Aim The protective effect and mechanism of TPG on PC12 cells in calcium overloading injury models were studied. Methods Two injury models induced by KCl and NMDA were used to assay the action of TPG in cultured PC12 cells. Results The morphological examination revealed that TPG possessed obvious protective effects on PC12 cells in injury models. MTT and LDH measurement indicated that TPG increased the number of live cells and reduced the extent of cell injury significantly. TPG also lessened the concentration of calcium ion in cytoplasm. Conclusion TPG protected rat PC12 cells against two calcium overloading injuries effectively in vitro. Its actions may deal with anti oxidation, inhibition of NO production and blocking of both types of calcium channel.

Objective To evaluate the anti-human respiratory syncytial viurs ( hRSV ) effect of aminoglucomannan ( AGM) in mice.Methods BALB/c mice infected with hRSV were randomly divided into AGM and konjac glucomannan ( KGM) groups with 54 in each.Then, mice in AGM group and KGM group were subdivided into three groups and treated with different doses of AGM or KGM as 2.5 μmol/L, 0.25 μmol/L or 0.025μmol/L.Each subgroup was further divided into 3 groups with 6 in each according to administration intervals of AGM or KGM as 8 h, 12 h or 24 h.All the mice are sacrificed at 72 h.The general condition of the mice was observed everyday.Reverse transcription-polymerase chain reaction ( RT-PCR) was used to detect the expression of hRSV fusion protein ( F) mRNA in the lung tissue of the mice. HE staining method and immunohistochemistry technique for intercellular cell adhesion molecule-1 ( ICAM-1) expression in the lung tissue were used to evaluate the inflammation in the lung tissue.ANOVA for randomized block design was used to examine the influence of dose and administration intervals on the expressions of hRSV F mRNA and ICAM-1.Results Reduced activity and asthma were observed in 16 mice in the KGM group, but not in the AGM group.And two mice in the KGM group died at d3, but there was no death in the AGM group.RT-PCR showed that the levels of hRSV F mRNA in lung tissues were 0.49 ±0.21 in AGM group and 0.88 ±0.06 in KGM group (t=6.71, P<0.05).HE staining of lung tissue showed that the inflammation in KGM group was more serious than that in AGM group.The expression of ICAM-1 in AGM group was much lower than that in KGM group, and there was statistical difference between two groups (t=13.88, P<0.05).In AGM group, the levels of RSV F mRNA and ICAM-1 decreased with the increase of AGM’s concentration and rised with longer administration intervals (P<0.05), but these were not observed in KGM group (P>0.05).Conclusion AGM can alleviate the inflammation of lung tissue in mice infected with hRSV in a dose and time-depandent manner.

Common mental health problems from the perspective of prevention among medical stu-dents were discussed in this activity which was learning from the concept and method of PBL. Participants had to find the solutions themselves by small-group discussion so as to improve their mental health. Results showed that most participants confirmed the innovation, interest and intellectuality of this activity. Moreover, students could not only learn knowledge related to mental health, but also improve their friendship as well as communication skills which were beneficial to medical students' mental health.

Objective To study the clinical and pathologic changes in gigantomast.Methods Tissue sections were prepared from 180 cases of breast hypertrophy and 45 cases of normal breast tissues.The morphological changes and the expression and localization of ERRγwere evaluated on the HE and immunohistochemistry stained sections between hypertrophy and normal breast tissues.Results Compared with normal breast,hypertrophic breast showed expended ducts and obvious hyperplasia of the duct epithelial papillary.Hypertrophic breast tissues demonstrated strong expression of ERR-γ in ducts and lobules.Conclusions Upregulated expression of ERRγ is identified in hypertrophic breast tissues that may associate with the development of gigantomastia.

