Objective To construct a short hairpin RNA (shRNA) adenovirus vector targeting P85 and protein kinase B1 (PKB1/Akt1) and study its effects on the growth of SGC-7901 human gastric adenocareinoma cells. Methods P85 and Aktl shRNA expression frames were subcloned to pGSadeno adenovirus vector with homologous recombination technology to construct pGSadeno-P85 + Akt1 (rAd5-P + A) vector. After screening and amplification, the recombinant adenovirus vector was digested with PacI and transfected into SGC-7901 cells and then its titer and transfection efficiency were detected with fluorescent microscope. P85 and Akt1 mRNA protein expression was identified with real-time PCR and Western blot. The proliferative activity of tumor cells was evaluated with MTr assay and flow cytometry in vitro, rAd5-HK and rAd5-P + A mediated by adenovirus were injected into the established subcutancous SGC-7901 gastric adenocarcinoma in nude mice. During the observation period of 21 days, tumor volume was measured every 3 days to further testify the anti-tumor effect of rAd5-P + A on the SGC-7901 gastric adenocarcinoma cells and cell in situ apoptosis was detected with TUNEL assay. Results The adenovirus vector rAd5-P + A was successfully constructed and it dramatically downregulated P85 and Akt1 mRNA expression in SGC-7901 gastric adenocarcinoma cells. Compared with a control group of SGC-7901 cells and cells transfected with general adenovirus rAd5-HK as control, P85 and Akt1 protein expression 48 h and 72 h after rAd5-P + A transfection was decreased by 57.5% and 63. 7%, 67. 8% and 75.6% with statistical significance(P = 0. 005, P = 0. 003). Cell proliferative activity in rAd5-P + A transfected cells was suppressed from the second day (P <0. 001) and the decreased P85 and Akt1 expression was accompanied by 5.9% -7. 1% decrease of S phase fraction and 12. 1% - 13.7% increase of G0/G1 phase. The tumor volume of rAd5-P + A treated group was smaller than that of the control and rAd.5-HK group with statistical significance (F = 9. 871, P = 0. 025) . Moreover, rAd5-P + A could induce cell in situ apoptosis. Conclusions Adenovirus-mediated targeting P85 and Akt1 shRNA can inhibit the growth of SGC-7901 human gastric adenocarcinoma cells and this may provide a new strategy of combination gene therapy in gastric adenocarcinoma.

Objective To investigate the anti-inflammatory and analgesic activities of different extracts from X. mongolicum compared with that of X. sibiricum. Methods Seventy male Kunming mices were randomly divided into seven groups:control group,aspirin group,the ethanol extracts of X. sibiricum group and X. mongolicum group,the extracts from X. mongolicum with ether,ethyl acetate,and n-butyl alcohol group(n=10 each). The mice were administrated with 500 mg·kg-1 ( p. o. ) of different extracts. The hot-plate tests and the xylene-induced ear edema in mice were performed to observe the analgesic and anti-inflammatory activities,respectively. Results In the hot-plate tests,the pain threshold from the extracts of X. sibiricum and the different extracts from X. mongolicum at 60,120,180 min were prolonged,and there was no statistically significant differences(P>0. 05) between the ethanol extracts from X. sibiricum and X. mongolicum. The best analgesic activity was the ether and butyl alcohol extracts from X. mongolicum. Meanwile,the ear edema from the extracts of X. sibiricum and the different extracts of X. mongolicum were inhibited significantly(P<0. 05). There was also no significant differences between the ethanol extracts of X. sibiricum and X. mongolicum(P>0. 05). The ether and butyl alcohol extracts from of X. mongolicum showed the best anti-inflammatory effect. Conclusion X. mongolicum exhibites significant anti-inflammatory and analgesic activities comparable to that of X. sibiricum. The best anti-inflammation and analgesic activities were from ether and butyl alcohol extracts from X. mongolicum.

