
Chinese Journal of Rehabilitation Theory and Practice 2011;17(1):51-53
Transplantation of embryonic stem cells could promote nerve repair by providing nutrients to the injured spinal cord, or differentiating into remyelinating oligodendrocytes or neurons integrating with the host neurons, which has a bright future in the treatment of spinal cord injury.
5.Expression of BMP4 mature peptide in eukaryotic cells and its differentiation-inhibiting effect in culturing induced pluripotent stem cells.
Daofang DING ; Zhen WANG ; Hao XU ; Leqin XU ; Qianqian LIANG ; Yongjian ZHAO ; Qi SHI ; Yongjun WANG
Journal of Southern Medical University 2012;32(10):1383-1388
OBJECTIVETo investigate the role of bone morphogenetic protein 4 (BMP4) in culturing induced pluripotent stem cells (iPSCs) and the related signal pathways.
METHODSWe amplified the mature peptide of BMP4 from the placenta through RT-PCR, and IgK secretion peptide was ligated to the N-terminal of BMP4 mature peptide. The recombinant plasmid pPYCAG-IgK-BMP4 was transfected into 293T cells and screened with puromycin, and the positive clones for expressing BMP4 were verified by cell immunofluorescence and Western blotting. To test the bioactivity of BMP4, iPSCs were cultured in the medium supplemented with leukemia inhibitory factor (LIF) plus the supernatant containing BMP4, and the cell phenotype, cell differentiation capacity into lineages of the 3 germ layers and expression levels of pluripotency-associated genes were investigated.
RESULTSSmad1 was phosphorylated by BMP4 from the culture medium. iPSCs cultured in the medium supplemented with LIF plus the supernatant containing BMP4 for 3 passages maintained the phenotype of stem cells with the expression levels of pluripotency-associated genes not affected. These iPSCs also maintained the capacity to differentiate into cell lineages of the 3 germ layers.
CONCLUSIONBMP4 can be efficiently expressed in mammalian cells to maintain the multipotent differentiation capacity of the iPSCs in in vitro culture.
Animals ; Bone Morphogenetic Protein 4 ; genetics ; Cell Differentiation ; genetics ; Cells, Cultured ; Female ; Gene Expression ; HEK293 Cells ; Humans ; Immunoglobulin kappa-Chains ; genetics ; Induced Pluripotent Stem Cells ; cytology ; Mice
6.Association analysis of serum complement C3 level with blood pressure and estimated glomerular filtration rate in patients with idiopathic membranous nephropathy
Daofang JIANG ; Xiaoyan MA ; Yi WANG ; Xinyu YANG ; Yishu WANG ; Shuchen MA ; Jun WANG ; Chao YU ; Lu FANG ; Na LIU
Chinese Journal of Nephrology 2024;40(6):475-480
It was a single-center cross-sectional study to investigate the association of serum C3 level with blood pressure and estimated glomerular filtration rate (eGFR) in patients with idiopathic membranous nephropathy (IMN). The clinical and pathological data of 98 patients with IMN diagnosed by renal biopsy in the Department of Nephrology of East Hospital Affiliated to Tongji University from August 1, 2018 to October 31, 2023 were retrospectively analyzed. The demographic characteristics, serum complement C3 and other clinical data were compared between the non-hypertension group ( n=37) and hypertension group ( n=61). Pearson or Spearman correlation analysis method was used to analyze the correlation between serum C3 and eGFR in IMN patients and IMN patients with hypertension. Multiple linear regression analysis was used to analyze the related factors of eGFR in IMN patients with chronic kidney disease stage 1-3 in hypertension group. The results showed that compared with the non-hypertension group, the patients in hypertension group were older, and had higher levels of serum creatinine, cystatin C, urinary microalbumin to creatinine ratio, serum C3 and C4, and lower eGFR (all P<0.05). The correlation analysis showed that there was no correlation between serum C3 level and eGFR in IMN patients ( r=0.118, P=0.247). However, serum C3 level was positively correlated with eGFR in IMN patients with hypertension ( r=0.325, P=0.011). Multiple linear regression analysis showed that eGFR was negatively correlated with age ( β=-0.328, P=0.013), and positively correlated with serum C3 level ( β=0.228, P=0.048). The study shows that serum C3 level in hypertension group is higher than that in non-hypertension group in IMN patients. Moreover, serum C3 is positively correlated with eGFR.
