Background Graves ophthalmopathy is generally considered to be an organ-speeifie autoimmune disease.It was reported that 99Tc-methylenediphosphonate (99Tc-M)P) has therapeutic effect on Graves ophthalmopathy,though the mechanism has not been completely delineated.Objective This study was to explore the effects of 99Tc-MDP on the proliferation of retrooeular fihrohlasts (RFs) hyaluronic acid (HA) synthesis,and the expression rate of human leucocyte antigen-DR (HLA-DR) and intercellular adhesion molecule-1 (ICAM-1) in cultured RFs.Methods Retroocular connective tissue was obtained from 2 eyes with Graves ophthalmopathy during the orbital decompression surgery.RFs were cultured with explant culture method in DMEM medium with 20％fetal bovine serum.The cells of 2-7 generations were used in the study.Interferon γ (IEN-γ),interleukin-1 (IL-1),tumor necrosis factor-α (TNF-α) was added in medium for 72 hours to stimulate the growth of the RFs,and then 10,100 and 1000 mg/L 99Tc-MDP was respectively used to co-culture the cells with IFN-γ,IL-1 or TNF-α.3H-TdR incorporation was used to detect the proliferation of RFs,and synthesis of HA,expression rate of HLA-DR and ICAM-1 in RFs were assayed by radio-immunoassay and flow cytometry,respeetively.Results Cultured cells showed the fibrous shape under the inverted microseope with the positive response for vimentin and absent response for cytokeratine.After addition of TNF-α,IFN-γ and IL-1,the proliferation value of RFs,HA level,the expression rate of HLA-DR and ICMA-1 in RFs was (4918.33±297.91) counts/min,(505.83±41.29)mg/L,(56.88±14.67)％ and (63.57± 14.11)％ respectively,showing significant differenee in comparison with normal control group(P＜0.01).However,after co-culture of ＞100 mg/L 99Tc-MDP with TNF-α,IFN-γ and IL-1,the proliferation value of RFs,HA level,the expressing rates of HLA-DR and ICMA-1 were significantly lower than those in the TNF-α,IFN-γ and IL-1 only culture groups(P＜0.01),but were still higher than those in the normal group.Conclusions 99Tc-MDP can suppress cytonike-induced activation and growth of RFs derived from Graves ophthalmopathy in vitro at a dosedependent manner.

Unicellular green alga,Dunaliella salina(D.salina),is a biflagellar alga without cell wall,which is a kind of very important eukaryotic microalga.In the previous study,the research of D.salina focus on the morphology,the mechanism of salt tolerance and ?-carotene,however,with the rapid development of microalgal biotechnology,a lot of work about D.salina was reported in recent years.In the area of molecular biology,the studies of D.salina mainly place emphasis on the cloning and analysis of important functional genes,regulatory sequences,and the expression of foreign genes using D.salina as host.The research advance in these aspects were reviewed.

The aerolysin genes (aerA) of BZ and NK isolates were cloned and sequenced. The sequence analysis showed that the partial aerA of BZ and NK isolates consisted of 1393 bp, encoding a protein of 464 amino acids. The similarity of nucleotide and amino acid sequences of aerA between BZ and NK isolates was 97.6% and 98.3% respectively. The nucleotide sequence of aerA of BZ strain exhibited 71.6% to 97.5% homology with other Aeromonas isolates, and the amino acid sequence exhibited 68.0% to 98.9% homology. The phylogenetic tree based on aerA nucleotide sequences from Aeromonas isolates was constructed with neighbor-joining method. It showed that there were three branches of aerolysin genes, and a close relation- ship among Aeromonas hydrophila isolates which were clustered into the same branch.

The human canstatin cDNA was amplified by RT-PCR and then directionally cloned into pU? expression vector. The recombinant pU?-Can vector was connected with the screening marker (bar box), to construct a eukaryotic expression vector called pU?-Can-Bar. This expression vector was introduced into the D.salina by glass beads method. The screening culture of transformants of D.salina was performed in solid media containing 5 ?g/ml PPT, and the analyses of transformants were carried out through PCR and Southern blot. PCR results revealed that specific 700 bp products were detected in the different transformants of D.salina but not in negative control. Southern blot analysis further demonstrated that human canstatin gene was integrated into the D.salina genome. Moreover, the results of genetic stability analyses of transformants demonstrated that canstatin gene was stably inherited in the D.salina transformants. The successful preparation of the D.salina transformants will provide the experimentation evidence for producing canstatin protein cosmically by using the D.salina bioreactor and give a better prophase work basis for clinic application of canstatin protein early.

Objective: To verify whether there exists melatoni n(Mel) receptor in human embryonic nervous system. Methods: Spec ific binding of Mel to embryonic brain and spinal cord was measured by radioliga nd binding assay. Results: 125　I-Mel binding s ites in optomeninx was the most, in eptochiasm and sniff ball was next; GTPγS d ose-de pendently inhibited the binding. Conclusion: The results demonst rate the presence of specific binding of Mel in human embryonic brain and spinal cord. GTPγS has some effect on 125　I-Mel specific binding,support ing the theory that Mel receptor is coupled to inhibitory G-proteins.

OBJECTIVETo investigate the growth activity of osteoblast on a novel strontium incorporated calcium sulfate and make comparison with normal calcium sulfate material.

METHODSOsteoblast was inoculated on samples and cell proliferation was measured on the 1st, 3rd, 5th days, and the activities of ALP and osteocalcin were observed on the 5th day. And microcosmic morphology of osteoblast was observed by scanning electron microscopy(SEM).

