Objective　To prevent early closure of growth plate and developmental deformities of limbs by allografts of cultured cartilages into growth plate defects of rabbits. Methods　Chondrocytes isolated from articular cartilage of 1-month rabbits formed cartilage after cultivation in centrifuge tubes. The cartilages cultured for two weeks were implanted into growth plate defects of proximal tibiae of 6-weeks rabbits. At 4th and 16th weeks, X-ray, histologic and immunohistochemical examination were performed. Results　The tibiae had no marked deformities after 4 weeks of operation. Histologic examinations showed that the defects were filled with cartilage. Immunohistochemical results of type Ⅱ collagen were positive. The tibiae with allografts of cultured cartilages had no evident deformities after 16 weeks of operation. Histologic examination showed nearly closure of growth plates. On the contrary, the tibiae on control side formed severe deformities and growth plate were closed. Conclusion　Allograft of cultured cartilages into growth plate defects may replace lost growth plate tissues, maintain normal growth of limbs and prevent developmental deformity.

Objective To explore the feasibility and reliability of laparoscopic fundoplication in the treatment of severe gastro-esophageal refulx disease. Methods Five cases of laparoscopic fundoplication were reviewed retrospectively from June 2001 to October 2001. Results Laparoscopic Nissen Fundoplication was performed in 3 cases, Laparoscopic Toupet Fundoplication in 2 cases. Preoperative symptoms were completely relieved. The postoperative esophageal manometry increased from (7 32?1 34)mmHg to (18 20?3 43)mmHg( t =12 23, P

Objective To explore the feasibility and rel ia bility of laparoscopic fundoplication in the treatment of severe gastro-esopha geal reflux disease.Methods Retrospectively forty-two cases of laparoscopic fundop lication were reviewed from June 2001 to March 2004.Results Laparoscopic Nissen fundoplication was performed in 33 cases, and laparoscopic Toupet fundoplication in 9 Symptom score was decreased from 11 56?1 32 to 2 04?1 36 (P

Objective To study on the effectiveness and reliability of endoscopic varicose vein ligation (EVL) combined with hand-assisted laparoscopic splenectomy (HLS) in portal hypertensive patients. Methods Retrospectively 15 cases of EVL combined with HLS were reviewed from June 2001 to October 2004. These patients with severe esophageal varicose and hypersplenism were adapted to perform endoscopic varicose vein ligation, then 1 -2 weeks after the varicose became milder or disappeared, hand-assisted laparoscopic splenectomy was conducted. Results The number of preoperative ligation was 2. 2 times in average. Esophageal varicose was completely disappeared in 11 and mild in 4. There were no postoperative complications, conversion and death in Hand-assisted laparoscopic splenectomy. The average postoperative count of platelets were (38-67) ? 106/L preoperatively raised to (89-310) ? 106/L postoperatively. In the follow up period (average 17. 6 months) , no varicose vein bleeding happened. Conclusions EVL combined with HLS is not only minimal invasive, but also a secure and effective measure for portal hypertension.

In order to adapt to the demand of culturing high quality talents and to strengthen student's comprehensive quality,pathophysiology department of medical college of Qinghai University carried on the reform on the pathophysiological experiment examination system.After researches and practices in recent years,we established a set of relatively sound examination system including experiment operating and experiment report in normal study,report for designable experiment and experiment skill and theory exam.Investigation results demonstrated that examination system after reform functioned well in training the students' operation ability,observation ability and ability to analyze and solve questions.

AIM:To provide evidence for the molecular mechanism of homocysteine (Hcy) as an independent risk factor in atherosclerosis (AS) by investigating the effect of Hcy on phenotype transformation and proliferation of vascular smooth muscle cells(VSMCs) in rats.METHODS:After treated with different concentrations of Hcy for 24 h,the cultured VSMCs were assayed for cell proliferation rate by MTT method,cell cycle by flow cytometry,the expression of SM22-? mRNA by semi-quantitative RT-PCR and the observation of morphological characteristics and the phenotype transformation by transmission electron microscopy.RESULTS:Hcy increased the cell proliferation rate and gradually reduced the proportion of the cells in G0 /G1 phase.Hcy down-regulated the expression of SM22? mRNA and the most significant effect was observed at concentration of 1 000 ?mol/L.The observations of transmission electron microscopy revealed an abundant endoplasmic reticulum,Golgi's complex,loose nucleus and puffy chromatin in VSMCs treated with high concentration of Hcy.CONCLUSION:Hcy promotes the proliferation and phenotype transformation of VSMCs simultaneously.

