Objective To observe the effects of electroacupuncture on genu recurvatum after stroke. Methods 80 stroke patients with genu recurvatum were randomly assigned to treatment group (n=40) and control group (n=40). The control group accepted routine rehabilitation, and the treatment group accepted electroacupuncture at Yanglingquan (GB34), Futu (ST32), Weizhong (BL40), Chengshan (BL57) and Zusanli (ST36) in addition, for 30 days. The incidence of effectiveness was compared between groups. All the patients were assessed with range of motion (ROM) of knee and Fugl-Meyer Assessment of lower limbs (FMA) before and after treatment. Results The incidence of effectiveness was 72.5% in the treatment group, which was more than 55% in the control group (P<0.05). The ROM and score of FMA improved more in the treatment group than in the control group (P<0.05). Conclusion The electroacupuncture can promote the recovery of genu recurvatum after stroke.

Objective To summarize the preliminary experience of endovascular stent-graft exclusion for aortic dissections. Methods From October 2003 to February 2007,121 patients[86 males,37 females,mean age(53.7?13.8)years,range 29～ 72 years]underwent endovascular stent-graft exclusion for aortic dissections,including Stanford B in 114 patients,Stanford A in 4, and traumatic aortic mptore in 3.An emergency operation was performed in 4 patients for acute aortic rapture.Results No primary conversion was needed.There was no postoperative death,no spinal cord iscbemic injury,or stent displacement or subclavian steal syndrome.Postoperative hospital stay time was(4.0?1.3)days.Complications included fever in 35 patients,type Ⅳ endoleak in 11,type Ⅰ endoleak in 1 and acute renal dysfunction in 1.Contusion Endovascular thoracic aorta repair is an effective,less inva- sire and safe surgery for patients with Stanford B or some Stanford A aortic dissection and traumatic aortic rupture.

Objective To study the relationship between the clinical manifestations and changes of pituitary magnetic resonance imaging(MRI) in short stature children with growth hormone deficiency(GHD).Methods The pituitary MRI finding in 38 cases of short stature children diagnosed as GHD(males 23,females 15;5-14 years old,10 children were in pubertas and Tanner Ⅱ-Ⅲ) were analyzed,and the pituitary morphology,size,signal and pituitary stalk's shape and location were observed.SPSS 12.0 soffware was used to analyze the data.Results The forms of pituitery were plaque in 20 children(53%),cupped in 17 children(45%),and carinate in 1 children.In the 22 cases of completely GHD,18 cases had different levels of anterior pituitary dysplasia,abnormal pituitary stalk and/or pituitary signal changes,5 cases without posterior lobe disappeared high signal and 4 cases with pituitary stalk interruption syndrome;the other 4 cases had completely normal pituitary.In the 16 cases of partially GHD,7 cases had varying degrees of pituitary size and/or abnormal pituitary stalk,8 cases had completely normal pituitary,and 1 case had pituitary adenoma.Conclusion Pituitary MRI could assist diagnosis and evaluate pituitary function in short stature children.

Objective To study apoptosis and antigen presentation changes of monocytes in HBV-related acute-on-chronic liver failure (ACLF) patients under immune dysfunction state.Methods Peripheral blood samples of 26 HBV-related ACLF patients (ACLF group),20 active chronic hepatitis B patients (CHB group) and 18 healthy individuals (control group) were collected.The changes of apoptosis and proliferation (Ki67) in monocytes and the expression of surface markers including human leukocyte antigen (HLA)-DR and B7 molecules (CD86) of monocytes were analyzed by flow cytometry. Results The percentage of Annexin V expressed monocytes of ACLF group (64％) was significantly higher than that of CHB group (28％) and control group (20％),and the difference was statistically significant (x2 value was 11.75 and 27.23 ; both P＜0.01),which indicated that monocytes apoptosis increased.The Ki67 expression in monocytes of ACLF group was lower than that of CHB group and control group,and the difference was statistically significant (x2 value was 4.71 and 4.83; both P＜ 0.05),which indicated that activated monocytes reduced. The mean fluorescence intensity (MFI) of HLA-DR and CD86 of monocytes in ACLF group was 22.85 and 11.63,which was significantly lower than that of CHB group and control group,indicating the antigen presentation ability of monocytes injured. The percentage of Annexin Ⅴ positive monocytes in survivals (62 ％ ) was significantly higher than that of dead patients (46 ％ ) in ACLF group.Conclusion In HBV-related ACLF patients under immune dysfunction state,the apoptosis of peripheral blood monocytes increased,and the quantity of activated cells reduced,resulting in the decline of the antigen presentation ability of monocytes.

