1.The effects of pulsed electromagnetic irradiation on neuropeptide Y regulation, the apoptosis of nucleus pulposus cells and degradation of the extracellular matrix in rats with intervertebral disc degeneration
Zhengkun WANG ; Zhi YAO ; Mengcheng WEI ; Shishuang ZHANG ; Junlong ZHOU ; Qingbo LI ; Lei CAI ; Chuankun ZHOU ; Bowen KOU ; Weijun LIU
Chinese Journal of Physical Medicine and Rehabilitation 2024;46(7):601-607
Objective:To document any effect of a pulsed electromagnetic field (PEMF) on the regulation of neuropeptide Y (NPY) in nucleus pulposus (NP) tissue, NP cell apoptosis and matrix degradation using rats with intervertebral disc degeneration (IDD).Methods:Eighteen Sprague-Dawley rats were randomly divided into a control group, an IDD model group (the model group), and a PEMF group. IDD was induced in both the model and PEMF groups. Right after the modeling, the PEMF group received 14 days of PEMF treatment, while the control group and model group were given no special treatment. Meanwhile, the primary rat nucleus pulposus cells (NPCs) were cultured using Dulbecco′s Modified Eagle Medium at 37℃ and 5% CO 2. When the fusion rate reached 90% after passage, the NPCs were divided into a control group, a TNF-α model group (referred to as model group) and TNF-α + PEMF group (referred to as PEMF group) and treated accordingly. Eight weeks after the modeling, safranin-o/fast green staining was used to assess any pathological morphology changes. The expression of NPY, neuropeptide Y receptor Y2 (NPY2R), bcl-2-associated X protein (Bax), B-cell lymphoma 2 (Bcl-2), collagen type II (Col-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral disc and the cultivated nucleus pulposus cells of the 3 groups were determined. Results:The intervertebral disc cells in the model group were ruptured and folded, with significantly increased polysaccharide and protein components, and significantly increased bone fibers. In the PEMF group the cell boundaries were clearer, with less fibrin fracture and increased cartilage tissue. NPY was expressed in the fibrous annulus and the nucleus pulposus of the intervertebral disc in the model group. The average expression levels of NPY and NPY2R were significantly higher than in the control group and the model group. Compared with the control group, there was a significant increase in the level of Bax and a significant decrease in the expression of Bcl-2 in the model group, and there was a significant decrease in the level of Bax in the PEMF group. Compared with the control group, there was a significant decrease in the Col-II level but a significant increase in the MMP3 protein expression in the model group. The average Col-II mRNA expression was significantly higher in the PEMF group compared with the model group, but the average MMP3 protein expression was significantly less. Those results are consistent with observations in vivo.Conclusion:PEMF may reverse the imbalance of ECM metabolism and delay IDD degeneration by up-regulating the expression of NPY and Bcl-2, as well as blocking the Bax/Bcl-2 signaling pathway to inhibit apoptosis of nucleus pulposus cells.
2.Lipoprotein glomerulopathy caused by mutation of apolipoprotein E gene in children: a report of two cases
Yaqin WEI ; Jun YAO ; Pei ZHANG ; Chunlin GAO ; Xu HE ; Lili JIA ; Xiaojie LI ; Zhengkun XIA
Chinese Journal of Nephrology 2023;39(3):215-219
The paper reports two cases of lipoprotein glomerulopathy (LPG) in children. The Sanger sequencing results in 2 cases indicated apolipoprotein E gene mutation[c.127 (exon3) C>T, p.R43C (p.Arg43Cys); c.494 (exon4) G>C, p.R165P (p.Arg165Pro),respectively]. Renal pathological presentation of two children showed that a large number of lipoprotein emboli were formed in the glomerular capillary loop, and the diagnosis of LPG was confirmed. The onset of LPG has no specific clinical manifestation, which is easy to be undiagnosed or misdiagnosed. Renal biopsy is a diagnostic means, glucocorticoid treatment is ineffective, and long-term lipid-lowering treatment may be required for LPG.
