Objective To investigate the proliferative characteristics of fibroblast-like synovial cells (FLS)in osteoarthritis in vitro and the mechanism of the immunosuppressive effect of total glucusides of paeony(TGP).Methods FLS of OA and non-inflamed synovium(NS)were cultured and identified in vitro in the presence or absence of TGP.After incubation,the survival fraction(SF)of FLS was evaluated by MTI' and the TNF-?,IFN-?and bFGF level in cultured FLS supernatant was measured by ELISA.The expression of FLS c-los mRNA and cell cycle of OA-FLS was observed by RT-PCR and flow eytometry respectively at the same time.Results No statistical significant differences were noted between the OA and NS FLS in pro- liferating double time.High doses of TGP suppressed FLS-SF more evidently in OA patients than in NS(P0.05).Conclusion High dose TGP can inhibit OA-FLS proliferation,modulate cy- tokine secretion and c-fos expression in OA.This suggests that TGP has immunosuppressive effect on OA syn- ovitis,probably by preventing the synovial hypertrophy in OA.

The aim of this study was to investigate the expression of matrix metalloproteinase 26 (MMP-26), tissue inhibitor of metalloproteinase-4 (TIMP-4) and matrix metalloproteinase 9 (MMP-9) in patients with diffuse large B cell lymphoma (DLBCL) and their correlations with pathogenesis and development of DLBCL. A total of 95 specimens excised from DLBCL patients were prepared. Expression of MMP-26, TIMP-4 and MMP-9 were tested by SABC immunohistochemistry method and its correlation to clinicopathology indexes were analyzed. The results showed that as compared with reactive hyperplasia of lymph nodes, the high expression of MMP-26, TIMP-4 and MMP-9 were found in different types of DLBCL. The positive expression rate of MMP-26 was related to immune typing (P < 0.05). The expression level of MMP-26 in GCB was lower than that in non-GCB, and did not relate to clinical staging, age, sex, diseased region (P > 0.05). The positive expression rate of MMP-9 was related to clinical staging, the positive expression rate of MMP-9 proteins in patient at III and IV stage was obviously higher than that in patients at I and II stage, but did not relate to immune type, age, sex and diseased region of DLBCL (P > 0.05). The expression of TIMP-4 did not relate to immune type, clinical stage, age, sex, disease region (P > 0.05). The expression of MMP-26 in pathologic tissue of DLBCL did not relate to expression of TIMP-4, but positively related to expression of MMP-9 protein (r = 0.486, P < 0.05). It is concluded that MMP-26 and MMP-9 synergically express in DLBCL. MMP-26 may be involve in pathogenesis and invasiveness of DLBCL, the expression of MMP-26 relates to subtypes of DLBCL. The MMP-26 may serve as an indicator for typing of DLBCL and contributes to predict the invasion and metastasis of DLBCL and itself may become a potential target for therapy.
Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Humans ; Lymphoma, Large B-Cell, Diffuse ; metabolism ; pathology ; Male ; Matrix Metalloproteinase 9 ; metabolism ; Matrix Metalloproteinases, Secreted ; metabolism ; Middle Aged ; Tissue Inhibitor of Metalloproteinases ; metabolism ; Young Adult

AIMTo develop simple but reliable intracellular labelling method for high-resolution visualization of the fine structure of single neurons in brain slice with thickness of 500 microm.

METHODSBiocytin was introduced into neurons in 500 microm-thickness brain slices while blind whole cell recording. Following processed for histochemistry using the avidin-biotin-complex method, stained slices were mounted in glycerol on special glass slides. Labelled cells were digital photomicrographed every 30 microm and reconstructed with Adobe Photoshop software.

RESULTSAfter histochemistry, limited background staining was produced. The resolution was so high that fine structure, including branching, termination of individual axons and even spines of neurons could be identified in exquisite detail with optic microscope. With the help of software, the neurons of interest could be reconstructed from a stack of photomicrographs.

CONCLUSIONThe modified method provides an easy and reliable approach to revealing the detailed morphological properties of single neurons in 500 microm-thickness brain slice. Without requisition of special equipment, it is suited to be broadly applied.

Animals ; Image Processing, Computer-Assisted ; Neurons ; cytology ; physiology ; Patch-Clamp Techniques ; Rats ; Rats, Sprague-Dawley ; Software ; Staining and Labeling ; methods

Adult ; Denervation ; methods ; Female ; Humans ; Male ; Middle Aged ; Nasal Provocation Tests ; Nose ; innervation ; Rhinitis, Allergic ; Rhinitis, Allergic, Perennial ; surgery ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the effect of nucleolin silencing on the differentiation of rat neural stem cells (NSCs) and the role of Wnt signaling pathway in mediating such effect.