The relationship between tyrosine phosphorylation (TP) and protein expression of insulin receptor (InsR) and insulin resistance (IR) in patients with gestational diabetes mellitus (GDM) was investigated. The InsR expression and TP in skeleton muscle tissue were determined by Western blotting and immunoprecipitation in women with GDM (GDM group, n=22), normal pregnant women (normal pregnancy group, n=22) and normal non-pregnant women (normal non-pregnant group, n=13). Fasting plasma glucose (FPG) and fasting insulin (FINS) were measured by oxidase assay and immunoradioassay. The results showed that the levels of FPG (5.61±0.78 mmol/L), FINS (15.42±5.13 mU/L) and Homeostasis model assessment-IR (HOMA-IR) (1.21±0.52) in GDM group were significantly higher than those in normal pregnancy group (4.43±0.46 mmol/L, 10.56±3.07 mU/L and 0.80±0.31 respectively) (P<0.01). The levels of FINS and HOMA-IR in normal pregnancy group were significantly higher than those in normal non-pregnant group (7.56±2.31 mU/L and 0.47±0.26 respectively) (P<0.01). There was no significant difference in the InsR expression level among the three groups (P>0.05). TP of InsR with insulin stimulation was significantly decreased in GDM group (0.20±0.05) as compared with normal pregnancy group (0.26±0.06) (P<0.01). TP of InsR with insulin stimulation in normal pregnancy group was lower than that in normal non-pregnant group (0.31±0.06) (P<0.01). TP of InsR with insulin stimulation was negatively related with HOMA-IR in GDM group (r=-0.525, P<0.01). There was no correlation between the protein expression of InsR and HOMA-IR in GDM group (r=-0.236, P>0.05). It was suggested that there is no significant correlation between the protein expression of InsR in skeletal muscle and IR in GDM, but changes in TP of InsR are associated with IR in GDM.

OBJECTIVETo study the mechanism underlying the inhibitory effects of peroxisome proliferator-activated receptor γ (PPARγ) agonists on transforming growth factor β1 (TGF-β(1))-induced scarring of skin.

METHODSFibroblasts isolated from healthy adult skin were cultured in vitro and divided into blank control group (serum-free DMEM culture), TGF-β(1) group (with stimulation of 10 ng/mL TGF-β(1) for 48 hours), troglitazone group (with the same treatment as in TGF-β(1) group after stimulation of 10 µmol/L troglitazone for 2 hours), and 15-dioxygen prostaglandin J2 (15d-PGJ2) group (with the same treatment as in TGF-β(1) group after stimulation of 10 µmol/L 15d-PGJ2 for 2 hours) according to the stimulation added into DMEM. The expression of connective tissue growth factor (CTGF) was determined with Western blot. The mRNA levels of CTGF, matrix metalloproteinase-1 (MMP-1) and platelet-derived growth factor (PDGF) were determined with real-time fluorescence RT-PCR. Data were processed with one-way analysis of variance.

RESULTSThe expression of CTGF at mRNA and protein levels in skin fibroblasts were significantly increased in TGF-β(1) group as compared with control group; while expression of CTGF at mRNA and protein levels in 15d-PGJ2 and troglitazone groups were significantly decreased as compared with that in TGF-β(1) group. The mRNA level of MMP-1 in TGF-β(1) group (0.193 ± 0.051) was obviously lower than that in blank control group (1.281 ± 0.195, F = 12.811, P < 0.01), while the mRNA levels of MMP-1 in troglitazone group (0.417 ± 0.043) and 15d-PGJ2 group (0.485 ± 0.027) were significantly increased as compared with that in TGF-β(1) group (F = 12.811, P values all below 0.01). The mRNA level of PDGF in TGF-β(1) group (1.044 ± 0.237) was obviously higher than that in control group (0.349 ± 0.057, F = 16.848, P < 0.01), while the levels in troglitazone group (0.677 ± 0.055) and 15d-PGJ2 group (0.511 ± 0.017) were significantly decreased as compared with that in TGF-β(1) group (F = 16.848, P values all below 0.01).