3.The application of antineutrophil cytoplasmic antibody renal risk score in children with antineutrophil cytoplasmic antibody-associated glomerulonephritis
Pei ZHANG ; Chao XU ; Chunlin GAO ; Xiang FANG ; Zhiqiang ZHANG ; Zhengkun XIA
Chinese Journal of Applied Clinical Pediatrics 2023;38(6):438-443
Objective:To investigate the potential of the antineutrophil cytoplasmic antibody (ANCA) renal risk score (ARRS) in predicting the prognosis of children with ANCA-associated glomerulonephritis (AAGN).Methods:Laboratory testing, renal pathology results, treatment and prognosis of 61 children with AAGN diagnosed by renal biopsy from June 2007 to May 2022 in General Hospital of Eastern Theater Command were retrospectively analyzed.The Kaplan-Meier method was used to evaluate the overall and renal survival of children with AAGN, and risk factors of progression to end stage renal disease (ESRD) were analyzed by Cox regression analysis. Results:Among the 61 children with AAGN, there were 14 males and 47 females with the age of (15.65±3.74) years.According to ARRS, AAGN children were assigned into low-risk group (27 cases), medium-risk group (21 cases) and high-risk group (13 cases). During a median follow-up duration of 46.36 (14.58, 95.62) months, the number of ESRD cases in the high-risk group (9 cases) was significantly higher than that of low-risk group (2 cases) and medium-risk group (3 cases) ( χ2=13.079, P<0.001). Kaplan-Meier survival analysis showed that AAGN children in the high-risk group had the worst renal prognosis ( χ2=5.796, P=0.016), while no significant difference was detected in the overall survival among the 3 groups ( χ2=2.883, P=0.237). Multivariate Cox regression showed that estimate glomerular filtration rate(eGFR)≤15 mL/(min·1.73 m 2) ( HR=9.574, 95% CI: 4.205-25.187, P=0.015) and ARRS ( HR=2.115, 95% CI: 1.206-4.174, P=0.012) were independent risk factors for children with AAGN progress to ESRD.Receiver operating characteristic (ROC) curve analysis results showed that the area under the curve of ARRS for predicting the risk of progressing to ESRD in AAGN children was 0.880 (95% CI: 0.759-1.000), and the optimal cutoff value of ARRS was 5.50, with the sensitivity and specificity of 85.71% and 82.98%, respectively. Conclusions:ARRS was an independent risk factor for children with AAGN progress to ESRD, which had a predictive value for the progression of AAGN to ESRD.
4.Diagnostic potential of magnetic resonance imaging in children with nutcracker syndrome
Ren WANG ; Meiqiu WANG ; Zhengkun XIA ; Pei ZHANG ; Chunlin GAO
Chinese Journal of Applied Clinical Pediatrics 2023;38(6):448-451
Objective:To explore the diagnostic potential of magnetic resonance imaging (MRI) in children with nutcracker syndrome (NCS).Methods:A retrospective analysis was performed in patients with suspected NCS(155 cases) diagnosed in the Department of Pediatrics, General Hospital of Eastern Theater Command from January 2017 to July 2020.Suspected NCS was diagnosed primarily based on clinical signs or symptoms, laboratory testing, and imaging reports, and other conditions that may cause hematuria and/or proteinuria were excluded.MRI examination was performed in all patients.According to the diagnostic criteria of NCS, patients diagnosed as NCS with the compression of the left renal vein (LRV) were included in the NCS group(58 cases), and those without the compression of the LRV or with the compression of the LRV but was not consistent with the diagnosis of NCS were included in the control group(97 cases). t test, Mann- Whitney U test and χ2 test were used to compare the baseline characteristics, clinical characteristics and imaging characteristics of the children in the nutcracker group and the control group.Receiver operating characteristic curves were plotted to explore the diagnostic potential of MRI in children with NCS. Results:(1)The area under curve of the angle between the superior mesenteric artery (SMA) and the aorta, compression ratio (CR) and beak sign in diagnosing NCS in children were 0.870, 0.895 and 0.878, respectively.(2)The optimal cut-off values of the angle between the SMA and the aorta and CR were 36.8° and 3.99, respectively.(3)The specificity of the angle between the SMA and the aorta<36.8°, beak sign, CR>3.99, the angle between the SMA and the aorta combined with beak sign, the angle between the SMA and the aorta<36.8° combined with CR>3.99, and beak sign combined with CR>3.99 in diagnosing NCS in children were 82.5%, 93.8%, 93.5%, 97.9%, 95.9% and 97.9%, respectively.Conclusions:Children with the angle between the SMA and the aorta<36.8°, beak sign and CR>3.99 suggested on MRI scans should be highly suspected of NCS.The beak sign has the highest specificity in the diagnosis of NCS in children, and the combination of any two parameters has a higher specificity than a single parameter.
