Objective To explore the curative effect and nursing of enhanced external counterpulsation (EECP) on patients with ischemic cerebral apoplexy. Methods In September 2013 to March 2013 in Shenzhen Futian District People′s Hospital neurology hospital, toally 286 patients with ischemic cerebral apoplexy were randomly divided into two groups, control group and treatment group with 143 cases in each group. The control group used conventional treatment and the treatment group used EECP. The score of clinical nerve function and defect improved Barthel index were compared. Result The score of clinical nerve function defect and improved Barthel index of the treatment group before the treatment were without difference (all P>0.05). The score of clinical nerve function defect and improved Barthel index of the treatment group were less than those of control group after the treatment (all P < 0.05). Conclusion Ischemic stroke patients with positive EECP can significantly increase clinical neural function and life ability , improve patient′s quality of life.

Objective To study the mechanism of traditional Chinese medicine Compound Xueniaoting (CXNT) in treating glomerulus diseases with the main pathological changes of proliferation in mesangial cells. Methods CXNT and rat liver microsomes were incubated together, and then the incubated CXNT was added into cultured glomerular mesangial cells (GMC) in vitro. The proliferation of GMC was observed by MTT assay, the levels of interleukin-6 (IL-6) and endothelin-1 (ET-1) were determined by radioimmunoassay, and content of lactic dehydrogenase (LDH) was determined by velocity assay. ResultsCXNT (2, 4, 8 mg/mL) metobolized by rat liver microsomes could all inhibite the proliferation of GMC and production of IL-6 and ET-1 in a dose-dependent manner. Conclusion CXNT can inhibite the proliferation of GMC and restrain the secretion of IL-6 and ET-1, this function may be one of CXNT mechanisms in treating some mesangial proliferative glomerulonephritis.

Objective To study the granulocyte colony stimulating factor ( G-CSF ) and granulocyte macro-phage colony stimulating factor ( GM-CSF) in complete remission of acute myeloid leukemia ( AML) patients with den-dritic cells(DC)changes in the function and its subsets .Methods 36 cases of complete remission according to AML patients were randomly divided into 3 groups,G group was given G-CSF 200g/d,subcutaneous injection ,GM group were given GM-CSF 200g/d,subcutaneous injection ,the control group were injected with normal saline .The curative effects were compared.Results The expression of G in group DC,CD83 and CD86 on surface of CD80 were lower than that in GM group and C group (t=4.34,5.43,4.54,4.54,5.25,3.54,all P<0.05),GM group,DC expression of CD11c was higher than that of G group and C group (t=4.54,4.27,all P<0.05).G group and GM group DC in promoting lymphocyte proliferation were higher than those in C group (t=4.54,5.64,all P<0.05);group GM DC to promote the ability of CD4 +T lymphocyte proliferation was higher than that of G group (t=3.54,P<0.05).ELISA assay,GM group DC secretion of IL-12 than that of group G and C group;group G DC secretion of IL-10 was higher than that of GM group and C group (t=3.54.4.23,4.32,3.87,all P<0.05).Conclusion G-CSF and GM-CSF can make the complete remission in patients with AML subgroup DC shift ,the bias of G-CSF DC2 and certain immuno-suppressive effect ,while GM-CSF DC subsets of DC 1 and promotes cell immune deviation .

Objective To report the treatment of 7 cases of carpal tunnel syndrome(CTS)with algesia.Methods One hundred and twenty eight cases of carpal tunnel syndrome within the period of March 2002 and March 2005 were retrospectively analyzed.There were 7 cases(4 female and 3 male)had algesia,4 cases were treated with endoscopic management of carpal tunnel release (ECTR) and 3 cases were treated with open management of carpal tunnel release(OCTR).These 7 cases were followed-up 1-4years(average 1.5 years)postoperatively.Results Two ECTR cases and 2 OCTR cases had bad therapeutic effect and the others had good effect.Both 2 bad-effect ECTR cases feel special pain when insert the catheter.Only inject Triamcinolone Acetonide-A within epineurium after completely release in the goodeffect OCTR ease.Conclusion CTS with algesia is a special type of CTS,the key to treat it is to protect epineurium.