CONCLUSIONSThe inhibitory effect of activated PPARγ on the expression of CTGF induced by TGF-β(1) may be the main mechanism of its inhibitory effect on TGF-β(1)-induced scarring on skin, and its influence on MMP-1 and PDGF may also be one of the underlying mechanisms.

Cell Line ; Connective Tissue Growth Factor ; metabolism ; Fibroblasts ; drug effects ; metabolism ; Humans ; Matrix Metalloproteinase 1 ; metabolism ; PPAR gamma ; agonists ; Receptors, Platelet-Derived Growth Factor ; metabolism ; Signal Transduction ; Transforming Growth Factor beta1 ; metabolism

OBJECTIVETo construct DNA methyltransferase 1 (DNMT1) low expression 16HBE cell line and observe the variation of cell cycle and global genomic DNA methylation.

METHODSThe method of Lenti-virus induced RNA interference was applied to introduce four different shRNA fragment into 16HBE cells. Flow cytometry and 5-mC immunofluorescence methods were used to observe the cell cycle and global DNA methylation status of DNMT1 low expression 16HBE cells.

RESULTSThe DNMT1 protein relative expression level of 16HBE-shDNMT1-4 cell line was down regulated about 44% (P < 0.05) compared with the control. No obvious differences of cell cycle and global genome DNA methylation status were observed between the 16HBE and 16HBE-shDNMT1.

CONCLUSIONThe DNMT1 gene low expression cell is successfully constructed, and there are no obvious changes happened on the cell cycle and global genomic DNA methylation.

Cell Cycle ; Cell Line ; DNA (Cytosine-5-)-Methyltransferase 1 ; DNA (Cytosine-5-)-Methyltransferases ; genetics ; metabolism ; DNA Methylation ; Down-Regulation ; Epithelial Cells ; metabolism ; Humans ; RNA Interference ; RNA, Small Interfering ; genetics

OBJECTIVETo examine the protective effect of ecdysterone (ECR) against beta-amyloid peptide fragment(25-35) (Abeta(25-35))-induced PC12 cells cytotoxicity, and to further explore its mechanism.

METHODSExperimental PC12 cells were divided into the Abeta group (treated by Abeta(25-35) 100 micromol/L), the blank group (untreated), the positive control group (treated by Vit E 100 micromol/L after induction) and the ECR treated groups (treated by ECR with different concentrations of 1, 50 and 100 micromol/L). The damaged and survival condition of PC12 cells in various groups was monitored by lactate dehydrogenase (LDH) release and MTT assay. The content of malondialdehyde (MDA) was measured by fluorometric assay to indicate the lipid peroxidation. And the antioxidant enzymes activities in PC12 cells, including superoxide dismutases (SOD), catalase (CAT) and glutathione peroxidase (GSH-Px), were detected respectively.

RESULTSAfter PC12 cells were treated with Abeta(25-35) (100 micromol/L) for 24 hrs, they revealed a great decrease in MTT absorbance and activity of antioxidant enzymes, including SOD, CAT and GSH-Px as well as a significant increase of LDH activity and MDA content in PC12 cells (P < 0.01). When the cells was pretreated with 1-100 micromol/L ECR for 24 hrs before Abeta(25-35) treatment, the above-mentioned cytotoxic effect of Abeta(25-35) could be significantly attenuated dose-dependently, for ECR 50 micromol/L, P < 0.05 and for ECR 100 micromol/L, P < 0.01. Moreover, ECR also showed significant inhibition on the Abeta(25-35) induced decrease of SOD and GSH-Px activity, but not on that of CAT.

CONCLUSIONECR could protect PC12 cells from cytotoxicity of Abeta(25-35), and the protective mechanism might be related to the increase of SOD and GSH-Px activities and the decrease of MDA resulting from the ECR-pretreatment.

Amyloid beta-Peptides ; toxicity ; Animals ; Catalase ; analysis ; Ecdysterone ; pharmacology ; Glutathione Peroxidase ; analysis ; L-Lactate Dehydrogenase ; analysis ; Malondialdehyde ; analysis ; PC12 Cells ; Peptide Fragments ; toxicity ; Rats

OBJECTIVETo explore the effects of soothing liver and invigorating spleen recipes on lipopolysaccharide(LPS) induced hepatocyte inflammation of rats and TLR4/p38MAPK signal pathway.

