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.

Arsenicals ; pharmacology ; DNA Methylation ; drug effects ; Humans ; Intercellular Signaling Peptides and Proteins ; genetics ; metabolism ; K562 Cells ; Membrane Proteins ; genetics ; metabolism ; Oxides ; pharmacology

Female ; Humans ; Hyperkeratosis, Epidermolytic ; pathology ; Infant, Newborn

Objective To investigate the effect of high altitude aviation on the pressure and diameter of the trachea cannula cuff after injecting air or water. Methods In the circumstance 5 km height and 795 hPa cabin pressure, the air injected cuffs were divided into two groups, one was under the ground circumstance, the other was in the high altitude aviation environment. The volumes of injected air were 5 ml, 7 ml, 9 ml, 11 ml, 13 ml, 15 ml, 17 ml, and the cuff pressure and its diameter were measured. The water injected cuffs were also divided into two groups of ground and high altitude aviation environment. The volumes of injected water were 10 ml, 12 ml, 14 ml, 15 ml, 16 ml, and the cuff pressure and its diameter were measured. The results were compared between the ground circumstance and high altitude aviation environment. Results The diameter of injection air group versus under the ground circumstance group had the statistical significance (t=5.000-9.449, P<0.05), when the injection air was larger than 15 ml, the pressure effect had statistical significance (t=5.000, 8.000, P<0.05). Water injection group had not statistically significant. Different water volume injection had no effect on pressure and diameter (P>0.05), while different air volume injection had significant effect on pressure and diameter (F=5.132, 5.980, P<0.01). When the water volume was 10 ml, the cuff pressure was (24.00±4.62) cmH2O (1 cmH2O=0.098 kPa) , which was appropriate to the range of cuff pressure (20-30 cmH2O). Conclusions In high altitude aviation environment the trachea cannula cuff should adopt water injection, and the best water volume is about 10 ml.

Objective To investigate the effects of artesunate (Art) on cell proliferation, apoptosis, nuclear factor (NF)-κB and monocyte chemotactic protein 1 (MCP-1) expressions induced by high glucose in rat renal mesangial cells (HBZY-1), and the mechanism thereof. Methods HBZY-1 cells were cultured and divided into normal glucose group (5.6 mmol/L), high glucose group (25 mmol/L) and high glucose with different concentrations of Art (10 mg/L, 20 mg/L, 30 mg/L) groups. MTT assay was used to detect the cell proliferation after 48 h. The apoptotic rate was evaluated by flow cytometry with Annexin V-FITC/PI double stains. The protein levels of NF-κB and MCP-1 in the cell culture supernatant were determined using ELISA. Results High glucose induced apoptosis and proliferation in HBZY-1 cells, and the expressions of NF-κB and MCP-1 in the supernatant were also increased (P<0.05). After treatment with Art, the proliferation was obviously abolished, and the apoptosis was increased, and the expressions of NF-κB and MCP-1 in the supernatant were decreased in HBZY-1 cells. The effects of Art showed a dose-dependent manner (P<0.05). Conclusion Artesunate treatment can reverse the effect of high glucose in HBZY-1 cells in a dose-dependent manner, which may provide a new therapeutic strategy for diabetic nephropathy.

AIM: To investigate the mechanism of basil polysaccharide in inhibiting the invasion and metastasis ability of ovarian cancer,and to evaluate the potency of hypoxia tumour microenvironment in affecting the action of basil polysaccharide.METHODS: The human ovarian cancer cell line SKOV3 was cultured with basil polysaccharide under normoxia(21% O2,5% CO2) and hypoxia(1% O2,5% CO2,94% N2),respectively.Morphology of this cell line in different groups under different oxygen concentrations was observed under inverted phase contrast microscope.The cell invasiveness was tested by transwell method and the activities of matrix metalloproteinase-2(MMP-2) among each group were analyzed by gelatin zymography.The expressions of OPN mRNA and protein levels were detected by RT-PCR and immunohistochemistry,respectively.RESULTS: Compared to the control group,basil polysaccharide decreased the invasiveness and migrating ability and inhibited the secretion of MMP-2 in SKOV3 cells.Basil polysaccharide also reduced the expression level of osteopontin(transcriptional level and protein level) of the SKOV3 cells in both normoxic and hypoxic conditions.Moreover,these functions of basil polysaccharide were more obviously in hypoxia.CONCLUSION: The migrating ability of the SKOV3 cells is enhanced under hypoxic condition.Basil polysaccharide may inhibit the invasiveness and migrating ability of SKOV3 cells through reducing osteopontin expression and MMP-2 secretion.

