Objective: To investigate the concentrations of secreted protein acidic and rich in cysteine(SPARC) in the serum and urine of patients with IgA nephropathy and its expression in the kidney tissues. Methods: The concentrations of SPARC, tumor necrosis factor-a (TNF-a),interleukin 1β (IL-1β), and interleukin 6 (IL-6) in the serum and urine were measured with enzyme-linked immunosorbent assay(ELISA). The contents of SPARC protein in the culture medium of human mesangial cell (HMC) and human renal tubular epithelial cell (HKC), which had been treated with IL-6, were determined by ELISA. The expression and distribution of SPARC in IgA nephropathy and normal kidney tissues were observed by immunohistochemistry assay. Results: The concentrations of SPARC in serum and urine of IgA nephropathy patients were higher than those of the normal control subjects ([2.43±±1.22] μg/ml vs [0.69±0.21] 2μg/ml, [7.73±2.81] μg/ml vs [1.17±1.03]ρg/ml, P<0.01). The serum levels of TNF-α, IL-1β and IL-6 in IgA nephropathy group were significantly higher than those in the control group (P< 0.05); the urinary levels of TNF-α and IL-6 in IgA nephropathy group were also higher than those in the controls (P<0.01). SPARC protein secreted by HKC was higher than that by HMC (P<0.01). SPARC was weakly positive in normal distal cortical tubules. SPARC protein expression in tubular epithelial cells of IgA nephropathy patients was obviously higher than that of the normal controls. Conclusion: The secretion of SPARC by renal tubular epithelial cells is increased in patients with IgA nephropathy, which results in elevation of serum SPARC and may have a protective feedback inhibitory effect on HMC proliferation.

Objective To investigate the expression of XIAP, Smac, HtrA2 and XAF1 in the hippocampus following SE in rats and to explore the pathophysiological mechanisms of expression of XIAP and its negative regulators after SE. Methods The lithium-pilocapine model of status epilepticus was established in SD rat. XIAP, Smac, HtrA2, XAF1 and activated caspase-3 protein were examined using immunohistochemistry. Western blot was used to detect the protein levels of XIAP, Smac, HtrA2 and activated easpase-3. Results XIAP immunoreactivity diffusely distributed within the neuron after SE. Compared with the control group, the expression of CA3 XIAP protein in the SE group was increased gradually since 2 hours (0.5503±0.0172 vs 0.1507±0.0165, t=115.87, P<0.01), peaking at 8 hours (0.6221±0.0238 vs 0.1507±0.0165, t=136.69, P<0.01). The expression of CA3 Smac, HtrA2, XAF1 and activated caspase-3 protein were increased generally following SE. Western blot analysis showed a significant increase in Stoat, HtrA2, activated caspase-3 protein levels from 2 to 72 hours following SE, but no significant differences were seen in XIAP protein levels between the control group and the SE group. Conclusions The XIAP, Smac, HtrA2 and XAF1 are involved in the regulation of neuronal apoptosis and implicated in pathophysiological mechanisms of neuronal damage after SE.

Objective To evaluate the application of measurement of T2*value,width of substantia nigra pars compacta (SNc) and the ratio of the width to the midbrain diameter in diagnosing Parkinson disease (PD) in early stage with susceptibility weighted imaging ( SWI) by 3T MR. Methods 59 patients with early stage idiopathic PD patients and 59 healthy controls,ranging in same ages and gender,had been scanned with routine sequences and SWI sequences by 3T MR. T2*value,width and the ratio of the width to the midbrain diameter of SNc were measured. The results of measurement were analyzed and compared. Results (1) The T2*values, width and the ratio of the width to the midbrain diameter was decreased in homolateral side SNc of symptoms of subjects with PD compared with the healthy controls ( <0.05) . (2) There was a significant reduction in the T2*values and the ratio of the width to the midbrain diameter in contralateral side SNc symptoms of subjects with PD compared with the healthy controls (P<0.05) . There was no differences in width of SNc ( >0.05) . Conclusion Measurement of T2*value, width and the ratio of the width to the midbrain diameter of SNc with SWI is reliable to diagnose PD.

