[Abstract] Objective: To investigate the mechanisms of carnitine palmitoyltransferase 1c (CPT1c) expression to affect the proliferation and apoptosis of human thyroid papillary cancer B-CPAP cells through the AMP-dependent/activated protein kinase (AMPK) pathway in the low glucose and hypoxic conditions. Methods: Firstly,humanthyroidpapillarycarcinomaB-CPAP cells were cultured under normal condition or low glucose and hypoxic condition respectively, followed with the treatment of AMPK inhibitor compound C. Western blotting was used to detect the expressions of AMPK, p-AMPK, peroxisome proliferator-activated receptor α (PPARα) and CPT1c; the proliferation and apoptosis were detected by CCK-8 and Flow cytometry, respectively. Then PPARα-siRNA was synthesized and transfected into B-CPAP cells to knock down PPARα, and then the cells were cultured under normal or low glucose and hypoxic condition respectively.Above indicators were also detected to verify the regulation of PPARα on CPT1c. Finally, the human luciferase reporter plasmid containing CPT1c gene promoter was constructed, and the effect of PPARα on the activity of CPT1c promoter luciferase activity was observed by immunofluorescence. Results: The expressions ofAMPK, p-AMPK, PPARα and CPT1c were significantly increased in B-CPAP cells under low glucose and hypoxia condition (P<0.05 or P<0.01), while cell proliferation and apoptosis rate did not change significantly (P>0.05). After the treatment of AMPK inhibitor compound C, the expressions of p-AMPK, PPARα and CPT1c in low glucose and hypoxia group were significantly decreased (P<0.05 or P<0.01), the inhibitory rate on cell proliferation and apoptosis rate were significantly increased (P<0.05). However, the change range was smaller than that in the normal culture + compound C group (P<0.05).After PPARα knockdown, the expressions ofAMPK, p-AMPK, PPARα and CPT1c in cancer cells cultured under normal conditions were significantly decreased (P<0.05 or P<0.01), and the inhibitory rate on cell proliferation and apoptosis rate were significantly increased (P<0.05). While under low glucose and hypoxia condition, the expression of CPT1c in cells after transfection was significantly decreased (P<0.05), and the inhibition rate on cell proliferation and the apoptosis rate were significantly increased (P<0.05); However, the change range was still lower than that of normal condition group after transfection (P<0.05).After PPARα overexpression, the ratio of fluorescence in the empty vector group was not significantly different from that of the blank group (P>0.05), and the ratio of fluorescence was significantly increased in PPARα over-expression group (P<0.05). Conclusions: AMPK can increase the expression of PPARα to promote the expression of CPT1c in thyroid cancer B-CPAP cells under low glucose and hypoxia conditions, thereby inhibiting cell apoptosis and maintaining cell proliferation ability.

Objective To analyze the clinic pathologic factors and survival rate by performing a retro-spective study on the early gastric carcinoma (EGC)with signet ring cell underwent curative gastrostomy (SRC) percentage less than 50%after operation .Methods A total of 424 patients was diagnosed as EGC from January 2008 to January 2010 in Changhai Hospital .The patients were divided into three groups according to the different percentage of SRC within tumor cells , SRC group ( with percentage of SCR more than 50%) , Mixed SRC group (with percentage of SRC less than 50%),and Non-SRC group(Classic adenocarcinoma without SRC ).Then we evaluated the clinic pathologic indicators and prognoses , and study on the relationship with histologic types . Results In ECG,the mixed-SRC group was similar to the SRC group on the phases of onset age and sex .On the other phases,the mixed-SRC group was more associated with mucosa -confined,lower lymph node metasta-sis(LNM),and 5-year survival rate was better than adenocarcinoma group (P<0.05).However,the mixed-SRC group showed more submucosal invasion ,and higher LNM than other groups ( P<0 .05 ) .The mixed-SRC component was one of the independent risk factors of LNM .Conclusion In EGC,mixed-SRC had a more favor-able risk factor of LNM ,showed more aggressive behavior than other groups and displayed poor prognosis .

Objective To investigate the relation of the peripheral blood C-reactive protein (CRP)and procalcitonin (PCT) and multiple organ dysfunction syndrome( MODS ) for patients with multiple trauma.Methods A total of 136 patients with multiple trauma were divided into MODS group (49 cases) and non MODS group (87 cases),and 50 healthy volunteers were chosen as control group. Peripheral blood CRP and PCT concentration were detected at different times. Results The highest concentration of peripheral blood CRP in MODS group was appeared on 48 h [(38.7 ± 2.7) mg/L], CRP concentrations in MODS group were significantly different with non MODS group and control group (P ＜0.05). The highest concentration of peripheral blood PCT in MODS group was appeared on 24 h [(20.3 ± 1.7)μ g/L], PCT concentrations in MODS group were significantly different with non MODS group and control group (P＜ 0.05). Conclusions CRP and PCT are relevant to the occurrence of MODS in trauma acute stage. The observation of peripheral blood CRP and PCT concentrations can predict the occurrence of MODS.

Objective To analyze the methods and strategy of treatment for multiple injuries in the use of damage control operation, and improve the treatment level and cure rate for multiple injuries.Method From March 2005 to March 2009, 21 patients with multiple injuries were treated by damage control strategy and performed the determinacy operation after the resuscitation in ICU. Results In 21 patients, 16 cases (76.2%) were cured and 5 cases (23.8%) were dead. The injury severity score of 5 dead cases were exceeded 35 points. Conclusions In the strategy of damage control operation, the surgical operation is considered to be a part of the total resuscitative procedure, not the ending of treatment. To those multiple injuries patients, it is safe and effective to perform the damage control operation. It has practical value in clinical treatment.

