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 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

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 observe the effect of Qi-strengthening and blood-activating Chinese patent medicine Qigui Ershen Granules on the carotid intima-media thickness(IMT ) , atheromatous plaque scores, serum fibroblast growth factor 23 (FGF23) and Klotho protein levels, and oxidation- and inflammation-associated indicators in carotid atherosclerosis patients. Methods Fifty-two carotid atherosclerosis patients were randomized into Chinese medicine group and western medicine group, 26 cases in each group. Chinese medicine group was treated with Qigui Ershen Granules orally, and western medicine group was treated with Atorvastatin Calcium Tablets orally. The mediation for the two groups lasted for 24 continuous weeks. Carotid ultrasonography was performed before and after treatment for the examination of carotid IMT and plaque Crouse scores. Double antibody sandwich enzyme-linked immunosorbent assay(ELISA) was applied for the detection of serum Klotho, FGF23, interleukin 1(IL-1) and tumor necrosis factorα(TNF-α) levels, and radio-immuno-precitation method was used for the assay of serum reactive oxygen species (ROS), malondialdehyde (MDA) and superoxide dismutase (SOD) levels. The clinical efficacy of the two groups was evaluated by the scores of Qi deficiency syndrome and blood stasis syndrome before and after treatment. Results (1) In western medicine group, 5 cases dropped out and were excluded, and a total of 21 cases completed the trial; in Chinese medicine group, 3 cases dropped out and were excluded, and a total of 23 cases completed the trial.(2) After treatment for 24 continuous weeks, IMT and Crouse scores of the plaque in the two groups were obviously reduced(P < 0.01 compared with those before treatment) , but the differences of IMT and the scores between the two groups were insignificant after treatment(P > 0.05). (3) Serum Klotho protein level was increased while FGF23 was decreased in Chinese medicine group after treatment (P < 0.01 compared with those before treatment); no obvious changes of serum Klotho protein and FGF23 levels were found in western medicine group before and after treatment(P > 0.05). The effects of Chinese medicine on increasing Klotho protein level and decreasing FGF23 level were superior to those of western medicine (P<0.01). (4) After treatment, serum IL-1, TNF-α, ROS and MDA levels were decreased and serum SOD level was increased in the two groups (P < 0.01 compared with those before treatment). The differences of the above indexes were insignificant between the two groups after treatment(P > 0.05).(5) The scores of Qi deficiency syndrome and blood stasis syndrome in Chinese medicine were decreased after treatment (P < 0.01), but showed no significant changes in western medicine group (P > 0.05). Chinese medicine group had better effect on improving the scores of Qi deficiency syndrome and blood stasis syndrome than western medicine group(P < 0.01).(6) After treatment, the total effective rate for improving Qi deficiency syndrome and blood stasis syndrome in Chinese medicine group was 82.61%, 78.26%, and that in western medicine group was 28.57%, 14.28%respectively, the difference being significant (P<0.01). Conclusion Qi-strengthening and blood-activating Qigui Ershen Granules have certain effects on counteracting atherosclerosis, inflammatory aging and oxidation.

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 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 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.