OBJECTIVETo observe clinical therapeutic effect of acupuncture combined with Yizhi Jiannao Granules for treatment of Alzheimer's disease and its effects on intelligence, daily life and social activity ability.

METHODSEighty-four cases were randomly divided into 3 groups, 28 cases in each group. The combined acupuncture and medication group was treated with acupuncture at Baihui (GV 20), Sishencong (EX-HN 1), Dazhui (GV 14), Guanyuan (CV 4), etc. and oral administration of Yizhi Jiannao Granules; the Chinese herb group was treated with Yizhi Jiannao Granules, and the western medicine group with oral administration of Aricept. The scores for the Mini-Mental State Examination (MMSE), Ability of Daily Life (ADL) and the therapeutic effects were assessed and compared before treatment and after treatment for 12 weeks among the groups.

RESULTSAfter treatment, the scores for MMSE and ADL were improved in the combined acupuncture and medication group, the Chinese herb group and the western medicine group, which were better in the combined acupuncture and medication group (P < 0.05). The total effective rate of 85.7% in the combined acupuncture and medication group was better than 71.4% in the Chinese herb group and 67.9% in the western medicine group.

CONCLUSIONAcupuncture combined with Yizhi Jiannao Granules has a significant therapeutic effect on Alzheimer's disease, which is better than that of Yizhi Jiannao Granules or Aricept.

Acupuncture Points ; Acupuncture Therapy ; methods ; Administration, Oral ; Aged ; Alzheimer Disease ; psychology ; therapy ; Combined Modality Therapy ; Drugs, Chinese Herbal ; administration & dosage ; therapeutic use ; Female ; Humans ; Indans ; administration & dosage ; therapeutic use ; Male ; Middle Aged ; Nootropic Agents ; administration & dosage ; therapeutic use ; Piperidines ; administration & dosage ; therapeutic use ; Quality of Life ; Treatment Outcome

OBJECTIVETo investigate the characteristics of the spontaneous isolated superior mesenteric artery dissection (SISMAD) on computed tomography angiography (CTA).

METHODSTwenty-five patients with unexplained acute abdominal pain received CTA.

RESULTSFour cases with the SISMAD were found and all were male with a mean age of (45.3±6.7) years. Two patients had hypertension history. CT showed enlarged diameter of the superior mesenteric artery with dissection in 4 cases, intimal flap and visible false lumen in 2 cases, ulcer-like laceration in 1 case, and intramural hematoma in 1 case. The proximal lacerations or entries were all at the proximal segment of the superior mesenteric artery. CTA classifications were type Ia (n=2), IIb (n=1), and III (n=1). Two patients underwent repeated CTA when discharged, and progressive changes were discovered.

CONCLUSIONCTA can clearly show the characteristics of the superior mesenteric artery dissection, confirm the diagnosis, and provide an important basis for the classification and follow-up observation.

Adult ; Aged ; Aged, 80 and over ; Aneurysm, Dissecting ; diagnostic imaging ; Angiography ; methods ; Female ; Humans ; Male ; Mesenteric Artery, Superior ; diagnostic imaging ; Middle Aged ; Retrospective Studies ; Tomography, X-Ray Computed ; Young Adult

Objective To establish a liquid chromatography-tandem mass spectrometry(LC-MS/MS)method for the determination concentration of duloxetine in human plasma and study on its pharmacokinetics in healthy human.Methods The separation of duloxetine was performed on Phenomenex C_(18) column with fluoxetine as the internal standard.The mobile phase was composed of 5 mmol L~(-1) ammonium acetate with 0.02% formic acid acetonitrile(55:45).Electrospray ionization source was applied and operated in positive ion mode.A single dose of 60 mg duloxetine hydrochloride was given to 5 male and 5 female healthy volunteers and the plasma was separated.The concentration of duloxetine was determined by HPLC-MS/MS and the pharmacokinetic parameters were calculated.Results The linear range of duloxetine was 0.89-106.80 ng·mL~(-1)(γ=0.9977).The methodological recovery and the extraction recovery ranged between 93.19%-107.27% and 72.81%-89.96%,respectively.Both the inter-day RSD and intra-day RSD were less than 11%.The main pharmacokinetic parameters after a single dose of 60mg duloxetine are as follows: C_(max) was(44.40 ±17.78)ng·mL~(-1),t_(max) was(6.10±1.29)h,t_(1/2) was(12.81 ±2.31)h;AUC_(0-60) and AUC_(0-∞) were (696.04±337.82),(733.82±343.40)ng·h·mL~(-1),respectively.Conclusion The method is simple,accurate and reliable,and suitable for the determination of duloxe-tine in therapeutic drug monitor and its pharmacokinetics study.