Objective To establish the young rabbit models with gastroesophageal reflux (GER) and observe the effect of erythromycin in different parts of muscle segment (fundus ventriculi and lower esophageal),so as to investigate the variation after destroying the interstitial cells of Cajal (ICC).Methods Forty young rabbits of 8 weeks old were randomly divided into control group(n =20) and experimental group(n =20).In the experimental group,the fun dus ventriculus was partly ligated combined with helicobacter restriction at a level near the pylorus to establish GER young rabbit models.In the control group,the fundus ventriculi and pylorus were dissociated then the abdomen was closed.Four weeks later,10 rabbits from the experimental group and 10 rabbits from the control group were taken,2 lower esophageal segments and 2 fundus ventriculi muscle segments were split in each animal,and the ICC were destroyed at 1 lower esophageal segment and 1 fundus ventriculi muscle segment.The amplitude and frequency of lower esophageal and fundus ventriculi muscle segments were measured by using electrophysiology in vitro.Results (1) After 4 weeks of operation,the GER young rabbit models were successfully established.The data were detected by esophageal pH monitoring,general morphology observation and HE stain.Immunohistochemical staining of specific receptors protein c-kit on ICC showed that the ICC in the experimental group was reduced remarkably,and the morphological characteristics changed.(2) In the control group,the frequency and amplitude of lower esophageal muscle segments were (2.60 ± 0.70) times/min and (3.03 ± 0.59) g,the frequency and amplitude of fundus ventriculi muscle segments were (3.50 ± 0.97) times/min and (5.33 ± 1.09) g;in the experimental group,the frequency and amplitude of lower esophageal muscle segments were reduced to (0.29 ± 0.49)times/min and (0.61 ±0.43) g;the frequency of fundus ventriculi muscle segmens in the experimental group turned to (0.43 ± 0.54) times/min and the amplitude was (1.13 ± 0.41) g,and there were statistically differences compared with those of the control group (t =9.86,8.15,8.29,12.55,all P ＜ 0.05).(3) The influence of erythromycin in the lower esophageal and fundus ventriculi muscle segments:the amplitude after injecting erythromycin of lower esophageal muscle segments in the experimental group was (1.16 ± 0.83) g,the amplitude was significantly different (t =2.35,P ＜ 0.05),while the frequency had no relevance (P ＞0.05);the amplitude of fundus ventriculi muscle segments in the experimental group was (4.20 ± 1.14) g,and the difference was statistically significant(t =4.76,P ＜ 0.05),and there was no statistically more differences in the frequency (P ＞ 0.05).In the control group,the frequency and amplitude of lower esophageal muscle segments were (4.50 ±0.84)times/min and (5.13 ± 0.94) g,the frequency and amplitude of fundus ventriculi muscle segments increased to(5.40 ± 1.43) times/min and(7.50 ± 1.28) g,and the differences were statistically significant (t =3.79,5.78,3.06,6.45,all P ＜ 0.05).(4) The contraction activity of muscle segments after adding the erythromycin was prevented in the lower esophageal muscle segments or fundus ventriculi muscle segmens.Conclusions ICC plays an important role in the animal models of GER;erythromycin promotes the motility of the muscle segments in GER young rabbit models;in the sick models,erythromycin accelerates the gastrointestinal motility via ICC on muscle segments.

Objective:To understand the rules and features of consultation service of outpatients by examining the outpatient consultation in clinical pharmaceutical-care practice.Method:The 1-year clinical charts for outpatient consulta- tion were monitored.The population and disease characteristics of outpatients,consulted problems and resolutions were re- viewed,Result:The counseling patient's median age was 32.And 56.7% of the patients were females.The disease charac- teristics were influenced by the distribution of beth regional diseases and hospital departments.Neither history of adverse drug reactions nor drug quantity of each prescription had any significant influence on the patient consultation behavior.The major problem(72%)concerned about was how to use drugs properly.Conclusion:The general disease information about different ages,genders,periods are indispensable for consultant pharmacists to acquire.More considerate and initiative service for drug uses should be provided for the outpatient counseling.

