OBJECTIVETo investigate clinical effects and mechanism of Tangweikang (TWK) in treating diabetic gastroparesis.

METHODSNinety diabetic gastroparesis patients were randomly assigned to 3 groups. Besides conventional hypoglycemic treatment, the 30 patients in the treated group were given TWK and the 30 in the control group were given Domperidone additionally, while to the 30 in the blank group, no additional drug was given. The clinical efficacy and the changes in level of motilin and gastric emptying rate were observed.

RESULTSTWK showed significant effects in improving clinical symptoms of patients, increasing gastric emptying rate, promoting gastrointestinal kinetics, shortening gastric emptying time and was beneficial to the control of blood sugar, including the 2 h post-prandial blood sugar and fructosamine. The curative rate and total effective rate in the treated group were 63.33% (19/30) and 93.33% (29/30) respectively, significantly different to those in the control group 26.67% (8/30) and 63.33%, also different to those in the blank group 23.33% (3/ 30) and 10.00%, respectively (P < 0.01). The clinical efficacy in the treated group was superior to that in the other two groups.

CONCLUSIONTWK has favorable therapeutic efficacy in treating DGP.

Aged ; Antiemetics ; therapeutic use ; Diabetes Complications ; drug therapy ; Domperidone ; therapeutic use ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Gastroparesis ; drug therapy ; etiology ; Humans ; Male ; Middle Aged ; Phytotherapy ; Treatment Outcome

Objective To explore the rehabilitation needs of people with disabilities and the related factors,so as to provide a scien-tific basis for the rehabilitation policy-making for people with disabilities. Methods From June,2014 to July,2015,based on the national special survey of basic services and needs of people with disabilities in Jiangsu,China,this paper clarified the factors related with the rehabilitation needs of the disabled into the individual characteristics, economic characteristics, environmental characteristics and so on by the de-scriptive statistics method. The Logit model was established to explore the factors related with rehabilitation needs. Results In Jiangsu,43% people with disabilities aged 16 and above had rehabilitation needs.Contingency table analysis and Chi square test showed that individual characteristics,economic characteristics and environmental character-istics had significant impact on the rehabilitation needs of the disabled.Multiple regression analysis showed that all the 18 variables were the significant related factors. Conclusion In order to meet the rehabilitation needs of the disabled, the refined and precise rehabilitation program should be designed in line with the different characteristics of the disabled.Through the effective docking of sup-plies and needs,we can provide accurate rehabilitation services for the disabled.

OBJECTIVEUndried silver-hydroxyapatite-titania (Ag-nHA-nTiO2) nanoparticles slurry was used to make membrane with polyamide 66 (PA66) by co-polymerization method. The purpose of this study is to test the physical and chemical characteristics and antibacterial ability.

METHODSThe morphology, chemical components and structures of the membrane were characterized by atomic absorption spectrometer (AAS), X-ray diffraction (XRD), scanning electron microscope (SEM) and energy-dispersive X-ray analysis (EDX). Staphylococcus aureus (S. aureus), Escherichia coli (E. coli), Porphyromonas gingivalis (P. gingivalis), Fusobacterium nucleatum (F. nucleatum) and Streptococcus mutans (S. mutans) were utilized to test the antibacterial effect.

RESULTSXRD results demonstrated that the membrane have characteristic diffraction peaks of pure hydroxyapatite (HA). A homogeneous distribution of the Ca, P, Ti and Ag element in the membrane was confirmed by EDX. Both surface and section showed porous structure which was confirmed by SEM and the average hole size was 20-30 microm. The bacteria assay reflected to the antibacterial effect, 50.10% of S. aureus and 56.31% of E. coli were killed. However, 91.84% of P. gingivalis, 90.64% of F. nucleatum and 90.49% of S. mutans were killed and pictures of SEM showed obviously fewer cells on the surface.

CONCLUSIONThe nanocomposite membrane could be one of the bioactive materials with antibacterial properties for oral guided bone regeneration technique.

Anti-Bacterial Agents ; Bone Regeneration ; Durapatite ; Escherichia coli ; Nanocomposites ; Nylons ; Silver ; Staphylococcus aureus ; Titanium ; X-Ray Diffraction

OBJECTIVETo investigate the effect of different magnitudes of tensile strain on human osteoblasts differentiation.

METHODSAccording to the strain amplification mechanism at cellular level and a data calculated by finite element analysis, the cellular level strain of 0.8%, 1.6%, 2.4% and 3.2% was respectively applied to human osteoblasts for 48 h at a frequency of 1 Hz. Alkaline phosphatase activity and the expression of osteoblast-related genes were detected by Semi-Quantitative RT-PCR and densitometric analysis.