Objective To summarize the experience in diagnosis and treatment of primary splenic tumor. Methods The clinical data of 43 cases of primary splenic tumor patients treated in our hospital between January 1990 to January 2003 were retrospectively analyzed. Results The detectable rate of B-US was 95.3% and that of CT was 96.6%. All the 43 patients included 28 with benign tumors and 15 with malignant ones. The median follow-up period was 6.8 years(2 months-15 years). The 5-year overall survival of benign tumor was 100% and The 1-,3-and 5-year overall survival rates of malignant tumor were 80%, 53.3 % and 26.7%, respectively. Conclusion The image methods such as B-US and CT are the main diagnostic methods for primary splenic tumor. Early diagnosis, radical operation and combined therapy are important for improving the prognosis of the tumor.

Objective To investigate the modulation of EPCs by interleukin 1β (IL-1β) and p38 mitogen activated protein kinase (p38MAPK) and the pathogenesis resulting from their dysdifferenfiation after trauma.Method Thirty pigs were divided into a control group (n = 15) and a multiple organ dysfimction syndrome (MODS) group (n = 15), the latter of which were subjected to a "two-hit" injury including hemon'hagic shock and endotoxemia. Phosphorylation of p38MAPK in peripheral blood mononuclear cells was monitored by western blotting. The concentration of IL-1β in peripheral blood plasma was determined by ELISA and the numbers of EPCs with FCM in peripheral blood plasma were monitored. The morbidity rates in the two groups were compared by chi square test. The levels of phosphorylation of p38MAPK in peripheral blood mononuclear cells, the concentmtions of IL-1β in peripheral blood plasma and the numbers of EPCs in the peripheral blood were compared between groups using with Student's t lest. Results The level of p38MAPK phosphorylation was more augmented and the concen-tration of IL-1β higher in peripheral blood mononuelear cells and plasma from MODS pigs compared with those from control pigs; nevertheless the mauler of EPC conspicuously decreased in the peripheral blood (P <0.01). The morbidity rate in the MODS group was much higher than that in the control group (P < 0.01). There were fewer EPCs in the peripheral blood of animals in group M than in the peripheral blood of animals in group C (P <0.01). Conclusions p38MAPK phosphorylation is important for the pathogenesis of MODS. p38MAPK phospho-rylation might cause the concentration of IL-1β in the peripheral blood plasma to rise and could cause a drop in the numbers of EPCs, thereby aggravating the inflanmmatory reaction in MODS.

Objective To analyze the etiology of duodenal trauma and improve the result of its diagnosis and treatment. Methods The data of 28 cases with duodenal trauma during March 1968 to September 2007 was analyzed retrospectively. Results Twenty-two (78.6%) patients had blunt injuries. The rate of duodenal trauma and associated injuries was 21.4%(6/28)and 89.3% (25/28) respectively. The easiest injury part on the deseending part constituting was 46.4% (13/28) ,the secondary on the horizontal part constituting was 21.4% (6/28).The postoperative complication rate was 28.6% (8/28), the mortality rate was 3.6% (1/28), and the cure rate was 96.4% (27/28). Conclusions The incidence of blunt injuries is high compared with other kinds of duodenal trauma in China. It suggests that the early diagnosis, exploration proper surgical management,and intensive postoperative care are emphasized to reduce the mortality and complication rate.

The regulation of a target gene expression is very important in gene therapy. However, constitutive or inappropriate expression of the genes with traditional expression system may interfere with the effect of the gene therapy, even may lead to lethal side effect. We constructed an RU486 inducible eukaryotic vector carrying DsRed protein and evaluated its regulatable effect in vitro. The single vector named PDC-RURED was constructed with molecular biological methods, which contained DsRed gene, promoter and RU486-inducible system. To minimize any potential interference, we spaced the two transcriptional elements with a 1.6 kb insulator. The vector was identified by different enzyme restrictions, sequencing analysis and PCR assay. We demonstrated the regulatable expression of this vector after transfection in HEK293 cells by fluorescence microscopy and flow cytometry. In the absence of RU486, no significant DsRed protein activation was observed, whereas in the presence of RU486 up to 40 fold activation of the DsRed protein was observed in cultured cells. The data show that the novel eukaryotic expression plasmid vector can be used to regulate the expression level of genes of interest in appropriate time under the control of RU486. This inducible expression vector provides a powerful tool for the research of gene regulation and gene therapy.
Cell Line ; Fluorescent Dyes ; metabolism ; Genetic Therapy ; methods ; Genetic Vectors ; genetics ; Humans ; Kidney ; cytology ; Luminescent Proteins ; biosynthesis ; genetics ; Mifepristone ; pharmacology ; Promoter Regions, Genetic ; genetics ; Transfection

This paper provides an insight into the "3+2" clinical teaching mode implemented from 2001,ponders on and explores related problems in the present mode and proposes the optimization strategy.

Objective:To investigate the effect of continuous blood purification on the expression of cytokines in pig lungs during MODS.Methods:Twenty-five pigs were randomly divided into model group,control group and high volume hemofiltration(HVHF) group.Blood bio-chemical and organ function indexes were determined.Real-time quantitative PCR was used to determine the expression level of IL-6 mRNA and IL-8 mRNA and TNF-? mRNA. Results:After HVHF,the serum chemical and organ-function indexes were improved and the morbidity and mortality of MODS were much lower than those in model group.The expression level of IL-6 mRNA and IL-8 mRNA and TNF-? mRNA in the lung were reduced significantly compared with model group.And the expression level of cytokines associated mRNA in the lungs was interrelated with the rate of respiratory failure. Conclusion:HVHF could decrease the over-expression of cytokine mRNA in the lung after trauma when used at early period so that pulmonary function would be improved.