Objective To investigate the expression of nestin, a type Ⅵ intermediate filament protein in the glomeruli with foot process effacement and the potential relationship between nestin expression in the kidney and the degree of proteinuria. Method Immunohistochemistry was used to determine the localization of nestin in the kidney samples obtained from needle biopsies of normal human and patients with minimal change disease (MCD). Puromycin aminonucleoside (PAN) nephrosis rat models were established by a single intraperitoneal injection of PAN. Both real time quatitative reverse PCR and Western blot methods were applied to evaluate the levels of nestin expression at day 1, 4, 10 and 20 after PAN injection. Results Immunohistochemistry showed that the expression of nestin in glomeruli of MCD patients was significantly reduced compared with normal samples (0.93±0.08 vs 1.65±0.12, P<0.05) . The mRNA and protein expressions of nestin in the rat kidney were transitorily increased by 1.23 folds and 1.48 folds of control group (NC) after 1 day of PAN injection (P<0.05), then decreased quickly in the following days. The mRNA levels of nestin in the kidney were 35.8% and 12.1% of NC after 4 days and 10 days of PAN injection, respectively, (P<0.01) as determined by real time PCR. After 20 days of PAN injury, nestin mRNA expression partly recovered to 65.8% of NC (P< 0.05 ). The protein levels of nestin detected by Western blot presented the similar trend, which were 77.0%, 58.0% and 83.4% of NC after 4 days, 10 days and 20 days of PAN injection, respectively (P<0.05). The degree of proteinuria in puromycin aminonucleoside nephrosis rats was negatively correlated with both mRNA and protein levels of nestin in the kidney(r=-0.667,P<0.05 and r=-0.621 ,P<0.05, respectively). Conclusions The expression of intermediate filament protein nestin is down-regnlated in the kidney characterized with foot process effacement and negatively correlated with the degree of proteinuria in puromycin aminonucleoside nephrosis rats. Nestin may play a potential role in modulating the structure and function of podocyte.

Objective To confirm the main pathway of chemokine-chemokine receptor which mediates the accumulation of regulatory T cell ( Treg) in pancreatic cancer .Methods The concentrations of protein of FOXP3 and chemokines of CCL2, CCL3, CCL5, CCL17, CXCL8 in human and mouse pancreatic cancer and adjacent normal pancreatic tissue were measured by the method of enzyme-linked immunosorbent assay (ELISA).The receptor of chemokine CCL5 (CCR5) in human and mouse pancreatic cancer were determined by the immunofluorescent stain .Results The concentration of FOXP 3 protein in human pancreatic cancer and adjacent normal pancreatic tissue as (487.5 ±534.1) and (162.6 ±42.0) pg/mg, respectively, while they were (84.6 ±54.1) and (14.4 ±7.6) pg/mg, respectively in mouse.The concentration of FOXP3 protein were significantly higher in pancreatic cancer than those in adjacent normal pancreatic tissue .The concentration of CCL2 in human pancreatic cancer and adjacent normal pancreatic tissue as (76.9 ±37.5), (40.8 ±25.5) pg/mg, and the concentration of CCL3 as (38.0 ±22.6), (21.3 ±16.5) pg/mg, and the concentration of CCL5 were (390.2 ±158.5), (59.1 ±22.8) pg/mg, and the concentration of CCL17 as (7.2 ±2.0), (4.1 ±2.4)pg/mg, and the concentration of CXCL8 as (9.3 ±5.5), (6.3 ±5.2)pg/mg.The concentration of CCL2, CCL5, CCL17 in pancreatic cancer was significantly higher than those in adjacent normal pancreatic tissue (P<0.05).The concentration of CCL2 in mouse pancreatic cancer and adjacent normal pancreatic tissue as (77.9 ±30.5), (43.6 ±16.6) pg/mg, and the concentration of CCL3 was (27.4 ±18.2), (14.0 ±4.5)pg/mg, and the concentration of CCL5 was (302.2 ±55.8), (64.5 ±30.3) pg/mg; and the concentration of CCL17 was (4.4 ±1.4), (2.2 ±1.0)pg/mg;and the concentration of CXCL8 was (55.1 ± 55.1), ( 93.4 ±7.3 ) pg/mg.The concentration of CCL2, CCL5, CCL17 in pancreatic cancer were significantly higher than those in adjacent normal pancreatic tissue , and the difference between the two groups was statistically significant (P<0.05).The level of FOXP3 in pancreatic cancer was positively correlated with the concentration of chemokine CCL 5 both in human and mouse pancreatic cancer .Immunofluorescent staining indicated that the FOXP3 +cells also expressed CCR5.Conclusions The CCL5-CCR5 is the main chemokine-chemokine receptor pathway mediating the accumulation of Treg cells in pancreatic cancer .

Objective To investigate the effects of 5-aza-2'-deoxycytidine (5-Aza-dC) alone or combined with trichostatin A(TSA) on cell proliferation, promoter methylation and mRNA expression level of PDX-1 gene in pancreatic β cells induced by high glucose toxicity. Method NIT-1 cells were treated in vitro by high glucose (33.3 mmol/L), then divided into five groups, control group, HG grpup, 5-Aza-dC treatment group, TSA interfere group and 5-Aza-dC + TSA group. Proliferation of NIT-1 cells, insulin secretion, promoter methylation and mRNA expression of PDX-1 gene were detected respectively. Results 5-Aza-dC and TSA alone or in combination could promote cell proliferation and recover insulin secretion in NIT-1 cells , could also reduce PDX-1 gene methylation and enhance expression of PDX-1 mRNA. Compared with single-treatment group , combined group was significantly different (all P < 0.05). Conclusion 5-Aza-dC and TSA could activate the expression of PDX-1 and, then recover insulin secretion in NIT-1 cells induced by high glucose. Combination of them had synergistic effect.