METHODSAdenovirus vectors expressing small interfering RNA (siRNA) against nucleolin were constructed, verified, and packaged in HEK293A cells. The adenovirus was then transfected into NSCs isolated from neonatal SD rats and the differentiation of the NSCs was examined by detecting the expressions of neuron specific encloase (NSE) and glial fibrillary acidic protein (GFAP) using immunocytochemistry. The expressions of nucleolin, nestin, Wnt3, and β-catenin in the cells were determined with Western blotting.

RESULTSRestriction endonuclease and sequencing analysis verified successful construction of the adenoviral vector expressing nucleolin siRNA (nucleolin-siRNA2). Infection of rat NSCs with nucleolin-siRNA2 significantly lowered nucleolin protein expression as compared with that in negative and blank control groups (P<0.05). The percentages of NSE-positive cells and GFAP-positive cells were significantly higher in NSCs infected with nucleolin-siRNA (P<0.01); the infection also resulted in obviously lowered expression of nestin protein and increased expressions of Wnt3 protein and β-catenin nucleoprotein in the cells.

CONCLUSIONSNucleolin silencing by adenovirus-mediated RNA interference induces the differentiation of NSCs into neurons and astrocytes, which is related with the activation of Wnt signaling pathway.

OBJECTIVETo investigate the effect of electro-acupuncture-anesthesia device (EAAD) as an auxiliary localizing method for brachiplex nerve blocking.

METHODSOne hundred and twenty patients scheduled for operation in upper limbs were randomly divided into group A and group B, 60 in each group. Bupivacaine 25 ml (0.375% in concentration) was used to block intermuscular plexus brachialis in the operation for anesthesia and EAAD was used for adjunctive localizing in group A, while in group B paresthesia elicitation was used.

RESULTSIn group A, the blocking was complete in 56 cases and incomplete in 4, the anesthesia effect was superior to that in group B, complete in 46, incomplete in 9, and of inefficient in 5 (P < 0.01).

CONCLUSIONEAAD provides a localizing method for plexus brachialis blocking more precise than paresthesia elicitation.

Acupuncture Analgesia ; Adolescent ; Adult ; Brachial Plexus ; Electroacupuncture ; Female ; Humans ; Humeral Fractures ; surgery ; Male ; Middle Aged ; Nerve Block ; methods ; Radius Fractures ; surgery ; Ulna Fractures ; surgery

AIMTo study the pharmacokinetics of bromotetrandrine (W198) in rats and beagle dogs.

METHODSThe concentrations of W198 in serum were determined using HPLC method with UV detection.

RESULTSThe pharmacokinetic parameters of W198 after single iv doses of W198 10, 20 and 40 mg x kg(-1) in rats were as follows: T1/2beta were 6.60, 7.36 and 6.77 h, AUC0-24 h were 3.797, 7.371 and 15.192 mg x h x L(-1), Vd were 7.14, 4.33 and 4.13 L x kg(-1), CL were 2.83, 2.60 and 2.71 L x (kg x h)(-1), respectively. The T1/2beta and AUCo-24 h of W198 after single im dose of W198 20 mg x kg(-1) in rats were 11.61 h and 12.646 mg x h x L(-1). The im bioavailability of W198 in rats was 56.9%. The T1/2beta, AUC0-24 h, Vd and CL of W198 after single iv dose of W198 5 mg x kg(-1) in beagle dogs were 11.72 h, 12.646 mg x h x L(-1), 0.70 L x kg(-1) and 0.46 L x (kg x h)(-1), respectively. The plasma protein binding ratio of W198 with human serum protein was 78.0%.

CONCLUSIONThe absorption of W198 in rats was of first order kinetics.

Alkaloids ; metabolism ; pharmacokinetics ; Animals ; Antineoplastic Agents ; metabolism ; pharmacokinetics ; Area Under Curve ; Benzylisoquinolines ; metabolism ; pharmacokinetics ; Biological Availability ; Blood Proteins ; metabolism ; Dogs ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Female ; Humans ; Male ; Protein Binding ; Rats ; Rats, Wistar ; Species Specificity

OBJECTIVEThe Jiangu decoction is used in the treatment of steroid-induced femoral head necrosis in clinical experiences, which has functions of tonifying kidney and activating blood, and invigorating spleen to remove phlegm. The decoction is mainly composed of Radix Polygoni Multiflori, Rhizoma alismatis Rhizoma Drynariae, haw, medlar, Radix Astragali, radix rehmanniae, angelica, Radix Codonopsis, radix salviae miltiorrhizae, Fructus Ligustri Lucidi, licorice, pharmaceutical composition. This study was designed to investigate the influence of Jiangu decoction on peroxisome proliferator-activated receptor gamma (PPARgamma) in the femoral head of rabbits with steroid-induced femoral head necrosis.

