Objective To explore the mechanism of the interaction between sulfur dioxide(SO2) and bovine serum albumin (BSA). Methods The spectrum characteristic of the interaction between SO2 and BSA was studied by fluorescence quenching spectrum and three dimensional fluorescence spectrum. Results The binding constants and thermodynamic parameters of SO2 with BSA were calculated at different temperatures. The quenching mechanism of BSA by SO2 was determined,the result showed that it did not belong to dynamic quenching but belonged to static quenching,which produced the complex. The hydrophobic interaction and electrostatic force played a main role in the binding of SO2 with BSA and only about one binding site occurred in the reaction. There was a certain influence to the conformation of BSA after adding SO2. Conclusion The quenching reaction of BSA by SO2 was weak. SO2 dissolved in body fluid easily and then SO32-and HSO3-are generated in vivo.

BACKGROUND: Based on the characteristics of cartilage tissue, such as consisting of single type of cells, the cartilage cells or chondrocyte, absence of blood vessel, rather low consumption level of oxygen and nutrition, low level of allo-immunocompetence and simple function in vivo, it seems to be easy for cartilage cell lines to be established for tissue and cell transplantation. We want to set up a cell line with the purpose of current use in tissue engineering in vitro. It will provide the basis for artificial tissue and organ that will become to be standardized and yielded in batch.OBJECTIVE: To explore the potential stimulatory effects of basic fibroblast growth factor(bFGF) and insulin on the proliferation and differentiation in primary culture mice chondrocytes in vitro. The effect and application of the cell factors will be evaluated for tissue engineering.DESIGN: A grouping controlled and repeated trial was conducted with the cells as the subjects.SETTING: Key laboratory of tissue transplantation and immunology of a college.MATERIAIS: The experiment was completed in the Key Laboratory of Tissue Transplantation and Immunology of the Ministry of Education, Jinan University from November 2002 to May 2003. Cultured cartilage cells at random were obtained as the study objects.METHODS: Mice cartilage cells were cultured in medium at the minimum concentrations of serum. The effects of different concentration of bFGF and insulin on the proliferation and differentiation in mice cartilage cells were observed with WST1 and immunofluorescence staining.MAIN OUTCOME MEASURES: Primary results: ① Effect of bFGF on proliferation of primary cultured mice cartilage cells. ② Effect of insulin on proliferation of primary cultured mice cartilage cells. Secondary results:morphological observation of cartilage cells RESULTS: Primary cultured mice cartilage cells were cultured in medium at the minimum concentration of serum(4 g/L fatal bovine serum). It was found that bFGF and insulin might play an important role on the proliferation and growth of mice cartilage cells in a dose-dependent manner. In addition, morphological observation of cartilage cells showed that both bFGF and insulin not only promoted the proliferation of the cells but also enhanced the matrix secretion of cartilage cells.CONCLUSION: Both bFGF and insulin can stimulate the proliferation of cartilage cells in vitro.

AIM: In order to understand the pathological changes, characteristic of degeneration in optic nerve and retina after strike of optic nerve. METHODS: According to methods of Allen's spinal injury, a 600gcm-strike power was put on the intraocular portion of the optic nerve and created a striking injury on optic nerve. After a survival interval of 48 h, 1 week, 2 weeks, 4 weeks, 3 months, the animal's optic nerves and retinas were collected and fixed for morphological examination. RESULTS: Forty-eight hours after nerve injuries, the optic nerves were slight enlargement and vacuolation. In 1 week, the optic nerve began to degenerate in injured part and the glia cell had proliferated, but the forms of retinal ganglion cells(RGCs) were normal. In 2 weeks, the vacuolation and focal necrosis were appeared between nerve fiber. The number of RGCs began to decrease. Condensed nuclei presented in the retina. In three month, the diameter of the optic nerve decreased in injury part and collo-scar was formed. The phenomenon mentioned above was more obviously. The internal nuclear neurons and outer nuclear neurons appeared rare. The thickness of retina decreased. The number of RGCs began to decrease in 48 hours and progressed thereafter. It decreased about 3.35%, 13.23%, 19.74%, 23.20%, 29.28% in 48 h, 1 week, 2 weeks, 1 month, 3 months compared with the number of normal RGCs. RGCs began to apoptosis in 48 h. CONCLUSION: The model in this experiment could make definite uncompleted optic nerve and retina injuries. The degree of neuron injuries decreased from RGC, internal nuclear neurons to outer nuclear neurons. The number of RGCs began to decrease in 48 hours, and most quickly periods from 48 hours to one week.