Objective To explore whether the stimulation effect of high phosphate on hyperplasia of human parathyroid cells and hyperparathyroidism through local cyclooxygenase 2 (COX2) up-regulation pathway. Methods Parathyroid glands were collected from 19 uremic patients undergoing parathyroidectomy.Expressions of COX1,COX2 and proliferative cell nuclear antigen (PCNA) of the glands were detected by immunohistochemistry.Primary parathyroid cells were cultured and treated with high or normal phosphate for 48 hours.Then expressions of COX2 and PCNA were detected by Western blotting and real-time PCR. Results Among 62 glands from above 19 patients,43 glands were nodular hyperplasia and 19 diffuse hyperplasia.Both high expressions of COX2 and PCNA were found in these blands.Expression of COX2 was found in both oxyphil and chief cells and was more in the diffuse hyperplasia glands than that in the nodular hyperplasia (P＜0.05).80.60％ and 85.20％ of COX2 positive cells in diffuse hyperplasia glands and nodular hyperplasia also expressed PCNA. High phosphate could stimulate iPTH secretion in vitro (P＜0.05).Expressions of COX2 and PCNA were higher in high phosphate group.(P＜0.05). Conclusion High phosphate may stimulate the hyperplasia of parathyroid cells by up-regulating the local COX2 expression.

Objective To observe the changes of renin-angiotensin system (RAS) in cultured mesangial cells by serum from 3/4 nephrectomized rats feeding with low protein diet and α-keto acid. Methods Thirty male SD rats received 3/4 nephrectomy (Nx) were placed on 18%normal protein diet (NPD,n=10),6% low protein diet(LPD,n=10) or 5% low protein plus 1%α-keto acid diet (LK,n=10) flor 12 weeks.Ten male SD sham-operated rats fed with 18% normal protein diet were used as control (sham group).In addition,mesangial cells were cultured in sera (10%) collected from above animals treated with or without losartan (0.02 mmol/L)for 48 hours.ELISA was applied to detect the level of Ang II,TGF-β1 and fibronectin (FN) in cell medium.Westem blotting was used to determine the protein level of ATI receptor (AT1R)and real-time PCR was used to detect the mRNA level of AT1R,TGF-β1 and FN. Results (1) Nutritional indices including body weight,total protein and albumin had no significant difference in each group. (2) Serum creatinine and 24 h pruteinuria were significantly inceased in nephrectomized groups compared to sham group(P<0.05,respectively).24 h proteinuria was greatly lower in LK group than that in NPD and LPD groups(P<0.05,respectively).(3)LK greatly decteased the level of Ang II[NPD(12.70±0.12)mg/g protein;sham(8.04±0.62)mg/g protein]in supernatant as well as the protein and mRNA expression of AT1R in cultured mesangial cells (P<0.05).(4)NPD serum directly induced higher secretion[FN:sham(20.58±0.46)g/g protein,NPD (39.84±0.06)g/g protein;TGF-β1:sham(10.12±O.56)mg/g protein,NPD(83.85±3.58)mg/g protein] and mRNA expression of FN and TGF-β1 compared with sham group (P<0.05).LPD decreased these increment (P<0.05) and LK showed stronger inhibitory effect (P<0.05). (5)Losartan application sharply reduced FN and TGF-β1 production both in supematant and in mRNA expression in NPD serum treated cells (P<0.05,respectively). Conclusion Low protein diet with α-keto acids supplement directly inhibits the RAS in mesangial cells which may contribute to its beneficial effect on the kidney.

Aim To explore the possibility of the multiple epitope DNA vaccines of hepatitis C virus (HCV). Methods A synthetic multiple epitope antigen gene PCX of HCV was cloned into vector pREP9(RSV promoter) and pcDNA3 (CMV promoter) to construct eukaryotic expression vectors pREP9/PCX and pcDNA3/PCX, then they were used to immunize mice and rabbits, the titer of specific humoral and cellular responses were detected and their safety were observed. Results In mice, specific anti-GZ-PCX antibody(IgG) was lower than 1∶ 1 000 and did not persist well. In rabbits, the highest titer of anti-GZ-PCX IgG reached at 1∶ 3 200 and remained for about one month. Delayed type hypersensitivity reactions (DTH)and proliferation response of peripheral lymphocytes were induced by GZ-PCX antigen. Body weights of immunized mice were normal and no obvious toxic reaction was observed. Conclusion The multiple epitope antigen gene of HCV could induce specific immune responses without obvious toxicity and it might be able to serve as an effective HCV vaccine candidate.