RESULTSAlkaline phosphatase activity significantly increased at 0.8% and 1.6%. The level of osteocalcin mRNA increased at 2.4% and 3.2%. Cbfa1/Runx2 gene expression only increased at 3.2%. Comparing to static control, mRNA level of type I collagen increased at every magnitude. The mRNA level decreased at 0.8% and increased at 3.2% when compared to the group with 1.6% elongation.

CONCLUSIONSHigher magnitudes of strain enhance expression of osteocalcin, type I collagen gene and Cbfa1/Runx2 in human osteoblasts, but lost the ability to increase ALP activity which is remained by lower magnitudes of strain. Type I collagen gene expression increases in a strain magnitude dependent manner.

Alkaline Phosphatase ; metabolism ; Cell Line ; Cell Proliferation ; Collagen Type I ; metabolism ; Core Binding Factor Alpha 1 Subunit ; metabolism ; Gene Expression Regulation ; Humans ; Osteoblasts ; cytology ; metabolism ; Osteocalcin ; metabolism ; Stress, Mechanical

OBJECTIVETo observe the distribution of epidermal stem cell (ESC) after soft tissue expansion, and to explore dynamic change in ESC under mechanical stress and kinetic mechanism of skin expansion.

METHODSSkin samples were collected from patients after expansion of the scalp. They were divided into three groups: A group (scalp harvested 3 cm away from the center of dilator), B group (scalp tissues at the edge of dilator), and control group (scalp without dilatation). The tissue structures were observed with optical microscope with HE staining. The distribution and differentiation characteristics of cell keratin 19 (CK19) positive cells were observed with inverted phase contrast microscope after immunohistochemistry staining.

RESULTSHE staining showed that the epidermis was thickened and distributed densely with uneven, rugged and increased layers in A, B groups. With immunohistochemistry staining, CK19 positive cells appeared in multilayers in basal membrane, a few of them were in cluster or dispersed , with" hollowing" structure formation. These phenomena were not seen in control group.

CONCLUSIONESC can proliferate with abnormal distribution and "hollowing" structure formation after mechanical dilatation, which may be related to dynamic changes in basal layer cells.

Adolescent ; Adult ; Cell Proliferation ; Cellular Structures ; Epithelial Cells ; cytology ; Humans ; Keratin-19 ; metabolism ; Male ; Stem Cells ; cytology ; Stress, Mechanical ; Tissue Expansion ; Young Adult

OBJECTIVETo investigate whether low molecular weight heparin (LMWH) may suppress the expression and secretion of vascular endothelial growth factor (VEGF) from tumor cells in vitro and inhibit the VEGF-induced proliferation of human tumor vascular endothelial cells.

METHODSHuman lung cancer cell line A549, human liver cancer cell line HepG2, human colon carcinoma cell lines HCT116 and HCT8 were used in this study. The expression levels of VEGF and TNF-alpha (tumor necrosis factor-alpha) in the tumor cells with or without pretreatment of LMWH/heparin were measured by standard sandwich ELISA technique. The VEGF mRNA level of HepG2 cells cultured with or without LMWH/heparin was determined by RT-PCR and real time PCR. Human umbilical vein endothelial cells (HUVEC) were cultured in tissue culture medium (TCM) with or without LMWH/heparin for 3 days. Then non-radioactive cell proliferation assay (MTS) kit and cell cycle assay by flow cytometry were performed to measure the proliferation of HUVEC.

RESULTSThe VEGF levels in the control, LMWH, and heparin groups of the pulmonary adenocarcinoma cell line A549 were (1045.89 +/- 165.30) pg/ml, (782.45 +/- 67.17) pg/ml and (916.54 +/- 71.25) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups of the colon adenocarcinoma cell line HCT116 were (955.76 +/- 51.14) pg/ml, (822.89 +/- 142.39) pg/ml and (951.77 +/- 188.22) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the colon adenocarcinoma cell line HCT8 were (1290.62 +/- 41.23) pg/ml, (1063.34 +/- 63.82) pg/ml and (1257.14 +/- 11.40) pg/ml, respectively. The VEGF levels in the control, LMWH, and heparin groups in the liver cancer cell line HepG2 were (1083.00 +/- 134.35) pg/ml, (758.00 +/- 84.85) pg/ml and (874.00 +/- 22.62) pg/ml, respectively. The VEGF expression levels in the above mentioned cell lines cultured in TCM were significantly reduced in the LMWH-treated groups compared with that of the control group (P < 0.05). But the level of TNF-alpha in TCM-cultured cells was unaffected by LMWH. The VEGF mRNA was reduced in the LMWH-treated HepG2 cell line. Moreover, TCM exhibited stimulating effect on proliferation of HUVEC and the effect was significantly impaired by LMWH treatment. Flow cytometric analysis revealed that LMWH treatment arrested HUVECs at the G1 phase of cell cycle.

