Objective To investigate the expressions and correlation between Survivin and PTEN proteins in astrocytoma.Methods The expressions of Survivin and PTEN proteins were examined by immunohistrochemistry in astrocytoma specimens from 65 patients with astrocytoma.Results The positive expression rate of Survivin in astrocytoma grade Ⅱ was significantly lower than that in grade Ⅲ and grade Ⅳ(all P

AIM: To establish an HPLC method of determining paeonol and cinnamaldehyde in Jiarong Tablet. METHODS: Column:Kromasil C_(18)(200 mm?4.6 mm I.D,5 ?m);mobile phase:MeOH-H_2O(65∶35);detection wavelength was at 288 nm;flow rate was 1.0 mL/min. RESULTS: The calibration curve showed a good linea-rity within the ranges of 0.003 6830.018 412 ?g for cinnamaldehyde,0.097 92-0.489 6 ?g for paeonol.(CONCLUSION:)The method is accurate and can be used for the quality control of Jiarong Tablet.

Objective To investigate the efficacy of umbilical cord mesenchymal stem cells (UC-MSCs) transplantation in the treatment of the MRL/lpr mice. Methods Twenty four 18-week-old MRL/lpr female mice were divided into 3 groups:group 1 (G1) were transplanted with 1×106 UC- MSCs through caudal vein, group 2 (G2) were transplanted with 1×106 UC- MSCs three times and group 3 (G3) were treated with 0.5 ml normal saline as controls. Enzyme linked immunosorbent assay (ELISA) was used to measure the levels of serum anti-dsDNA antibodies. Twenty-four hours proteinuria and body weight were assessed every two weeks. The histopathology changes of the kidneys and lungs were observed. Results ① At the 25th weeks, the 24 hours proteinuria in group G1 (2.3±1.9) mg and G2 (1.8±1.4) mg was decreased than that in the control group (3.8±2.1) mg (P<0.05), and at the 27th weeks, that of groups G1 (2.5±1.5) mg and G2 (1.9±1.2) mg was also significantly decreased than in the control group (5.4±2.4) mg (P<0.01); ② From the 24th week, the body weight of groups G1 and G2 increased significantly than that of the control group (P< 0.05). At week 29, serum creatinine decreased significantly in both groups G1 (7.2±3.2) μmol/L and G2 (6.2±2.8) μmol/L than in the control group (12.5±2.3 ) μmol/L (P<0.05); ③One week after transplantation, the levels of anti-dsDNA antibodies in group G1 (46±11)×102 U/ml and G2(49×43)×102 U/ml were bothsignificantly decreased than those of the control groups (99±42)×102 U/ml (P<0.05) and the difference between group G2 (36±15)×102 U/ml and the controls (68±32)×102 U/ml was statistically significant; ④The nephron crescent formation in group G1 (0.12±0.07) and G2 (0.08±0.02) was significantly lower that of the control group (0.20±0.06) (P<0.05) and that of group G2 was significantly less that of froup G1 (P<0.05); ⑤ The interstitial pneumonitis was singnificantly milder in group G1 than group G2. Conclusions UC- MSCs is very effective in treating MRL/lpr mice. It is safe and free of rejection reactions.