METHODSEighteen adult SPF healthy New Zealand rabbits were divided into 3 groups: control group, model group, Jiangu decoction group. The rabbits of Jiangu decoction group orally received Jiangu decoction suspension with a dose of 10 ml/kg each day and the drug content was 0.719 g/ml. The rabbits in control and model groups were given saline with a dose of 10 ml/kg. The methylprednisolone sodium succinate was injected intramuscularly into left leg with a dose of 40 mg/kg. Then the rabbits were fed continuously for 3 weeks. The glucocorticoid levels, PPARgamma and plasma glucocorticoid levels in the femoral head were measured before and after modeling.

RESULTSBefore model established, the plasma glucocorticoid levels had no significant difference among three groups (P=0.301). At 3 weeks after model established,the plasma glucocorticoid level of rabbits in model group increased compared to the control group (P=0.001); and the plasma glucocorticoid level of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The glucocorticoid level in the local femoral head of rabbits in model group increased compared to the control group (P=0.001); and the glucocorticoid level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.001). The PPARgamma level in the local femoral head of rabbits in model group increased compared to the control group (P=0.018);and the PPARgamma level in the local femoral head of rabbits in Jiangu decoction group decreased compared with model group (P=0.033).

CONCLUSIONThe Jiangu decoction is effective to inhibit the femoral head adipogenic differentiation by decrease the PPAR content, so as to prevent and treat steroid-induced femoral head necrosis.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Femur Head Necrosis ; chemically induced ; drug therapy ; pathology ; Glucocorticoids ; blood ; toxicity ; Male ; Medicine, Chinese Traditional ; PPAR gamma ; analysis ; Rabbits

Objective:To verify the dose of static and dynamic intensity modulated radiotherapy (IMRT) through 2-dimension ionization chamber matrix after external electrically-driven multi-leaf collimator (MLC) was installed in KB1800 medical linear accelerator. Methods: 40 patients who need receive IMRT were divided into static IMRT group (20cases) and dynamic IMRT group (20cases) as the random number table. The ionization chamber was applied to implement scale dose, and solid water and 2-D ionization chamber matrix was applied to verify dose. Besides, the feasibility of static and dynamic IMRT were compared and researched.Results: The verification results of plane dosage of static IMRT plan and dynamic IMRT plan showed that the passing rates both of them were above 90.0%, and the verification of intensity modulated dosimetry of medical electric linear accelerator KB1800 with external electric MLC was consistent with standard.Conclusion:After the external electrically-driven MLC is installed on the accelerator, the dosage of IMRT can achieve the verification standard, and it can be applied to clinical treatment and can satisfy the requirement of clinical radiotherapy.

Twenty Newcastle disease virus (NDV) strains were isolated from chickens and geese in the field outbreaks during 2005 and 2006 in some regions of Jiangsu and Guangxi province. Assessment of the virulence by MDT and ICPI, RT-PCR and sequence analysis of fusion protein gene were used to compare the properties of NDV isolates. The results indicated that MDT and ICPI of the isolates were 45.3h - 58.2h and 1.61 - 2.00 respectively, which confirmed that the all NDV isolates were highly virulent. And their hemagglutinin were not resistant to heat and belonged to fast pattern of elution. The results of nucleotide sequencing and phylogentic analysis of fusion protein gene showed that the twenty strains shared homology from 79.7% to 100% among themselves, from 78.1% to 83.4% and from 80.2% to 90.1% with NDV LaSota, F48E8, respectively. The putative amino acid sequences of fusion protein at the cleavage sites of all the isolates were 112R-R-Q-R/K-R-F117, with the motif characteristics of the virulent NDV strain, which was in accordant with the results of assessment of the pathogenicity. The phylogentic tree based on sequences of fusion protein gene variable regions (47-420nt) revealed that the 18 strains belonged to sub-genotype VIId and the others belonged to an old genotype III of NDV, revealing that subgenotype VIId virus was responsible for the NDV outbreaks in some regions of Jiangsu and Guangxi promince recently.
Amino Acid Motifs ; Amino Acid Sequence ; Animals ; Base Sequence ; Chickens ; virology ; China ; epidemiology ; Disease Outbreaks ; Geese ; virology ; Molecular Epidemiology ; Newcastle Disease ; epidemiology ; genetics ; Newcastle disease virus ; genetics ; pathogenicity ; Phylogeny ; Poultry Diseases ; epidemiology ; genetics ; virology ; Reverse Transcriptase Polymerase Chain Reaction ; Viral Fusion Proteins ; genetics