Objective To construct a short hairpin RNA (shRNA) adenovirus vector targeting Aktl (protein kinase B1, PKBI/Aktl) and cyclooxygenase-2 (COX-2) and study its effects on the invasion and metastasis of SGC-7901 human gastric adenocarcinoma cells. Methods Aktl and COX-2 shRNA expression frames were subcloned to pGSadeno adenovirus vector by homologous recombination technology to construct pGSadeno-Aktl+COX-2 (rAdS-A+C) vector. After screening and amplification, the recombinant adenovirus vector was digested with PacI and transfected into SGC-7901 cells, the titer and transfection efficiency were detected by fluorescent microscopy. Aktl and COX-2 mRNA and protein expression was identified by real-time PCR and Western blot. MMP-2 and MMP-9 contents in control group SGC-7901、rAd5-HK and treatment group rAdS-A+C were detected by ELISA assay and transwell assay analyzed cell invasion and metastasis ability. Results Adenovirus vector rAdS-A+C was successfully constructed and it dramatically down-regulated Aktl and COX-2 mRNA and protein expression in SGC-7901 gastric cancer cells. MMP-2 and MMP-9 contents in treatment group rAd5-A+C were respectively (39.7± 1.7) ng/ml, (31.3±3.6) ng/ml, and they were lower than those in control group SGC-7901 (278.4± 15.5) ng/ml, (225.4±15.1) ng/ml and rAd5-HK (275.5±2.1) ng/ml, (226.0±23.3) ng/ml (P= 0.01, P=0.021). Transwell assay showed treatment group rAd5-A+C significantly inhibited the invasion and metastasis of SGC-7901 gastric adenocacinoma cells. Conclusions Adenovirus-mediated targeting Aktl and COX-2 shRNA can inhibit the invasion and metastasis of SGC-7901 human gastric adenocarcinoma cells.

Objective To investigate the effect of electroacupuncture on acupoint local extracellular ionized atom concentrations under physiological status and provide a basis for exploring the mechanism of action of electroacupuncture. Method Twenty male SD rats were selected. Rat point Zusanli (ST36) was given electroacupuncture (1 mA, 0.2 ms and 2 Hz) for 60 min. Meanwhile, local tissue fluid was collected at point Zusanli and non-acupoints using a microdialyzer. The collection by molecular probe membrane sampling lasted 4 hrs: 60 min physiological status before electroacupuncture, 60 min electroacupuncture, 60 min after electroacupuncture and 120 min after electroacupuncture. Real-time analysis of the sample was made by electrolyte analysis to observe local changes in concentrations of Ca﹢﹢, K﹢, Na﹢and Cl- at point Zusanli. Result Local Ca﹢﹢concentrations at point Zusanli increased significantly during electroacupuncture (P=0.003, vs before electroacupuncture), rose gradually afterwards and reached the peak at 60 min after electroacupuncture (P=0.75, vs during electroacupuncture). Ca﹢﹢concentrations decreased at 120 min after electroacupuncture; there was a statistically significant difference comparedwith during electroacupuncture (P=0.04). Acupoint local extracellular concentrations of Na ﹢ and Cl- also increased significantly during electroacupuncture (P<0.001, P=0.007, vs before electroacupuncture) but decreased gradually during 60 min after electroacupuncture and to (71.81±15.09) mmol/L and (57.42±14.30) mmol/L, respectively, at 60 min after electroacupuncture (P=0.09, P=0.07 vs during electroacupuncture). Acupoint extracellular K ﹢concentrations had a tendency similar to those of Na﹢and Cl- but there was no statistically significant difference. Non-point electroacupuncture slightly increased extracellular concentrations of Ca﹢﹢, K﹢, Na﹢and Cl- but there were no statistically significant differences compared with before electroacupuncture (P>0.05). Conclusion Rat point Zusanli electroacupuncture can induce significant increases in acupoint local extracellular concentrations of Ca﹢﹢, K﹢, Na﹢and Cl- . Ionized atom concentrations decrease in different degrees after electroacupuncture. These provide an experimental basis for studying the physiological mechanism of electroacupuncture treatment.

Objective To evaluate the effects of extremity ischemic preconditioning on liver injury after cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Sixty ASA physical status Ⅱ or Ⅲ patients (NYHA Ⅱ or Ⅲ), weighing 44-72 kg, with ejection fraction≥40％, schedule for elective cardiac valve replacement with CPB, were randomly divided into 2 groups (n =30 each) using a random number table: control group (group C) and extremity ischemic preconditioning group (group E).Immediately after anesthesia induction, an auto-inflatable cuff was placed under the right lower extremity, and was inflated to 300 mmHg to induce ischemia (oxygen saturation at the finger tip undetected or foot pulse untouched on the operated side) which was preceded by 3 cycles of 5 min ischemia-5 min reperfusion in group E.Before anesthesia induction (T0) , and at 2, 12, 24, and 72 h, and 7 days after operation (T1-5) , blood samples were taken from the right internal jugular vein for determination of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities.The expression of serum miR-122 was detected at T0-4 by real-time reverse transcriptase polymerase chain reaction.Results Compared with the value at T0, the serum ALT and AST activities were significantly increased at T2-4, and the expression of miR-122 was up-regulated at T1-3 in the two groups (P＜0.05).Compared with group C, the serum ALT and AST activities were significantly decreased at T2-4, and the expression of miR-122 was down-regulated at T1-3in group E (P＜0.05).Conclusion Extremity ischemic preconditioning can mitigate liver injury after cardiac valve replacement with CPB.