Objective To investigate the effect of combined double-stranded RNA (dsRNA) and imiquimod stimulation on uterine immune cells. Methods In BALB/c × C57BL/6 mice and non-obese dia-betic (NOD) × C57BI/6 mice, embryo resorption rate was detected in the presence or absence of Toll-like receptor 3 (TLR3) agonist dsRNA [poly( 1: C)], TLR7 agonist imiquimod ( R837), or their combination, respectively. In in vivo system, the status of intracellular cytokine production in uterine CD45 + cells was de-tected by flow cytometry. To identify the CD45 + cells, uterine CD3+ T cells and CD49b + NK cells derived from placenta and decidua basalis were stimulated with dsRNA and imiquimod in in vitro systems, and the status of intracellular cytokine production was detected. Mitogen-activated protein kinase (MAPK) antago- nists SP600125 and PD98059 were used to block the increase of cytokine production. Results A synergistic increase of embryo resorption was observed after the induction of dsRNA and imiquimod combination. Mean-while, a synergistic increase of TNF-α and IFN-γ production was detected after the induction in CD45 + cells. Further study found that although synergistic effect can be detected in both CD3 + cells and CD49b + cells in BALB/c mice, the status was different in NOD mice. The cytokine increase should mainly be attrib-uted to CD3 + T cells, since no such increase was detected among the CD49b + NK cells in the NOD mice. The synergistic effect of combined agonists was partially inhibited by Jun N-terminal kinase (JNK) MAPK inhibitor SP600125 and almost completely abrogated by extracellular signal-regulated kinase (ERK) MAPK inhibitor PD98059. Conclusion Boosted TLR3 and TLR7 signal may be transmitted via Thl-type T cells, rather than NK cells in NOD mice. ERK MAPK pathway may be critical in TLR3 and TLR7 involved signa- ling.

AIM:To investigate the effect of doublecortin-like kinase 1(DCLK1)on the biological properties of gastric cancer stem cells,and to explore its possible mechanism.METHODS:Serum-free suspension culture of gastric cancer stem cells and targeted inhibition of DCLK1 activity in gastric cancer stem cells with DCLK1 inhibitor DCLK1-IN-1 were performed.The expression levels of DCLK1,stemness-related proteins(SOX2 and OCT4),proliferation-related pro-teins(cyclin D1 and c-MYC),drug resistance-related proteins(ABCG2 and TOP2A),epithelial-mesenchymal transition-related proteins(E-cadherin,vimentin and Snail),and PI3K/AKT/mTOR signaling pathway-related proteins in gastric cancer stem cells were examined by Western blot.The effects of DCLK1 on viability and drug resistance of gastric cancer stem cells were determined by CCK-8 assay,and the effects of DCLK1 on self-renewal of gastric cancer stem cells were de-termined by methylcellulose spheroid-forming assay.Wound-healing and Transwell assays were performed to assess the ef-fect of DCLK1 on the migration and invasion of gastric cancer stem cells.RESULTS:The expression levels of DCLK1 and stemness-related proteins SOX2 and OCT4 in gastric cancer stem cells were significantly higher than those in parental cells(P＜0.01).The proliferation,drug resistance,migration and invasion of gastric cancer stem cells in DCLK1 inhibi-tion group were significantly lower than those in Sphere cell group(P＜0.01).The expression levels of proliferation-related proteins(c-MYC and cyclin D1)and drug resistance-related proteins(TOP2A and ABCG2)were down-regulated,the ex-pression of epithelial marker E-cadherin was up-regulated,the expression of mesenchymal markers vimentin and Snail was down-regulated,and the expression levels of PI3K/AKT/mTOR signaling pathway-related proteins and their phosphoryla-tion levels were reduced in DCLK1 inhibition group(P＜0.05).CONCLUSION:DCLK1 is highly expressed in gastric cancer stem cells,which may be involved in the proliferation,drug resistance and invasion of gastric cancer stem cells by regulating PI3K/AKT/mTOR signaling pathway.It suggests that DCLK1 can be used as a potential target for gastric cancer stem cells.

