Objective The present study aims to explore the optimal combination of growth factors, including TGF-?_1,IGF-I and BMP-2,for the induction of ehondrogenesis of MSCs in vitro monolayer culture. Methods MSCs were induced to differentiate into ehondrocytes in the medium with the following growth fac- tors:TGF-?_1,BMP-2 and IGF-I,either alone or in combination.We detected two cartilage specific markers in differentiated cells.Alcian blue stain was used for aggreean and RT-PCR for collagen type II mRNA.Results The level of Collagen typeⅡmRNA of group BMP-2+IGF-I,TGF-?_1,TGF-?_1+IGF-I,TGF-?_1+BMP-2 and TGF-?_1+IGF-I+BMP-2 was 0.147?0.062,0.478?0.069,0.489?0.022,0.653?0.013,0.807?0.051 respectively. Alcian blue stain in all gruups were positive.Conclusion The TGF-?_1+IGF-I+BMP-2 group is the most ef- fective combination in chondrogenesis induction.

Adult ; Diagnostic Errors ; Humans ; Hypophosphatemia ; diagnosis ; etiology ; therapy ; Male ; Middle Aged ; Neoplasms ; complications ; Osteomalacia ; diagnosis ; etiology ; therapy ; Young Adult

OBJECTIVETo investigate the application and mechanism of tissue-engineered skin with mouse embryonic fibroblasts (MEFs) for the full-thickness skin defects on mice.

METHODSThe MEFs and fibroblasts were cultured and seeded in scaffold made of rat tail collage. ELISA method was used for detection of secretory function. The full-thickness skin defects were created on mice and covered by MEFs-scaffold complex (experimental group), or FBs-scaffold complex (control group 1), or scaffold only (control group 2). The process of wound healing was evaluated by observation of the re-epithelization rate. Microvessel density (MVD) and vimentin within the wound sites were also detected with immunohistochemistry staining technique to describe the characteristics of wound healing. Hoechst 33342 staining was performed to trace MEFs'fate.

RESULTSMEFs scaffold group had higher level secretion of IL-6 and lower of TGF-beta1 than FBs scaffold group (P<0.05). Compared with wounds in control groups, the wounds in MEFs group healed markedly fast (P<0.05) and the MVD was significantly higher (P <0.05). The fibroblasts in the wounds of MEFs group were arranged regularly and the MEFs decreased during the healing process.

CONCLUSIONSThe MEFs-scaffold complex can promote wound healing with less scar formation. MEFs may have an inducing effect on the wound healing.

Animals ; Female ; Fibroblasts ; cytology ; Interleukin-6 ; metabolism ; Pregnancy ; Rats ; Rats, Sprague-Dawley ; Skin, Artificial ; Tissue Engineering ; Tissue Scaffolds ; Transforming Growth Factor beta1 ; metabolism ; Wound Healing

Objective To investigate plasma levels of total carnitine (TC) and free camitine (FC) in patients undergoing hemodialysis and peritoneal dialysis.Methods 200 cases of normal group came from physical examination in this hospital,all testing cases were the in-hospital patients in the department of nephropathy.TC and FC were determined by use of an enzymatic cycle assay on Hitachi 7170 automatic biochemical analyzer.Results In 200 cases of normal group,TC level was (56.52?9.61) ?mol/L,and FC was (46.60?8.23) ?mol/L.In 37 hemodialysis patients,TC and FC levels were (41.47?13.22) ?mol/L and (24.58?8.91)?mol/L before dialysis,a statistic difference was observed against the control group (P0.05).Conclusions Carnitine deficiency was seen in most patients undergoing hemodialysis and peritoneal dialysis.Furthermore,the deficiency status got worse along with the dialysis course in hemodialysis patients.Carnitine infusion can effectively improve the status of these patients.

Aim To explore the inhibitory effect of Euonymus alatus on hepatic fibrosis induced by carbon tetrachloride (CCl4) in mice and its mechanism. Methods Eighty C57BL/6 male mice were randomly divided into eight groups: normal group, CCl4model group, Euonymus alatus(EA) ethanol extracts groups in early stage(EAE), EA ethanol extracts groups in later stage(EAL),two drug groups with low/medium/high dose(EAE-L/M/H, EAL-L/M/H), with 10 mice in each group. Fibrosis model was established by injecting CCl4in peritoneal cavity,and the study lasted for 30 days. Different doses of drugs were given from 1 st day to 15 th day in EAE while from 16 th day to 30 th day in EAL,then all mice were sacrificed to for the observation of the morphological changes and collage-nous fiber by HE and Masson staining. Liver index, ALT,AST and TNF-α were tested by ELISA. The ex-pressions of α-SMA and CollagenⅠwere measured by immunohistochemistry and Western blot. Results Compared to normal group, liver index, ALT, AST, TNF-α, α-SMA and CollagenⅠ in EA groups were lower than those in model group in a dose-dependent manner(P<0.01 or P<0.05). Liver morphology and collagenous fiber in EAE and EAL were better than those in model group in a dose-dependent manner. The effect of EAE were superior to that of the EAL in HE, Masson, α-SMA, Collagen Ⅰ indexes(P <0.05). Conclusions Euonymus alatus may inhibit the process of hepatic fibrosis in mice with dose-effect de-pendence, and drug treatment in early stage performs better,which may be related to the decrease of TNF-α that affects the expression of α-SMA and Collagen Ⅰ.

