Background:Gastric cancer (GC) is one of the most common malignancies,and intestinal-type GC is the main histopathologic type of GC in China.We previously reported that casein kinase 2 interacting protein 1 (CKIP-1) acts as a candidate tumor suppressor in intestinal-type GC.CKIP-1 participates in the regulation of multiple signaling pathways,including the Wnt/β-catenin pathway,of which caudal-related homeobox 1 (CDX1) may be a downstream target gene.The purpose of this study was to investigate the relationship between CKIP-1 and CDX1 in intestinal-type GC.Methods:Sixty-seven gastroscopy biopsy specimens and surgically resected gastric specimens were divided into four groups:gastric mucosa group,intestinal metaplasia (IM) group,dysplasia group,and intestinal-type GC group.The expression levels of CKIP-1 and CDX1 were detected in these groups and GC cell lines,and the correlations between these expression levels were analyzed.SGC7901 and BGC823 cells were divided into CKIP-1 shRNA groups and CKIP-1 over-expression groups,and CDX1 expression was detected.β-Catenin expression was detected in intestinal-type GC tissue samples and CKIP-1 shRNA and CKIP-1 over-expression SGC7901 cells,and its correlation with CKIP-1 expression in intestinal-type GC tissue was analyzed.The Wnt/β-catenin pathway inhibitor DKK-1 and activator LiCl were incubated with SGC7901 cells,BGC823 cells,and CKIP-1 shRNA and CKIP-1 over-expression SGC7901 and BGC823 cells,following which CDX1 and Ki-67 expression were detected.Results:The expression levels of CKIP-1 and CDX1 were lower in patients with intestinal-type GC than in patients with IM and dysplasia (both P ＜ 0.05).CKIP-1 and CDX1 expression levels were positively correlated in IM,dysplasia,and intestinal-type GC tissue and cell lines (r =0.771,P ＜ 0.01;r =0.597,P ＜ 0.01;r =0.654,P ＜ 0.01;r =0.811,P ＜ 0.01,respectively).CDX1 expression was decreased in the CKIP-1 shRNA groups and increased in the CKIP-1 over-expression groups of SGC7901 and BGC823 cells compared to that in the corresponding control groups (both P ＜ 0.05).CKIP-1 expression was negatively correlated with β-catenin expression in intestinal-type GC patients (r =-0.458,P ＜ 0.01).Compared to the control group,β-catenin expression was increased in the CKIP-1 shRNA SGC7901 cell group and decreased in the CKIP-1 over-expression SGC7901 cell group (P ＜ 0.05).CDX1 expression was increased in SGC7901 and BGC823 cells treated with DKK-1,DKK-1 increased CDX1 expression and decreased Ki-67 expression in the CKIP-1 shRNA group;the opposite result was observed in SGC7901 and BGC823 cells treated with LiCl,and LiCl decreased CDX1 expression and increased Ki-67 expression in the CKIP-1 over-expression group (both P ＜ 0.05).Conclusions:Through the Wnt/p-catenin signaling pathway,CKIP-1 may positively regulate CDX1 in intestinal-type GC.

The recombinaut porcine insulin precursor(PIP)produced by Pichia pastoris in shake-flask and 501.fermenter was investigated respectively.The results indicated that 60h induction time length and 2.0%～2.5% methanol addition every day was optimum in shake- flask.The process in 50L fermenter was consisted of batch,feed-batch and induction phases.The relationship between dry cell weight(y) and culture time (t) in growth phase(batch and feed-batch phase)could be described by model y=0.6525e~(0.1907t).Glycerol and ammonia were almost used for cell growth and maintain,and no by-product was observed in batch and fed-batch phase Only 80% ammonia and 70% methanol were used by cell in induction phase.By comparison the results of shake-flask and 50L fermenter,it was concluded that the limit- ing factor in the fermentation of shake-flask and 50L fermenter was dissolved oxygen(DO)and.carbon source,respectively.When scaling the result of shake-flask to 501.fermenter,the control strategy was adapted for 50L fermenter by increasing the feed rate of methanol and the maximum PIP concentration reached 1.72 g/L.