Objective To establish a management mode for patients with chronic kidney disease (CKD) of stage 3 to 4 by continuous quality improvement (CQI), and to observe the effect of CQI on renal function in CKD patients. Methods A total of 86 patients with CKD (50 in stage 3 and 36 in stage 4) were enrolled in this study, and they were regularly followed-up in the CKD outpatient of the Department of Nephrology of Jing’an District Zhabei Centre Hospital of Shanghai. The patients were randomly divided into observation group and control group, with 43 cases in each group. In the observation group, we used the management mode combining medical intervention and health education by plan-do-check-act (PDCA) four-step method; in the control group, we used the traditional management mode of medical intervention. All the patients were followed up once a month for one year. The end points included doubling serum creatinine (Scr) or entering end-stage renal disease, and occurence of cardiovascular and cerebrovascular events. The follow-up on time rate (%), Scr level, and estimated glomerular filtration rate (eGFR) were compared between the two groups. Results In the observation group, the average follow-up times were 10.7±2.8 and the follow-up on time rate was (89.9±12.8)%; while those were 4.1±2.2 and (34.2±4.9)% in the control group, and there were significant differences between the two groups (all P0.01). During the 1-year follow-up period, two cases had end-stage renal disease and one case had acute angina in the control group, while no end point was found in the observation group. Before the implementation of CQI, there were no significant differences in eGFR or Scr level between the two groups (all P0.05). The eGFR of the observation group after implementation of CQI was (39.35±12.23) mL/(min • 1.73 m2), which was significantly higher than those of the observation group before implementation ([37.22±11.02] mL/[min • 1.73 m2], P0.05) and the control group after implementation ([35.04±12.31] mL/[min • 1.73 m2], P0.05). The Scr level of the observation group after implementation of CQI was (139.25±14.15) µmol/L, which was significantly lower than those of the observation group before implementation ([145.16±15.41] µmol/L, P0.05) and the control group after the implementation ([148.06±15.63] µmol/L, P0.05). Conclusion CQI management method with the combination of medical intervention and health education can improve the renal function of patients with CKD stages 3-4, and reduce the incidence of end-stage renal disease and cardiovascular and cerevascular events.

OBJECTIVETo investigate the effect and mechanism of Compound Salvia injection (CSI) on nitrate ester tolerance.

METHODSEighty-four patients with coronary heart disease (CHD) were randomly divided into three groups, Group A treated with isosorbide dinitrate (ISD, 15 mg, 4 times per day) alone, Group B with ISD plus CSI and Group C with ISD plus vitamin C. The therapeutic course for all groups was 10 days. The tolerance to nitrate ester and blood pressure were monitored. Before and after treatment, the color Doppler ultrasonic apparatus was used to detect the baseline value of humeral arterial internal diameters (D0), the humeral arterial dilatory response under compression [D1, that is, the flow-mediated vasodilation (FMD)] and the vasodilatory response after sucking of nitroglycerin (D2). And the blood levels of endothelin-1 (ET-1), endothelial nitric oxide synthase (eNOS) mRNA expression were determined. The endothelial-dependent vasodilation (EDD) was expressed by (D1 - D0)/D0 x 100%, and the endothelial-independent vasodilation (EID) was expressed by (D2 - D0)/D0 x 100%.

RESULTS(1) The occurrence rate of nitrate tolerance in Group B and C (28.57% and 35.7%) was lower than that in Group A (64.29%), but insignificant difference was found between the former two. (2) After treatment, blood pressure increased in Group A to the level of pre-treatment, that in Group C also increased but still lower than that of pre-treatment, while insignificant increase was observed in Group B, comparison between Group B and C showed significant difference (P < 0.05). (3) After treatment, EID lowered in Group A, EDD increased in Group B and C (P < 0.05), EDD and EID in Group B and C were higher than those in Group A (P < 0.05), and EDD was higher in Group B than in Group C (P < 0.05). (4) After treatment, ET-1 mRNA expression lowered in Group B, eNOS mRNA expression increased in Group B and C, with significant difference as compared with those before treatment and those in Group A (P < 0.05), and eNOS mRNA expression in Group C was lower than that in Group B (P < 0.05).

CONCLUSIONCSI could partially prevent the occurrence of tolerance to nitrate ester, with the effect better than vitamin C, the mechanism might be related with its regulation on eNOS, ET-1 mRNA expression and protection on vascular endothelial function.

Adult ; Aged ; Coronary Disease ; drug therapy ; Drug Resistance ; Drugs, Chinese Herbal ; administration & dosage ; Endothelin-1 ; biosynthesis ; genetics ; Female ; Humans ; Injections, Intravenous ; Isosorbide Dinitrate ; therapeutic use ; Male ; Middle Aged ; Nitric Oxide Synthase ; biosynthesis ; genetics ; Nitric Oxide Synthase Type III ; Phytotherapy ; RNA, Messenger ; biosynthesis ; genetics ; Salvia miltiorrhiza ; Vasodilator Agents ; therapeutic use

OBJECTIVETo observe and compare the clinical effect of Dingweiban and Xieban manipulation, and to compare the change of the deviation of spinous processes between two methods.

METHODSOne hundred and twenty-two cases were divided into two groups. Sixty-two cases were treated with Dingweiban manipulation method and 60 cases by Xieban manipulation. The changes of Fairbank scores, the clinical effects and the difference of the deviation of the spinous processes (L3, L4, L5) from the lumbar posterior-anterior X-ray were compared.

RESULTSThe scores before and after treatment and 3 months after treatment were compared. There were significant differences between two groups (P < 0.05) by nonparametric test. The result of Dingweiban manipulation group: 53 cases cured, 5 cases better, 3 cases effective and 1 case no effect. The result of clinical Xieban manipulation group: 43 cases cured, 6 cases better, 7 cases effective and 4 cases no effect. The clinical effects had significant differences after treatment and 3 months after treatment (P < 0.05, P < 0.0l) by nonparametric test. After the first treating, there was clear difference of the deviations' distance of the L4 spinous process compared with the Xieban manipulation group (P < 0.05). After the last treating, there were clear differences of the deviation distance of the L4 and L5 spinous processes compared with the Xieban manipulation group (P < 0.05, P < 0.01).

