Objective To study the effects of repetitive transcranial magnetic stimulation(rTMS) of the supplementary motor area(SMA)on the cortical excitability in patients with Parkinson's disease(PD).Methods Sixteen patients with PD were included in this study.The motor evoked potentials(MEP)and the N30 component of somatosensory evoked potentials(SEP) were assessed for each patient before and after 1200 pulses of rTMS of the SMA at 5 Hz and an intensity of 100% of relaxed motor threshold (RMT) for the abductor pollicis brevis.Results Ten minutes after the rTMS intervention,the peak-to-peak amplitude of the SEP component P20-N30 increased significantly(P<0.05),with the P/F index decreased simultaneously(P<0.05).The MEP amplitude increased significantly,and reached the highest value at 10min after the rTMS intervention. Conclusion 5 Hz rTMS of the SMA can improve the excitability of the SMA itself temporarily.Meanwhile,it can induce a short-lasting facilitation of the excitability of M1 connected with SMA.

Objective To investigate the adjustment effect of microRNA-155 on CD4+CD25+ regulative T cell (Treg) in peripheral blood of sepsis patients,and to elucidate the role of miR-155 in the pathogenesis of sepsis.Methods A retrospective study was corducted.60 sepsis patients (mild n =20,moderate n=20,and severe n =20)from emergency room or intensive care unit (ICU) of the Fourth Hospital of Jiangsu University were enrolled.20 healthy volunteers were enrolled as controls.Real-time fluorescent quantitation polymerase chain reaction (qRT-PCR) was used to detect the levels of miR-155 and Foxp3 mRNA expressions in peripheral blood.CD4+CD25+ Treg cells in peripheral blood were identified by flow cytometry.Peripheral interleukin-10 (IL-10) was measured by enzyme-linked immunosorbent assay (ELISA).Results The expressions of miR-155,Treg,Foxp3 mRNA and the level of IL-10were higher in the patients with sepsis than those in healthy control group [Treg:(2.89 ± 1.13)％ vs.(2.32 ± 0.91)％,t=10.540,P=0.002; miR-155:1.19 ±0.48 vs.0.80 ±0.33,t=8.605,P=0.006; Foxp3 mRNA:0.18 ±0.08 vs.0.13 ±0.03,t=6.862,P=0.008; IL-10 (ng/L):56.89 ± 17.28 vs.33.24 ± 11.93,t=12.742,P=0.001].These variables were elevated gradually with the elevation of acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ)score.The expressions of Treg,miR-155,Foxp3 mRNA and the level of IL-10 were (3.05 ± 1.21)％,1.36 ± 0.79,0.21 ±0.10,(62.82 ±21.38) ng/L in severe sepsis group; they were (2.86 ±0.88)％,1.25 ±0.56,0.17 ±0.08,(56.38 ± 19.65) ng/L in moderate group; they were (2.61 ± 0.87)％,0.94 ± 0.52,0.15 ± 0.05,(45.43 ± 14.40) ng/L in mild group.The values showed significant statistical difference among the mild,moderate and severe sepsis groups (all P＜0.01).Above values were significandy higher in non-survival group than those in survival group [Treg:(3.46 ±1.53)％ vs.(2.85 ± 1.03)％,t=14.250,P=0.005; miR-155:1.41 ±0.85 vs.1.16 ±0.76,t=11.875,P=0.006;Foxp3 mRNA:0.24 ± 0.11 vs.0.17 ± 0.09,t=8.795,P=0.001 ; IL-10 (ng/L):65.47 ± 23.58 vs.51.70 ± 16.86,t=16.313,P=0.001].The expression of miR-155 was positively correlated with the expression of CD4+CD25+ Treg and Foxp3 mRNA (r1=0.635,P1=0.007; r2=0.671,P2=0.005).Conclusion The result of this study suggest that miR-155 is involved in the cell proliferation regulation of CD4 +CD25 + Treg cells,and play some role in the immunological dissonance in sepsis.

