OBJECTIVETo observe beta-catenin expression of Wnt signaling pathway in rats with knee osteoarthritis, and influence of Bushen Huoxue decoction on beta-catenin and MMP-7 expression of synoviocytes in rats with knee osteoarthritis (OA).

METHODSRats model with knee osteoarthritis were established by Hulth method. Primary synoviocytes and OA synoviocytes were cultured with collagenase digestion method. The cultured synoviocytes were divided into normal group, OA model group and Bushen Huoxue decoction group. Western blotting method was used to detect beta-catenin, MMP-7 protein expression of synoviocytes after acting by Bushen Huoxue decoction for 48 h; ELISA method was used to detect MMP-7 expression of synovial supernatant.

RESULTSOA synoviocytes were cultured successfully. Western blotting showed that beta-catenin, MMP-7 expression in OA synoviocytes was significantly higher than normal group, Bushen Huoxue decoction could significantly reduce beta-catenin, MMP-7 expression; ELISA results showed that MMP-7 expression of OA synovial supernatant was significantly higher than normal synoviocytes supernatant, Bushen Huoxue decoction significantly regulated the level MMP-7 down.

CONCLUSION(1) High expression of beta-catenin in OA synoviocytes indicates that Wnt classical signal pathway is activated in rat with knee osteoarthritis; (2) High expression of MMP-7 expression in OA synoviocytes confirms the MMP-7 is downstream genes of Wnt/beta-catenin signal pathway; (3) Activation of Wnt signal pathway and increase of MMP-7 may cause degradation of articular cartilage, and promote the formation of osteoarthritis; (4) Bushen Huoxue decoction can reduce expression of MMP-7, and promote cartilage repair, which may be one of mechanisms of osteoarthritis.

Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; Enzyme-Linked Immunosorbent Assay ; Male ; Matrix Metalloproteinase 7 ; analysis ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Rats ; Rats, Wistar ; Synovial Fluid ; chemistry ; cytology ; beta Catenin ; analysis

Osteoarthritis is a degenerative joint disease,with the characters of degradation of articular cartilage, the formation of the joint marginal osteophyte and synovium lesions. Previous studies have focused on the treatment of articular cartilage lesions. In recent years, new research in shows synovial inflammation plays an important role in OA. Synovium lesions and synovial inflammation-related factors induced the degradation and destruction of articular cartilage, and promoted the development of osteoarthritis. The role of synovial lesions in osteoarthritis is increasingly prominent, and the treatment for synovial lesions will become a new target. So this paper reviews the various manifestations of synovial in osteoarthritis.
Humans ; Osteoarthritis ; pathology ; Synovial Membrane ; pathology

OBJECTIVETo observe the long-term efficacy and safety of warm needling therapy combined with rehabilitation training in the treatment of knee osteoarthritis (KOA) for the patients living in simple room after earthquake.

METHODSEighty-eight cases of KOA were randomized into a warm needling group and an acupuncture group, 44 cases in each one. In the warm needling group, the warm needling therapy was applied at Neixiyan (EX LE 4) and Dubi (ST 35), in combination with rehabilitation training. In the acupuncture group, the same rehabilitation training was received, and the same acupoints were selected as the warm needling group, but only stimulated with acupuncture without moxibustion applied. The clinical efficacy was observed after 20 treatments and in 1-2 months of follow-up.

RESULTSThe total effective rate was 100.0% (41/41) in the warm needling group, which was apparently superior to 85.4% (35/41) in the acupuncture group (P < 0.05). The symptom integral, pain and knee joint function score were all improved significantly after 20 treatments in both groups (all P < 0.05). The improvements in symptoms and pain in the warm needling group were superior apparently to the acupuncture group (all P < 0.05). The efficacy remained well within 2 months follow-up (all P < 0.05). But the difference in knee joint function score was not statistically significant between the two groups (all P > 0.05).

CONCLUSIONThe combined therapy of warm needling and rehabilitation training achieves the superior efficacy and good safety in the treatment of KOA as compared with the combined therapy of simple acupuncture and rehabilitation training for the patients living in simple room after earthquake.

Acupuncture Points ; Acupuncture Therapy ; instrumentation ; Adult ; Aged ; Female ; Humans ; Knee Joint ; physiopathology ; Male ; Middle Aged ; Osteoarthritis, Knee ; physiopathology ; rehabilitation ; therapy ; Treatment Outcome

OBJECTIVES: To detect diatom in the organs of drowners by enzyme combined with strong acid digestion method, and evaluate its application value. METHODS: A total of 40 cases which have been identified as drowning in local region were collected. Samples of the lung, liver, kidney, and the water of the scene were also gathered from each case. Strong acid digestion method, enzyme combined with strong acid digestion method, and enzymic digestion method were respectively performed to detect the diatom in the samples. The comparative analysis was made on digestion time, digestive power and detection rate of diatom, etc. RESULTS: Enzyme combined with strong acid digestion method was significantly better than enzymic digestion method on digestion time and digestive power; enzyme combined with strong acid digestion method were obviously superior to strong acid digestion method on the detection rate of diatom. CONCLUSIONS Enzyme combined with strong acid digestion method combines the advantages of strong acid digestion method and enzymic digestion method. It has the characters of operation safety with little pollution to environment, which is worthy of further popularization and practice.
Diatoms/isolation & purification* ; Drowning ; Forensic Pathology/methods* ; Humans ; Kidney/metabolism* ; Liver/metabolism* ; Lung/metabolism*