Results With the prolonged exposure to heat , the mice exhibited swollen and disorderly arranged neurons , shrunken cells , and contracted and deeply stained nuclei , with significantly higher scores on nerve pathological injury evaluation at 6, 12, and 24 h (2.78 ± 0.71, 3.21 ±0.56, and 3.36 ±0.63) than the control mice (0.43 ±0.10) (P<0.05).ELISA showed remarkably elevated levels of UCH-L1 in the serum (F=147.7, P=0.05) and brain tissue (F=145.7, P=0.05) in the heat stress group as compared with the con-trol, and Western blot also revealed a markedly higher expression of UCH-L1 in the brain tissue in the former group than in the latterObjective The abnormal expression of ubiquitin C-terminal hydrolase-L1 ( UCH-L1 ) has an important role in the diagnosis and prognosis of brain damage .This study was to investigate the changes of UCH-L1 in the serum and brain tissue in the mouse model of heat stress . Methods Twelve BALB/c mice were randomly divided into a control and a heat stress group of equal number, the former placed at a temperature of (25.0 ±0.5)℃and a relative humidity of (35 ±5)%and the latter in a simulated in-cubator at (35.5 ±0.5)℃and a relative humidity of (60 ±5)%.When the rectal temperature reached 42℃, the animals were re-moved from the incubator and cooled at an ambient temperature of (25.0 ±0.5)℃and a humidity of (35 ±5)%for 0, 6, 12, and 24 h.Then the brain tissues of all the animals were harvested for HE staining , evaluation of neuronal injury under the light microscope , measurement of the UCH-L1 levels in the serum and brain tissue by ELISA , Western blot, and immunohistochemistry , respectively. (F=261.2, P=0.01).Immunohistochemistry manifested that , with the prolonged exposure to heat , the UCH-L1 expression in the brain tissue was characterized by gradually increased light brown of the neurons at staining . Conclusion Severe heatstroke causes brain injury in a time-dependent manner , and the abnormally elevated levels of UCH-L1 in the serum and brain tissue can be a marker of heatstroke-induced brain injury .

Objective To observe the effect of heat stress-induced reactive oxygen species (ROS) burst on the regulation of expression of Bcl-2 and Bax in human umbilical vein endothelial cell (HUVEC) apoptosis induced by heat stress,and explore the pathogenesis of vascular endothelial damage caused by severe heat stroke.Methods HUVEC heat stress model was reproduced.Cells of heat stress group were incubated at either 39,41,or 43 ℃ for 2 hours,then all the cells were further incubated at 37 ℃ and 5％ CO2 for 24 hours.Before heat stress,cells of 43 ℃ heat stress group were pretreated with 10 μmol/L MnTMPyP,which was a specific scavenger of ROS,for 1 hour.Cells of control group were incubated at 37 ℃ and 5％ CO2.The amount of ROS was assayed with 2',7'-dichlorofluorescin diacetate (DCFH-DA) and dihydroethidium (DHE) staining.Apoptosis was determined by using staining with Hoechst33258.The mRNA expressions of Bcl-2 and Bax were determined by reverse transcription-polymerase chain reaction (RT-PCR).The protein levels of Bcl-2,Bax,caspase-3 were analyzed by Western Blot.In addition,the effect of MnTMPyP on heat stress-induced apoptosis was also studied.Results Compared with control group,there was no obvious change in cells after 39 ℃ heat stress.With the increase in heat stress temperature up to 41 ℃ and 43 ℃,viability of cells showed a lowering trend,with a burst of ROS,and an increase of mRNA and protein of Bax,and the protein of caspase-3 was significantly increased,the mRNA and protein of Bcl-2 were significantly decreased in a temperature-dependent manner.These changes were marked in 43 ℃ heat stress group as compared with those of the control group [cell viability:(46.00 ±4.00)％ vs.(96.33 ± 1.53)％,t=20.164,P=0.001; ROS (fluorescence relative value):400.67 ± 12.10 vs.99.33 ±4.04,t=32.909,P=0.001; Bax mRNA (A value):3.03 ±0.15 vs.1.00 ± 0.00,t=23.056,P=0.001; Bax protein (gray value):3.97 ±0.21 vs.1.00 ± 0.00,t=24.684,P=0.001; caspase-3 protein (gray value):4.80 ± 0.20 vs.1.00 ± 0.00,t=32.909,P=0.001 ; Bcl-2 mRNA(A value):0.42 ± 0.30 vs.1.00 ± 0.00,t=33.072,P=0.001 ; Bcl-2 protein (gray value):0.39 ± 0.25 vs.1.00 ± 0.00,t=42.212,P=0.001].It was shown that pre-condition with the antioxidant MnTMPyP significantly decreased the heat stress-induced expression of Bax,caspase-3,and apoptosis,and the expression of Bcl-2 was elevated [Bax mRNA (A value):2.00 ± 0.20 vs.3.33 ±0.25,t=7.184,P=0.002; Bax protein (gray value):2.03 ±0.25 vs.3.23 ±0.25,t=5.840,P=0.004; caspase-3 protein (gray value):2.07 ± 0.21 vs.5.00 ± 0.20,t=17.600,P=0.001 ; Bcl-2 mRNA(A value):0.71 ± 0.40 vs.0.42 ± 0.26,t=8.126,P=0.002; Bcl-2 protein (gray value):0.57 ± 0.31 vs.0.40 ± 0.06,t=5.091,P=0.007].Conclusions A burst in an increase of ROS plays an important role on heat stress-induced HUVEC apoptosis,and the mechanism is probably related to the expressions of Bcl-2 and Bax.The vascular endothelial cells apoptosis may be one of the pathogenetic factor in severe heat stroke.

