OBJECTIVETo investigate the effect of different kinds of mechanical stress on the mRNA expression of integrin beta1 subunit in cultured human periodontal ligament fibroblasts (hPDLF).

METHODSTo scalp and remove the periodontal ligament attached to the mid-third part of the fresh root of young premolars extracted for the cause of orthodontics. Cultured hPDLF by the method of digesting by I-type collagenase combining with tissue adhering. Then hPDLF was isolated and purified by cells passage. The sixth passage's cells were selected to be loaded. A new cyclic strain loading apparatus. Forcel four point bending device was used for mechanically loading. Cells were loaded by three levels (1000, 2000, 4000 microstrain) of tensional and compressive forces and collected at different times (0, 0.5, 1, 4, 8, 12 h) course after strain loading. The quantity of integrin beta1 mRNA in every group was analyzed by means of quantitative real-time PCR with the special primers of up- and down-regulated genes.

RESULTSDynamic mechanical forces down-regulated the expression of integrin beta1 subunit mRNA in hPDLF and the difference in groups by different magnitude, different kinds, and different time of mechanical forces loading were statistically significant. The stronger stimulated forces, the more down-regulated expression. Compression down-regulated the expression of integrin beta1 subunit mRNA more than tension did.

CONCLUSIONDynamic mechanical forces could regulate the expression of integrin beta1 subunit mRNA. The difference among all the groups by different magnitudes, different kinds, and different time of mechanical forces loading were statistically significant.

Cells, Cultured ; Fibroblasts ; Humans ; Integrin beta1 ; Periodontal Ligament ; RNA, Messenger ; Stress, Mechanical

Objective To investigate the effect of different dynamic tensional and compressive stress on the mRNA expression of collagen type Ⅰ and fibronectin in cultured human periodontal ligament fibroblasts (hPDLF), and explore the regularity of functional change in hPDLF. Methods A new cyclic strain loading apparatus was used for mechanically loading, Cells cultured in vitro were loaded with three levels (1000 μstrain, 2000 μstrain, 4000 μstrain) of tensional and compressive forces and collected at different time (0 h,0. 5 h,1 h,4 h,8 h,12 h) course after strain loading. The quantity of collagen type Ⅰ and fibronectin mRNA was analyzed by means of quantitative real-time PCR with special primers of up- and down-regulated genes. Data were analyzed using SPSS version 10. 0 software. Results Different magnitude and different kinds of mechanical forces as well as the force application time significantly changed the expression of collagen type Ⅰ and fibronectin mRNA in hPDLF. Conclusions Dynamic mechanical forces could regulate the expression of collagen type Ⅰ and fibronectin mRNA in hPDLF. Collagen type Ⅰ and fibronectin participated in the mechanical signal transduction in human periodontal ligament fibmblasts.

Objectives To investigate the effect of focal adhesion kinase (FAK) on the mRNA expression of transcription factors c-fos/c-jun in cultured human periodontal ligament fibroblasts (hPDLF)loaded with tensional force.Methods The periodontal ligament,which was attached to the mid-third part of the fresh root of young premolars extracted for orthodontic purposes,was removed.The hPDLF was cultured using the method of digesting by Ⅰ-type collagenase combined with tissue adhering.Then hPDLF was isolated and purified by cells passage and PDLF were used in the experiments at passage six.A four-point bending device named " Forcel" was used for force loading.Cells cultured in vitro were loaded with 2000 μstrain tensional force and collected at different time (0,0.5 h,1 h,2 h,or 4 h) after strain loading.Cells were divided into two groups (with or without FAK antibody).Aherations in the mRNA expression of transcription factors c-fos/c-jun due to cyclic tensional force were investigated using real-time polymerase chain reaction (PCR).ANOVA and LSD were used to process the data obtained.Results After strain loading the c-fos/c-jun mRNA expression was upregulated,with a peak at 0.5 h,and then downregulated after 0.5 h.The levels of gene expression in cells treated with the FAK antibody were greater than those in cells untreated with the FAK antibody (P ＜0.05).Conclusions These results provided evidence that transcription fac tors c-fos/c-jun probably played a critical part in the response of periodontal tissue (periodontal ligament fibroblasts) to mechanical stimulation and that FAK probably played an important role in the mechanical signaling pathway of hPDLF.

OBJECTIVETo explore possible correlations between renal Th1/Th2 ratio and renal microvascular injury in children with Henoch-Sch-nlein purpura nephritis (HSPN).

METHODSThirty-two children with HSPN were enrolled. They were classified into four groups by renal pathology: HSPN class II (n=8), HSPN class IIIa (n=7), HSPN class IIIb (n=10) and HSPN class IV/V (n=7). Five patients undergoing nephrectomy due to trauma were used as the controls. INFγ, IL-4 and CD34 in the renal tissues were measured by immunohistochemical analysis. INFγ was used as a marker of Th1, IL-4 was used as a marker of Th2 and CD34 was used as a marker of microvessel. The renal microvessel density was evaluated according to the Weidner standard. The relationships among the local Th1/Th2 ratio, renal pathological grade, microvessel score and microvessel density were studied.