7.Establishment of a chondrocyte degeneration model by over-expression of human Wnt7b gene in 293ft cell line.
Xiaojun WANG ; Hao ZHANG ; Jie ZHENG ; Yuxin ZHENG ; Yuelong CAO ; Hongsheng ZHAN ; Yinyu SHI ; Daofang DING
Journal of Southern Medical University 2015;35(3):370-374
OBJECTIVETo investigate the role of human Wnt7b gene in rat chondrocyte degeneration.
METHODSWnt7b gene obtained by PCR was cloned to PCDH-GFP. 293ft cell line was transfected with PCDH-GFP and PCDH-Wnt7b, and the supernatant and transfected cells were collected. The expression level of Wnt7b in 293ft cells was detected by Western blotting. The first passage of chondrocytes were isolated from articular cartilages of newborn born (within 24 h) SD rats were cultured in the supernatants from the transfected cells (at 10- and 50-fold dilutions). The cell morphology of the rat chondrocytes was observed under inverted microscope, and the protein expressions of MMP13, MMP3 and type II collagen and mRNA expressions of A-can, ADAMTS5, Col X and Sox9 were examined by Western blotting or real-time PCR.
RESULTSHuman Wnt7b gene cloned to PCDH-GFP was expressed efficiently in 293ft cell line. Rat chndrocytes cultured for 24 h in the supernatants from PCDH-Wnt7b-transfected 293ft cells underwent changes from a polygonal to a spindle-shaped morphology. The protein expression levels of MMP13 and MMP3 increased while type II collagen decreased significantly, and the mRNA levels of A-can and Sox9 were down-regulated while Col X and ADMATS5 up-regulated in ratchondrocytes after incubation in supernatants from PCDH-Wnt7b-transfected 293ft cells.
CONCLUSIONHuman Wnt7b gene can be expressed efficiently in 293ft cell line and can induce rat chondrocyte degeneration in vitro.
Animals ; Cartilage, Articular ; cytology ; Cell Line ; Chondrocytes ; pathology ; Humans ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction ; Transfection ; Wnt Proteins ; genetics
8.Establishment of a chondrocyte degeneration model by over-expression of human Wnt7b gene in 293ft cell line
Xiaojun WANG ; Hao ZHANG ; Jie ZHENG ; Yuxin ZHENG ; Yuelong CAO ; Hongsheng ZHAN ; Yinyu SHI ; Daofang DING
Journal of Southern Medical University 2015;(3):370-374
Objective To investigate the role of human Wnt7b gene in rat chondrocyte degeneration. Methods Wnt7b gene obtained by PCR was cloned to PCDH-GFP. 293ft cell line was transfected with PCDH-GFP and PCDH-Wnt7b, and the supernatant and transfected cells were collected. The expression level of Wnt7b in 293ft cells was detected by Western blotting. The first passage of chondrocytes were isolated from articular cartilages of newborn born (within 24 h) SD rats were cultured in the supernatants from the transfected cells (at 10-and 50-fold dilutions). The cell morphology of the rat chondrocytes was observed under inverted microscope, and the protein expressions of MMP13, MMP3 and type II collagen and mRNA expressions of A-can, ADAMTS5, Col X and Sox9 were examined by Western blotting or real-time PCR. Results Human Wnt7b gene cloned to PCDH-GFP was expressed efficiently in 293ft cell line. Rat chndrocytes cultured for 24 h in the supernatants from PCDH-Wnt7b-transfected 293ft cells underwent changes from a polygonal to a spindle-shaped morphology. The protein expression levels of MMP13 and MMP3 increased while type II collagen decreased significantly, and the mRNA levels of A-can and Sox9 were down-regulated while Col X and ADMATS5 up-regulated in ratchondrocytes after incubation in supernatants from PCDH-Wnt7b-transfected 293ft cells. Conclusion Human Wnt7b gene can be expressed efficiently in 293ft cell line and can induce rat chondrocyte degeneration in vitro.
