Endothelial monocyte-activating polypeptide-Ⅱ (EMAP-Ⅱ ) is a novel proinflammatory cytokine with proinflammatory,proapoptotic and antiangiogenic properties.It is associated with many tumorassociated proteins,such as tumor necrosis factor,vascular endothelial growth factor,hypoxia inducible factor-1α,arginyl-tRNA synthetase,and insulin-like growth factor- Ⅰ.EMAP-Ⅱ has anti-tumor properties and has a good prospect in cancer prevention and treatment.

0.05). The data proved that PSMA7 was overexpressed in pcDNA3.1(-)/PSMA7-transfected A549 cell line, both in mRNA level and in protein level. Conclusion Eukaryotic expression plasmid pcDNA3.1(-)/PSMA7 has been successfully constructed, and the PSMA7 is stably expressed in A549 cell line. This would pave the way for further study of PSMA7.

OBJECTIVE:To study the effective portion of the ultrafiltration membrane-separated ethanol extract of Pae-onia lactiflora and its antineoplastic activity in vitro. METHODS:The content of peoniflorin in ethanol extract of P.lactiflora was determined by HPLC. The drug-containing serum of mice was used to cultivate tumor cells such as HT29,HL60 and S180,then the inhibitory effect of P.lactiflora ultrafiltrate on the tumor cells and the association between dosage and administration time of the P.lactiflora ultrafiltrate were observed. RESULTS:P.lactiflora(10～100 g?kg-1) showed marked inhibition on all kinds of the above-mentioned tumor cells in a dose-dependent manner; the drug-containing serum also showed marked inhibition on all of kinds of tumor cells at different time in a time-dependent manner. CONCLUSION:The effective portion of ultrafiltration membrane-separated ethanol extract of P.lactiflora has antineoplastic activity in vitro.

Objective:To investigate the effect of HSPC 238 overexpression or low expression on the regulation of the target protein ( HMOX1, MT2A, UBB, RPS27a ) and the regulation pathways.Methods: To construct the interference vector and overexpression vector of HSPC238 respectively,transfected the HEPG2 cell lines by the liposome method,detect the expression of mRNA and protein of the HSPC 238 and the target proteins by the RT-PCR and Western blot , further to transfer the overexpression plasmids of the target proteins into the HEPG 2 cell lines which had been transfected with interference vector and overexpression vector of HSPC238,the experimental group cell lines were treated with proteasome inhibitor MG 132,to purify the target proteins with nickel column,do immunoblotting with HSPC238 to detect the accumulation situation of the target proteins.Results: The HMOX1,MT2A, RPS27a were downregulated obviously when the HSPC 238 was interfered exogenous;and the MT2A,UBB,RPS27a were up-regulated after the HSPC238 overexpressed.The overexpression plasmid of target proteins were transfected into the HEPG 2 cell lines which have been transfected with interference vector and overexpression vector of HSPC 238 ,compared with the control group ,the target protein band in the experimental group was significantly increased after treatment with the proteasome inhibitor MG 132.Conclusion:The HSPC238 overexpression can upregulate the expression of MT 2A,UBB and RPS27a,and interfering HSPC238 can downregulate the expression of HMOX1,MT2A and RPS27a;the HSPC238 target protein may play a regulatory role through the UPP pathway;the HSPC238 may play a regulatory role on the target proteins through the UPP pathway.

Objective:To construct a bait vector for HSPC238, and to screen the target proteins which interact with the HSPC238.Methods:Gene synthesis method was used to synthetic gene HSPC238, then connected with the pGBKT7 vector after digesting by the sfiIA and sfiIB,to obtain the bait plasmid pGBKT7-HSPC238,then transferred into the yeast strains AH109 with the empty plasmid pGBKT7after sequencing,to observe its self-activating effect in the nutrient deficiencies medium,and further to screen the target proteins which interact with HSPC238 from the human fetal liver cDNA library.Results:The bait vector pGBKT7-HSPC238 was successfully constructed,and it had no self-activating effect through the phenotypic screening,after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5(RPL5) may be the one of the target proteins which interacted with HSPC238 from the human fetal liver cDNA library.Conclusion: We successfully constructed the bait plasmid vector PGBKT7-HSPC238,and after the yeast two-hybrid technology with literature analysis,we preliminary screened and found that the ribosomal protein L5( RPL5) may be the one of the target proteins which interacted with HSPC238.

Objective To investigate the effect of up‐regulation and down‐regulation of HSPC238 on the RB gene mRNA and pRb protein .Methods PcDNA3 .1‐HSPC238 vector and PLL3 .7‐HSPC238 vector was transiently transfected into HepG2 cells re‐spectively .The level of RB mRNA was detected by real‐time PCR and pRb was detected by Western blotting .Results The level of RB mRNA and pRb in up‐regulation group both increased compared with control grops respectively .Conversely ,the level of RB mRNA and pRb in up‐regulation group both decreased compared with control grops respectively .Above results were all statistically significant(P<0 .05) .Conclusion HSPC238 could have a positive effect on the expression of the RB gene .