Immobilized penicillin acylase was used for bioconversion of penicillin G into 6-APA in aqueous two-phase systems consisted of a light-sensitive polymer PNBC and a pH-sensitive polymer PADB.Partition coefficients of 6-APA was found to be:about 5.78,in the presence of 1% NaCl.Enzyme kinetic showed that reaction reached equilibrium at 7h or so.The 6-APA mole yields were 85.3%(pH 7.8,and 20 ℃) and this value was about 20%higher than control in reaction of single aqueous phase buffer.Partition coefficient of penicillin G(Na) washardly changeable,while partition coefficient of product,6-APA and phenylacetate acid was significantly changeable.Reason is due to Donnan effect of phase systems andhydrophobicity of products.The change of partition coefficients of products also affects bioconversion yield of products.In the aqueous two-phase systems,substrate,penicillin G,products 6-APA and phenylacetate acid are biased in top phase,while immobilized penicillin acylase is completely partitioned in bottom.Substrate,penicillin G enters into bottom phase,and it is catalyzed into 6-APA and phenylacetate acid,then the products enter into top phase.Finally,inhibition of substrate and products is removed to result in improvement of products yield.Moreover,immobilized enzymehashigher efficiency than immobilized cells and occupy smaller volume.Comparing with free enzyme,immobilized enzymehashigher stability,longer use life,completely partitioned in bottom phase and recycle.Bioconversion in two-phase systems using immobilized penicillin acylase showed outstanding advantage.The light-sensitive copolymer forming aqueous two-phase systems could be recovered by laser radiation at 488 nm or filtrated 450 nm light,while pH-sensitive polymer PADB could be recovered by isoelectric point(pH 4.1).The recovery of the two copolymers was 95%~99%.

13C metabolic flux analysis(13CMFA)have been the research hotspots of metabolic engineering internationally due to its accuracy and applicability.It is vital that the measurement of 13C labelling pattern of proteinogenic amino acids for 13C metabolic flux analysis.To acquire 13C-labelling proteinogenic amino acids,Pseudomonas denitrifican which products Vitamin B12 was firstly fed with mimimal culture medium contained 20% U-13C and 80% natural glucose,after the culture reached a steady state,then about 20 mg biomass was hydrolyzed by 1 ml of 6 mol/L hydrochloric acid for 24h at 110℃.Then amino acids was separated,concentrated,evaporated in a vacuum,and derivatized with MBDSTFA,TBDMS-derivatized amino acids can be observed by GC-MS last we get 13C labelling pattern of fifteen aminio acids through mass spectrum.The experimental methods and sample preparation offers referential value for the development of 13C metabolic flux analysis in our courtry.

Objective - （ ） To explore the influence on the diagnosis of occupational noise induced deafness ONID using three ， Methods versions of diagnostic criteria in 2002 2007 and 2014. A total of 1 766 workers who asked for ONID diagnosis were selected as the research subjects using judgment sampling method. The results of pure tone audiometry were collected. GBZ 49-2002Diagnostic Criteria of Occupational Noise-inducedHearing Loss（ The ONID was diagnosed using hereinafter referred to as GBZ 49-2002），GBZ 49-2007Diagnostic Criteria of Occupational Noise-induced Deafness（ GBZ 49-2007） hereinafter referred to as GBZ 49-2014 Diagnostic of Occupational Noise-induced Deafness（ GBZ 49-2014）， and hereinafter referred to as and the Results - - ， - diagnostic results were compared. Compared with GBZ 49 2002 and GBZ 49 2007 diagnosis with GBZ 49 2014 had （ vs ， vs ， P ）， （ vs ， a higher rate of ONID 57.9% 66.0% 44.8% 66.0% both <0.01 and had a higher rate of mild ONID 47.3% 54.6% vs ， P ） - - 36.0% 54.6% both <0.01 . The diagnostic rate for ONID using GBZ 492014 was higher than those using GBZ 49 2002 and - （ P ）Conclusion - GBZ 49 2007 in each age groups all <0.01 . GBZ 49 2014 improved the diagnostic rate of ONID compared - - with GBZ 49 2002 and GBZ 49 2007. The reason is related to the inclusion of 4 000 Hz hearing threshold with a weight of 0.1 - as the diagnostic hearing threshold and the use of a new age and gender correction method in GBZ 49 2014.

