Background Hypertension,ischemia and hypoxia induce the increase of glutamate level in eye tissue and furthermore produce excitatory damage and apoptosis of retinal ganglion cells(RGCs).At present,the study on the protection of traditional Chinese medicine on glutamate-induced retinal excitatory damage is lack.objective Present study was to explore the protective effects of Huoxue huayu decoction,a Chinese herbal recipe,on RGCs in acute ocular hypertension model. Methods Fony-five healthy clean Wistar rats were divided into three groups randomly.The acute ocular hypertension models were established in 40 Wistar rats by injecting the normal saline solution into the anterior chamber to elevate the intraocular pressure for 60 minutes.Huoxue huayu decoction of 4 ml was administered intragastrically once per day in 20 model rats.Other matched 5 normal Wistar rats served as normal control group.The rats were sacrificed on 1,3,7 and 14 days after modeling and the retinas were isolated for the histopathologieal and uhrastructure examination.Expression of glutamate transporter in the retina of acute ocular hypertension and normal rat retina were detected using quantitative real-time polymerase chain reaction.The utilization of animals followed the Association for Research in vision and Ophthalmology. Results After acute ocular hypertension.the retina thickness attenuated and the numbers of RGCs decreased under the light microscope in 1,3,7 and 14 days after modeling in comparison with normal control rats.The degradation of the organelle and edema as well as changes of cell nuclei were seen in model rats under the transmission electron microscopy.The expression of glutamate transporter mRNA in model group and glutamate transporter group was rapidly elevated in the first day (P<0.05),descended at the third days(P<0.01)and returned to the normal level 7 days later(P>0.05).Conclusion Huoxue huayu decoction can protect the retina against RGCs damage in the acute ocular hypertension by elevating the expression of glutamate transporters in retina.

OBJECTIVETo investigate the role of EPHA2 in regulating apoptosis, proliferation and vasculogenic mimicry of osteosarcoma cells, by gene silencing through RNA interference.

METHODSEPHA2-siRNA plasmids were achieved by gene cloning. The plasmids were transfected into human osteosarcoma cells (MG63). The expression level of EPHA2 protein was measured by Western blotting. The proliferation, apoptosis and vasculogenic mimicry features of osteosarcoma MG63cells were assessed by light microscopy, MTIP assay, flow cytometry, annexin V-FITC/PI and HE staining, respectively.

RESULTSThe EPHA2-siRNA plasmid was confirmed by DNA sequencing. After treatment with Sequence-specific siRNA targeted EPHA2, the protein level of the transfected group decreased significantly. As compared to non-siRNA transfected cells, the transfected group showed lower proliferation, higher and earlier apoptosis and less osteosarcoma-generated vasculogenic mimicry.

CONCLUSIONEPHA2 gene may be involoved in apoptosis and proliferation of osteosarcoma cells, and may be necessary for vasculogenic mimicry. Down-regulation of EPHA2 expression by sequence-specific siRNA may be considered as a new option in the treatment of EPHA2 over-expressing cancer including osteosarcoma in future.

Apoptosis ; Bone Neoplasms ; metabolism ; pathology ; Cell Line, Tumor ; Cell Proliferation ; Humans ; Neovascularization, Pathologic ; pathology ; Osteosarcoma ; metabolism ; pathology ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Receptor, EphA2 ; genetics ; metabolism ; Transfection

Objective To evaluate the advantages and short term clinical results of arthroscopic release with radiofrequeney vaporization for knee joint stiffness.Methods Arthroscopic release with ra- diofrequency vaporization was done in 32 cases with knee joint stiffness resulted from various causes be- tween January 2002 and June 2005.According to degree of stiffness,32 cases were divided into three groups,ie,Group A(18 cases with suprapatellar bursa adhesions)treated with confectioning at suprapa- tellar bursa and lateral grooves;Group B(eight cases with joint compartment adhesions)managed with debridement at suprapatellar bursa and lateral groove and intereondyloid fossa,bending the knee with pressure and abscising the adhesion part in the joint compartment with radiofrequency vaporization;and Group C(six cases with most part of the joint adhered)treated by making small incision at the lateral or medial upper pole of the patellar as well as a space at the suprapatellar bursa by blunt dissection when it was difficult to insert arthroscope.After that,release combined with massage manipulation and thorough hemostasis by radiofrequency vaporization were carried out.There was no drainage after operation.Pas- sive and active flexion and extension exercises with pressure began two days after operation.Results Pre-operative limitation of flexion was 35?-75?that was increased to 115?-125?three weeks after opera- tion,with mean improvement of 78?in Group A.Pre-operative limitation of flexion was 40?-60?that was increased to 95?-120?three weeks after operation,with mean improvement of 72?in Group B.Pre-opera- tive limitation of flexion was 25?-45?that was increased to 90?-110?three weeks after operation,with mean improvement of 64?in Group C.No haematocele occurred postoperatively,with only slight swelling of the knee.Conclusion Arthroscopic release with radiofrequency vaporization for knee joint stiffness is characterized by less bleeding,minor trauma and reliable release and can be applicable especially for the eases with only adhesion at suprapatellar bursa,lateral groove or joint compartment.