5.Interpretation of 2022 IPNA clinical practice recommendation for the diagnosis and management in children with steroid-sensitive nephrotic syndrome and comparison with 2016 guideline of China
Pei ZHANG ; Chunlin GAO ; Jun YAO ; Zhengkun XIA
Chinese Journal of Nephrology 2023;39(11):872-878
Relapsing is the clinical characteristic of nephrotic syndrome (NS) in children, and some steroid-sensitive NS (SSNS) children have frequently relapsing NS and steroid-dependent NS. The long-term and frequent uses of glucocorticoids, as well as the adverse effects of immunosuppressants, seriously affect the quality of life in children with NS. The International Pediatric Nephrology Association released the clinical practice recommendation for SSNS in children in 2022. Based on the guideline for the diagnosis and treatment of steroid-sensitive, frequently- relapsing/dependent NS in children (2016) formulated by the Nephrology Group of Pediatrics Society of Chinese Medical Association, we interpreted the guideline on clinical treatment and management suggestions of SSNS, to provide references for the diagnosis and treatment of SSNS in children.
6.Pulsed electromagnetic field activation of the A2A adenosine receptors can retard intervertebral disc degeneration
Qingbo LI ; Lei CAI ; Zhengkun WANG ; Weizhi FANG ; Chuankun ZHOU ; Yichi ZHOU ; Zhi YAO ; Mengcheng WEI ; Shishuang ZHANG ; Weijun LIU
Chinese Journal of Physical Medicine and Rehabilitation 2023;45(9):769-775
Objective:To explore any effect of pulsed electromagnetic field (PEMF) stimulation on intervertebral disc degeneration (IDD).Methods:Forty Sprague-Dawley rats were randomly divided into a control group, an IDD model group, a PEMF group and an observation group. An IDD model was induced in all except those in the control group. Both the PEMF and observation groups were given PEMF stimulation, while the latter was additionally injected with the A2AR agonist CGS-21680. Eight weeks after the modelling any pathological changes in the morphology of the rats′ intervertebral disc tissues were evaluated using saffron solid green staining. The expression of A2AR, cyclic adenosine phosphate (cAMP), protein kinase A (PKA), cysteine aspartate proteolytic enzyme-3 (Caspase-3), type II collagen (COL-II) and matrix metalloproteinase-3 (MMP3) in the intervertebral discs were evaluated.Results:The nucleus pulposus had shrunk, while fibrous tissues and chondrocytes had increased in the IDD model group. In the observation group the nucleus pulposus was intact and of basically normal shape. A2AR mRNA and protein levels were higher in the intervertebral disc tissue of the model group than among the control group, on average, while the levels in the observation group were significantly higher than in the other groups. In the PEMF and observation groups cAMP and PKA mRNA were significantly higher than in the IDD model group. The p38 MAPK and P-P38 MAPK levels of the IDD model group and its average P-P38 MAPK/p38 MAPK ratio were significantly higher than in the control group. In the PEMF and observation groups those indices had decreased to varying degrees, with those of the observation group significantly lower than among the model and PEMF groups on average, except for the p38 MAPK values. Caspase-3 and its mRNA were significantly higher in the model group than in the control group, on average, and those values were significantly lower in the PEMF and observation groups than in the IDD model group. The average MMP3 contents of the IDD model group had increased significantly compared with the control group, while the Col-Ⅱ level had decreased significantly. Compared with the IDD model group, the MMP3 level had decreased but Col-Ⅱ expression had increased in both the PEMF and observation groups, with significant differences between the IDD model and observation groups.Conclusions:The activation of the p38 MAPK signaling pathway by inflammatory factors to induce apoptosis is one of the important reasons for the aggravation of IDD lesions. PEMF combined with A2AR agonists can activate the A2AR/cAMP/PKA signaling pathway, inhibit p38 MAPK phosphorylation, reduce apoptosis of nucleus pulposus cells, and relieve IDD damage.