RESULTSOsteoblast grows robustly on tested material. Cell quantity on the surface of novel material was obviously higher than normal calcium sulfate material (P < 0.05). The activity of ALP and osteocalcin on novel material was 57.8% and 40.2% higher than on normal calcium sulfate material respectively (P < 0.05). On strontium incorporated surface, osteoblast spread well. Cells were polygonal with abundant cytoplasm and the morphology was active.

CONCLUSIONStrontium incorporated calcium sulfate can sustain robust growth activity of osteoblast, which is promising to be used for bone substitute materials.

3T3 Cells ; Alkaline Phosphatase ; metabolism ; Animals ; Bone Substitutes ; chemistry ; pharmacology ; Calcium Sulfate ; chemistry ; pharmacology ; Cell Proliferation ; drug effects ; Mice ; Osteoblasts ; cytology ; drug effects ; metabolism ; Osteocalcin ; metabolism ; Strontium ; chemistry

At present,the unrealistically high drug prices become an indisputable fact to cause the chaos in the medical field.They are the main reasons of "difficult and too expensive to see doctors".And they are deeply loathed by the people in the country.In addition,the unrealistically high drug prices have become the shackles in deepening medical and health reform.Therefore,a determined effort must be made to resolve the serious problems.Through analyzing the status of high drug prices,the main reasons of the unrealistically high drug prices,and the hazards of high drug prices,the article puts forward countermeasures of state monopoly in national essential drugs to solve the unrealistically high drug prices problems for structural reform of supply side.

A novel transformation method was firstly established using glass beads in Dunaliella salina (D. salina). The results showed that the GUS gene, a reporter gene, was successfully expressed in D. salina. Cells of D. salina presented blue color under the microscope after stained. In addition, different factors which influenced transformation were optimized including the transformation consecutive time, rotate speed, concentration of the plasmid and PEG 6000. The experiment indicated that this fit together can obtain the best results for D. salina transformation: adding 150 microL PEG and 90 microL plasmid DNA to 800 microL culture of D. salina (10(6) cells/mL) containing 300 mg glass beads, swirling 12 seconds under the rotate speed 2400r/min. This newly method can be used as a potential tool in the research of D. salina gene engineering with the advantage of more simpleness, convenience, quickness and less expense.
Chlorophyta ; genetics ; DNA ; chemistry ; genetics ; Genetic Engineering ; methods ; Glass ; Glucuronidase ; genetics ; metabolism ; Histocytochemistry ; Microspheres ; Plasmids ; genetics ; Polyethylene Glycols ; chemistry ; Time Factors ; Transformation, Genetic ; genetics

OBJECTIVETo determine the effects of the autologous nucleus pulposus on the pain-related behaviors of hind paws in rats.

METHODSThe nucleus pulposus harvested from autologous coccygeal vertebra was applied beside the unilateral L4 and L5 nerve roots without compression. The mechanical withdrawal threshold of both paws were measured in different times after surgery. And hematoxylin and eosin (HE) staining was applied to observe the changes of nucleus pulposus and nerve roots.

RESULTSMechanical sensitivity of the operated side in paws obviously increased after application of autologous nucleus pulposus beside the lumbar nerve roots without compression. And HE staining showed obviously inflammatory changes in the nucleus pulposus and vacuolation in the nerve roots.

CONCLUSIONSInflammation resulted from nucleus pulposus may contribute to the development of mechanical hyperalgesia. The results suggest that in addition to mechanical compression, inflammation may be an important factor in the pathogenesis of sciatica.

Animals ; Disease Models, Animal ; Hyperalgesia ; etiology ; pathology ; physiopathology ; Intervertebral Disc Displacement ; complications ; pathology ; physiopathology ; Male ; Radiculopathy ; complications ; pathology ; physiopathology ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo study the effect of silicosis alveolar macrophages (AM) restimulated by SiO(2) on expression of c-myc oncogene in human embryo lung fibroblasts.

METHODSThe bronchoalveolar lavage of silicosis patients was collected. AMs were divided into 2 groups: (1) SiO(2): AMs were stimulated with SiO(2) (30 microg/ml) for 1, 2, 6, 12, 24 and 36 h; (2) control: treated for the same time without SiO(2). Fibroblasts were cultured with different AMs supernatants for 2 h or 7 h respectively. The expression of c-myc mRNA was determined by RT-PCR and protein by Western Blot.

RESULTSThere was no c-myc expression when fibroblasts were static. The supernatants in the S6 group stimulated expression of c-myc mRNA and protein, with the peak expression at 2 h and 7 h respectively. In the control group, AMs supernatants cultured in different time stimulated expression of c-myc mRNA and protein with the most evident expression at 12 h. The ratios were 0.749 +/- 0.088 and 0.759 +/- 0.101 respectively. Compared with control in the same period, c-myc mRNA and protein expression were significantly stronger treated with the supernatants in which AMs were stimulated for 1 h, 2 h and 6 h by SiO(2) (P < 0.05 or P < 0.01).

CONCLUSIONAMs stimulated with SiO(2) has the ability to induce c-myc oncogene expression in human embryo lung fibroblasts.

Cells, Cultured ; Fibroblasts ; drug effects ; metabolism ; Humans ; Lung ; drug effects ; metabolism ; pathology ; Macrophages, Alveolar ; drug effects ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; RNA, Messenger ; genetics ; Silicon Dioxide ; toxicity ; Silicosis ; metabolism ; pathology