OBJECTIVETo compare the effects of Bushen Jiedu Recipe (BJR) and Jianpi Jiedu Recipe (JJR) containing plasma on dendritic cells (DCs) of chronic hepatitis B virus (HBV) infection patients under different immune states.

METHODSRecruited were 36 chronic HBV infection outpatients from First Affiliated Hospital of Hunan University of Traditional Chinese Medicine from April 2010 to January 2011. They were assigned to the immune tolerance group (18 cases) and the immune clearance group (18 cases).Another 10 healthy subjects were recruited as the healthy control group. Their anticoagulated peripheral venous blood was respectively collected. The peripheral blood mononuclear cells (PBMCs) were isolated and further extracted for incubating DCs. The DCs were intervened by BJR and JJR containing plasma. The morphology of DCs was identified. The expressions of CD1alpha, CD80, CD86, and HLA-DR were detected. The level of interferon-alpha (IFN-alpha) in the supernatant was observed by ELISA.

RESULTSThe CD80 expression level was lower in the immune clear group than in the healthy control group before intervention (P < 0.05). The expression levels of CD80, CD86, and HLA-DR were lower in the immune tolerance group than in the healthy control group before intervention (P < 0.05).The IFN-alpha expression level was lower in the immune tolerance group and the immune clearance group than in the healthy control group before intervention (P < 0.05). The expression levels of CD80, HLA-DR, and IFN-alpha were lower in the immune tolerance group than in the immune clearance group before intervention (P < 0.05). Compared with the same group before intervention, the CD80 expression significantly increased in each treatment group (P < 0.05). After intervention the expression levels of CD80 and HLA-DR were higher in the immune tolerance group than in the immune clearance group in the same time phase, and the CD86 expression level was higher in the BJR group than in the immune clearance group in the same time phase, showing statistical difference (P < 0.05).

CONCLUSIONSThe middle dose BJR and the small dose JJR both could promote the recovery of DCs in chronic HBV infection patients. Besides, BJR showed more prominent effects on the function of DCs in chronic HBV infection patients in the immune tolerance stage.

Adult ; B7-1 Antigen ; metabolism ; B7-2 Antigen ; metabolism ; Case-Control Studies ; Dendritic Cells ; drug effects ; immunology ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Female ; HLA-DR Antigens ; metabolism ; Hepatitis B, Chronic ; blood ; drug therapy ; immunology ; Humans ; Immune Tolerance ; drug effects ; Interferon-alpha ; metabolism ; Male ; Phytotherapy ; Plasma ; Young Adult

OBJECTIVETo investigate the relation between p16 gene expression and the carcinogenesis and progress of hepatitis B virus (HBV) related hepatocellular carcinoma (HCC).

METHODSIn 35 specimens of HCC tissue and the adjacent liver tissue, the integration of HBV X gene was detected by polymerase chain reaction (PCR) and Southern blot. The point mutation of exon-1alpha, 2 and 3 of p16 gene were detected by PCR-single strand conformation polymorphism (SSCP). The expression of p16 mRNA and p16 protein was detected by RT-PCR and Western blot.

RESULTSThe integration of X gene correlated with the expression loss of p16 mRNA and p16 protein in HCC (P < 0.05). The expression loss rates of p16 protein in HCC and adjacent tissues were 62.9% (22/35) and 40.0% (14/35) with significant difference (P < 0.05). The expression loss of p16 protein in HCC correlated with the differentiation degrees of HCC and the infiltration of tumor cells (P < 0.05).

CONCLUSIONThe integration of X gene correlates with the expression loss of p16 protein. The alteration of p16 gene, playing an important role in all stages of hepatocarcinogenesis, correlates with the progress and invasion of hepatocellular carcinoma.

Carcinoma, Hepatocellular ; genetics ; metabolism ; virology ; Cyclin-Dependent Kinase Inhibitor p16 ; biosynthesis ; genetics ; Exons ; Female ; Genes, p16 ; Hepatitis B Antigens ; metabolism ; Hepatitis B virus ; genetics ; Humans ; Liver Neoplasms ; genetics ; metabolism ; virology ; Male ; Middle Aged ; Point Mutation ; RNA, Messenger ; biosynthesis ; genetics ; Trans-Activators ; genetics ; Virus Integration

OBJECTIVETo investigate the characteristics of lymphatic metastasis in different types of adenocarcinoma of the esophagogastric junction (AEG) and provide guidance for surgical approach adoption.

METHODSClinical data of 228 patients with AEG undergoing surgery were analyzed retrospectively. According to Siewert classification, there were 9 cases of type I (3.9%) who all underwent left thoracoabdominal approach procedures. A total of 121 patients belonged to type II (53.1%), of whom 12 underwent left transthoracic approach, 48 left thoracoabdominal approach, and 61 transabdominal approach. Ninety-eight patients belonged to type III (43%), of whom 22 underwent left thoracoabdominal approach procedures and 76 transabdominal. The pattern of lymph node metastasis was analyzed and the association between surgical approach and oncological clearance was examined.