No abstract available.
Irritable Bowel Syndrome*

BACKGROUNDVoltage-gated K+ channel (Kv) plays a critical role in the modulation of detrusor contraction. This study was conducted to investigate the expressions of Kv2.1 and Kv2.2 in rat bladder with detrusor hyperreflexia (DH).

METHODSThirty adult female Sprague-Dawley rats (200-220 g) were randomly divided into the control group and the experimental group. The experimental group was subjected to spinal cord injury (SCI). In the controls, the surgical procedure was identical with the exception that dura and spinal cord were transected. Four weeks after SCI, in vivo cystometry and mechanical pulling tests of isolated detrusor strips were performed. mRNA was extracted from the detrusors of normal and DH rats for the detection of expression of Kv2.1 and Kv2.2 by RT-PCR. Differences in expression between normal and overactive detrusors were identified by gel imaging.

RESULTSFourteen rats in the experimental group exhibited uninhibited bladder contraction (>8 cmH2O) before voiding after SCI. One rat died from infection. The frequency of DH in the experimental group was significantly different from that in the control group with or without treatment with 4-aminopyridine (4-AP) (P < 0.05), while the amplitude of DH did not change markedly. The rates of variation of the automatic contractile frequency and amplitude were (66.8 +/- 12.4)% and (42.6 +/- 12.6)% respectively in the control group, and (38.4 +/- 9.8)% and (28.0 +/- 4.6)% respectively in the DH group. 4-AP increased the automatic contractile frequency apart from the automatic contractile amplitude in both the control and DH groups (P < 0.05). 4-AP increased the rate of variation of the automatic contractile frequency more markedly in the control group than in the DH group (P < 0.05). Significant expression of Kv2.2 was not detected in bladders in the control group. Compared to the mRNA levels of beta-actin, the mRNA level of Kv2.1 was 1.26 +/- 0.12 in the control group and 0.66 +/- 0.08 in the DH group. SCI significantly reduced the mRNA level of Kv2.1 in rat bladders with DH (P < 0.05).

CONCLUSIONSOur study showed that the mRNA level of Kv2.1 decreased significantly in rat bladder with DH, which was one of the important pathogenetic mechanisms for DH, and suggested that Kv2.1 might be one of the therapeutic targets for bladder overactivity.

Animals ; Female ; In Vitro Techniques ; Muscle Contraction ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Reverse Transcriptase Polymerase Chain Reaction ; Shab Potassium Channels ; genetics ; physiology ; Urinary Bladder ; metabolism ; Urinary Bladder, Overactive ; etiology ; metabolism

OBJECTIVETo study the mechanism of microelement including Cu and Zn on the accumulation of three danshinones in Salviae miltiorrhizae root and build a theory basis for its good quality and high yield.

METHODSand culture experiments were conducted to study the effect of Cu and Zn on the accumulation of three danshinones and oxidase including peroxidase and polyphenol oxidase activity in the plant root. The correlation between available Cu and Zn contents in matrix and oxidase activity in the plant root and, the correlation between available Cu and Zn contents in matrix and contents of tanshinones in the root were discussed.

RESULTContents of danshinones in the root increased with the increasing of Cu and Zn concentration. Dynamic monitoring on contents of dan-shinones of the plant roots growing in the pots with different Cu and Zn concentration in the whole growing season showed that the contents of danshinones for 60 days were the lowest, for 120 days the highest and then dropped for 150 days. In addition, among available Cu and Zn contents of matrix, oxidase including peroxidase and polyphenol oxidase activity and contents of tanshinones in the root,the correlation between two factors were significant difference (P < 0.05 or P < 0.01).

CONCLUSIONThe mechanism of Cu and Zn on the accumulation of danshinones may be that Cu and Zn improve the activity of peroxidase and polyphenol oxidase, which promote transformation of phenolic compounds to terpenes and therefore to increase contents of danshinones.