7.Clinical and pathological features in children diagnosed with primary focal segmental glomerulosclerosis after repeated renal biopsy
Zhiqiang ZHANG ; Yingchao PENG ; Lili JIA ; Chunlin GAO ; Zhengkun XIA
Chinese Journal of Nephrology 2022;38(8):657-663
Objective:To analyze the clinical and pathological characteristics in children diagnosed with primary focal segmental glomerulosclerosis (FSGS) after repeated renal biopsy.Methods:The clinicopathological data of children who ever experienced renal biopsy in Jinling Hospital from January 1, 2000 to December 31, 2020 were retrospectively reviewed. Clinical manifestations, pathological characteristics and treatment responses were analyzed.Results:Of the 34 enrolled patients, there were 22 males and 12 females. The median age of the first renal biopsy was 14 years old (1-18 years old), and the median interval between repeat renal biopsy and first renal biopsy was 6 months (1-151 months). Thirty-one showed nephrotic syndrome, of which 22 had microscopic hematuria, and 4 had elevated serum creatinine. Among the other 3 patients, 2 had hematuria and proteinuria, and 1 had proteinuria. In the first renal biopsy, 16 cases were diagnosed as minimal change disease, 14 cases were diagnosed as mesangial proliferative glomerulonephritis, 2 cases were diagnosed as IgA nephropathy, and 2 cases were diagnosed as IgM nephropathy. All 34 children showed poor responses to hormone and immunosuppressive therapies. The pathological features of the first renal biopsy in some patients were adhesion (2/34), decreased loop podocyte attachment (2/34), peripheral loop extension to the urinary pole (2/34), renal tubular reflux (4/34), capillary thrombosis (2/34) and IgM deposition (12/34).Conclusions:The initial diagnosis of FSGS is difficult, and the lesions are atypical and easily misdiagnosed. The patients have poor responses to hormone and immunosuppressive therapies. For patients with the pathological changes of adhesion, decreased loop podocyte attachment, peripheral loop extension to the urinary pole, renal tubular reflux, capillary thrombosis and IgM deposition, follow-up is required, and if necessary, repeat renal biopsy needs be performed to determine whether it is FSGS.
8.Effects of a pulsed electromagnetic field on intervertebral disc degeneration and A2A adenosine receptor-mediated reactive oxygen signaling
Lei CAI ; Weizhi FANG ; Qingbo LI ; Zhengkun WANG ; Zhi YAO ; Mengcheng WEI ; Shishuang ZHANG ; Hongbo JIAO ; Weijun LIU
Chinese Journal of Physical Medicine and Rehabilitation 2022;44(11):966-972
Objective:To observe any regulatory effect of a pulsed electromagnetic field (PEMF) on A2A adenosine receptors (A2ARs) in the nucleus pulposus of rats with intervertebral disc degeneration (IDD), and to explore any combination with the A2ARs′ agonist-mediating ROS/PI3K/Akt signaling pathway.Methods:Fifty Sprague-Dawley rats were randomly divided into a control group, an intervertebral disc degeneration group (the model group), an A2AR agonist CGS-21680 treatment group (the agonist group), a PEMF group and a PEMF combined with CGS-21680 treatment group (the observation group). IDD was modeled in all except the rats in the control group. 100μL of CGS-21680 (100μg/kg) was injected into the L 5-6 intervertebral discs of the agonist group, while the PEMF group was given 30 minutes of PEMF intervention daily for 14 days at 1.5mT and 75Hz with a pulse width of 150μs. The observation group was injected with CGS-21680 and then given the same PEMF intervention. Primary nucleus pulposus cells from each group (50ng/mL) were cultured and the expressions of 8-OHDG, SOD, MDA and ROS were detected by immunohistochemistry, immunofluorescence or with an ELISA kit. The A2AR, PI3K, AKT and p-AKT protein levels were detected using western blotting. Results:The nucleus pulposus cells and the annulus fibrosus were obviously wrinkled, necrotic and broken in the model group but the annulus fibrosus was intact and the nucleus pulposus was almost normal in the observation group. Compared with the model group, the levels of SOD and A2AR, PI3K, p-AKT and AKT protein were higher in the agonist, PEMF and observation groups, while the expressions of MDA, ROS and 8-OHDG were weaker. The ROS level in the observation group was significantly lower than that in the agonist and PEMF groups, and the phosphorylation level of p-AKT in the observation group was significantly higher than in the agonist and PEMF groups. The average levels of SOD, A2AR, PI3K, p-AKT and AKT protein in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly higher than the IL-1β group′s average, while the average levels of MDA, ROS and 8-OHDG were significantly lower. The ROS levels in the observation group were significantly lower than in the agonist and PEMF groups, while the A2AR protein content and p-AKT phosphorylation in the observation group were significantly greater. The average Bax levels in the nucleus pulposus cells of the agonist, PEMF and observation groups were significantly lower than that in the IL-1β group, and the expression of Bcl-2 was significantly increased. There was significantly less apoptosis observed in the observation group than in the agonist and PEMF groups, while the expression of Bcl-2 was significantly higher.Conclusions:PEMF plays an anti-oxidative stress role by up-regulating A2AR activity and reducing ROS generation. Treatment with PEMF and A2AR agonist could further activate the phosphorylation of PI3K/Akt, down-regulate Bax and up-regulate Bcl-2, thus inhibiting the apoptosis of nucleus pulposus cells and alleviating the malignant progression of IDD.