9.Establishment of a chondrocyte degeneration model by over-expression of human Wnt7b gene in 293ft cell line
Xiaojun WANG ; Hao ZHANG ; Jie ZHENG ; Yuxin ZHENG ; Yuelong CAO ; Hongsheng ZHAN ; Yinyu SHI ; Daofang DING
Journal of Southern Medical University 2015;(3):370-374
Objective To investigate the role of human Wnt7b gene in rat chondrocyte degeneration. Methods Wnt7b gene obtained by PCR was cloned to PCDH-GFP. 293ft cell line was transfected with PCDH-GFP and PCDH-Wnt7b, and the supernatant and transfected cells were collected. The expression level of Wnt7b in 293ft cells was detected by Western blotting. The first passage of chondrocytes were isolated from articular cartilages of newborn born (within 24 h) SD rats were cultured in the supernatants from the transfected cells (at 10-and 50-fold dilutions). The cell morphology of the rat chondrocytes was observed under inverted microscope, and the protein expressions of MMP13, MMP3 and type II collagen and mRNA expressions of A-can, ADAMTS5, Col X and Sox9 were examined by Western blotting or real-time PCR. Results Human Wnt7b gene cloned to PCDH-GFP was expressed efficiently in 293ft cell line. Rat chndrocytes cultured for 24 h in the supernatants from PCDH-Wnt7b-transfected 293ft cells underwent changes from a polygonal to a spindle-shaped morphology. The protein expression levels of MMP13 and MMP3 increased while type II collagen decreased significantly, and the mRNA levels of A-can and Sox9 were down-regulated while Col X and ADMATS5 up-regulated in ratchondrocytes after incubation in supernatants from PCDH-Wnt7b-transfected 293ft cells. Conclusion Human Wnt7b gene can be expressed efficiently in 293ft cell line and can induce rat chondrocyte degeneration in vitro.
10.Celecoxib inhibits thrombin-induced chondrocyte degeneration in rats
Zhiheng ZHU ; Jiaying DING ; Yangshuo GE ; Chunmeng HUANG ; Jun SHEN ; Xuezong WANG ; Yuxin ZHENG ; Daofang DING
Chinese Journal of Tissue Engineering Research 2024;28(34):5446-5451
BACKGROUND:The content of serum thrombin in patients with osteoarthritis is significantly higher than that in normal individuals,and thrombin can induce inflammatory degeneration of rat chondrocytes,suggesting that inhibiting the function of thrombin may become a method for treating osteoarthritis.Celecoxib is a common therapeutic drug for the clinical treatment of osteoarthritis.It is not yet known whether it improves chondrocyte degeneration by inhibiting the activity of thrombin. OBJECTIVE:To investigate the effect of celecoxib on thrombin-induced degeneration of rat chondrocytes. METHODS:Thrombin levels in the serum of osteoarthritis patients and normal individuals were detected by an ELISA kit.Primary chondrocytes of neonatal Sprague-Dawley rats were isolated,and all experiments were performed with cells from passage one.Chondrocytes were randomly divided into three groups:control group,thrombin group,and celecoxib group.The cell morphology of the three groups was observed under an inverted microscope,and an Edu kit was used to detect the cell proliferation.qRT-PCR was used to detect the expression of extracellular matrix components(aggrecan,elastin,cartilage oligomeric matrix proteins),inflammatory factors(interleukin-1,interleukin-6,and tumor necrosis factor-α),and chemokines(monocyte chemotactic protein 2,monocyte chemotactic protein 7,granulocyte chemotactic protein 6).The expression of type 2 collagen α1 was detected by immunofluorescence.Western blot method was used to detect the expression of catabolic metabolism genes,such as matrix metalloproteinase 9,matrix metalloproteinase 13,and cyclooxygenase 2. RESULTS AND CONCLUSION:Patients with osteoarthritis had higher levels of thrombin in the serum compared with normal individuals.Under the microscope,celecoxib was found to significantly inhibit fibroid changes in chondrocytes.Compared with the thrombin group,celecoxib inhibited the proliferation of chondrocytes.The downregulation of extracellular matrix gene expression,such as type II collagen α1,in the thrombin group was inhibited by celecoxib(P<0.05).Thrombin promoted the expression of inflammatory factors(interleukin-1,interleukin-6,and tumor necrosis factor-α),chemokines(monocyte chemotactic protein 2,monocyte chemotactic protein 7,granulocyte chemotactic protein 6),as well as catabolic genes(matrix metalloproteinase 9,matrix metalloproteinase 13,and cyclooxygenase 2),and under the intervention of celecoxib,the expression of these genes could be downregulated(P<0.05).Overall,these findings indicate that celecoxib inhibits the pro-inflammatory effects of thrombin and thereby ameliorates chondrocyte degeneration in rats.