Objective To investigate the diagnosis and disease assessment of serum pancreatic stone protein(PSP/reg) in ventilator-associated pneumonia(VAP) by a prospective study.Methods The blood sample was collected in patients with mechanical ventilation(MV) from September 2012 to October 2015.Then the concentration of PSP/reg was detected by ELISA method and the time of MV and the outcome of VAP were recorded,while the patients who did not have VAP occurred in the same period regarded as control group.Results Compared with the control group,there was no significant difference in the serum concentration of PSP/reg in VAP group(P>0.05).From continuous monitoring,compared with the start time of MV,there was a significant increase in the concentration of PSP/reg in VAP group(P<0.05),and reached the highest peak in the fourth day.That of the seventh day was significantly decreased,but still higher than the first day(P<0.05).It decreased further in the fourteenth day,but compared to the start time of MV was still higher(P<0.05).In this study,38 cases were successfully evacuated in VAP group.Compared with the fourth day,the concentration of PSP/reg in the 38 cases in the stop of MV had significantly decreased(P<0.05),close to that of in the start of MV,but still a difference between them(P<0.05),and higher than that of the control group in the stop of MV(P<0.05).By Spearman analysis,we found that PSP/reg concentration in the first day had a significant correlation with CRP(r=0.570,P<0.05),but WBC and PCT were not(P>0.05).Conclusion PSP/reg can be used as one of the indicators for diagnosis of VAP,and may be related to the severity of VAP.

Objective: To investigate the expression of HSPC238 in cervical intraepithelial neoplasia and cervical cancer.Methods:We collected 76 cases of cervical cancer,105 cases of CIN and 28 cases of normal cervical epithelial.Then we inves-tigated the expression of HSPC238 by using immunohistochemistry and compared the significant differences between them.Results:There was no significant difference in the expression of HSPC238 between the cervical cancer and normal cervical epithelial ( Z=-0.242,P>0.05).However,there was significant difference between the cervical intraepithelial neoplasia and normal cervical epithelial (χ2=19.159,P<0.01) and the expression of HSPC238 was correlated with the grades of CIN.The expression of HSPC238 decreased when the grade of CIN was increasing.( rs=-0.327,P<0.01 ).Conclusion:The low expression of HSPC238 might be correlated with the development of cervical neoplasia.

Objective To investigate the diagnosis and prognosis evaluation value of neutrophil elastase (NE) in ventilator-associated pneumonia (VAP).Methods A retrospective analysis was conducted.The data of patients undergoing mechanical ventilation admitted to Department of Central Intensive Care Unit (ICU) of Boai Hospital of Zhongshan City Affiliated to Southern Medical University from September 2012 to October 2015 were enrolled.The patients were divided into two groups according to whether they suffered from VAP or not.The content of NE in serum and bronchoalveolar lavage fluid (BALF) at the time of mechanical ventilation start,VAP diagnosis (the worst value from 48 hours after mechanical ventilation start to weaning in non-VAP patients),and at the time before mechanical ventilation weaning,as well as inflammation parameters,clinical pulmonary infection score (CPIS),duration of mechanical ventilation and prognosis were recorded.Receiver operating characteristic curve (ROC) was used to analyze the predictive value of NE on VAP diagnosis and prognosis.Results Finally 38 patients were enrolled in the VAP group,and 40 in non-VAP group,and baseline data was similar between the two groups.There was no significant difference in the content of NE in serum and BALF between VAP group and non-VAP group [serum NE (μg/L):67.04 (63.00,75.75) vs.69.00 (63.75,75.00),BALF NE (μg/L):96.26 (85.26,176.01) vs.95.26 (86.76,107.11),both P ＞ 0.05].From continuous monitoring,no significant change in the content of NE in serum and BALF during mechanical ventilation was found in the non-VAP group,but the content of NE in serum and BALF at the time of VAP diagnosis in VAP group was significantly higher than that at mechanical ventilation start [μg/L:157.00 (153.04,165.75) vs.67.04 (63.00,75.75),178.04 (153.00,188.25) vs.96.26 (85.26,176.01),both P ＜ 0.05],and NE content in serum and BALF was significantly decreased at the time after VAP clinical recovery and before mechanical ventilation weaning [μg/L:75.67 (64.51,110.55) vs.157.00 (153.04,165.79),95.50 (66.56,183.02) vs.178.04 (153.00,188.25),both P ＜ 0.05].The NE in the start time of VAP in VAP group was divided into four groups according to quartile,it was found that with the increase of NE content in serum and BALF,the CPIS was increased,the duration of mechanical ventilation was prolonged,and the prognosis was poor (all P ＜ 0.01).Compared with non-VAP group,white blood cell count (WBC),neutrocyte proportion,C-reactive protein (CRP),and procalcitonin (PCT) in VAP group were significantly increased (all P ＜ 0.01).NE in BALF was significantly positively correlated with WBC,neutrocyte proportion,CRP and PCT (r value was 0.507,0.432,0.779,and 0.519,respectively,all P =0.000),among which the highest correlation was CRP.NE in BALF used for VAP diagnosis has good accuracy,with sensitivity of 87.4％,and specificity of 90.6％,and sensitivity and specificity of NE in serum for VAP diagnosis was 78.6％ and 79.2％ respectively.Conclusion NE can be used as one of the indicators for VAP diagnosis,and it is related to the prognosis of VAP.

[ ABSTRACT] AIM:To observe the effect of high glucose on the protein expression of calreticulin ( CRT) and its association with cell apoptosis and mitochondrial dysfunction in the cardiomyocytes.METHODS: AC-16 cardiomyocytes were randomly divided into normal glucose group, high glucose group, high glucose+CRT siRNA group and isotonic con-trol group.The cell apoptotic rate, reactive oxygen species (ROS), mitochondrial membrane potential level, respiratory enzyme activity, and protein expression of CRT were observed.RESULTS: Compared with the cardiomyocytes in normal glucose group, the apoptotic rate and ROS production of cardiomyocytes increased in high glucose group, accompanying with the decreases in the mitochondrial membrane potential level and enzyme activitiy of the respiratory chain.The protein expression of CRT was significantly increased in high glucose group.However, compared with high glucose group, high glucose+CRT siRNA decreased the expression of CRT and attenuated the damage of mitochondria, but CRT siRNA did not reduce the ROS level in cardiomyocytes.CONCLUSION:High glucose brings about CRT over-expression to induce mito-chondrial injury, thus increasing myocardial apoptosis.