Objective To investigate the release and intracellular localization of high mobility group box chromosomal protein 1(HMGBl)in the peripheral blood monocytes of rheumatoid arthritis(RA) patients and the inhibitive effect of thaiidomide.Methods 19 RA patients and 20 healthy controls were included in the study.Monocytes were separated from peripheral blood with Ficoll density gradient centrifugation.Monocytes were treated with 100 ng/ml tumor necrosis factor α(TNFa)or 100 ng/ml TNFα plus 40 μg/ml thalidomide and grown in an incubator at 37℃ with 5%CO,for 24 hours.The cuIture supernatants of the monocytes were collected.HMGB1 level in the culture medium was detected with Western blot.In addition,the intraceUular localization of HMGB1 in the fflonocytes was investigated with immunocytochemical analysis. Results Without stimulation. the release of HMGBl protein was significantly increased in the culture supernatants of peripheral blood monocytes from RA patients as compared with that from healthy controls(P<0.05).TNFα(100 ng/ml)did not further increase the release of HMGBl in the monocytes from the patients with RA.Thalidomide(40 μg/ml)could inhibit the release of HMGB1 in the monocytes from RA patients stimulated with TNFα(P<0.05).In the monocytes from RA patients,HMGBl was mainly localized in the nucleus.Treatment with TNFOL(100 ng/ml)for 24 hour resulted in a cytoplasmic translocation of HMGB1,which was inhibited significantly by thalidomide. Conclusion TNFα induces the release and cytoplasmic translocation of HMGBI in the peTipheral blood monocytes of RA patients and thalidomide inhibits the release and translocation of HMGB1.

Granular corneal dystrophy is a rare autosomal dominant genetic disease in clinic.Due to the TGFBI mutation on the 5q31 chromosome,the TGFBIp abnormally aggregates in the Bowman layer and the matrix layer and metabolic disorders,patients' bilateral cornea arise opacity,making visual acuity Progressive impairment.At present,there are at least 66 TGFBI mutations,at least 10 of which are related to granular corneal dystrophy,due to variation in genotype and the difference between homozygous and heterozygous,the patients' phenotype shows a significant difference.Along with the improvement of people's cognition,and the application of laser scanning confocal microscope and the gene diagnosis,More and more patients get the correct diagnosis,Current treatment methods mainly include corneal transplantation and laser ablation,patients are not satisfied because of the postoperative recurrence and aggravated.Due to the establishment of granular corneal dystrophy animal model,lithium and gene therapy will show a good application prospects.

Objective:To investigate the breakthrough points and methods of pharmaceutical care performed by clinical pharma-cists for chemotherapy-induced Ⅳ degree myelosuppression. Methods: One advanced lung adenocarcinoma patient suffering from IV degree myolosuppression after being treated with pemetrexed combined with nedaplatin was selected as the example, and the chemother-apy regimen, the cause and treatment of IV degree myolosuppression and the pharmaceutical service could be carried out were ana-lyzed. Results: With the help of clinical pharmacists, the patient conquered chemotherapy-induced myelosuppression, and clinical pharmacists enhanced the awareness of pharmaceutical care and played a positive role in the safe and effective drug use. Conclusion:The participation of clinical pharmacists in clinical pharmaceutical care through providing pharmaceutical service is beneficial to safer and more effective drug therapy.

Objective To investigate the influences of heart in patients with hepatocellular carcinoma closed to heart after high intensity focused ultrasound ablation(HIFU). Methods Thirty four patients with hepatocellular carcinoma closed to heart received detecting the values of periphery blood aspartate transarninase(AST), creatine kinase (CK), CK-MB, α-hydroxybutyrate dehydrogenase (α-HBDH), lactate dehydrogenase(LDH), myoglobin(Myo) and troponin I (cTnI) before and 1,3, 7 days after HIFU respectively. ECG was monitored in all patients simultaneously. Results The values of AST,CK,CK-MB,α-HBDH, LDH and Myo increased 1 day after HIFU significantly (P<0.05), which decreased greatly 3 days after HIFU and nearly recovered well (P>0.05) 7 days after HIFU. However, the value of cTnI and ECG were normal before and after HIFU. Conclusions No obvious myocardium injury was found after HIFU in patients with hepatocellular carcinoma dosed to heart, cTnI may be an important marker to evaluate myocardium injury after HIFU.