This study aims to establish a method to determine the serum acetaminophen concentration based on diazo reaction, and apply it in the gastric emptying evaluation. Theoretically, acetaminophen could take hydrolysis reaction in hydrochloric acid solution to produce p-aminophenol, which could then take diazo reaction resulting in a product with special absorption peak at 312 nm. Then the serum acetaminophen concentration and recovery rate were calculated according to the standard curve drawn with absorbance at 312 nm. ICR mice were given a dose of acetaminophen (500 mg x kg(-1)) by gavage and the serum acetaminophen was dynamically measured through the diazo reaction. Besides, ICR mice were subcutaneously injected with the long-acting GLP-1 analog GW002 before the gavage of acetaminophen, and serum acetaminophen concentration was measured as above to study how GW002 could influence the gastric emptying. The data showed acetaminophen ranging from 0 to 160 μg x mL(-1) could take diazo reaction with excellent linear relationship, and the regression equation was y = 0.0181 x +0.0104, R2 = 0.9997. The serum acetaminophen was also measured with good linear relationship (y = 0.0045 x + 0.0462, R = 0.9982) and the recovery rate was 97.4%-116.7%. The serum concentration of acetaminophen reached peak at about 0.5 h after gavage, and then gradually decreased. GW002 could significantly lower the serum acetaminophen concentration and make the area under the concentration-time curve (AUC) decrease by 28.4%. In conclusion, a method for the determination of serum acetaminophen based on the diazo reaction was established with good accuracy and could be used in the evaluation of gastric emptying.
Acetaminophen ; blood ; pharmacokinetics ; Aminophenols ; Animals ; Gastric Emptying ; Mice ; Mice, Inbred ICR

BACKGROUND:Dendritic cel s can regulate the immunological reaction in the intestinal tract, this functional deficit may induce inflammatory bowel disease. Tol-like receptor-4/nuclear factor-κB pathway is highly involved in this reaction. OBJECTIVE:To establish experimental colitis model in rats, to observe effects of resina draconis on dendritic cel s and Tol-like receptor-4/nuclear factor-κB expression in rats with experimental colitis, and to explore its action mechanism. METHODS:A total of 44 male Sprague-Dawley rats were randomly assigned to four groups (n=11):blank control group, model group, resina draconis group, 5-aminosalicylic acid treatment group. With the exception of blank control group, 2,4,6-trinitrobenzenesulfonic acid-induced ulcerative colitis models were established in the model group, resina draconis group and 5-aminosalicylic acid treatment group. After the models were successful y established, the rats in the resina draconis and 5-aminosalicylic acid treatment groups were intragastrical y treated with resina draconis [(0.75 g(kg·d)] and 5-aminosalicylic acid [100 mg(kg·d)] respectively for 10 days. RESULTS AND CONCLUSION:Disease activity index, macroscopic colonic damage score and histopathological score were significantly decreased in the resina draconis group compared with the model group (P<0.05). Symptoms and tissue damages were obviously lessened in the 5-aminosalicylic acid treatment and resina draconis groups compared with the model group. Expression rates of CD80 and CD86, as wel as expression levels of Tol-like receptor-4 and nuclear factor-κB were significantly higher in the model group compared with the blank control group, resina draconis group and 5-aminosalicylic acid treatment group (P<0.05, P<0.01). Tol-like receptor-4 and nuclear factor-κB expression was significantly lower in the resina draconis group than that in the 5-aminosalicylic acid treatment group. Experimental findings indicate that, resina draconis can partial y relieve experimental colitis symptoms in rats and effectively inhibit the activation of dendritic cel s in the mesenteric lymph node. Resina draconis can relieve enteric inflammatory reaction by suppressing the expression of Tol-like receptor-4 and nuclear factor-κB in rats.