OBJECTIVE: Postoperative delirium (POD) is a highly prevalent complex neuropsychiatric syndrome in elderly patients. However, its pathophysiology is currently unknown. Early detection and prevention of POD is important; therefore, the aim of this study was to investigate the link between preoperative insulin growth factor 1 (IGF-1) levels in the serum and POD in the Chinese elderly patients. METHODS: One hundred and three patients who were undergoing an orthopedic operation took part in the study. Preoperative serum IGF-1 levels were measured. POD was determined daily using the Confusion Assessment Method (CAM) and DSM-IV TR. Baseline serum IGF-1 levels were compared between patients who did and did not develop POD. Correlation coefficients were calculated to evaluate relationship between baseline characteristics and serum IGF-1 levels. The relationship between baseline biomarkers and delirium status was investigated using logistic regression analysis, adjusting for potential confounding variables. RESULTS: Twenty-three patients developed POD. The POD group had lower MMSE scores and higher CCI scores and proportions of acute admission. Preoperative serum IGF-1 levels were correlated with MMSE scores and age (MMSE: r=0.230, p<0.05; age: r=-0.419, p<0.001). Baseline serum IGF-1 levels did not differ between patients who did and did not develop POD, even after adjusting for potential confounding factors, MMSE score, and age. CONCLUSION: No association was found between preoperative IGF-1 levels and POD, suggesting that they are not direct biomarkers of the incidence of POD among the Chinese elderly population. Further research with larger sample sizes is warranted to clarify the relationship.
Aged* ; Asian Continental Ancestry Group* ; Biomarkers ; Confounding Factors (Epidemiology) ; Delirium* ; Diagnostic and Statistical Manual of Mental Disorders ; Humans ; Incidence ; Insulin ; Insulin-Like Growth Factor I ; Logistic Models ; Orthopedics ; Sample Size

AIM:To explore the effect of genistein on ammonia-induced nuclear factor-κB (NF-κB) activation and the underlying mechanism.METHODS:Primary astrocyte cultures were prepared and challenged with NH4Cl to establish a hyperammonemic model.The activation of ERK,Akt and NF-κB was examined by Western blot.RESULTS:AG1478 and genistein significantly inhibited ammonia-induced activation of ERK and Akt.Ammonia-induced NF-κB nuclear translocation was significantly inhibited by the pretreatment of LY294002,genistein and AG1478.CONCLUSION:Genistein significantly inhibited ammonia-induced ERK activation and Akt-mediated NF-κB activation,which might represent the important mechanism by which this naturally occurring substance exerts its swelling-inhibiting effect.

Objective: To explore influence of continuous thoracic paravertebral block(CTPVB)under ultrasound-guidance combined with general anesthesia on inflammatory reaction and homodynamic of patients who underwent thoracotomy.Methods: 138 patients who underwent thoracotomy were prospectively selected and were divided into observation group(69 cases)and control group(69 cases)as random number table.Patients of observation group received CTPVB under ultrasound-guidance combined with general anesthesia and that of control group received general anesthesia.The changes of IL-6,IL-10 and TNF-α postoperative 48h and postoperative VAS score between the two groups were compared.And the VAS scores postoperative 2h,12h,24h and 48h also were observed.Besides,the mean arterial pressure and heart rate between the two groups at various time point included tranquillization time(T0),the 15 min(T1)after CTPVB combined with general anesthesia,the time post induced intubation(T2),5 min post skin incision(T3)and the time pre extubation(T4)were compared and analyzed so as to research its influence on hemodynamic.Results: The IL-6 and TNF-α postoperative 48h of observation group were significantly lower than that of control group(t=15.95,t=46.99,P＜0.05).And postoperative IL-10 of observation group was significantly higher than that of control group(t=134.3,P＜0.05).And the VAS scores of observation group at postoperative 2h,12h,24h and 48h were significantly lower than that of control group(t=13.91,t=17.1,t=2.321,t=9.231,P＜0.05),respectively.Conclusion: CTPVB under ultrasound-guidance combined with general anesthesia can improve postoperatively analgesic effect for patients,and it don't affect patients' blood pressure and heart rate,and it can reduce inflammatory reaction of patients who underwent thoracotomy and promote rehabilitation of patients.