Objective To set up a method of stools protein extraction,analysis and identification in order to get the new nonin-vasive indicators of human digestive diseases.Methods The stools proteins,collected from healthy persons,the patients with atrophic gastritis,those who suffed from gastric carcinoma and postoperative patients with gastric carcinoma respectively, were extracted in three different ways including saline,Tris-HCl buffer and Urea buffer,the best way was selected by using SDS-PAGE,then a preliminary analysis of stools proteins was performed by 1D LC-MS/MS.Results The methods of saline and Tris-HCl buffer could get more stools proteins than the method of urea.The proteins in stools from the healthy persons, the patients with atrophic gastritis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma were all abundant and more than one hundred.There was a significant difference in stools protein maps among the various populations.Alpha1-antitrypsin,a number of immunoglobulin and keratin were identified in the stools from patients with gastric carcinoma but not postoperative patients with gastric carcinoma and the healthy persons.Conclusions In this re-search,there was a significant difference in stools protein maps among the healthy persons,the patients with atrophic gastri-tis,the patients with gastric carcinoma and postoperative patients with gastric carcinoma,not only the composition of stools proteins,but also the abundance of same proteins.Therefore,using proteomics technologies to screening of the noninvasive indicators in human stools is viable.The study recommended that the noninvasive indicators in human stools should be iden-tified with quantitative differences analysis combination of quality of mass spectrometer method in the future research.

Objective To explore the effect and mechanism of Cyclosporin A (CsA) and cholic acid on reducing the human liver cell damage induced by α-amanitin (AMA).Methods According to the previous research results,the minimum hepatocellular survival concentration against αt-AMA (1.4 g/L),the experiment was conducted in 5 groups:control group,damage group,glycochenodeoxycholic acid group,CsA group,and CsA combined with cholic acid group (CsA+ taurocholic acid,CsA+ chenodeoxycholic acid,CsA+ glycocholic acid,CsA+ glycochenodeoxycholic acid,and CsA+ taurochenodeoxycholic acid).For each group,there were 3 time points for observation (24 h,48 h and 72 h after attacking),CsA and CsA+ glycochenodeoxycholic acid was used to protect hepatocytes,respectively,and morphological changes in cells were observed with inverted phase contrast microscope,and the live cells were counted by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) method,and aspertate aminot ransferase (AST) and alanine aminotransferase (ALT) in the culture supernatant were detected by biochemical method.Results Compared with the control group,hepatocellular growth in the injury group was obviously suppressed,with progressive cellular apoptosis and significantly decreased absorbance value of MTIT (0.345 ± 0.021);the activity of AST and ALT increased gradually,reaching the highest after 72 h [(98.4 ± 6.7) U/L and (116.2 ± 9.5) U/L,respectively].Compared with injury group,broken organelles decreased significantly and absorbance value elevated in glycochenodeoxycholic acid group,CsA group and CsA combined with cholic acid group,and at each time point,the highest absorbance value in the CsA+ taurochenodeoxycholic acid group [the highest was (0.656 ± 0.014),P ＜ 0.05];at the same time,the activity of AST and ALT didn't increase obviously,at each time point,the lowest in CsA+ taurochenodeoxycholic acid group [the lowest was (22.3 ± 6.2) U/L and (20.2 ± 5.4) U/L,P ＜ 0.05,respectively].Conclusions CsA,as well as cholic acid,can protect human liver cells from the attack of α-AMA.The mechanism may be CsA,as an organic anion transfer peptide in humans (OATP1B1 and OATP1B3) suppressant,inhibits the absorption of α-AMA.The joint application of CsA and taurochenodeoxycholic acid is superior to the single OATP substrate or inhibitor.