CONCLUSIONDingweiban manipulation is better than Xieban manipulation in effects and has influence on the deviation of spinous processes, especially for the L5 spinous process.

Adult ; Female ; Humans ; Intervertebral Disc Displacement ; therapy ; Lumbar Vertebrae ; Male ; Manipulation, Spinal ; methods ; Middle Aged

ADP Ribose Transferases ; chemistry ; pharmacology ; Animals ; Antineoplastic Agents ; pharmacology ; Bacterial Toxins ; chemistry ; pharmacology ; Exotoxins ; chemistry ; pharmacology ; Female ; HMGN2 Protein ; chemistry ; pharmacology ; HeLa Cells ; Humans ; Immunotoxins ; pharmacology ; Mice ; Mice, Inbred BALB C ; Protein Structure, Tertiary ; Recombinant Fusion Proteins ; biosynthesis ; pharmacology ; Virulence Factors ; chemistry ; pharmacology

To optimize the belt drying process conditions optimization of Gardeniae Fructus extract from Reduning injection by Box-Behnken design-response surface methodology, on the basis of single factor experiment, a three-factor and three-level Box-Behnken experimental design was employed to optimize the drying technology of Gardeniae Fructus extract from Reduning injection. With drying temperature, drying time, feeding speed as independent variables and the content of geniposide as dependent variable, the experimental data were fitted to a second order polynomial equation, establishing the mathematical relationship between the content of geniposide and respective variables. With the experimental data analyzed by Design-Expert 8. 0. 6, the optimal drying parameter was as follows: the drying temperature was 98.5 degrees C , the drying time was 89 min, the feeding speed was 99.8 r x min(-1). Three verification experiments were taked under this technology and the measured average content of geniposide was 564. 108 mg x g(-1), which was close to the model prediction: 563. 307 mg x g(-1). According to the verification test, the Gardeniae Fructus belt drying process is steady and feasible. So single factor experiments combined with response surface method (RSM) could be used to optimize the drying technology of Reduning injection Gardenia extract.
Chemistry, Pharmaceutical ; instrumentation ; methods ; Desiccation ; instrumentation ; methods ; Drugs, Chinese Herbal ; chemistry ; Fruit ; chemistry ; Gardenia ; chemistry ; Research Design ; Vacuum

OBJECTIVETo explore the in vitro methods of isolation and culture of human fetal epidermal stem cells (HFESCs) and the feasibility of the cultured cells as the target cells for gene transfection.

METHODSThe HFESCs were isolated by means of type IV collagen rapid adhering method. The culture medium for HFESCs was prepared according to that for human fetal fibroblasts. The cultured cells were identified by immunohistochemistry staining of keratin-19 and integrin-beta1, cell cycle analysis and clone forming rate determination. Then the cultured cells were gene transfected in vitro by liposome mediating method in which eukaryon expression vector pcDNA3.1/VEGF165 containing vascular endothelial growth factor 165 (VEGF165) were transfected into cultured cells, or by virus vector mediating method in which recombinant adenovirus accompanied vector (raav) containing green fluorescent protein (GFP) (raav/GFP) were transfected into the cultured cells, respectively. The results of in vitro gene transfection of HFESCs were observed by immunohistochemisty staining and fluorescence microscope.

RESULTSHFESCs grew well and formed large clones with higher cloning efficiency and higher ratio of G1 cells than keratinocytes. The cultured cells were strongly positive with immunohistochemistry staining of keratin-19 and integrin-beta1. After being gene-transfected by pcDNA3.1/VEGF165, the VEGF165 of HFESCs showed positive immunohistochemistry staining property, while the HFESCs transfected by raav/GFP exhibited strong fluorescence.

CONCLUSIONHFESCs could be isolated and cultured in vitro by means of rapid adherence to type IV collagen. It seemed feasible that HFESCs were gene transfected with liposome or adeno-associated virus as the vector.

Cell Adhesion ; Cell Cycle ; physiology ; Cells, Cultured ; Endothelial Growth Factors ; genetics ; metabolism ; Epidermis ; Fetus ; G1 Phase ; Green Fluorescent Proteins ; Humans ; Immunohistochemistry ; Integrin beta1 ; analysis ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; Keratinocytes ; cytology ; Keratins ; analysis ; Luminescent Proteins ; genetics ; metabolism ; Lymphokines ; genetics ; metabolism ; Microscopy, Fluorescence ; Plasmids ; genetics ; Stem Cells ; chemistry ; cytology ; metabolism ; Transfection ; Vascular Endothelial Growth Factor A ; Vascular Endothelial Growth Factors

Adolescent ; Adult ; Arteries ; injuries ; pathology ; surgery ; Child ; Child, Preschool ; Extremities ; blood supply ; pathology ; physiopathology ; Female ; Follow-Up Studies ; Humans ; Male ; Microsurgery ; methods ; Middle Aged ; Recovery of Function