This paper aimed at investigating the effects of Xinfukang Oral Liquid on mitochondrial proteomic alterations in rats with heart failure after myocardial infarction and exploring its possible mechanisms. Heart failure models were established by ligating the left anterior descending coronary artery of SD rats. Rats were randomly divided into sham group, model group, Xinfukang group. 8 weeks after drug intervention, the mitochondrial proteomic alterations of myocardial tissue were detected by two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) . Furthermore, expression levels of part of the differently expressed proteins were verified by western blot. Compared with model group, 20 differentially expressed protein spots were detected in Xinfukang group, 13 of which showed increased protein expression and 7 decreased; 13 differentially expressed protein spots were successfully identified by MALDI-TOF-MS. Bioinformatics analysis showed that these differential proteins were mainly associated with energy metabolism, stress reaction, oxidative damage, cyto-skeleton, cell differentiation and proliferation. Western blot results showed that the expression of Stress-70 protein and Nucleophosmin increased and the expression of ATP-αdecreased in model group. The expression of Stress-70 protein and Nucleophosmin decreased and the expression of ATP-αincreased in Xinfukang group, which shows the same results in proteomics. Xinfukang Oral Liquid can partly adjust proteomic alterations of myocardial mitochondrial in HF rats, and its intervention mechanism may involve improving energy metabolism, reliving stress reaction and oxidative damage, as well as regulating cell differentiation and proliferation. The results of comparative proteomic analysis performed by using2-DE and MALDI-TOF-MS are acurate, stable and reliable.

OBJECTIVETo study the effects of chrysotile on the activities of some enzymes for xenobiotics metabolism.

METHODSUICC chrysotile (UC) and China Mangya chrysotile (MC) asbestos fibers were used at different doses (0, 25, 50, 100, 200 mg/L) to study its effects on the activities of cytochrome P4501A1 and glutathione S-transferase (GST) in A549 cell line. Also, the effects of chrysotile on the activities of CYP1A1 and GST induced by benzo(a)pyrene were studied.

RESULTSThe activity of EROD increased slowly with the increasing dose of UC, as A549 cells were incubated with UC for 24 h, and EROD activity increased by 40% at a dose of 200 mg/L UC. However, activity of EROD decreased by 32% with 48 h incubation at the same dose, indicating that lower dose of UC and short time could induce the activity of EROD in A549 cells, whereas higher doses and long time could inhibit its activity. MC exhibited a multiphasic effects on the activity of EROD, whether at a dose of 25 mg/L for 24 h or 48 h all gave the strongest induction, 1.86 times or 1.28 times as controls, respectively. However, EROD activity decreased with the increases in MC doses and incubation time, with the lowest as 35% of the controls. The effect of UC on GST activity was not so obvious, with the highest as the increase in its activity by 20%. The highest induction of MC to GST activity was at a dose of 25 mg/L, 2, 5 times as that in controls. With the increases in its doses, effects of MC on GTS activity became inhibition from induction, like that on EROD activity. MC at a dose of 200 mg/L could lower the activity of GST by 18.7%. A549 cells were incubated with chrysotile fiber for 24 h firstly, and then incubated with benzo(a)pyrene to induce the activities of EROD and GST. The results showed that neither UC nor MC could affect the activity of EROD induced by benzo(a)pyrene. However, UC at a dose of 200 mg/L and MC at 100 mg/L could increase the activity of GST induced by benzo(a)pyrene.

CONCLUSIONChrysotile at different doses or its different types showed varied effects on the activities of EROD and GST in A549 cell lines, probably because of different physicochemical characteristics and surface activity of two kinds of asbestos.

Asbestos, Serpentine ; toxicity ; Cytochrome P-450 CYP1A1 ; metabolism ; Dose-Response Relationship, Drug ; Enzyme Induction ; Glutathione Transferase ; metabolism ; Humans ; Lung Neoplasms ; enzymology ; Tumor Cells, Cultured