Objective To investigate the effect of high mobility group box chromosomal protein 1 (HMGB-1) on the proliferation and inflammatory phenotype of human fibroblast like synoviocytes (FLS).Methods FLSs were isolated from the synovial tissues of rheumatoid arthritis (RA) patients undergoing joint replacement surgery.All the experiments described here utilize the FLSs between the third and sixth passages.FLS were incubated with HMGB1 at (100,500,2 000 ng/ml) or interleukin (IL)-1β (0.5 ng/ml) or lipopolysaccharide (LPS) (100 ng/ml) alone or HMGB1/IL-1β complexes or HMGB1/LPS complexes.Cell proliferation assay were used by CCK-8,IL-6,IL-8 and matrix metalloproteinase-3 (MMP-3) in culture supernatants were measured using enzyme-linked immunosorbent assays.The measurement data were compared with single factor analysis of variance.Results Stimulation with all concentrations of HMGB1,IL-1β and LPS alone did not affect the cell proliferation of FLS.HMGB1/IL-1β complexes and HMGB1/LPS complexes did not affect the cell proliferation of FLS,neither (F=0.415.P=0.915).Except high concentration of HMGB1 (2 000 ng/ml) could significantly stimulate the secretion of IL-6 from FLS [(23.0±1.1) ng/ml],HMGB1,IL-1β and LPS alone did not affect the production of IL-6,IL-8 and MMP-3.However,HMGB1/IL-1β complexes and HMGB1/LPS complexes increased IL-6,IL-8 and MMP-3 production from FLS (F=97.804,117.383,70.179,P=0.000).Conclusion Neither HMGB1,IL-1β,LPS alone nor HMGB1/IL-1β complexes or HMGB1/LPS complexes affect the cell proliferation of FLS.HMGB1 in complex with LPS or IL-1β boost IL-6,IL-8 and MMP-3 production in synovial fibroblasts from RA patients.

Objective To demonstrate high mobility group box chromosomal protein 1(HMGBI) expression in synovium and joint,and to identify the role of HMGB1 in the pathogenesis of synovitis and joint destruction in adjuvant-induced arthritis(AA).Methods AA of 15 male rats were induced in SD rats by intradermal injection of 100?l Freud's complete adjuvant in the foot pad of the left hind paw.All rats were killed at the 18th day.Synovium and joints were collected for histopathology studies and determining the expression of HMGB1 by immunohistochemistry,and serum was collected for determining the expression of HMGB1 by western blotting analysis.Results Immunostaining of specimens from normal rats showed that HMGB1 was primarily confined to the nucleus of synoviocytes with occasional cytoplasmic staining.In contrast, inflammatory synovial tissues from AA rats showed a distinctly different HMGB1 staining pattern.Nuclear HMGBI expression was accompanied by a cytoplasmic staining in many mononuclear cells.The cytoplasmic HMGB1 expression in synovium of AA rats is significantly higher than that of normal rats.Additionally,HMGBI was highly expressed in the nuclei and cytoplasm of the subchondral chondrocytes and inflammatory cells in bone erosion in AA rats(P＜0.01),while fewer positive cytoplasmic staining of HMGB1 was found in chondrocytes and fewer positive nuclear staining was found in bone cells in normal rats.HMGB1 concentration was significantly higher in serum of AA rats than that in normal rats(P＜0.001).Conclusion The cytoplasmic HMGBI expression in synovium and joints is greatly upregulated;the level of HMGB1 in serum is increased in AA rats which suggests a patbogenetic role of HMGB1 in synovitis and bone destruction of adjuvant-induced arthritis.

Objective To detect the in vitro effect of the traditional Chinese medicine on the tachyzoites of Toxoplasma gondii. Methods Supernatant (1^5 ml) of different doses of the traditional Chinese medicine (Changqing capsule) was collected by normal saline immersion and 2^5?10+4 Toxoplasma gondii tachyzoites were added in each paste well for 8 hours. Spiramycin, pyrimethamine and azithromycin in different doses were used as controls. Normal saline was used as negative control. Mice were inoculated with drug-treated tachyzoites intraperitoneally or intragastrically. The normal mice were subcultured after 8 days for 3 generations. Results The incident number of the infected mice was significantly different among groups with different drugs and doses: 2/60, 16/60, 10/60 and 10/60 in the groups of Changqing capsule, spiramycin, pyrimethamine and azithromycin respectively (P