OBJECTIVE: To investigate the effect of high glucose on the expression of c-met in human kidney fibroblasts in vitro, and to explore the regulation of Radix Astragali. METHODS: A cell culture system of human kidney fibroblasts was developed in vitro. The human kidney fibroblasts were divided into normal control group, high glucose group and mannitol group. Expressions of c-met and transforming growth factor-beta1 (TGF-beta1) mRNAs were detected by reverse transcription polymerase chain reaction (RT-PCR) and the expressions of c-met protein were analyzed by Western blot method after 6-, 12-, 24-, 48- and 96-hour culture. The human kidney fibroblasts were also cultured with 10% Radix Astragali containing serum; the expressions of c-met mRNA and protein were detected after 24- and 48-hour culture. RESULTS: Compared with the normal control group, expression of c-met mRNA in the high glucose group was significantly increased after 12-hour culture (P<0.05), arriving at the peak after 24-hour culture (P<0.01). The level of TGF-beta1 mRNA was higher in the high glucose group than that in the normal control group after 24-hour culture (P<0.05), arriving at the peak after 96-hour culture (P<0.01). Forty-eight hours after treating with 10% Radix Astragali containing serum, the levels of c-met mRNA and protein in fibroblasts were increased, and were higher than those in the high glucose group (P<0.01, P<0.05). CONCLUSION: High glucose can induce the expressions of c-met mRNA and protein in earlier period, and then inhibit the expressions. Radix Astragali can up-regulate the expressions of c-met mRNA and protein of human kidney fibroblasts, which may be one of its action mechanisms in delaying the progression of diabetic nephropathy.

Objective To explore the protective effect of high density lipoprotein on the liver function of rats with severe burns. Methods 135 Wistar rats employed in present study were randomly divided into control group (n=15, without treatment), burn group (n=60, with 30%TBSA full-thickness burn on the back) and experimental group (n=60, with the injection of 80mg/kg HDL via the caudal vein immediately after burns). The rats in the groups with burns injury were resuscitated with intraperitoneal isotonic saline (50ml/kg) 30 minutes after burn. The serum content of AST, ALT, ICAM-1 and TNF-? of the rats were determined with corresponding methods. The histological changes in the liver tissue of the rats in all groups were observed under light microscope and electronic microscope. Results The serum content of AST, ALT, ICAM-1 and TNF-? in the rats of control group were significantly lower than those in burn group (P

Objective To examine the expression of hepatocyte growth factor(HGF) receptor (c-met) and investigate the changes in activity of the HGF/c-met system in human kidney fibroblast by high glucose. Methods HGF and c-met mRNA levels in fibroblast induced by high glucose were detected by RT-PCR. C-met protein was examined by Western blotting. At the same time, the expression of TGF-? and c-met in the exogenous HGF and treating with anti-c-met antibody in vitro were measured. Results Extremely rapid induction of HGF and c-met mRNA was observed at the first six hours by high glucose. On the other hand, both c-met mRNA and c-met protein were markedly increased. HGF (50 ng/ml) induced the expression of c-met ( P

BACKGROUND:Previous studies show that the fracture resistant force of endodonticaly treated teeth can be improved by post. But this idea has been controversial in recent years. Many scholars believe that the root canal preparation might weaken dental root the post increases the risk of tooth fracture. OBJECTIVE: To evaluate the fracture resistance ability of severe wedge-shaped-defect premolar to oblique loading, which was restored with fiber reinforced composite posts by different therapy methods. METHODS:A total of 50 human maxilary premolars were randomly divided into five groups, with ten teeth in each group. They were given folowing treatments: Group A: 10 untreated premolars; Rest 40 premolars of Groups B, C, D and E were prepared 1/3 bucco-lingual distance for artificial severe wedge-shaped defects at the buccal cervix. Group B: untreated severe wedge-shaped defects premolars; Group C: severe wedge-shaped defects premolars were endodonticaly treated, remaining dentin over hang above the wedge shaped defect, LuxaPost posts reinforced in buccal canal and lingual canal, LuxaCore composite resin restored dentin defect; Group D: severe wedge-shaped defects premolars were endodonticaly treated, LuxaPost posts reinforced in buccal canal and lingual canal, LuxaCore composite resin restored dentin defect, and then covered with ful metal crown; Group E: severe wedge-shaped defects premolars were endodonticaly treated, removing dentin over hang above the wedge shaped defect, LuxaPost posts reinforced in buccal canal and lingual canal, restored with LuxaCore composite resin, and then covered with ful metal crown. Fracture resistance of each specimen was measured in each group. RESULTS AND CONCLUSION: The fracture strength of each group were Group A (1 002.69±147.62) N, Group B (439.28±66.34) N, Group C (958.30±101.23) N, Group D (1 207.09±143.48) N, and Group E (1 056.44±139.30) N. Group D had the highest fracture strength (P < 0.01), while Group B had the lowest fracture strength (P < 0.01). There were no significant difference among the fracture strength of Group A, Group C and Group E. Our findings indicated that the fracture resistance of the severe wedge-shaped defected premolar can be improved by fiber reinforced composite post and dentin above wedge shaped defect remained.