Objectiye To study the mechanism by which transcranial magnetic stimulation (rTMS) affects cognitive dysfunction in vascular dementia (VD). Methods Thirty-six male Wistar rats were randomly divided into a control group, a VD group, a low frequency rTMS group and a high frequency rTMS group. Two-vessel occlusion was employed to induce VD models. Low frequency rTMS group rats were given 0.5 Hz rTMS for six weeks. High frequency rTMS group rats were given 5 Hz rTMS for six weeks. Morris' water maze test was used to measure their spatial learning ability and memory. The ultrastructures of the synapses in the four groups were detected with transmission electron microscopy. The expression of synaptophysin (SYN), brain derived neurotrophic factor (BDNF) and Nmethyl-D-aspartate receptor 1 ( NMDAR1 ) mRNA and protein in the hippocampus were determined by real-time PCR and Western blotting. Results The behavior and morphology of the rats treated with rTMS improved. The average expression of SYN, BDNF and NMDAR1 mRNA and protein in the low frequency rTMS group and the high frequency rTMS group were significantly higher than in the VD group. Conclusion rTMS can provide a rehabilitative effect for VD. The mechanism might be associated with enhancing the expression of SYN, BDNF and NMDAR1 in the hippocampus.

OBJECTIVE To investigate the cytotoxic activity of Arca subcrenata Lischke anticancer protein(ASAP)constituents on human myeloid leukemia K562 cells in vitro and analyze its anticancer mechanisms. METHODS ASAP was extracted by low temperature water and ammonium sulfate precipitation. Protein concentration of ASAP was detected by Bradford method. Morphological changes of cultured K562 cells treated with ASAP were observed under the inverted phase-contrast micro?scope. The cell and nucleus changes were analyzed by Giemsa staining. The cytotoxicity of ASAP on K562 cells was detected by MTT assay. Flow cytometry was used to detect apoptosis and cell cycle of K562 cells treated with ASAP. The expression of apoptosis and cell cycle related proteins procaspase 3, caspase 3,P53 and programmed cell death 4(PDCD4)were analyzed by Western blotting. RESULTS ASAP exhibited significant cytotoxic effect on K562 cells in a time- and concentration-dependent manner. The concentration-effect correlation coefficient of ASAP 50,100 and 200 mg · L-1 on K562 cells for 24, 48 and 72 h was 0.851,0.8977 and 0.8997,respectively. Under an optical microscope,K562 cells showed cytomorphosis,or nuclear fragmentation after treatment with ASAP 200 mg · L-1 for 48 h. Flow cytometry analysis and Giemsa staining assay indicated that apoptotic cells increased and G2/M phase cells accumulated significantly with the increase of ASAP concentration. After treatment with ASAP 200 mg · L-1 for 48 h,the early and late apoptosis cell rate increased to(32.8 ± 0.1)%and(31.2 ± 2.2)%vs control group(3.7 ± 1.1)% and (9.9 ± 0.8)%(P<0.01),respectively,and the G2/M phase cells increased to (55.2 ± 1.7)% vs (15.3 ± 0.8)% in control group(P<0.01). After treatment with ASAP 200 mg · L-1 for 0-40 h,the expression of apoptotic protein procaspase 3 was down-regulated and its active form caspase 3 was significantly up-regulated at 32 h,while PDCD4 and P53 protein expression was down-regulated significantly in 0-40 h. CONCLUSION Apoptosis and cell cycle arrest induced in G2/M phase may account for ASAP cytotoxic activity to K562 cells. K562 cell apoptosis induced by ASAP depends on caspase 3 signal pathway. Down-regulated expression of PDCD4 and P53 proteins may be related to K562 cell apoptosis and cell cycle arrest in G2/M phase by ASAP.

Objective To analyze the clinical and immunological features of 45 pediatric patients with lupus nephritis (LN). Methods Forty-five LN patients were included in this study. Clinical, pathological data and immunological parameters were retrospectively analyzed. Results Forty-five LN patients had 6 males and 39 females, with the mean onset age of (10.9 ± 2.8) years. Acute nephritis was the most common type, accounting for 42.2%. Nephrotic syndrome accounted for 31.1%. Renal biopsy showed class II (17.8%), III (4.4%), IV (48.9%), V (2.2%), V+III (6.7%)and V+IV (13.3%)in 42 cases. The remis-sion rate reached 91.1%in the early therapeutic stage, and 15.0%patients recurred after 24-month follow-up. Conclusions The clinical manifestations of LN children are diverse. The renal pathology is complex. The clinical manifestations in part of the chil-dren are not consistent with renal pathology.