RESULTSImmunohistochemical analysis showed a lower expression of INFγ and a higher expression of IL-4 in the HSPN groups than in the control group. The local Th1/Th2 ratio in the HSPN groups decreased and correlated significantly with the renal pathological grade. There were significant differences among four HSPN subgroups (P<0.05). Compared with the control group, the renal microvessel density in the HSPN class II and class IIIa groups increased significantly (P<0.05), but it decreased in the HSPN class IV/V group (P<0.05). The renal microvessel scores in the HSPN class IIIa, class IIIb and class IV/V groups increased significantly compared with those in the control and the HSPN classⅡ. The increased renal microvessel scores were associated with more severe renal pathological changes. A negative correlation was found between the local Th1/Th2 ratio and the microvessel density in kidneys (r=-0.921, P<0.01).

CONCLUSIONSThe decrease of Th1/Th2 ratio in kidneys might be responsible for renal microvascular injury in children with HSPN.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Kidney ; blood supply ; pathology ; Male ; Microvessels ; pathology ; Nephritis ; immunology ; pathology ; Purpura, Schoenlein-Henoch ; immunology ; pathology ; Th1 Cells ; immunology ; Th2 Cells ; immunology

The study was purposed to investigate the polymorphism of killer cell immunoglobulin-like receptor (KIR) gene of the patients with leukemia and to explore the correlation between the KIR gene and susceptibility of leukemia. The KIR genotype of 50 patients with leukemia and 60 healthy controls in northern. Hans were analyzed by PCR-SSP. The results indicated that the present known 18 KIR genes were detected and identified. The frequencies of KIR 3DL3, 3DL2 and 2DL4 were 100% in all subjects, with the most frequent genotype KIR 3DP1 (0.86) followed by 2DP1, 2DL3, 3DL1, 2DL1, 3DS1, 2DL5, 2DS4, 2DS2, 1D, 2DS5, 2DL2, 2DS1, 2DS3 and 3DP1v in leukemia successively. Compared with the control, the KIR 3DL1 (0.60) and 2DL1 (0.57) were significantly lower in the leukemia patient group than that in the control group (1.00) (P < 0.01). It is concluded that the polymorphism of KIR gene is associated with susceptibility of leukemia in Hans. There may be a negative correlation between pathogenesis of leukemia and KIR 3DL1, KIR 3DS1, KIR 2DL1, KIR 2DL5 genes.
Adolescent ; Adult ; Child, Preschool ; Female ; Genetic Predisposition to Disease ; genetics ; Genotype ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; genetics ; Male ; Middle Aged ; Polymorphism, Genetic ; Precursor Cell Lymphoblastic Leukemia-Lymphoma ; genetics ; Receptors, Immunologic ; genetics ; Receptors, KIR ; Receptors, KIR2DL1 ; Receptors, KIR2DL3 ; Receptors, KIR2DL4 ; Receptors, KIR3DL1 ; Receptors, KIR3DL2 ; Receptors, KIR3DS1