9.Two decomposition algorithms of dual-energy cone beam CT and their dependence on the phantom sizes
Chenguang LI ; Tianye NIU ; Li ZHOU ; Jun DENG ; Chengyuan ZOU ; Sha LI ; Hongjia LIU ; Zhengkun DONG ; Ling HUA ; Yichen PU ; Liangzi QU ; Qiao LI ; Yibao ZHANG
Chinese Journal of Radiological Medicine and Protection 2022;42(4):269-276
Objective:To analyze the effects of two decomposition algorithms of dual-energy cone beam CT (DECBCT) (direct decomposition and iterative decomposition) on the image quality and material decomposition accuracy of different sizes of phantoms.Methods:Different sizes of imaging parts of patients were simulated using the combination of CatPhan604 phantoms and customized annuluses. CBCT with high energy of 140 kVp and low energy of 100 kVp were acquired using the Varian Edge CBCT system. Then the material decomposition of DECBCT images was performed using the two algorithms. The electron density (ED) and contrast-to-noise ratio (CNR) of each material in the CTP682 module were calculated. They were used to assess the decomposition accuracy and image quality of the two algorithms.Results:Based on the values in the Catphan604 manual, both algorithms have high ED accuracy. Only the ED accuracy of four materials of the smallest sized phantom showed statistical difference ( z = -4.21, 4.30, 2.87, 5.45, P < 0.05), but the average relative error was less than 1%. The CNR of the iterative decomposition algorithm was significantly higher than that of the direct decomposition, increasing by 51.8%-703.47%. The increase in the phantom size significantly reduced the accuracy of ED, and the increased amplitude of the relative error was up to a maximum of 2.52%. The large phantom size also reduced the image quality of iterative decomposition, and the decreased amplitude of CNR was up to a maximum of 39.71. Conclusions:Compared with the direct decomposition, the iterative decomposition algorithm can significantly reduce the image noise and improve the contrast without losing the accuracy of electron density in the DECBCT construction of different sizes of phantoms.
10.A five-year follow-up study on the clinicopathology of 130 children with Henoch-Sch?nlein purpura nephritis
Meiqiu WANG ; Ren WANG ; Xu HE ; Pei ZHANG ; Zhengkun XIA ; Chunlin GAO
Chinese Journal of Nephrology 2022;38(3):168-176
Objective:To analyze the clinicopathologic features and prognosis of children with Henoch-Sch?nlein purpura nephritis (HSPN).Methods:The clinicopathological data of children with HSPN who were followed up for more than 5 years and underwent renal biopsy in Jinling Hospital affiliated to Medical School of Nanjing University from January 2001 to June 2015 were retrospectively analyzed. The follow-up endpoint event was defined as estimated glomerular filtration rate (eGFR)<90 ml·min -1·(1.73 m 2) -1. Participants were divided into two groups according to whether the children had reached the primary endpoint event or not. Cox proportional hazards model was used to analyze the influencing factors of renal poor prognosis in children with HSPN. Kaplan-Meier survival curve method was used for survival analysis, and log-rank test was used to compare the difference of renal cumulative survival rate between segmental sclerosis/adhesion (S1) group and non-segmental sclerosis/adhesion (S0) group. Receiver operating characteristic curve (ROC curve) and area under the curve ( AUC) were used to evaluate the diagnostic value. Results:A total of 130 children with HSPN were enrolled in the study. The median onset age was 11.7(8.6, 13.3) years old, of whom 71 cases were males (54.6%). At a median follow-up time of 100.0(75.8, 119.0) months, 12 cases (9.23%) with HSPN reached the primary endpoint event. Compared with the non-endpoint event group, the endpoint event group had higher proportion of hypertension, higher levels of 24-hour urinary protein, serum cholesterol, serum uric acid, and serum creatinine, and lower levels of serum albumin (all P<0.05). There was no statistical difference in treatment between the two groups (all P>0.05). In terms of pathological features, compared with the non-endpoint event group, the endpoint event group had higher proportion of mesangial hyperplasia (M1), S1, tubular atrophy/interstitial fibrosis (T1/T2) and Glomerulus-Bowman's capsule adhesion (all P<0.05). Multivariate Cox regression model showed that S1 was significantly correlated with renal poor prognosis ( HR=7.739, 95% CI 1.422-42.114, P=0.018). As was revealed in a Kaplan-Meier plot, renal cumulative survival rate in the S1 group was significantly lower than that in the S0 group (log-rank χ2=17.069, P<0.001). The ROC curve showed S1 accurately predicted the outcome ( AUC=0.710, 95% CI 0.549-0.872) with specificity of 0.667(95% CI 0.349-0.901) and specificity of 0.754(95% CI 0.667-0.829). Conclusions:S1 is an independent risk factor affecting renal poor prognosis and has a diagnostic value.

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