Objective To investigate whether emodin combined with 5AzA-cdR can enhance the demethylation of tumor suppressor genes p16,RASSF1A and ppENK in nude mice with subcutaneously transplanted pancreatic cancer.Methods Pancreatic cancer cells Panc1 burdened subcutaneous xenograft nude mice model was established,which were randomly divided into control group,emodin group,5AzA-cdR group and emodin combined 5AzA-cdR group (combined group).The growth of transplanted tumors wasobserved in each group.Methylation specific PCR (MSP) was used to detect the methylation levels of p16,RASSF1A and ppENK in the xenograft tumor tissue among three groups.The mRNA and protein expression of three tumor suppressor genes were detected by FQ-PCR and Western blotting,respectively.Results The weight of xenografts in the control group,emodin group,5AzA-cdR group,and combination group were (0.28 ±0.01),(0.17 ± 0.01),(0.12 ± 0.02),(0.08 ± 0.01)g,respectively.The tumor volume was (517 ±0.02),(382 ± 0.01),(232 ± 0.03),(169 ± 0.01) mm3.The methylation levels of p16 were 1.00 ± 0.00,0.89 ± 0.02,0.63 ± 0.02,and 0.19 ± 0.01;the methylation levels of RASSF1A were 1.00 ± 0.00,0.88 ± 0.02,0.51 ± 0.01,and 0.32 ± 0.01;the methylation degree of ppENK was 1.00 ± 0.00,0.92 ± 0.02,0.77 ± 0.02 and 0.31 ± 0.01,respectively.The expression of p16 mRNA was 1.00 ± 0.00,1.71 ±0.02,2.67 ± 0.02,3.81 ± 0.01.The expression of RASSF1A mRNA was 1.00 ± 0.00,1.92 ±0.02,2.73 ± 0.03,3.77 ± 0.01.The expression of ppENK mRNA was 1.00 ± 0.00,1.69 ± 0.03,2.17 ± 0.02 and 4.28 ± 0.01.The expression of p16 protein was 1.00 ± 0.00,1.71 ± 0.02,2.67 ± 0.02,3.81 ± 0.01;the expression of RASSF1A protein was 1.00 ± 0.00,1.92 ± 0.02,2.73 ± 0.03.3.77 ± 0.01;ppENK protein expression levels were 1.00 ±0.00,1.69 ±0.03,2.17 ±0.02,4.28 ±0.01.The weight and volume of xenografts in the three treatment groups were significantly smaller than those in the control group.The methylation of three tumor suppressor genes was lower than that of the control group,and the expression of tumor suppressor mRNA and protein was all significantly higher than the control group,which the combination drug group was also significantly stronger than that in emodin group and 5AzA-cdR group,and the differences were statistically significant (P ＜ 0.05 or ＜ 0.01).Conclusions The combination of emodin and 5AzA-cdR can enhance the demethylation effect of 5A6A-cdR on the tumor suppressor genes p16,RASSF1A and ppENK in the tumor tissue of pancreatic cancer xenograft model.

Objective To investigate the effects of 3 different ventilation methods,including conventional mechanical ventilation (CMV),high frequency oscillatory ventilation(HFOV) and partial liquid ventilation (PLV),on the changes of inflammatory factors and pulmonary surfactant associated protein A (SP-A) in bronchoalveolar lavage fluid (BALF) in newborn piglets with acute lung injury(ALI).Methods Twenty-four newborn piglets,no more than 3 days old,were enrolled.After ALI made with saline lavage(38 ℃,35 mL/kg),newborn piglets were randomly assigned to 4 groups:control group (n =6,no ventilation),CMV group(n =6),HFOV group(n =6),and PLV group(n =6).Piglets were sacrificed after being ventilated for 24 h.Tumor necrosis factor-α (TNF-α),interleukin-8 (IL-8),interleukin-1 (IL-1) and SP-A in BALF were measured quantitatively by using enzyme-linked immunosorbent assay.Results In 3 groups using different ventilation methods,the population mean of TNF-o,IL-8,IL-1 and SP-A were statistically different (all P =0.000).SP-A in PLV group and HFOV group were higher than that in CMV group (all P ＜ 0.05),while IL-8,IL-1 and TNF-α in PLV group were lower than those in CMV group (all P ＜ 0.05),IL-8 and TNF-α in PLV group were lower than those in HFOV group (all P ＜ 0.05),IL-8 and TNF-α in HFOV group were lower than those in CMV group (all P ＜ 0.05).Conclusions Pulmonary inflammatory reaction was different in 3 ventilation groups.Compared with CMV and HFOV,PLV attenuated inflammatory reaction,so it could increase the expression of SP-A and decrease the degradation of SP-A.