OBJECTIVE: To investigate the difference of Pax2 and P53 expressions in children with primary nephritic syndrome (PNS) and the effect of Pax2 on glucocorsteroid (GC)-resistance. METHODS: Renal Pax2 and P53 expressions in children with PNS (40 patients) were detected by immunohistochemistry. A semiquantitative score was used to evaluate the injury degree of the glomeruli and the tubulointerstitium, and correlation analysis was done among Pax2, P53 and pathologic score. RESULTS: Pax2 and P53 expressions were not found in the control group. Pax2 expression of renal tubule epithelia exsisted in children with PNS and there was weak or no expression of Pax2 in the podocytes. Pax2 expressions in the proximal tubule and the distal tubule in the GC-resistant group were more intense than those in the GC-intensive group (P <0.01). The more the Pax2 expression in the tubule, the more abnormal structure such as dilation and atrophy. Pax2 expression in the tubule epithelia was positively correlated with pathologic score of tubulointerstitium (P < 0.01). There was no P53 expression in the GC-intensive group, but there exsisted P53 expression in parts of the patients from the GC-resistant group, mainly distributing in the renal tubular epithelia. P53 expression was positively correlated with P53 expression and the pathologic score of tubulointerstitium (P < 0.01). CONCLUSION Over-expression of Pax2 in the renal tubule epithelia may improve P53 expression to a certain degree, which may aggravate the lesion of the renal tubule. It may be one of the mechanisms resulting in GC-resistant in children with PNS.
Adolescent ; Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Male ; Nephrotic Syndrome ; drug therapy ; metabolism ; PAX2 Transcription Factor ; biosynthesis ; genetics ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVE: To evaluate the benefits and toxicities of different corticosteroid regimes in preventing relapse in children with steroid sensitive nephrotic syndrome (SSNS). METHODS: MEDLINE (Jan. 1963-Mar. 2007), elsevier (Jan. 1997-Aug. 2006), OVID databank (Jan. 1993-Aug. 2006), Springer databank (Jan. 1994-March 2007), the Cochrane Controlled Trials Register (Cochrane Library, Issue Feb. 2006), Cochrane Renal Group Specialised Register (Jul. 2006), EMBASE (Jan. 1980-Mar. 2007) and CNKI (Jan. 1994-Mar. 2007) etc, were searched by the terms primary nephrotic syndrome, glucocorticoid, corticosteroid, steroid, prednisone, methylprednisolone, dexamethasone and children etc for the human clinical trials about glucocorticoid (GC) administration in primary nephrotic syndrome (PNS) (aged 3 months to 18 years), controlled or semi-controlled ones, including unpublished documents from scientific meetings and theses, and similar documents listed in the references of the above documents were also included. All the studies were evaluated strictly according to Jadad Standard, and the Meta-analysis were adopted. Review manager 4.2 software was used to analyze the data. The odds ratio was calculated for the relapse rate and side effect from the initial episode to the end of follow-up between the patients treated with corticosteroids and the controls. RESULTS: Totally 12 trials with 868 subjects meeting the criteria were included in this review. A Meta-analysis of 7 trials, which compared between 2 months of prednisone and 3 months or more in the first episode, showed that longer treatment duration significantly reduced the risk of relapse at 12-24 months (RR=0.70,95% CI:0.60-0.89),without an increase of side effect. There was a negative linear relationship between the duration of treatment and risk of relapse (r2 =0.66, P=0.05). CONCLUSION (1) Children in their first episode of SSNS should be treated for at least 3 months of GC. The therapeutic effect of patients in the primary nephrotic syndrome treated with GC for 12 weeks was prior to that for 8 weeks, compared with that in the controls. It could reduce the relapse rate of half year, the longer treatment duration in the NS patients at the first relapse was, the lower relapse risk was.(2) Compared with alternative GC administration, standard GC administration can reduce the side effects; Course more than 1 year of GC administration can reduce the 2-year relapse rate. Hence in children who relapse frequently, multicentre, well-designed experiments should be adopted.
Child ; Drug Resistance ; Glucocorticoids ; pharmacology ; therapeutic use ; Humans ; Nephrotic Syndrome ; drug therapy

OBJECTIVE: To investigate the efficacy and adverse effect of mycophenolate mofetil (MMF) in the treatment of frequently relapsing nephrotic syndrome in children. METHODS: The study population consisted of 37 children (24 simple nephrotic syndrome and 13 nephritis-type syndrome) suffering from frequently relapsing nephrotic syndrome. Patients received 20-30 mg/(kg d) of MMF in conjunction with 1 mg/(kg d) prednisone for 3-6 months. RESULTS: Out of 24 patients suffered from simple nephrotic syndrome, 17 patients (70.8%) with complete relief, 4 patients (16.7%) with partial relief and 3 patients (12.5%) with non-relief, whereas out of 13 patients suffered from nephritis-type syndrome 6 patients (46.2%) with complete relief, 3 patients (23.1%) with partial relief and 4 patients (30.7%) with non-relief. Eight patients with Minimal Change Disease (MCD) achieved complete relief. Of 23 patients with Mesangial Proliferative Glomerulonephritis (MsPGN) or Membranoproliferative Glomerulonephritis (MPGN), complete relief was observed in 17 patients (73.9%), partial relief in 4 patients (17.4%) and non-relief in 2 patients. CONCLUSION These Results suggest that MMF has better efficacy against simple renal disease than against nephritis-type syndrome, and MMF may be more suitable for the treatment of frequently relapsing nephrotic syndrome characterized by proliferative lesions.
Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Immunosuppressive Agents ; adverse effects ; therapeutic use ; Male ; Mycophenolic Acid ; adverse effects ; analogs & derivatives ; therapeutic use ; Nephrotic Syndrome ; drug therapy ; Recurrence ; Treatment Outcome

OBJECTIVEGlucocorticoid (GC) is the first therapeutic choice of primary nephrotic syndrome (PNS). The response to GC treatment is an important indicator for the outcome of PNS children. Children with GC-resistant PNS present with incomplete or no response to GC, and may herald the progression to end-stage renal failure. However, the detailed mechanism of GC-resistance or GC-sensitive effect in these PNS children has not been clearly elucidated. The previous study by the authors indicated that there was increased expression of GR beta in PBMCs in GC-resistant children with PNS, and the over expression of GR beta resulted in GC resistance via influencing the ability of GR alpha nuclear translocation. To elucidate the relationship between GR beta expression in renal and in PBMCs and the effect of glucocorticoid on glucocorticoid-resistance children with PNS, the expression of GR alpha and GR beta in renal tissue and in PBMCs were detected by immunohistochemistry.