BACKGROUND: Endothelin(ET) -1 is a peptide with potent actions on blood vessels and nerve system. Its expression increases in the central nervous system(CNS) in a variety of pathological conditions, inducing harmful effects on the nervous tissue. However it is not clearly elucidated whether the over-expressed ET-1 can directly induce neuronal apoptosis.OBJECTIVE: To investigate whether ET-1 can directly induce apoptosis in primarily cultured brain neurons of rat, and which ET receptor subtype(s) is involved in this action.DESIGN: Completely randomized and controlled experimental study based on cells.SETTING: Neurological department in a university hospital, pathological department of a university and laboratory center of tissue transplantation and immunology, life science and technology college.MATERIALS: This study was completed in the Pathology Department, the Institute of Tissue Transplantation and Immunology, the Life Science and Technology College of Jinan University. The subjects were primarily-cultured neurons obtained from cerebral cortex of newborn rats that were provided by the Experimental Animal Center of the Medical College, Sun Yat-sen University.INTERVENTIONS: After culturing for five days, the neurons were treated with ET-1 (0. 2 nmol/L and 20 nmol/L) for 24 hours. Apoptotic neurons were semi-quantitatively measured with Annexin V and Hoechst 33258 staining respectively. ET-1(20 nmol/L), with BQ123(a selective antagonist for ET receptor A, 1 mmol/L) or with BQ788(a selective antagonist for ET receptor B, 1 mmol/L), was added respectively into the cultures simultaneously. And the apoptotic neurons were quantitatively measured with flow cytometry 24 hours later. Equal amount of PBS, instead of ET-1, waw added into the control subjects.MAIN OUTCOME MEASURES: The effect of ET-1 on apoptosis rate of cultured rat cortical neurons, and the ET receptor subtypes involved in this action.RESULTS: Twenty-four hours after treated with 0.2 nmol/L ET-1, the Annexin-V, and Hoechest 33258 positive stained cell rates[ (23.00 ± 9.96)%,(9.82 ±0.95)% ] were of no difference as compared with those of the controls[ (13.50 ± 3.35)%, (8.21 ± 2. 17)% ]. By contrast, after incubation with the higher dose of ET-1 (20 nmol/L), significant higher rate of apoptosis was measured in Annexin V staining[(50.50 ± 10.78)%, P=0.01, n=4] and Hoechest 33258 staining[(13.78±1.52)%, P= 0. 000, n = 8] . Analyzed with flow cytometry, the apoptosis rate was (0.20±0. 15)% in the control group, (26. 11 ±3.28)% in 20 nmol/LET-1 group, and(13.58 ±4. 92)% in BQ123 +ET-1 and(9.99 ±3.30)% in BQ788 +ET-1 respectively, indicating that BQ123 and BQ788 partially-blocked the apoptosis effect of ET-1 on. cultured neurons(BQ123 + ET-1 vs ET-1, P = 0. 005; BQ788 + ET-1 vs ET-1, P = 0. 001, n = 4, respectively).CONCLUSION: The higher dose of ET-1 (20 nmol/L) can directly induce apoptosis of primarily-cultured cerebral neurons of rats. The effect of ET-1 inducing neuronal apoptosis may be mediated via both ET receptors A and B.