Objective:To describe the clinical,radiological and pathological characteristics of idiopa-thic pulmonary haemosiderosis(IPH) in adults and to evaluate the methods of diagnosis and treatment.Methods:Two patients were successfully diagnosed and treated in our hospital and the literature on the subject was reviewed.Results:Two adult patients(19 and 34 years old) diagnosed in our hospital had 5 and 10 years of history of hemoptysis respectively,and chest CT showed bilateral diffuse alveolar opacities over mid and lower zones.Tests of antinuclear antibodies(ANAs),rheumatoid factor(RF),antineutrophilic cytopasmic antibodies(ANCA) and Anti-glomerular basement membrane(anti-GBM) antibody were negative.Haemosiderin-laden macrophages were found in bronchoalveolar lavage fluid(BALF) whose color was yellow.Microscopic examination of the lung tissue specimens obtained by transbronchial lung biopsy(TBLB) revealed hemorrhage and numerous hemosiderin-laden macrophages in the alveoli and no vasculitis or capillaritis were seen.These findings were consistent with a diagnosis of IPH.Steroid therapy had good effects.Conclusion:IPH is a diagnosis of exclusion of other causes of diffuse alveolar hemorrhage(DAH).IPH adults have relatively good drug responses and relatively good prognoses.

Background Allograft rejection is a main cause of failure of penetrating keratoplasty,especially in the patient with high risk of rejection condition.Previous study on allograft rejection mechanism focused on limbal and corneal neovascularization,but these factors did not explain all the phenomena of allograft rejection.Research found that immune cells appeared in iris and ciliary body when rejection occurred,but the relationship between these immune cells and allograft rejection is unclear Objective This study was to evaluate the relationship between diversity of vascular permeability in the iris and ciliary body and allograft rejection after penetrating keratoplasty.Methods Seventy clean eight-week-old BALB/c mice were divided into allogeneic corneal transplantation group (60 mice) and blank control group (10 mice).Allogeneic corneal transplantation was performed with the same age of C57BL/6 mice as donor and BALB/c mice as the recipients.The grafts were examined under the slit lamp microscope and scored based on the criteria of Hegde.The mice were sacrificed and iris and ciliary tissue were obtained 5,10 days and rejection after surgery.Immunohistochemistry and reverse transcription PCR (RT-PCR) was used respectively to detect the expression diversities of occludin,zonula occludens protein-1 (ZO-1),matrix metalloproteinase-9(MMP-9),major histocompatibility complex-Ⅱ (MHC-Ⅱ),and CCR5,CCR7 and their mRNA in iris and ciliary body.Image-J image analysis software was used to calculate the quantity of positive cells on iris wholemount,and absorbance of target genes (A values).The use and care of the experimental animals complied the ARVO Resolution on the Use of Animals in Research.Results The mean survival time of corneal gratts was (17±3) days after operation.The mean score was 0.6 in 5 days and 0.5 in 10 days,and 3.3 in 18 days after operation.Expression of ZO-1 reduced significantly,and that of MMP-9 increased obviously at the time of rejection.MHC Ⅱ + cells were scattered in iris and ciliary body in normal mice,and the number of the positive cells (cells/field) was increased after operation with a peak value when rejection occurred.A significant difference was seen between normal mice and rejection mice (1559.67±350.29 vs.4021.83±495.18) (P=0.000).The expressions of occludin mRNA and ZO-1 mRNA in the iris and ciliary body decreased obviously in the rejection mice.Compared with normal mice,theA value of ZO-1 and occluding were 36.74±3.13 vs.110.11±11.88 and 57.54±3.41 vs.59.90±3.50respectively,with significant differences between them (all P＜0.05).The expressions of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA in the iris and ciliary body increased gradually with the time lapse after operation and peaked when the rejection appeared.The A value of MMP-9 mRNA,CCR5 mRNA and CCR7 mRNA were significantly higher than those of normal mice (20.29±1.19 vs.2.77±0.85 for MMP-9 mRNA; 35.43±2.56 vs.9.11±0.29 for CCR5 mRNA,and 60.83±0.87 vs.0.89 ±0.95 for CCR7 mRNA) respectively (all P＜0.05).Conclusions The permeability of vascules in the iris and ciliary body increase during the allograft rejection after penetrating keratoplasty.Increased antigen presenting cells were also detected.