Objective To observe the expressions of cytochrome C,Caspase-9,Caspase-3 and their relationships,and investigate apoptosis signal transduction mechanism after heat stress-induction in human umbilical vein endothelial cell (HUVEC),and explore pathogenesis of vascular endothelial damage in the wake of severe heat stroke.Methods Human umbilical vein endothelial cell heat stress model was set up.Control group were incubated at 37℃,while heat stress group of cells were incubated at 39℃,41℃,and 43℃ for 2h,then all the cells were further incubated at 37℃ for 24 h.Mitochondria of human umbilical vein endothelial cell were examined with electron microscopy.Apoptosis was analyzed by flow cytometry using Annexin V-FITC/PI staining,and protein levels of cytochrome C,caspase-9,caspase-3 were analyzed by western blot.Results In the control group (37℃),the structure of mitochondrial was intact in HUVEC under transmission electron microscope.In contrast,mitochondrial swelling was found in the group of 43℃ heat stress.Compared with control group,as increasing in heat stress temperature,the rates of induced apoptosis were 17.8％ at 41℃ and 25.6％ at 43℃,and the levels of cytochrome C,Caspase-9,and caspase-3 were significantly increased (P ＜0.05).There was no obvious change at 39℃ heat stress (P ＞ 0.05).Conclusions Mitochondrial apoptosis pathway is involved in apoptosis of human umbilical vein endothelial cells in the wake of heat stress.The vascular endothelial cells apoptosis may be associated with the occurrence of severe heat stroke.

Objective To investigate the effects of artesunate (ART) on interstitial pneumonia and sialadenitis in MRL/lpr mice.Methods A total of 18 MRL/lpr mice were randomly allocated to a hydroxychloroquine sulfate (HCQ) group,a ART group and a control group.At the age of 18 weeks,the mice in the HCQ group and ART group were given HCQ 150 mg/kg daily and ART 50 mg/kg daily for 12 weeks,respectively.The histopathological changes of pneumonitis and submaxillaritis were assessed by hematoxylin and eosin staining.The levels of monocyte chemoattractant protein-1 (MCP-1) in the serum and urine were measured by the enzyme-linked immunosorbent assay.Results At the age of 30 weeks,the index of peribronchiolar lesion (1.62 ± 0.19,1.52 ± 0.30 vs.1.95 ± 0.34;all P＜0.05),the index of perivascular lesion (1.23 ± 0.18,1.28 ± 0.12 vs.1.57 ± 0.33;all P＜0.05),the alveolar lesions index (1.35 ± 0.16,1.05 ± 0.15 vs.1.72 ± 0.34;all P＜0.05) and the submaxillaritis index (1.48 ± 0.22,1.43 ± 0.15 vs.1.84 ± 0.34;all P＜0.05) in the HCQ group and the ART group were significantly decreased than those in the control group.The MCP-1 levels in the serum (1 103.02 ± 185.56 pg/ml,1 072.37 ± 242.43 pg/ml vs.1 490.67 ± 329.43 pg/ml;all P＜0.05) and urine (189.16 ± 70.85 pg/ml,198.79 ± 113.47 pg/ml vs.446.79 ± 192.31 pg/ml;all P＜0.05) in the HCQ group and the ART group were significantly lower than those in the control group.Conclusion ART can decrease the MCP-1 level,and ameliorate interstitial pneumonitis and sialadenitis in MRL/lpr mice.