Objective To investigate the change of expression patterns of miRNA in the serum and peripheral blood mononuclear cells (PBMC) of lung cancer and its significance.Methods Clinical case control study was employed.Establish the method of microRNA(miRNA) detection by real time quantitative PCR (RT-qPCR).peripheral blood of the study subjects were collected in First affiliated hospital of SooChow University from November 2011 to September 2012.Gender and age matched subjects whose median age was 64(40-85) included 61 lung cancer cases,48 healthy control and benign lung diseases.We used quantitative RT-PCR to assess miRNA expression pattern of-miR-20a,21,-25,-29,-31,-126,-129,-145 and -205 in peripheral blood.U6 was taken as reference,and the expression of miRNA were indicated as F =2-△Ct,ACt =CtmiRNA--CtU6.F represents relative change of miRNA expression compared to U6 in the same sample.SPSS 19.0 was used as statistical software; t test was used for comparison of two sets of samples One way ANOVA was used for multiple groups' comparison,and make multiple comparison by the S-N-K method if the result with a significant difference.Pearson correlation analysis were used for the relationship between two variables,Brown-Forsythe test was used for Ct value equality testing among multiple samples.P ＜ 0.05were regarded as statistically significant.Results miR-20a (F =271.64,P ＜ 0.01),miR-21 (F =2232.51,P＜0.01),miR-205 (F=45.13,P＜0.01),miR-29a (F=19.98,P ＜0.01),miR-25 (F=313.19,P ＜ 0.01) and miR-126 (F =32.38,P ＜ 0.01) were differently expressed in the serum of lung cancer patients and healthy control or benign disease control.miR-29a,miR-25,miR-126 was down regulated in the development of malignant lung disease; miR-31 elevated in lung cancer compared with healthy control,while miR-145 fell; miR-31 expression changed with various differentiation of lung cancer (F =5.22,P ＜ 0.01)itwas significantly increased in the moderate-differentiated cancer,but decreased when distant metastasis existed (especially bone metastasis).But in PBMC paired with the serum samples above,statistically significance was shown in lung cancer and healthy control group and benign lung diseases group in miR-126 (F=690.58,P＜0.01),miR-129 (F=26.66,P＜0.01),miR-145 (F=48.57,P＜0.01),miR-205 (F=308.61,P＜0.01).miR-25 (F=218.57,P＜0.01) and miR-31 (F=48.05,P＜0.01),were down regulated in the development of malignant lung disease.miR-20a,miR-29a were elevated in lung cancer compared to healthy controls,miR-21 was up-regulated when distant metastasis existed; the expression of miR-31 in serum and PBMC was negatively correlated (r =-0.369,P ＜ 0.05).Areas under ROC curve of miR-25 (S =0.906,P ＜ 0.01) and miR-126 (S =0.969,P ＜ 0.01) were statistically different.Conclusions miRNA may contribute to several steps of metastasis,including local invasion,extravasation or initial survival at a distant site,and metastatic colonization,or can affect the prognosis of lung cancer.The detection of miRNA in lung cancer provides a new clue to the research of its chronic progress.

AIM: To study the effect of trichosanthin (TCS), a type I ribosome-inactivating protein, on the induction of apoptosis in human leukemic cell line HL-60 cells and the influence of cycloheximide (CHX) on TCS-induced apoptosis. METHODS: Flow cytometry together with fluorescent microscopy were adopted to investigate the apoptotic cell death in HL-60 cells treated with TCS. RESULTS: Flow cytometric analysis indicated that TCS was able to induce significant apoptosis in HL-60 cells. The rates of apoptotic cells in HL-60 cells treated with TCS (20 mg/L) for 48 h was 48.7%±2.3%(±s), which was significantly higher than that of control (6.3%±1.0%)(P＜0.05). Under the same condition, the rate of apoptosis caused by CHX (5 mg/L) was 65.3%±3.9%. TCS-induced apoptosis was further confirmed by fluorescent microscopy observation and DNA gel electrophoresis, in which typical nuclear morphological changes such as chromatin condensation, nuclear fragmentation, were observed in many of the cells treated with TCS， and DNA extracted from these cells displayed typical ladder pattern. Furthermore, the effect of TCS was significantly enhanced with the pretreatment of CHX (0.2 mg/L) which did not induce any significant apoptosis when used at 0.2 mg/L seperately. TCS-induced apoptosis was time- and dose-dependent. CONCLUSION: TCS was able to induce apoptosis in HL-60 cells, which was enhanced by CHX. It was suggested that TCS-induced apoptosis was independent of new protein synthesis.