Objective To summarize the clinical characteristics and laboratory test results of children with Henoch - Schonlein purpura(HSP),and further to analyze the risk factors for HSP combined with cardiac damage. Methods The clinical and laboratory tests findings from 707 children diagnosed as HSP at Nanjing Children's Hospi-tal were retrospectively analyzed,who were recruited from November 2011 to December 2012. The possible risk factors for HSP with cardiac damage in children were recorded,including gender,age,predisposing causes,gastrointestinal symptoms,joint pain,kidney disorders,serum electrolytes,anti - streptolysin 〝O〝 test,erythrocyte sedimentation rate, and complement level were summarized. Chi - square test and Logistic regression were performed to analyze the risk fac-tors of cardiac damage in children with HSP. Results Among 707 cases,192(27. 2% )patients were combined with car-diac damage,115 male and 77 female,and the proportion of men to women was 1. 00: 0. 67;age ranged from 11 months to 15 years and 4 months(6 years and 5 months for median age),6 patients ﹤ 3 years old occupying 3. 1% ,103 patients≥3 - 7 years old occupying 53. 7% ,82 patients≥7 - 14 years old occupying 42. 7% ,1 patient≥14 years old occupying 0. 5% ,and the age of onset in preschool and school age. Electrocardiogram(ECG)abnormalities were found in 190 patients,the main manifestations including long Q - T interval,ST - T segment falling down and sinus bradycar-dia,and one or more items of abnormal myocardial enzymes existed in 24 cases;echocardiography was performed in 35 cases of children,but no abnormality was detected,no obvious symptoms such as flustered or chest tightness or precor-dial distress. Statistical analysis showed that gender,predisposing causes,mixed HSP,complement level were related to the incidence of cardiac damage in children with HSP(P ﹤ 0. 05). Furthermore binary Logistic regression identified that in male patients,the ratio of X1 vs OR ratio was 0. 654(95% CI 0. 462 - 0. 926,P ﹤ 0. 05),for predisposing causes,the ratio of X2 vs OR ratio was 2. 63(95% CI 1. 838 - 3. 765,P ﹤ 0. 001),for mixed HSP,the ratio of X3 vs OR ratio was 2. 452(95% CI 1. 301 - 4. 621,P ﹤ 0. 01),which were independent factors for cardiac damage in chil-dren with HSP. Conclusions ECG and/ or myocardial enzyme spectrum abnormalities are the main clinical ma-nifestations of cardiac damage in children with HSP. Male patients,predisposing causes of the respiratory tract infec-tion,mixed HSP and hypocomplementemia were high risk factors in the development of cardiac damage,which require special consideration clinically,and earlier ECG and myocardial enzymes examination,early diagnosis and treatment are necessary to avoid the occurrence of severe cases.

Objective To investigate the radiobiological effects of VPA-BSANPs on C6 and U87 glioma cells in vitro.Methods C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h and 24 h,and MTT assay was used to determine cell viability.C6 and U87 cells were treated with different concentrations of VPA and VPA-BSANPs conbined with X-ray irradiation (0,2,4,6,and 8 Gy),and colony formation assay was used to determine plating efficiency (PE).C6 and U87 glioma cells were treated with different concentrations of VPA and VPA-BSANPs for 12 h,followed by X-ray irradiation (0,4,and 8 Gy),and flow cytometry using Annexin V-FITC/PI staining was used to examine cell apoptosis.Western blot was used to evaluate the effects of VPA and VPA-BSANPs on radiation-induced apoptosis protein expression.One-way ANOVA was used for comparison of means with homogeneity of variance between multiple groups,and the t-test was used for comparison of means between two groups.Results Without irradiation,VPA and VPA-BSANPs had no significant inhibitory effects on the proliferation of C6 and U87 cells (P=0.328,0.920).The PE of cells treated with VPA-BSANPs combined with irradiation was significantly lower than that of cells treated with VPA combined with irradiation (P=0.000).In C6 and U87 cells,VPA-BSANPs combined with irradiation increased the expression of p53 and Bax (P =0.000,0.000 and P =0.010,0.002),but reduced the expression of Bcl-2 (P =0.008,0.000).Active caspase-3 fragments were only found in the cells treated with VPA-BSANPs combined with irradiation and VPA combined with irradiation,but were less in the former cells than in the latter cells (P=0.004).The active fragments of peroxisome proliferator-activated receptor were only found in the cells treated with VPA-BSANPs combined with irradiation.Conclusions VPA-BSANPs can increase the radiosensitivity of C6 and U87 glioma cells in vitro,possibly by promoting the apoptosis of tumor cells induced by radiation.