RESULTSThe resection margin was positive in 20(8.8%) patients, including 10 with type II (8.3%) and 10 with type III (10.2%), and the difference was not statistically significant (P>0.05). The rate of positive resection margin was 12.4%(17/137) in the transabdominal group and 16.7%(2/12) in the left transthoracic group, both significantly higher than the left thoracoabdominal group (1.1%, 1/88) (both P<0.05). Lymph node metastasis was found in 159(69.7%) patients. The metastasis was found in 4 of 9 patients with type I cancer and two were thoracic metastasis, no metastasis was found in the upper mediastinum. For type II cancer, the rate of lymph node metastasis was 66.9%(81/121), including thoracic metastasis ( n=32, 26.4%) and abdominal metastasis (n=81, 66.9%). For type III cancer, the rate of lymph node metastasis was 66.9%(81/121), including thoracic metastasis (n=15, 15.3%) and abdominal metastasis (n=69, 70.4%).

CONCLUSIONSFor type I AEG, left thoracoabdominal approach should be used because the pattern of lymph node metastasis is similar to that of the distal esophageal carcinoma. For type II , left thoracoabdominal approach should be used to ensure adequate resection of the tumor and clearance of lymph node in the lower esophagus and upper mediastinum because of high rate of intrathoracic lymph node metastasis. For type III cancer, transabdominal incision offers better benefit with less impact on respiratory function. However, thoracic incision should be used to ensure adequate clearance for tumors of larger size and significant external invasion.

Adenocarcinoma ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Esophagectomy ; methods ; Esophagogastric Junction ; pathology ; surgery ; Female ; Humans ; Lymph Node Excision ; methods ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

BACKGROUNDStroke volume variation (SVV) is a robust indicator of fluid responsiveness during volume change. We compared the sensibility of SVV by Vigileo/Flotrac to central venous pressure (CVP) when volume changes in patients undergoing intraoperative acute normovolemic hemodilution (ANH) and acute hypervolemic hemodilution (AHH).

METHODSForty patients were randomly divided into an ANH group (n = 20) and an AHH group (n = 20). All patients received general anesthesia and were mechanically ventilated. Data were collected from 7 different time-points in the ANH group: baseline, after withdrawal of 5%, 10%, and 15% of the estimated blood volume (EBV) and after replacement with an equal volume of 6% hydroxyethyl starch 130/0.4 (HES) in 5% EBV increments to baseline. There were four time points in the AHH group: baseline, after 5%, 10%, and 15% expansion of the EBV with 6% HES. At each time-point, CVP, SVV and other hemodynamic parameters measurements were obtained.

RESULTSAfter removal of 10% and 15% EBV, SVV significantly increased from 10.9 ± 3.0 to 14.1 ± 3.4 and 10.9 ± 3.0 to 16.0 ± 3.3 (P < 0.01), and returned to a final value of 10.6 ± 3.4 after volume replacement. The CVP value was unchanged after removal and replacement of 15% of the EBV. There were no significant changes in SVV after 5%, 10% whereas there was a significant reduction after 15% (8.2 ± 1.7) expansion of the EBV compared with baseline (9.9 ± 1.8) (P = 0.033). However, there was a significant increase in CVP after 10% (10.3 ± 2.4), 15% (11.3 ± 2.2) expansion of the EBV compared with baseline (8.2 ± 2.7) (P < 0.01).

CONCLUSIONSVV is a more sensitive parameter for volume than CVP during hypovolemia, on the contrary CVP is more sensitive than SVV during hypervolemia.

Anesthesia ; Central Venous Pressure ; physiology ; Hemodilution ; Humans ; Hypovolemia ; physiopathology ; Stroke Volume ; physiology

OBJECTIVETo establish the model of bone mesenchymal stem cell-derived smooth muscle cells (BMSC-SMCs) and investigate the role of BMSC-SMCs in the development and progression of artherosclerosis.

METHODSBMSCs were isolated from the femoral bone of SD rats by adherent tissue culture method, and vascular smooth muscle cells (VSMCs) were obtained from the thoracic aorta. The differentiation of BMSCs into BMSC-SMCs was induced in the conditioned medium. The specific markers of BMSCs and BMSC-SMCs were identified by immunofluorescence (IF) staining. After treatment with 80 mg/L oxidative low-density lipoprotein (ox-LDL) for 72 h, the growth characteristics of BMSC-SMCs and VSMCs were observed. Flow cytometry was applied to analyze the cell cycle of BMSC-SMCs and VSMCs.