OBJECTIVE To provide the clinical anatomy data of the lingual artery for clinical application concerning about the lingual artery. METHODS CTA examination of the carotid artery was performed in 80 adult volunteers. The 3D reconstruction images of the carotid artery with hyoid bone were obtained by using 64-slices spiral CT. At the same time, 20 extraoral dissections of the submandibular region were performed on 10 human cadavers. The origin, pathway, and anatomic relations of the lingual artery in CTA and cadavers were studied. The distance from the origin of the lingual artery to the bifurcation of the common carotid artery and tip of the greater horn of hyoid bone were separately measured, and the distance between the segment of the greater horn of hyoid bone of lingual artery and the middle of greater horn of hyoid bone were also measured. And the contrast analysis was carried on the result. RESULTS The lingual artery was mainly come from the external carotid artery (74.4 % in the CTA; 80% in the Cadavers), the next origin was come from the facial artery (23.1 % in the CTA; 20 % in the Cadavers), and it was few to found that the lingual artery had origin in the superior thyroid artery, which was 2.5 % in CTA and none in Cadavers. The path of the lingual artery had high variation, but the position between the segment of the greater horn of hyoid bone of lingual artery and the greater horn of hyoid bone had constancy relatively. The lingual artery run forward with approximation parallel to the greater horn of hyoid bone into tongue,and which located in thesuperior to the greater horn of hyoid bone (2.32?1.29 mm) or in the inferior to the greater horn of hyoid bone (2.00?1.68 mm). The distances from the origin of the lingual artery to the bifurcation of the common carotid artery and tip of the greater horn of hyoid bone in CTA and cadavers were (12.93?7.36) mm, (10.40?5.75) mmand (14.80?6.18) mm, (8.35?5.44) mm respectively. CONCLUSION The lingual artery can be show clearly in CTA and can get the anatomy data in physiological condition with CTA.

AIM: To investigate the influences of protein kinase C(PKC) inhibitors on the expression of interleukin-2(IL-2) and interferon-?(IFN-?) by in vitro activated T-lymphocytes. METHODS: Double fluorescent staining together with flow cytometry was adopted to detect intracellular cytokines and to analyze the effects of H7 and gossypol on IL-2 and IFN-? expression levels of T-lymphocytes stimulated with phorbol ester (PDB)+ionomycin(I) in the presence of monensin.RESULTS: The expression rates of IL-2 and IFN-? of CD3 + T cells stimulated with PDB+I for 4 h were 16.64?2.04 and 25.81?3.53( ?s ), respectively, which were significantly higher than that of control (1.06?0.22 and 3.12?0.77)( P

Objective To investigate the status of cellular immunity from Th cell polarization in pa-tients with different courses of condyloma acuminatum(CA).Methods The isolated PBMC were polarized by PHA and self-plasma for72hours,then followed by two-color immunofluorescent staining with anti-CD4-PE and anti-CCR5-FITC,or with anti-CD4-FITC and anti-CCR3-PE.Finally the stained cells were analyzed by flow-cytometry.Results The percentages of Th1/Th2cells of the short-course group and long-course group were(25.82?2.22)%/(14.80?1.14)%and(12.20?1.37)%/(13.74?0.99)%,respectively;the differ-ences between normal control and two CA groups were significant(P

Objective To explore the expression of interleukin -17A(IL-17A),interleukin-10(IL-10), interleukin-6 ( IL-6 ) in the maternal plasma , umbilical plasma , neonatal plasma from the pregnant women who accepted the Telbivudine for the prevention of perinatal transmission of hepatitis B virus infection .Methods 85 preg-nant women who were delivered in the hospital were chosen [ Telbivudine group 25 cases,HBVDNA high viral level group,HBVDNA negative group,non-infection group(health pregnant),each group included 20 cases].The level of the serum of IL-17A,IL-10,IL-6 in the maternal plasma,umbilical plasma,neonatal plasma from the different group was prospectively analyzed by the CBA method .Results The levels of IL-17A in the maternal plasma,um-bilical plasma,neonatal plasma of the Telbivudine group [(15.29 ±4.80)pg/mL,(19.45 ±4.80)pg/mL,(17.32 ± 4.18)pg/mL]were lower than the HBVDNA high viral level group [(26.03 ±4.88)pg/mL,(34.07 ±5.59)pg/mL, (24.12 ±4.76)pg/mL],there were significant differences(P=0.000,0.001,0.019);The level of IL -10 in the maternal plasma,umbilical plasma,neonatal plasma of the Telbivudine group [(6.08 ±1.32) pg/mL,(2.33 ± 0.68)pg/mL,(3.74 ±0.61)pg/mL]were higher than the HBVDNA high viral level group [(3.95 ±1.21)pg/mL, (1.71 ±0.53)pg/mL,(2.22 ±0.55)pg/mL],there were significant differences(P=0.023,0.015,0.012);The level of IL-6 in the maternal plasma ,umbilical plasma ,neonatal plasma of the Telbivudine group were lower than the HBVDNA high viral level group,there were significant differences (all P <0.05).Conclusion Telbivudine can cause the inhibition of the replication of HBV virus , partially reverse the immunologic dissonance of the pregnant women with HBV virus ,which is conducive to maintaining pregnancy .