Abstract?AIM: To investigate mechanism of bradykinin ( BK) on inflammations of retinal pigment epithelium ( RPE) cells.?METHODS: ARPE -19 cells were cultured in vitro, stimulated by 100nM BK for 24h. Cell morphology changes were observed by microscope, and BK receptor localization was detected through cell immunofluorescence. Changes of Ca2+in BK and BR antagonist stimuli were detected by laser scanning confocal microscopy.The expressions of COX-1, COX-2, eNOS and iNOS protein in control group and BK group were detected by Western Blot.?RESULTS: After the stimulation of BK, there was no significant changes of ARPE-19 cells in morphology.Kinin B1 receptors ( B1R ) and B2 receptors ( B2R ) could be detected in ARPE-19 cells.Compared with control group, Ca2+concentrations significantly increased in BK group; in B1R antagonist group and B2R antagonist group Ca2+concentrations increased less than BK group; B1R and B2R antagonist group showed no obvious changes in Ca2+concentrations.Compared with control group, COX-2 and iNOS protein concentrations were significantly increased in BK group (P<0.001).?CONCLUSION:BK induces the increasing expression of COX-2 and iNOS in the cultured ARPE cells through binding with either B1R or B2R.

The attenuating effect of daidzein (DAI) on oxidative toxicity induced by Aroclor 1254 (A1254) was investigated in mouse testicular cells. Cells were exposed to A1254 alone or with DAI. The oxidative damage was estimated by measuring malondialdehyde (MDA) formation, superoxide dismutase (SOD) activity and glutathione (GSH) content. Results show that A1254 induced a decrease of germ cell number, an elevation in thiobarbituric acid reactive substances (TBARS) but a decrease in SOD activity and GSH content. However, simultaneous supplementation with DAI decreased TBARS level and increased SOD activity and GSH content. Consequently, dietary DAI may restore the intracellular antioxidant system to attenuate the oxidative toxicity of A1254 in testicular cells.
Animals ; Chlorodiphenyl (54% Chlorine) ; toxicity ; Hypoxanthine ; toxicity ; Isoflavones ; pharmacology ; Male ; Malondialdehyde ; metabolism ; Mice ; Mice, Inbred ICR ; Oxidation-Reduction ; Testis ; drug effects ; metabolism ; Xanthine Oxidase ; toxicity

OBJECTIVETo investigate the position change of upper molars and incisors in order to evaluate the stability of posterior anchorage with the application of micro-screw implant.

METHODSEight adult patients with severe maxillary protrusion were included. Upper first premolars were extracted and upper posterior anchorage was reinforced with micro-screw implant in all patients. Cephalometric and cast analyses were carried out to record the position change of molar and micro-screw.

RESULTSDuring treatment the micro-screw implants kept stable in sagittal and vertical plane. Neither the mesial-distal movement nor the tipping of the upper molars during the treatment was statistically significant (P > 0.05). The edge of upper incisors was retracted by 6.86 mm and the tipping was reduced by 18.03 degrees . The center of resistance was intruded by 3.28 mm on average. Significant change was observed (P < 0.01).

CONCLUSIONSMicro-screw implant could provide good anchorage control in the orthodontic treatment.

Adolescent ; Adult ; Female ; Humans ; Male ; Malocclusion ; therapy ; Orthodontic Anchorage Procedures ; Treatment Outcome ; Young Adult

OBJECTIVETo investigate the expression of gastrin in human gastric cancer cell line SGC-7901 and the effects of gastrin-17 and anti-gastrin mAb on its growth.

METHODSThe expression of gastrin was determined by immunohistochemistry with anti-gastrin mAb prepared by our group. In a series of experiments, the growth of SGC-7901 cells was evaluated by MTT assay on cells grown in serum-free medium and treated with gastrin-17 and/or anti-gastrin mAb.