CONCLUSIONLMWH can suppress the expression and secretion of VEGF by tumor cell lines and therefore have a potential inhibiting effect on angiogenesis induced by VEGF.

Adenocarcinoma ; metabolism ; pathology ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Culture Media, Conditioned ; Endothelial Cells ; cytology ; HCT116 Cells ; Hep G2 Cells ; Heparin ; pharmacology ; Heparin, Low-Molecular-Weight ; pharmacology ; Humans ; Lung Neoplasms ; metabolism ; pathology ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Umbilical Veins ; cytology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; secretion

OBJECTIVETo investigate the effect of nano-TiO(2) intratracheal instillation on the progression of dyslipidemia and atherosclerosis in apolipoprotein E-knockout mice.

METHODSThe nano-TiO(2) was ultrasound with phosphate-buffered saline solutions (PBS) into its suspension for exposure. A total of 46 specific pathogen free (SPF) level of 11-week-old male apolipoprotein E-knockout mice were randomly divided into groups by their body weights: non-treatment group (8 mice), PBS control group (9 mice), high dose group (1.0 mg/ml, 10 mice), medium dose group (0.5 mg/ml, 10 mice), and low dose group (0.1 mg/ml, 9 mice). Except the non-treatment group, mice from other groups were intratracheally instilled with 0.05 ml each time, twice a week. After exposure of 6 weeks, viscera index, blood TC, TG, HDL-C, LDL-C, and organic lipid ratio were assessed as biomarkers. Artery and aortic root issues were assessed by histopathology.

RESULTSAfter 5 weeks exposure, mice body weights in high dose group ((29.7 ± 1.9) g) started to drop, compared to PBS control ((31.3 ± 1.9) g, t = -1.58, P < 0.05) and low dose group ((31.4 ± 1.4) g, t = -1.17, P < 0.05); after 6 weeks, high dose group ((28.8 ± 1.5) g) was lower than PBS control ((30.4 ± 1.9) g, t = -1.60, P < 0.05), non-treatment group ((30.2 ± 1.3) g, t = -1.43, P < 0.05) and low dose group ((30.6 ± 1.0) g, t = -1.83, P < 0.05). TC levels of non-treatment, PBS control, high dose group, medium dose group and low dose group were (2.92 ± 1.18), (3.12 ± 0.73), (4.19 ± 1.86), (3.46 ± 0.72) and (2.57 ± 0.64) mmol/L, respectively; TG levels were (0.39 ± 0.13), (0.39 ± 0.08), (0.60 ± 0.21), (0.55 ± 0.19) and (0.41 ± 0.11) mmol/L, respectively; HDL-C levels were (1.67 ± 0.45), (1.54 ± 0.67), (0.93 ± 0.50), (1.02 ± 0.48) and (1.31 ± 0.64) mmol/L; TG levels of high dose group were higher than that of non-treatment group (t = 1.27, P = 0.03) and low dose group (t = 1.62, P = 0.01); TG levels of medium dose group was higher than PBS control (t = 0.16, P = 0.04), and TC levels of high dose group were higher than PBS control (t = 0.22, P = 0.01), non-treatment group (t = 0.22, P = 0.04) and low dose group (t = 0.20, P = 0.03), and HDL-C levels of high dose group were lower than PBS control (t = -0.61, P = 0.04) and non-treatment group (t = -0.74, P = 0.04); organic lipid ratio of each group were (2.27 ± 0.51)%, (2.06 ± 0.53)%, (2.90 ± 0.50)%, (2.60 ± 0.23)%, (2.24 ± 0.45)%; high dose group were higher than PBS control (t = 0.85, P = 0.00), non-treatment group (t = 0.64, P = 0.03) and low dose group (t = 0.67, P = 0.01); medium dose group was higher than PBS control (t = 0.54, P = 0.02). The plaque lipid content and calcium content which showed the progression of atherosclerosis and plaque rupture were elevated in medium and high dose groups.

CONCLUSIONIntratracheal instillation of nano-TiO(2) can induce dyslipidemia and accelerate the development of atherosclerosis and plaque rupture in ApoE-/-mice.