Catechol Oxidase ; metabolism ; Copper ; metabolism ; Diterpenes, Abietane ; Drugs, Chinese Herbal ; analysis ; metabolism ; Peroxidase ; metabolism ; Phenanthrenes ; metabolism ; Plant Proteins ; metabolism ; Salvia miltiorrhiza ; enzymology ; metabolism ; Zinc ; metabolism

OBJECTIVETo observe the impact on lower limbs balance function in treatment of yin-yang meridians acupuncture with respiratory reinforcing and reducing manipulation involved in the patients of stroke by applying B-PHY balance function test training system so as to provide the objective evidence in treatment of stroke; with acupuncture.

METHODSOne hundred patients were randomized into an observation group and a control group, 50 cases in each one. In the control group, the basic treatment was applied, without other relevant rehabilitation therapies associated. In the observation group, with the basic treatment as the control group's, the therapy of the yin-yang meridians acupuncutre with respiratory reinforcing and reducing manipulation was adopted. On the yin meridians, Zuwuli (LR 10), Xuehai (SP 10), Yinlingquan (SP 9), Sanyinjiao (SP 6) and the others were selected and stimulated with reducing manipulation achieved by the coordination of patient's respiration. On the yang meridians, Biguan (ST 31), Liangqiu (ST 34), Yanglingquan (GB 34) and the others were selected and stimulated with reinforcing manipulation achieved by the coordination of patient's respiration. The treatment was given once a day and for 28 days totally. Before treatment and in 28 days of treatment, B-PHY balance function test training system was used to determine the weight shift track parameters (track length, peripheral square, track length of per unit square, left-right offset and rectangle square), the weight shift track distance parameters [mean of X axle weight shift distance (Mean-X), mean of Y axle weight shift distance (Mean-Y), maximum of X axle weight shift distance (Max-X), maximum of Y axle weight shift distance (Max-Y), weight shift distance (LSKG), weight shift square (SSKG), square ratio of weight shift (LFS)], stability coefficient (SI) and weight distribution coefficient (WDI).

RESULTSAfter treatment, the differences in the weight shift track parameters, SI and WDI were significant as compared with those before treatment in the patients of the two groups (all P<0.01); while the differences in the weight shift distance parameters in the observation group were improved obviously after treatment as compared with those before treatment (all P<0.01), the differences of Mean-X, Max-Y and LFS in the control group were improved after treatment as compared with those before treatment (all P<0.01). Except SSKG, the improvements after treatment in the rest indices in the observation group were better than those in the control group (all P<0.05).

CONCLUSIONThe yin-yang meridians acupuncture with respiratry reinforcing and re- ducing manipulation effectively improves the lower limbs balance function in the patients of stroke.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Lower Extremity ; physiopathology ; Male ; Meridians ; Middle Aged ; Postural Balance ; Respiratory System ; physiopathology ; Stroke ; physiopathology ; therapy ; Treatment Outcome ; Yin-Yang

OBJECTIVETo construct the human/mouse chimeric antibody of a functional anti-death receptor 5 (DR5) antibody. Methods The viable region of light chain (VL) and viable region of heavy chain (VH) genes of anti-DR5 antibody were amplified and cloned into the light- and heavy-chain expression vectors respectively, then the recombinant plasmids were co-transfected into dihydrofolate reductase(-) Chinese hamster ovary cell (CHO-dhfr(-)) for expression. The positive clone was screened by the two selective genes (neo and dhfr). The humanization and specificity of chimeric antibody was identified by ELISA and Western blotting, and the tumoricidal activity of the expressed chimeric antibody was detected by tetrazolium salt phenazine methosulfate assay.

RESULTSThe expression vectors stably expressed chimeric antibody in CHO-dhfr(-). In the cell supernatant of the F4' clone, the human IgG heavy constant region and light constant region were identified. Moreover, the secreted chimeric antibody retained the binding capacity to the antigen (DR5) and decreased the cell viability of Jurkat and HCT116 cells to 73.15% and 77.30% in vitro respectively.

CONCLUSIONThe human/mouse anti-DR5 chimeric antibody has been successfully expressed in eukaryotic cells and shows tumoricidal activity, which establishes a foundation for the future research of humanized antibody medicine.

Animals ; Antibodies ; genetics ; immunology ; pharmacology ; Antineoplastic Agents ; immunology ; pharmacology ; CHO Cells ; Cell Survival ; drug effects ; Cricetinae ; Cricetulus ; Gene Expression ; Humans ; Mice ; Protein Engineering ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; genetics ; immunology ; Recombinant Fusion Proteins ; genetics ; immunology ; pharmacology