METHODSForty children with PNS were divided into two groups, GC-resistant group(20) and GC-sensitive group(20), the expression of GR alpha and GR beta in renal intrinsic cells and in PBMCs were measured with the immunohistochemistry technique. A semiquantitative score was used to evaluate the injury degree of the glomeruli and tubulointerstitium.

RESULTSCompared with GC-sensitive group, the glomerular pathologic scores (6.91 +/- 1.98) and renal tubular pathologic scores (7.12 +/- 1.62) in GC- resistant group were significantly different (P < 0.01, respectively). GR alpha expressions of renal tissue and PBMCs were higher in the control group (58.3 +/- 2.6, 59.1 +/- 7.2) than those in the GC-sensitive group (40.2 +/- 7.2 and 36.6 +/- 5.1, P < 0.01, respectively) and GC-resistant group (35.0 +/- 8.2 and 36.4 +/- 6.6, P < 0.01, respectively). GR beta expressions of renal tissue and PBMCs were higher in the GC-resistant group (13.8 +/- 3.0 and 12.1 +/- 4.1) and in the GC-sensitive group (6.5 +/- 1.9 and 5.9 +/- 1.0) than that in control group (2.3 +/- 0.4 and 3.2 +/- 1.1, P < 0.01, respectively). GR beta expressions in renal tissue and PBMCs were higher in the GC-resistant group than that in the GC-sensitive group (P < 0.01). Compared with control group, GR beta expressions in PBMCs and in renal tissue were lower than those in mild renal lesion group (5.4 +/- 2.8, 6.46 +/- 2.50), midmedium renal lesion group (8.7 +/- 2.4 and 11.4 +/- 3.7) and (17.1 +/- 0.4 and 18.7 +/- 0.7) in severe renal lesion group (F = 5.8, 15.6, P < 0.01, respectively). GR beta expression of PBMCs had a positive correlation with GR beta expression of renal intrinsic cells (r = 0.651, P < 0.01). GR beta expressions by PBMCs and renal intrinsic cells were positively correlated with renal pathologic scores (r = 0.579 and 0.623, P < 0.01, respectively).

CONCLUSIONGC-resistant children with PNS were related to the increased GR beta expression in PBMCs and renal intrinsic cells. There was no correlation between the GR alpha expressions in PBMCs and in renal intrinsic cells. Increased GR beta expression might decrease the effect of GC via inhibiting the activity of GR alpha.

Adolescent ; Child ; Child, Preschool ; Drug Resistance ; Female ; Glucocorticoids ; therapeutic use ; Humans ; Kidney Glomerulus ; pathology ; Kidney Tubules ; pathology ; Male ; Nephrotic Syndrome ; drug therapy ; pathology ; Receptors, Glucocorticoid ; analysis

OBJECTIVETo investigate the clinicopathologic characteristics of childhood renal diseases.

METHODSA retrospective analysis of 1316 renal biopsies performed over the past 20 years was performed.

RESULTSOf the 1316 patients, 383 (29.09% ) were diagnosed as nephrotic syndrome, 291 (22.00%) as acute nephritis syndrome, 224 (17.21%) as isolated hematuria, 209(15.87%) as purpura nephritis, and 96 (7.30% ) as hepatitis B virus-associated nephritis . Mesangial proliferation was the most common pathological change (756 cases; 57.45%), followed by IgA nephropathy (113 cases; 8.59%), endothelial capillary proliferation(112 cases; 8.51%), membranous nephropathy (66 cases; 5.02%), and various minor and minimal changes (59 cases; 4.48%). Alport syndrome, congenital nephrotic syndrome, thin basement membrane nephropathy, fibrillary glomerulopathy disease, and Fabry disease were confirmed by electronic microscopy. IgA, IgM and C1q nephropathy were definitely diagnosed using immune histochemistry or immunofluorescent. A diagnosis of primary glomerular disease was made in 69.53% of the cases (915 cases); secondary glomerular disease was noted in 26.14% (344 cases). Of the 915 cases of primary glomerular disease, 375 (41.0%) had nephrotic syndrome. Secondary glomerular disease due to purura nephritis was common (209/344; 60.8%).

CONCLUSIONSPrimiary glomerular disease predominates in children. Nephrotic syndrome is the most common clinical diagnosis. Mesangial proliferation is the most common pathological patterns in children with renal disease.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Kidney ; pathology ; Kidney Diseases ; pathology ; Kidney Glomerulus ; pathology ; Male ; Renal Insufficiency ; pathology ; Retrospective Studies