Objective To observe high glucose induced expression of hepatocyte growth factor (HGF) and c met in human kidney fibroblast. Methods The effects of glucose concentrations on expression of HGF, c met and plasminogen activator inhibitor (PAI) 1 in cultured human kidney fibroblasts were observed by RT PCR. In the same system, the effect of exogenous HGF on the expression of PAI 1 was investigated. Results Human kidney fibroblasts cultured in high glucose concentration (25 mmol/L) showed higher HGF and c met expressions in the early stage and then manifested a gradient decrease of HGF and c met expressions, but PAI 1 expression was gradiently increased. Exogenous HGF resulted in inhibiting PAI 1 expression. Conclusion HGF is a potential anti fibrogenic factor and activates matrix degradation pathways in diabetic kidney by reducing PAI 1 expression.

Objective To explore the outcome, recurrence rate and postoperative MRI of the modified microendoscopy discectomy (MED) and the conventional MED. Method A total of 107 patients with disc herniation treated from January 2005 to December 2009 were randomly divided into Group A (54 patients) and Group B (53 patients). The patients in Group A were treated with the modified MED that removed the prominent nucleus pulposus but did not access into the annulus. The patients in Group B were treated with traditional MED that removed the prominent nucleus pulposus and cut annulus fibrosus and removed the nucleus pulposus within the annulus fibrosus. The operation outcome, recurrence rate and lumbar spine MRI were compared for observing the morphological changes of intervertebral disc and its surrounding structures. Results All patients were followed up for 1-4 years ( average 2 years),which showed no recurrence in Group A but four patients with recurrence Group B. According to Japanese orthopedic association (JOA) scoring of low back pain, the efficacy was excellent in 51 patients (94%)and good in three (6%), with excellence rate of 100% in Group A; while the efficacy was excellent in 42 patients (79%), good in seven (13%) and poor in four (8%), with excellence rate of 92% in Group B. Videman semi-quantitative assessment of disc signal showed no significant difference of MRI in aspects of signal intensity, thickness, diameter of the spinal canal in Group A before and after operation but highlighted significant differences in protruding degree,spinal canal diameter and lateral recess diameter. Substantial differences of MRI in aspects of signal intensity, thickness, protruding degree, spinal canal diameter and width were observed before and after operation in Group B. There was no significant difference in spinal canal diameter. Conclusions Both methods have sound clinical efficacy. However, the modified MED procedures take advantages of minimizing the damage of disc structure, maintaining the relative stability of the spine and delaying the disc degeneration, which contributes to lower recurrence rate.

Objective To establish a real‐time fluorescent polymerase chain reaction and detect 16S rRNA gene of Legionella strains isolated from sputum specimens of patients with pulmonary infection by using this method .Methods 16s rRNA gene of Le‐gionella was used to design primers and probes .The reaction system and reaction conditions were optimized and the specificity ,sen‐sitivity and repeatability of this method were verified by detecting Legionella pneumophila ,non‐Legionella pneumophila and other bacteria .A total of 557 sputum specimens of patients with pulmonary infection were detected ,and PCR‐digestion identification method was carried out as control .Otherwise ,sequences of 16S rRNA were verified in patients with positive detection results .Re‐sults The results showed that all reference strains of Legionella were positive ,while all of other bacteria were negative ,and the sensitivity was 102 CFU/mL .Among sputum specimens collected from 577 cases of patients with pulmonary infection ,the positive rate of Legionella detected by using real‐time fluorescent PCR and PCR‐digestion identification method was 23 .1% and 19 .9% re‐spectively ,while the positive rate was 17 .2% by verifying the sequences of 16s rRNA .There were no statistically significant differ‐ences of positive rate among the three methods(P>0 .05) .Conclusion The real‐time fluorescent PCR is fast and convenient in de‐tection of Legionella strains isolated from sputum specimens of patients ,which could be an assisted method for clinically diagnosing Legionella infection .