RESULTSBMCS-SMCs transformed into foam cells after treatment with ox-LDL, which was more obvious in comparison with VSMCs. The growth curve of BMSC-SMCs and VSMCs presented with an S-shape pattern with the cell doubling time of 20 and 32 h, which was reduced to 15 and 28 h after treatment with 80 mg/L ox-LDL, respectively. Flow cytometry showed that exposure to 80 mg/L ox-LDL significantly increased G(0)/G(1) and decreased S and G(2)/M phase cells in both BMSC-SMCs (P<0.01, n=3) and VSMCs (P<0.05, n=3) in comparison with the control cells.

CONCLUSIONBMSC-SMC might be involved in the formation of fatty core and accelerate the development of atherosclerosis.

Animals ; Atherosclerosis ; etiology ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cell Proliferation ; drug effects ; Cells, Cultured ; Female ; Foam Cells ; cytology ; Lipoproteins, LDL ; pharmacology ; Male ; Mesenchymal Stromal Cells ; cytology ; Muscle, Smooth, Vascular ; cytology ; Rats ; Rats, Sprague-Dawley

Based on site-specific transposition of an expression cassette into a baculovirus shuttle vector (Bacmid) which propagated in Escherichia coli, the Bac-to-Bac System provides a rapid and efficient method to generate recombinant baculoviruses and is widely used for high level expression of heterologous proteins. And the efficiency of recombinant baculovirus infecting cells plays an important role on the protein expression. In this study, we introduced an EGFP expression cassette driven by polyhedrin promoter into the p74 locus of Bacmid by homologous recombination. The target Bacmid-egfp was then transformed into E. coli DH10B containing the transposition helper plasmid to gain a new transposition receipt strain E. coli DH10Bac-egfp. Because of the intact attTn7 sites and lacZ', target gene cloned in a pFastBac vector can be transposed into the Bacmid-egfp shutter vector to construct recombinant baculovirus, which would allow the tracing of the target protein expression and the recombinant Bacmid transfection or recombinant baculoviral infection under fluorescence microscopes. Recombinant virus Bac-egfp-DsRed was constructed by transposing DsRed into the Bacmid-egfp in E. coliDHl0Bac-egfp, and the Sf9 cells infected with the recombinant virus expressed DsRed and EGFP efficiently. Another protein IL-6 fused with 6 x his tag was expressed and purified sucessfully from Sf9 cells infected with recombinant virus Bac-egfp-6 x his-IL6 constructed by the improved Bac-to-Bac system.
Animals ; Baculoviridae ; genetics ; Green Fluorescent Proteins ; genetics ; Interleukin-6 ; biosynthesis ; genetics ; Plasmids ; Polymerase Chain Reaction ; Recombinant Proteins ; biosynthesis ; Spodoptera

OBJECTIVETo explore anti-inflammation and mechanism of Qinghuachang Decoction (QD) by using LPS stimulated differentiated colon cancer Caco-2 cells (as an inflammation model of human enterocytes).

METHODSQD was prepared. Human colonic epithelial Caco-2 cells were cultured. Expressions of TNF-alpha and IL-8 were determined using ELISA. Expressions of inhibitory Kaba protein (IkappaB-alpha), phosphorylated inhibitory Kaba protein (p-lkappaB-alpha), nuclear transcription factor p50 (p50), and nuclear transcription factor ReIA (ReIA) protein were determined by Western blot.

RESULTSCompared with the negative control group (without LPS stimulation), LPS stimulated the release of IL-8 and TNF-alpha in Caco-2 cells (P < 0.05). QD treatment could reduce the secretion of TNF-alpha and IL-8 induced by LPS in a dose dependent manner (P < 0.05). QD at 0, 5, 10, and 50 microg/mL had no significant effect on Caco-2 cell survival rates (P > 0.05), with no statistical difference among various concentrations (P > 0.05). QD could significantly suppress nuclear factor-kappa B (NF-kappaB) phosphorylation stimulated by LPS. The expression of p-IKappaB-alpha was decreased with increasing concentrations of QD (P < 0.05). There was no obvious change in IKB-alphaB expressions (P > 0.05). Expressions of p50 and ReIA decreased with increasing concentrations of QD (P < 0.05). Both of them were in a dose dependent manner.

CONCLUSIONQD inhibited LPS mediated NF-kappaB activation, which might be one of its mechanisms for treating inflammatory bowel disease (IBD).

Caco-2 Cells ; Colon ; Drugs, Chinese Herbal ; pharmacology ; Enterocytes ; Humans ; I-kappa B Proteins ; metabolism ; Inflammation ; Interleukin-8 ; Lipopolysaccharides ; NF-KappaB Inhibitor alpha ; NF-kappa B ; metabolism ; Phosphorylation ; Tumor Necrosis Factor-alpha ; metabolism