METHODThe hepatocytes of SD rats were cultured and identified in vitro. The medicated serum of soothing liver and invigorating spleen recipes was prepared. The hepatocytes were treated with soothing liver and invigorating spleen recipes. Then Interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) expression in cultural supernatants were assayed by ELISA. The expressions of Toll-Like 4 (TLR4), p38 mitogen activated protein kinases (p38MAPK) and p-p38 mitogen-activated protein kinase (p-p38MAPK) were detected by Western blot.

RESULTThe rat medicated serum of soothing liver and invigorating spleen recipes was extracted for 2-3 mL. The purified rat hepatocytes were 1.5 x 10(8)-2.0 x 10(8). The cell viability was above 95% detected by Typan blue staining. The hepatocytes were identified by immumofluorescence assay. The detection of hepatocyte cultural supernatants: compared with that of the control group, IL-6 and TNF-α expression were increased in the LPS group (P < 0.01). While compared with that of the LPS group, the expressions of IL-6 and TNF-α were decreased after soothing liver and invigorating spleen recipes intervention (P < 0.01). The detection of hepatocyte proteins: compared with that of the control group, the protein expressions of p38MAPK, p-p38MAPK and TLR4 were all increased significantly in the LPS group (P < 0.01). Compared with that of the LPS group, the protein expressions of p38MAPK was decreased significantly in SB239063 group and it was also decreased in the soothing liver and invigorating spleen recipes group, but with no significant difference. Compared with that of the LPS group, p38MAPK expression was reduced significantly in the soothing liver and invigorating spleen recipes group and the SB239063 (p38MAPK pathway inhibitor) group (P < 0.01). TLR4 protein expression was decreased markedly in the soothing liver and invigorating spleen recipes group (P < 0.01) but had no difference between the SB239063 group and the LPS group.

CONCLUSIONThe soothing liver and invigorating spleen recipes may regulate hepatocyte inflammatory injury of rats through TLR4/p38MAPK signaling pathway.

Animals ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hepatocytes ; drug effects ; metabolism ; Humans ; Lipopolysaccharides ; adverse effects ; Liver ; drug effects ; injuries ; metabolism ; Male ; Non-alcoholic Fatty Liver Disease ; drug therapy ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction ; drug effects ; Spleen ; drug effects ; metabolism ; Toll-Like Receptor 4 ; genetics ; metabolism ; p38 Mitogen-Activated Protein Kinases ; genetics ; metabolism

OBJECTIVETo investigate the effects of hydroquinone (HQ) on expression of ubiquitin-ligating enzyme Rad18 in human hepatic cells (L-02), and to explore the role and possible mechanism of Rad18 involved in toxicity of HQ to hepatic cells.

METHODSAfter L-02 hepatic cells were exposed to HQ with various concentrations (0, 5, 10, 20, 40, 80 and 160 micromol/L) for 24 h, cell survival rate was measured by MTT assay; DNA impairment was evaluated by single cell gel electrophoresis (SCGE); The expression levels of Rad18 mRNA and protein were detected by Real-time fluorescent quantitative polymerase chain reaction (QPCR) technique and Western blot method respectively.

RESULTSHQ with concentration from 0 to 80 micromol/L had little effect on survival rate of L-02 (P > 0.05); Whereas the survival rate in the group of 160 micromol/L was significantly lower than in the control with the significant difference (P < 0.01) after treated with HQ for 24 h; The higher dose of HQ presented, the more degrees of olive tail moment (OTM) were produced and a dose-dependent relationship was shown. HQ in a low concentration (0 to approximately 40 micromol/L) could induce increase in the expression of Rad18 mRNA and protein which was in proportion to the increment of HQ concentration; the expression of Rad18 mRNA was enhanced increasingly, while the expression of Rad18 protein unchanged basically once the concentration of HQ exceeded 40 micromol/L; Besides, there was a positive correlation between OTM and the expression level of Rad18 mRNA (r = 0.919, P < 0.01).

CONCLUSIONHQ could regulate up the expression of Rad18 in L-02 hepatic cells.

Cell Survival ; drug effects ; Cells, Cultured ; DNA Damage ; drug effects ; DNA-Binding Proteins ; metabolism ; Hepatocytes ; drug effects ; enzymology ; Humans ; Hydroquinones ; toxicity ; Ubiquitin-Protein Ligases