Objective To investigate the role of oxidative stress-dependent Rasextracellular signal-regulated kinase (ERK1/2) signaling in aldosterone (ALDO)-induced mesangial cell proliferation. Methods The incorporation of 3H-thymidine (3H-TdR) and cell count were used as the measure of mesangial cell (MC) proliferation.Western blotting was used to detect the activation of Ki-RasA,c-Raf,MEK1/2,ERK1/2 and PI3K. Results Aldosterone significantly induced human mesangial cell proliferation,and anti-oxidant N-Acetylcysteine (NAC),catalase,and super oxide dismutase (SOD) significantly inhibited ALDO-induced mesangial cell proliferation (P＜0.01,respectively).Stimulation by ALDO for 3 h,Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activity increased by 4.05-, 3.62-, 4.52-, and 3.40-fold compared with control group (P ＜0.01,respectively).NAC almost completely blocked ALDO-induced Ki-RasA,c-Raf,MEK1/2,and ERK1/2 activation (P＜0.01,respectively).Ki-RasA siRNA dose-dependently inhibited Ki-RasA expression, ALDO-induced Ki-RasA activation, and mesangial cell proliferation (P ＜0.01,respectively).c-Raf inhibitor GW5074 and MEK1/2 inhibitor PD98059 also reduced ALDO-induced mesangial cell proliferation by 65％ respectvely (P＜0.01).Ki-RasA siRNA had no effect on ALDO-induced PI3K phosphorylation.Combining LY294002 and PD98059 completely blocked ALDO-induced mesangial cell proliferation (P＜0.01). Conclusions ALDO-induced Ki-RasA-c-Raf-MEK-ERK signaling activation is dependent on reactive oxygen species (ROS) production,which mediates ALDO-induced mesangial cell proliferation.Inhibition of both ERK1/2 and PI3K signaling simultaneously completely blocks ALDO-induced mesangial cell proliferation.

Objective To investigate the clinicopathological characteristics and prognosis of Henoch-Schonlein purpura nephritis with diffused endothelial cell proliferation (DEP-HSPN) in children. Methods Data of 8 DEP-HSPN cases in Nanjing Children's Hospital within recent ten years were retrospectively reviewed. The clinicopathological features, efficacy and prognosis were compared between DEP-HSPN cases and 48 cases of non-DEP-HSPN. Non-DEP-HSPN cases were divided into two groups according to the clinical classification or the pathological classification.Results (1) In DEP-HSPN, HSP developed nephritis within 4 to 15 days after the initial onset of purpuric rashes. Hematuria was present in all the 8 patients. The main clinical manifestation of DEP-HSPN was nephritic-nephrotic syndrome (4 cases), nephrotic level proteinuria (3 cases) and acute nephritic syndrome (1 case). Four cases had macrohematuria. Six cases had abdominal symptoms and two cases had arthritis. Pathology of all the cases showed grade Ⅲ-b lesion with diffused endocapillary proliferation and segmental necrotizing lesion of the capillary wall, always accompanied with intraglomerular inflammatory cell infiltration. Crescent was found in 4 cases. (2)Compared to non-DEP-HSPN grades Ⅲ, DEP-HSPN showed a shorter course of disease.Macrohematuria, heavy proteinuria, nephritic-nephrotic syndrome, and segmental necrotizing lesion of capillary wall were more common in DEP-HSPN. Compared to non-DEP-HSPN with nephrotic level proteinuria, DEP-HSPN had a lower rate of crescent. (3) Methylprednisolone pulse therapy in early stage, then prednisone combined with cyclophosphamide were used in the treatment of DEP-HSPN.After an average follow-up period of seven months, one patient showed complete remission, five showed persistent microhematuria, and two showed persistent microhematuria accompanied with minor proteinuria. No significant difference of prognosis was found between DEP-HSPN and nonDEP-HSPN. Conclusions DEP-HSPN has an acute onset. The main clinical manifestation of DEP-HSPN is nephritic-nephrotic syndrome and nephrotic level proteinuria, always accompanied with macrohematuria. Immunosuppressant treatment in the early stage of disease is effective for a short-term outcome.