Aim To evaluate the endothelial dysfunction level of different arteries at different stages of SHR,and the recovery after administration.Methods SHR model was used,captopril(3.375 g·kg~(-1)·d~(-1)) was administered from week 7 to week 24 and the effects were observed continuously until 8 weeks post treatment.Pathological changes of aorta,mesenteric and apex cordis arteries were examined at the time points of 6,18,24,32 wk,and the endothelial-dependent relaxation of the former two preparations were tested by acetylcholine(ACh)(n=6).Results There were pathological changes in the thoracic aorta,mesenteric artery and arteriole at 18 wk,and aggravated along the age.The thoracic aorta demonstrated the most severe pathological changes appearing endothelial cells ablated and tunica media thickening.The significant decline of endothelium-dependent relaxation in aorta,and mesenteric arteries of SHR reflected an aging dependent change of vascular function with the most severe situation in the aorta(P=0.10,18 wk,P<0.01 24,32 wk);captopril increased the aorta vasodilatation of SHR at 18 wk time point,without the effect in mesenteric artery(P<0.05 vs SHR).Conclusions During the progress of SHR,endothelial damages have been observed in all three kinds of vasculatures together with the reduced endothelial-dependent relaxation.The aorta presents earlier and deeper damage than middle and small size vessels,and is sensitive towards the antihypertensive therapy such as the angiotensin converting enzyme inhibitor.

Objective To investigate the correlation between invasion ability and cytoskeleton remodeling of hepatocarcinoma cell by atomic force microscopy (AFM) and to explore mechanical properties during genesis,development and invasion of hepatocellular carcinoma (HCC).Methods Four HCC cell lines (MHCC-97H,MHCC-97L,SMMC-7721 ,Huh-7 )with different invasive ability were studied.Mechanical parameter (Young′s modulus)was measured by AFM.The pattern of cytoskeleton remodeling of HCC cell lines with different invasive ability was detected by immunofluorescent staining. The difference of cell invasive ability was tested by cell scratch experiment in other to verify mechanical data.Kruskal-Wallis H test was used to compare the differences between groups.Results The results of AFM indicated that Young′s modulus of cytoplasma area and nucleus area decreased gradually as cell invasion ability increased (χ2 =472.78,622.43,both P <0.01).According to invasive ability from low to high,Young′s modulus of cell cytoplasm area of Huh-7,SMMC-7721 ,MHCC-97L and MHCC-97H were 1 602.43 (845 .48,3 317.25)Pa,1 055 .28 (367.48,2 280.77)Pa,1 026.78 (369.20,2 019.96)Pa and 503.12 (366.11 ,700.31)Pa,respectively.Young′s modulus of cell nucleus area of Huh-7,SMMC-7721 ,MHCC-97L and MHCC-97H were 2 823.98 (1 262.78,4 440.07 )Pa,1 313.43 (590.71 , 2 678.62)Pa,1 285 .17 (583.29,1 961 .19)Pa and 655 .57 (441 .29,943.39)Pa,respectively.The results of immunofluorescent staining showed that the stronger the cell invasive ability,the worse cytoskeletal integrity and more irregular cell microfilament distribution. In cell scratch assay, the migration rate of MHCC-97H was 46.67% in 24 h and 86.47% in 48 h,that of MHCC-97L was 45 .70%in 24 h and 82.86% in 48 h,that of SMMC-7721 was 39.41 % in 24 h and 79.85 % in 48 h and that of Huh-7 was 34.60% in 24 h and 72.09% in 48 h,which showed that the cell migration orderly in creased as the cell invasion ability increased.Conclusions It seemed that HCC with higher invasive ability had lower Young′s modulus,softer cell,stronger deformability,worse cytoskeleton integrity and more irregular cell structure,and vice versa.