Animals ; Apolipoproteins E ; genetics ; Atherosclerosis ; blood ; chemically induced ; Dyslipidemias ; blood ; chemically induced ; Instillation, Drug ; Lipid Metabolism ; Lipids ; blood ; Male ; Mice ; Mice, Knockout ; Nanoparticles ; Specific Pathogen-Free Organisms ; Titanium ; administration & dosage ; pharmacology

Transgenic Atropa belladonna with high levels of scopolamine was developed by metabolic engineering. A functional gene involved in the rate limiting enzyme of h6h involved in the biosynthetic pathway of scopolamine was over expressed in A. belladonna via Agrobacterium-mediation. The transgenic plants were culturing till fruiting through micropropogating and acclimating. The integration of the h6h genes into the genomic DNA of transgenic plants were confirmed by genomic polymerase chain reaction (PCR) analysis. Analysis of the difference of plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight was carried out using SPSS software. The content of hyoscyamine and scopolamine in roots, stems, leaves and fruits was determined by HPLC. The investigation of the expression levels of Hnh6h by qPCR. Both Kan(r) and Hnh6h genes were detected in five transgenic lines of A. belladonna plants (A8, A11, A12, C8 and C19), but were not detected in the controls. The plant height, crown width, stem diameter, leaf length, leaf width, branch number and fresh weight of transgenic plants did not decrease by comparison with the non-transgenic ones, and furthermore some agronomic characters of transgenic plants were better than those of the controls. The highest level of scopolamine was found in leaves of transgenic A. belladonna, and the content of scopolamine was also higher than that of hyoscyamine in leaves. The contents of scopolamine of leaves in different transgenic lines were listed in order: C8 > A12 > C19 > A11 > A8, especially, the content of scopolamine in transgenic line C8 was 2.17 mg x g(-1) DW that was 4.2 folds of the non-transgenic ones (0.42 mg x g(-1) DW). The expression of transgenic Hnh6h was detected in all the transgenic plants but not in the control. The highest level of Hnh6h expression was found in transgenic leaves. Overexpression of Hnh6h is able to break the rate limiting steps involved in the downstream pathway of scopolamine biosynthesis, and thus promotes the metabolic flux flowing toward biosynthesis of scopolamine to improve the capacity of scopolamine biosynthesis in transgenic plants. As a result, transgenic plants of A. belladonna with higher level of scopolamine were developed.
Atropa belladonna ; genetics ; metabolism ; Atropine ; metabolism ; Gene Expression ; Mixed Function Oxygenases ; genetics ; metabolism ; Plant Proteins ; genetics ; metabolism ; Plants, Genetically Modified ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Solanaceae ; enzymology ; genetics

Objective To explore the siRNA as a new antiviral therapy,evaluate the inhibition effect of siRNA based on vector on the HBV of HepG2.2.15 cell,and observe the side effect and toxicity of siRNA vector on cells and the off-target effect of siRNA.Methods Three pairs of siRNA duplexes targeting HBV C gene were designed as double strands,and the duplex were annealed and ligated into the p-Silencer-Cmv 4.1-hygro vector.The ligation products were used to transform JM109 cells.The clones with shRNA were obtained,and the vectors were purified.After the initial identification of the vector with agarose gel and the size of the inserted sequence got examined by native polyacrylamide gel electrophoresis,furthermore the sequencing was further carried out.The recombinant plasmids were purified with ultrapure Midipreps DNA Purification System.Then HepG2.2.15 cells were transfected with the plasmid mixed with siPort XP-1.The expression of HBsAg and HBeAg were detected by immunofluorescence and Western blot,and the HBV RNA was investigated by RT-PCR.Furthermore the real-time quantitive PCR was carried out to detect the changes of HBV DNA.In order to evaluate the toxicity of the shRNA,MTT was used to examine the growth rate and curve of cells.The ELISA was performed to detect the changes of interferon-? (IFN-?).Results The Western blot showed that the HBsAg and HBeAg protein were suppressed with (81.15?0.69)%,(88.12?0.92)% respectively by vector p-C2 on the third day of post-transfection.It had the similar result indicated by immunofluorescence.And the RT-PCR showed that the specific siRNA targeting HBV C gene could markedly suppress the expression of HBV mRNA and the HBV C gene mRNA was inhibited with 96.9%.The real-time quantitive PCR showed that the specific functional siRNA could markedly suppress HBV DNA copy with two orders of magnitude,while the siRNA vector had no effect on the growth of cell showed by MTT detection.Compared with the non-transfected group and p-NC group,the IFN-? level was almost the same with siRNA p-C1,p-C2,p-C3 groups.Conclusions The siRNA based on the expression vector can suppress the expression and replication of HBV in HepG2.2.15 cell.The inhibition effect was specific and had a certain dependency on siRNA concentration.No toxicity effect was found in the study.And the drug resistance wouldn′t happen because the silence was based on the split of gene.