Objective: To determine the diagnostic value of serum cystatin C (Cys C) for acute kidney injury (AKI) in patients with liver cirrhosis. Methods: Serum Cys C levels in 150 liver cirrhosis patients (88 AKI and 62 non-AKI patients) were measured by the Particle-Enhanced Nephelometric Immuno-Assay. The accuracy of serum Cys C for the diagnosis of AKI in liver cirrhosis was evaluated by the ROC curve. Results: Liver cirrhosis patients with AKI had significantly higher serum Cys C levels [2.37 (1.75-2.83) mg/L] than those without AKI [0.97 (0.85-1.09) g/L] (P <0.001). Serum Cys C level was highest in the acute tubular necrosis group [5.41 (2.77-6.19) mg/L], followed by the hepatorenal syndrome group [2.55 (2.28-3.59) mg/L] and prerenal azotemia group [2.07 (1.70-2.41) mg/L], and the serum Cys C level was significantly different between the three groups (P <0.001). In addition, patients with AKI were further divided into infection group and non-infection group. Serum Cys C level was significantly higher in the infection group than in the non-infection group (P <0.05). The area under the ROC curve of serum Cys C for the diagnosis of AKI in liver cirrhosis was 0.99 (0.98-1.00) at a cut-off value of 1.36 mg/L, and the sensitivity and specificity were 97% and 95%, respectively. Conclusion Serum Cys C is a good marker for detecting AKI in liver cirrhosis, and the different levels of increase in Cys C may be useful in differentiating the different types of AKI.

Objective To investigate the influencing factors for rebleeding after gastroscopy in patients with liver cirrhosis and esophagogastric variceal bleeding. Methods A retrospective analysis was performed for the clinical data of the patients with liver cirrhosis and esophagogastric variceal bleeding who were hospitalized in Tianjin Third Central Hospital from January 1, 2017 to December 31, 2018, and according to the presence or absence of rebleeding and bleeding time, the patients were divided into non-bleeding group ( n =148) and bleeding group ( n =119). The risk factors for rebleeding after gastroscopy were analyzed. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. The Cox regression model was used for univariate and multivariate analyses. The receiver operating characteristic (ROC) curve was used to evaluate the accuracy of Child-Turcotte-Pugh (CTP), fibrosis-4 (FIB-4), and albumin-bilirubin (ALBI) scores in predicting rebleeding after gastroscopy, and MedCalc was used to compare the area under the ROC curve (AUC). Results A total of 267 patients with liver cirrhosis and esophagogastric variceal bleeding were enrolled, among whom 53 (19.9%) had liver cancer. A total of 119 patients suffered from rebleeding, with an overall rebleeding rate of 44.6% and a median time to rebleeding of 11.0 (0-39.0) months. The univariate Cox regression analysis showed that liver cancer (hazard ratio [ HR ]=0.377, P < 0.001), aspartate aminotransferase (AST) ( HR =1.002, P =0.025), serum Na ( HR =0.935, P =0.004), and FIB-4 ( HR =1.030, P =0.049) were associated with rebleeding, and the multivariate Cox regression analysis showed that liver cancer ( HR =0.357, P < 0.001), AST ( HR =1.003, P =0.030), prothrombin time (PT) ( HR =0.196, P =0.001), CTP score ( HR =1.289, P =0.014), FIB-4 ( HR =1.062, P =0.033), and ALBI score ( HR =0.433, P =0.011) were independent risk factors for rebleeding. CTP, FIB-4, and ALBI scores had an AUC of 0.711 (95% confidence interval [ CI ]: 0.647-0.776), 0.705 (95% CI : 0.640-0.770), and 0.730 (95% CI : 0.667-0.793), respectively, in predicting rebleeding. There was no significant difference in AUC between CTP, FIB-4, and ALBI scores ( P > 0.05). Conclusion Liver cancer, AST, PT, CTP score, FIB-4 score, and ALBI score are associated with rebleeding after gastroscopy in patients with liver cirrhosis and esophagogastric variceal bleeding, among which CTP, FIB-4, and ALBI scores have a good value in predicting rebleeding outcome, while there is no significant difference in predictive ability between them.