Objective To establish information management systems of drinking water defluoridation project in water-related endemic fluorosis areas and investigate the status of drinking water defluoridation project in Liaoning, provide the basis for the development of control measures. Methods Global positioning systems (GPS)and geographic information systems(GIS) were used in the study in August 2006 - July 2008. Water defluoridation projects of 1234 in 48 counties(cities, districts) in drinking water type of fluorosis areas were positioned. Latitude and longitude, water samples, water fluoride content were collected or tested. GIS was used to establish information management system of water defluoridation projects. Results We have established information management system for the facilities of decreasing water fluorine in drinking water type of endemic fluorosis regions in Liaoning. One thousand two hundred and thirty four defluoridation facilities distributed in east longitude between 39.39° - 43.37°,north latitude between 119.25° - 125.50°, and altitude between - 6.60 and 801.14 meter in 48 endemic fluorosis counties in 13 cities. Nine hundred and twenty seven facilities for decreasing fluorine were able to supply water regularly, accounting for 75.1% of investigated projects;29 facilities was .not yet completely rebuilt, accounting for 2.4%;278 facilities(supply water for 344 villages) were out of order or discarded for 22.5% of investigated projects.Water fluorine contents of 63 facilities were greater than 1.2 mg/L, accounting for 6.8% of investigation project.Facilities working regularly and water fluorine was in accord with hygienic standard for drinking water facilities were 70.0%. Conclusions The establishment of Liaoning province defluoridation project information management system in the whole province of drinking water type of fluorosis areas provides scientific basis for accurate decision-making on prevention and control of the disease.

Adult ; Diagnostic Errors ; Female ; Glioma ; diagnosis ; Humans ; Male ; Middle Aged

OBJECTIVETo silence the expression of Raf-1 gene in HNE1 cells using vector-based RNA interference (RNAi) technique.

METHODSThe vector containing the human U6 promoter was used for targeted gene silencing when a dsDNA oligonucleotide encoding an appropriate shRNA was ligated into the vector, and 67nt oligonucleotide fragment was inserted into the downstream of the U6 promoter. Plasmids containing different Raf-1 target sequences [ (1) pshuttle-Raf-1-a( 225), (2) pshattle-Raf-1-b ( 358) and (3) pshuttle-Raf-1-c(474)], were transfected into HNE1 cells. Expression of Raf-1 mRNA was assayed by RT-PCR. Apoptosis were determined by cytometry.

RESULTSVector-based RNAi had advantages over antisense RNA because it could be delivered to the target cell more efficiently, and effect could last longer. Raf-1 expression could be inhibited by plasmid-expressed shRNA. Three different targeting sequences were selected from Raf-1 gene, and the inhibitory effect of pSIREN shuttle-Raf-1-b (358) was biggest.

CONCLUSIONRaf-1 expression in HNE1 cells can be inhibited significantly using plasmid-based RNAi.

Apoptosis ; genetics ; Cell Line, Tumor ; Gene Expression ; Genetic Vectors ; Humans ; Nasopharyngeal Neoplasms ; genetics ; metabolism ; pathology ; Promoter Regions, Genetic ; Proto-Oncogene Proteins c-raf ; genetics ; RNA Interference ; RNA, Messenger ; genetics ; Transfection

OBJECTIVEThe complex of the cyclic AMP receptor protein (CRP) and cAMP is an important transcriptional regulator of numerous genes in prokaryotes. The transport of mannitol through the phosphotransferase systems (PTS) is regulated by the CRP-cAMP complex. The aim of the study is to investigate how the CRP-cAMP complex acting on the mannitol PTS operon mtl of the Vibrio cholerae El Tor biotype.

METHODSThe crp mutant strain was generated by homologous recombination to assess the need of CRP to activate the mannitol PTS operon of V. cholerae El Tor. Electrophoretic mobility shift assays (EMSA) and the reporter plasmid pBBRlux were used to confirm the role that the CRP-cAMP complex playing on the mannitol PTS operon mtl.

RESULTSIn this study, we confirmed that CRP is strictly needed for the activation of the mtl operon. We further experimentally identified five CRP binding sites within the promoter region upstream of the mannitol PTS operon mtl of the Vibrio cholerae El Tor biotype and found that these sites display different affinities for CRP and provide different contributions to the activation of the operon.

CONCLUSIONThe five binding sites collectively confer the strong activation of mannitol transfer by CRP in V. cholerae, indicating an elaborate and subtle CRP activation mechanism.

Bacterial Proteins ; genetics ; Base Sequence ; Cyclic AMP ; metabolism ; Cyclic AMP Receptor Protein ; Gene Expression Regulation, Bacterial ; Mannitol ; Molecular Sequence Data ; Operon ; Phosphotransferases ; Vibrio cholerae

OBJECTIVETo construct a recombinant adenovirus vector expressing hCD40L gene and explore it in the use of anti-tumor gene therapy.

METHODS1,900 bp gene fragment was obtained form plasmid pORF-hCD40L by Xho I/Swa I cutting and then cloned directionally into the pShuttle plasmid, finally, the resultant plasmid was digested by restriction endonnuclease PmeI and subsequently cotransformtion into BJ5183 cells with the adenoviral backbone pAdEasy-1 to obtain the homologous recombinant and then the recombinant was packaged in the 293 cells. Some methods such as PCR and endonulease digestion were employed to identify the recombinant adenovirus.

RESULTSThe evidences of endonulease digestion and PCR analysis confirmed that recombinant hCD40L gene was correctly inserted into adenovirus vector.

CONCLUSIONThe adenoviral vector which expressed hCD40L gene was constructed. It provides an experimental basis for studies on it expression in the mammalian cells and in tumor gene therapy.

Adenoviridae ; Adenoviruses, Human ; Animals ; CD40 Ligand ; Genetic Therapy ; Genetic Vectors ; Humans ; Plasmids ; Polymerase Chain Reaction