The gene expression of both the mouse plasmocytoma (SP2/0) and the hybrid cells crossed with rat nucleated erythroblasts were detected by in situ hybridization technique using the probes of mouse ?-globin gene and 7 oncogenes (v-Ki-ras, v-H-ras, v-sis, erb-B, v-abl, v-fos, c-myc). After plasmic amplification, DNA was isolated by alkali lysis, purified and recovered, the DNA containing gene fragments were labelled with ~(32)P to become high activity ~(32)P-cDNA probes through nick translation, and the labelled probes were used to detect the gene transcripts in cellular level. The results indicated that: (1) no mouse ?-globin gene transcripts could be detected in the cytoplasm of SP2/0 cells, as well as in hybrid cells within 72 hours after cell fusion, but transcript signals could be observed in hybrid cells from 4th to 26th passages. (2) Active expression of multioncogenes in SP2/0 cells was demonstrated, all the 7 oneogenes tested, except v-sis, were expressed more strongly. On the other hand, the expression of oncogenes in hybrid cells was found to be dramatically decreased, among them, the oncogenes of c-myc and Ki-ras been suppressed completely. After long term of passages (26th subcultures), the expression of c-myc and Ki-ras was still lower than that of SP2/o ceils although in some cases other oncogenes increased in their expression levels. These results confirmed that the multistep carcinogenesis involved multi-oncogenes expression and that the decancerization of tumor cells may be due to the suppression of multi-oncogenes activity as well as to activate the expression of differentiation genes.

Objective To explore the changes of gene and protein expressions of nuclear factor-κB(NF-κB) and tumor necrosis factor-α (TNF-α) in immature rats with pilocarpine-induced epilepsy treated with ghrelin. Methods The pllocarpine-induced epilepsy model in immatured rats were built, then the rats were divided into three groups: Ghrelin-treated group, saline-treated group and model group, meanwhile the normal control group was set. The NF-κB and TNF-α levels of gene and protein in the cerebral cortex of immature rats were detected. Results The expression levels of gene and protein of NF-κB and TNF-α were increased in model group,but decreased in the normal control group;NF-κB and TNF-α levels in Ghrelin treated group were obviously lower than those of saline-treated group and model group(P < 0. 05). Conclusion The protective mechanism of Ghrelin for nerve cell is cutting down the expressions of NF-κB and TNF-α in the cerebral cortex of immature rats with epilepsy and lessening inflammatory reaction in neurocytes.

Objective To investigate the heterogeneity of immunomodulatory function of exosomes secreted from human umbilical cord-derived mesenchymal stem cells(hUC-MSC).Methods Five different hUC-MSCs lines were isolated and cultured.Exosomes were isolated from the supernatant.The expression of specific surface markers CD9,CD63,CD81 and CD44 was detected by flow cytometry.The protein concentration of hUC-MSCs exosomes(hUC-MSCs-ex)was evaluated by the BCA assay.Concanavalin A and rhIL-2 stimulated umbilical cord blood mononuclear cells (UBMCs) from healthy donor were co-cultured at different concentrations of hUC-MSCs-ex from different cell lines for 72 h.The growth rate of UBMCs was detected by MTT assay.ELISA was used to test the levels of IFN-γ and TNF-α of the supernatants.The UBMCs co-cultured with hUC-MSCs were co-cultured with K562 cells at the ratio of 5∶1.The cytotoxic activity was calculated by MTT assay.Results hUC-MSCs-ex expressed CD9,CD63,CD81and CD44.Different hUC-MSCs lines had different regulating activities on the proliferation,secretion and killing ability of UBMCs.Conclusion hUC-MSCs-ex has heterogeneity of immunomodulatory function on UBMCs.