Objective To evaluate the contrast enhanced perfusion pattern of PTC micro-vascular imaging (MVI) quantitatively.Investigate the correlation between PTC MVI features and CD34 micro-vascular density (MVD).Methods Thirty-nine pathological and clinical confirmed sporadic PTCs were evaluated with real-time gray-scale contrast-ernhanced micro-vascular imaging under a low mechanical index.The micro-bubble agent was SoneVue.Of the 39 PTCs,33 were classical PTCs,6 were PTC with follicular variant (FVPTC).The △ ROI,which is the subtraction of peak echo intensity between the lesion region of interest (ROI) and normal thyroid parenchyma ROI,was used to evaluate the perfusion characteristics of PTC MVI quantitatively.The paraffined specimens were selected for immunohistochemical (IHC) staining for CD3,and the correlation between △ ROI and the CD34 were evaluated.Results △ ROI was strongly correlated with the CD34 expression (P=0.000),significant differences were detected in the distribution pattern of △ ROI value among different CD34 expression levels,no overlapping of the mean △ ROI values and the 95％ confidence intervals was found among the 3 CD34 expression levels.The PTC MVI perfusion was classified into 3 patterns,low perfusion,focal perfiusion and high perfusion,on the basis of combining△ ROI values with the peak ehco pattem in time-intensity curve.Conclusions The △ ROI is strongly correlated with the CD34 expression in papillary thyroid cancer.It can be used for the quantitative evaluation ofPTC MVI pattem and intensity as an objective indicator.

AIM: To optimize the conditions for in vitro culture of retinal pigment epithelium (RPE) cells, we characterized expressions of various growth factors in RPE cells, including tumor necrosis factor (TNF-α), vascular endothelial growth factor (VEGF), β fibroblast growth factor (βFGF), transforming growth factor β2 (TGFβ2), and interferon-γ (IFN-γ). We also studied expressions of caspase-3 under different concentrations of fetal bovine serum (FBS) with insulin-transferrin-sodium selenite (ITS) supplement. METHODS: First, we investigated if expressions of TNF-α, VEGF, βFGF, TGFβ2, IFN-γ, and caspase-3 in FBS and ITS with of concentration. Second, we cultured primary RPE cells from eyes of forty C57 BL/6 mice in standard dulbecco's modified eagle's medium (DMEM) containing 20,40,100mL/L FBS and 20,40,100mL/L FBS together with 10g/L ITS. Immunohisto-chemical staining and cell counting were performed to verify the existence and growth condition of RPE cells. Expressions of TNF-α, VEGF, βFGF, TGFβ2 and IFN-γ were determined using cells and supernatant from passage-3 to -4 primary RPE cell after 48 hours of culture with RT-PCR and enzyme-linked immunosorbent assays (ELISA). The expression of casepase-3 was determined via Western blotting. The major outcome measurement is the expression level of growth factors in cultured RPE cells and the experiment design is to expose the RPE cells to different culture medium. RESULTS: TNF-α, VEGF, βFGF, TGFβ2, but not IFN-γ, were expressed and the expressions increased with concentration. No expression of the aforementioned genes was detected in presence of ITS. The primary cultures of RPE cells were successfully established. TNF-α, VEGF, βFGF, TGFβ2 (but no IFN-γ) and the active caspase-3 were detected in 20,40,100mL/L FBS or 20,40,100mL/L FBS combined with 10g/L ITS; the expressions were upregulated with increasing concentration of FBS. There is no significant difference in the expression of growth factors between these groups. However, significant differences were shown among different concentration of FBS (P＜0.01）. The lowest expression was observed in 20mL/L FBS or 20mL/L FBS combined with 10g/L ITS medium with RPE cells. But RPE cells were shown in better growth condition in 20mL/L FBS combined with 10g/L ITS.CONCLUSION: TNF-α, VEGF, βFGF, TGFβ2 and caspase-3 were expressed in RPE cells and supernatants. The production of above 20mL/L FBS combined with 10g/L ITS in DMEM may be the ideal cell culture medium that supports the normal growth of RPE cells.