Objective To explore the effect of activation of caspase-3 on apoptosis of human umbilical vein endothelial cells induced by heat stress. Methods A model of human umbilical vein endothelial cell heat stress was established. The endothelial cells in the control group were incubated at 37 ℃, while the cells in the heat stress group were incubated at 39 ℃, 41 ℃, or 43 ℃ for 2 h , then all the cells were further incubated at 37 ℃ for 24 h. Apoptosis was detected using staining with Hoechst33258; protein levels of caspase-3 were analyzed by Western blot; and the effect of caspase-3 inhibitor Z-DEVD-FMK on heat stress-induced apoptosis was determined. Results As compared with the control group, with an elevation in heat stress temperature (41℃ to 43 ℃), apoptosis was markedly increased and level of caspase-3 was significantly increased. In addition , caspase-3 inhibitor Z-DEVD-FMK significantly decreased the heat stress-induced apoptosis and levels of caspase-3. Conclusions Caspase-3 mediates apoptosis of human umbilical vein endothelial cells can be induced by heat stress.

Objective To investigate the therapeutic effects and mechanisms of hydroxychloroquine (HCQ) in the MRL/lpr mice. Methods MRL/lpr mice were divided into HCQ, the artesunate (ART) and proteinuria was detected with Coomassi Brilliant blue method. Enzyme linked immunosorbent assay (ELISA) was used to measure the anti-doubM-stranded DNA (ds-DNA) antibody. Renal tissue sections were dyed By PAS methods. The percentage of CD4+ Foxp3+ T cells in the spleen and lymph nodes were detected by flow 2.0) mg groups were decreased than in the control group (4.8±3.2) mg (P<0.05). And it was also lower in the HCQ (2.8±1.1) mg and ART (2.4±1.9) mg group than in the control group (6.4±1.9) mg (P<0.01) at 30 in the control group (37.1±1.0) g (P<0.01), while serum creatinine decreased significantly (7.8±4.0) μmol/L than in the control group (12.5±2.3) μmol/L (P<0.05), and the serum anti ds-DNA antibodies levels (3047±renal damage in the HCQ group and in the ART group was Both significantly improved than that in the entages of CD4+ Foxp3+ T cells in spleen when compared with the control group (1.5±0.5)% (P<0.05). The mice in the HCQ group (0.68±0.33)% and in the ART group (0.97±0.28)% had higher percentages of CD4+ Foxp3+ T cells in lymph nodes as compared with control group (2.15±0.72)%(P<0.01). Conclusion HCQ is effective in treating MRL/lpr lupus mice. It can improve the pathologic lesions of lupus nephritis, reduce proteinuria and antibody production. Both HCQ and ART can up-regulate the percentage of CD4+ Foxp3+ T cells in spleen of MRL/lpr mice.

Objective To investigate the expression of slit diaphragm proteins of glomerular podocyte,such as NEPH1 and Nephrin in type Ⅴ lupus nephritis (V-LN). Methods Twenty-five patients with V-LN and 18 patients with idiopathic membranous nephritis (IMN) were enrolled into the study, and 5 normal renal samples were the normal control group. Twenty-four hours urine protein excretion, serum albumin, creatinine, triglyceride, total cholesterol, serum C3, C4, urine C3 and NAG were tested respectively.Glomerular lesions were measured by light microscopy. The expressions of NEPH1 and Nephrin were determined by indirect immunofluorescent staining. The statistical treatment was used t-test. Results Compared to the IMN group, the 24 hours urine protein excretion and the concentrations of serum albumin, creatinine, urine C3 were not significantly different while the triglyceride, total cholestorel, serum C3, C4 were significantly decrease in the V-LN group (P<0.05). Urine NAG was increased in the V-LN group (P<0.01). By indirect immuno-fluorescent histochemitry examination, the glomerular expressions of NEPH1 and Nephrin were significantly decreased in both V-LN and IMN. Compared with the IMN group, the decrease of NEPH1 and Nephrin expression was more remarkable in the V-LN group. Conclusion The expression changes of NEPH1 and Nephrin may play an important pathogenic role in proteinuria of Ⅴ lupus nephritis. Renal tubular epithelial cell damage may play a role in proteinuria of V-LN.