RESULTSImmunohistochemical examination of SGC-7901 cells revealed a specific gastrin immunoreactivity. Gastrin-17 significantly stimulated cell growth at the concentrations of 1 x 10(-9) mol/L approximately 1 x 10(-5) mol/L in a dose-dependent manner. The growth of SGC-7901 cells treated with anti-gastrin mAb, either alone or in combination with gastrin-17 (1 x 10(-7) mol/L), was significantly inhibited.

CONCLUSIONGrowth of human gastric cancer cells SGC-7901 can be stimulated in an autocrine fashion. The gastrin-stimulated growth of gastric cancer cells can be blocked by anti-gastrin mAb bound specifically with gastrin. Further study on the significance of anti-gastrin mAb in designing immunotherapy targeting to gastrin or gastrin receptor is warranted.

Antibodies, Monoclonal ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; Gastrins ; biosynthesis ; genetics ; immunology ; Humans ; Receptor, Cholecystokinin B ; biosynthesis ; genetics ; Stomach Neoplasms ; metabolism ; pathology

Professor LUO Cai-gui's experience of acupuncture at acupoint "Baliao" with twisting manipulation for treatment of low back pain is introduced. This method has significant efficacy on improving low back pain and numbness of lower extremities, which is characterized with short-time manipulation, quick de-qi and long effective time. The acupuncture methods, manipulations, precautions, etc. are elaborated in details. A typical case is added.
Acupuncture Points ; Acupuncture Therapy ; instrumentation ; methods ; Humans ; Low Back Pain ; therapy ; Male ; Middle Aged

OBJECTIVETo investigate transient expression of fusion protein with a chimeric HBsAg-HSP70 construct in HepG2 cells.

METHODSEnkaryotic expression plasmids inserted HBsAg gene or chimeric HBsAg-HSP70 gene were prepared and transfected into HepG2 cells by means of cationic liposome. mRNA were detected by RT-PCR and proteins expressed in the cells were detected by immunocytochemistry 48 hours later. HBsAg in cultured supernatants and cell lysates were assayed by ELISA.

RESULTSFusion protein (HBsAg-HSP70) transient expression in HepG2 cells were confirmed by RT-PCR, immunocytochemistry or ELISA, but fusion protein was not assayed in cell cultured supernatants by ELISA.

CONCLUSIONSTransfection of HepG2 cells with a chimeric HBsAg-HSP70 construct leads to express fusion protein, but it does not secrete into cell cultured supernatants.

Bacterial Proteins ; Enzyme-Linked Immunosorbent Assay ; HSP70 Heat-Shock Proteins ; genetics ; Hepatitis B Surface Antigens ; genetics ; Hepatitis B Vaccines ; immunology ; Humans ; Immunohistochemistry ; Plasmids ; Recombinant Fusion Proteins ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Vaccines, DNA ; immunology

OBJECTIVETo investigate whether Candida albicans-native phospholipomannan (PLM) induce an inflammation response through Toll-like receptor(TLRé2 in human acute monocytic leukemia cell line (THP-1) cells.

METHODSHuman THP-1 monocytes were challenged with PLM in vitro. The mRNA expressions of TLR2, TLR4, proinflammatory cytokine [interleukin(IL)-6], and chemokine (IL-8) were assayed by real time reverse transcription polymerase chain reaction. The secretions of IL-6 and IL-8 were measured by enzyme-linked immunosorbent assay. The expression of TLR2 was analyzed with Western blot.

RESULTSPLM increased the mRNA expressions and secretions of proinflammatory cytokines (IL-6) and chemokines (IL-8) in THP-1 cells (all P=0.0000). PLM up-regulated the mRNA and protein levels of TLR2 (P=0.0000), whereas the mRNA level of TLR4 was not altered. PLM hydrolyzed with β-D-mannoside manno hydrolase failed to induce gene and protein expressions of TLR2, IL-6, and IL-8. Anti-TLRS-neutralizing antibody blocked the PLM-induced secretions of IL-6 and IL-8 in THP-1 cells (P = 0.0003, P = 0.0010).

CONCLUSIONCanidada albicans-native PLM may contribute to the inflammatory responses during Candida infection in a TLR2-dependent manner.

Candida albicans ; chemistry ; Cells, Cultured ; Glycolipids ; pharmacology ; Humans ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Monocytes ; drug effects ; immunology ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism