Objective To investigate the relationship between the expression of glucose regulated protein 78 (GRP78) in the placental trophoblast cells and the pathogenesis of gestational diabetes mellitus (GDM).Methods All the patients were recruited from Qingdao Municipal Hospital from May 2013 to May 2014.Among them, fifty women with GDM were assigned to the GDM group, and fifty healthy women were defined as the control group.All of them received cesarean section because of breech presentation, contracted pelvis, scarred uterus or on mother's demand.Real-time PCR was conducted to analyze the expression of GRP78 mRNA in the trophoblasts.Immunohistochemistry was performed to detect the localization of GRP78 protein in the placentasl trophoblast cells.Results (1) GRP78 mRNA expressed in the cytoplasm of trophoblasts of both the GDM group and the control group.The GRP78 mRNA levels in the GDM group and the control group were 15.6±0.4 and 6.0±0.7, respectively.The relative expression level of GRP78 mRNA in the GDM group was 2.6 times of that in the control group, with statistically significant difference (P＜0.01).(2) The expression of GRP78 protein was found in the cytoplasm of the trophoblasts of the GDM group.It showed in deep, light brown or yellow after staining, according to the expression degree.The expression of GRP78 protein was also found in the cytoplasm of the trophoblasts of the control group, but it mainly showed yellow color (38/50).The strong positive rate of GRP78 protein in the GDM group (96％, 48/50) was higher than that in the control group (22％, 11/50;P＜0.01).Conclusion The expression of GRP78 increased in the placental trophoblast cells of GDM patients.It might suggest that GRP78 had some effect on the pathogenesis of GDM.

It is significantly important but difficult to establish healthcare payment standard system in case our country has cancelled government and government guidance prices. In 1989, Germany was the first country to intro-duce the medicine reference price system in Statutory Health Insurance. It does not only aim to regulate the medicine prices, but also defines a reimbursement level for a cluster of products considered to be therapeutically equivalent, and has a good influence on lowering the price of medicine. This paper systematically reviews the medicine reference price system in Germany, including four aspects:the reference price system overview, the reference price groups’ de-termination procedure, the reference prices calculation and the co-payment mechanisms. Based on the Germany refer-ence price system, we can establish healthcare payment standard in over-the-counter or chronic market first, then sci-entifically divide the reference price groups, formulate and adjust payment standard based on the market price, and perfect the co-payment mechanism to avoid moral hazard.

Objective To investigate the down regulation of phosphatase and tensin homolog deleted on chromosome 10 ( PTEN) gene by adenovirus mediated short hairpin RNA ( shRNA) on the adhesion in activated hepatic stellate cells( HSC ) in vitro and the related signal transduction mechanism .Methods The recombinant adenovirus ( Ad-shRNA/PTEN) with shRNA targeting PTEN and expressing green fluorescent protein ( GFP) were transient trans-fected into the cultural activated HSC in vitro.The experimental group as follows:1)Control group, viral medium was replaced by DMEM at virus transfection step .2 ) Ad-GFP group , HSC were infected with adenovirus expressing GFP alone.3)Ad-shRNA/PTEN group, HSC were infected with adenovirus both taking shRNA targeting PTEN and expressing GFP.PTEN mRNA expression was detected by RT-qPCR, and western blot was used for detecting pro-tein expressions of PTEN , focal adhesion kinase ( FAK) and phosphorylated FAK ( Thr397 ) [ p-FAK( Tyr397 ) ] in HSC.The toluidine blue stain method and MTT colorimetric method were used to determine the adhesion ability of HSC.Results When HSC were infected by adenovirus for 48 hours, PTEN protein and mRNA expressions in Ad-shRNA/PTEN group significantly decreased ( P<0.05 ) , compared to control group and Ad-GFP group, and the expressions of p-FAK ( Tyr397 ) in Ad-shRNA/PTEN group were significantly higher than those in control group and Ad-GFP group ( P<0.05 ) .The adhesion cell counting and the adhesion rate of HSC in Ad-shRNA/PTEN group significantly increased as compared with control group and Ad-GFP group ( P<0.05 ) .Conclusions The down-regulation of PTEN expression can promote the adhesion by increasing the activation of FAK signaling trans -duction in activated HSC in vitro.

Objective: To investigate the role of adenovirus-mediated short hairpin RNA (shRNA) in down-regulating the expression of phosphatase and tensin homologue deleted on chromosome ten (PTEN) on p130Crk-related substrates(p130Cas) and paxillin signal transduction to activate hepatic stellate cell (HSC) in vitro. Methods: The rat hepatic stellate cell line, HSC-T6 was cultured and activated in vitro. The adenovirus was used as a vector to transiently transfect shRNA targeting PTEN to activate HSC in vitro, and then PTEN low expression model of activated HSC in vitro was established. Western blot and real-time fluorescence quantitative PCR were used to detect the protein and mRNA expression of PTEN, p130cas and paxillin in activated HSC. The experiment was divided into control group (HSC were transfected with DMEM medium instead of adenovirus), Ad-GFP group (HSC were infected with empty the adenovirus expressing green fluorescent protein (GFP) alone), and Ad-shRNA/PTEN group (HSC were infected with the recombinant adenovirus containing both shRNA targeting PTEN and GFP gene). One-way analysis of variance was used for comparison of multiple groups, and LSD test was used for inter-group comparison. Results: shRNA targeting PTEN was successfully transfected and significantly down-regulated the PTEN protein and mRNA expression of HSC in vitro (P < 0.05), and the PTEN low expression model of HSC in vitro was successfully constructed. Compared with the expression of p130cas mRNA in the three groups of HSC, the expression fold of p130cas mRNA in the Ad-GFP group and the Ad-shRNA / PTEN group was 1.01 times and 1.52 times, respectively. The expression of p130cas mRNA in HSC of the Ad-shRNA / PTEN group was significantly higher than control group and Ad-GFP group (P < 0.05), but there was no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). The expression of p130cas protein in the three groups was higher than that in the control group (0.74 ± 0.07) and the Ad-GFP group (0.72 ± 0.02); P < 0.05, but there was no statistically significant difference between the Ad-GFP group and the control group (P > 0.05). The expression of paxillin mRNA in the three groups of HSCs was compared with the expression of paxillin mRNA in the control group of HSC being 1, the expression folds of paxillin mRNA in the Ad-GFP group and Ad-shRNA / PTEN group were 0.97 times and 1.58 times, respectively. The expression of paxillin mRNA in the Ad-shRNA / PTEN group was higher than that in the control group and the Ad-GFP group (P < 0.05), and there was no statistically significant difference between the control group and the Ad-GFP group (P > 0.05). The expression of paxillin protein in the three groups of HSCs was higher in the Ad-shRNA / PTEN group (0.91 ± 0.05) than control group (0.46 ± 0.03) and Ad-GFP group (0.50 ± 0.04), P < 0.05, and there was no statistically significant difference between the Ad-GFP group and the control group (P > 0.05). Conclusion Down-regulation of PTEN expression can significantly boost p130cas and signal transduction activity of paxillin protein in activated HSC in vitro.

OBJECTIVE: To investigate the correlation of hemoglobin (Hb) level with prognosis of elderly patients diagnosed as sepsis. METHODS: A retrospective cohort study was conducted. Information on the cases of elderly patients with sepsis in the Medical Information Mart for Intensive Care-IV (MIMIC-IV), including basic information, blood pressure, routine blood test results [the Hb level of a patient was defined as his/her maximum Hb level from 6 hours before admission to intensive care unit (ICU) and 24 hours after admission to ICU], blood biochemical indexes, coagulation function, vital signs, severity score and outcome indicators were extracted. The curves of Hb level vs. 28-day mortality risk were developed by using the restricted cubic spline model based on the Cox regression analysis. The patients were divided into four groups (Hb < 100 g/L, 100 g/L ≤ Hb < 130 g/L, 130 g/L ≤ Hb < 150 g/L, Hb ≥ 150 g/L groups) based on these curves. The outcome indicators of patients in each group were analyzed, and the 28-day Kaplan-Meier survival curve was drawn. Logistic regression model and Cox regression model were used to analyze the relationship between Hb level and 28-day mortality risk in different groups. RESULTS: A total of 7 473 elderly patients with sepsis were included. There was a "U" curve relationship between Hb levels within 24 hours after ICU admission and the risk of 28-day mortality in patients with sepsis. The patients with 100 g/L ≤ Hb < 130 g/L had a lower risk of 28-day mortality. When Hb level was less than 100 g/L, the risk of death decreased gradually with the increase of Hb level. When Hb level was ≥ 130 g/L, the risk of death gradually increased with the increase of Hb level. Multivariate Logistic regression analysis revealed that the mortality risks of patients with Hb < 100 g/L [odds ratio (OR) = 1.44, 95% confidence interval (95%CI) was 1.23-1.70, P < 0.001] and Hb ≥ 150 g/L (OR = 1.77, 95%CI was 1.26-2.49, P = 0.001) increased significantly in the model involving all confounding factors; the mortality risks of patients with 130 g/L ≤ Hb < 150 g/L increased, while the difference was not statistically significant (OR = 1.21, 95%CI was 0.99-1.48, P = 0.057). The multivariate Cox regression analysis suggested that the mortality risks of patients with Hb < 100 g/L [hazard ratio (HR) = 1.27, 95%CI was 1.12-1.44, P < 0.001] and Hb ≥ 150 g/L (HR = 1.49, 95%CI was 1.16-1.93, P = 0.002) increased significantly in the model involving all confounding factors; the mortality risks of patients with 130 g/L ≤ Hb < 150 g/L increased, while the difference was not statistically significant (HR = 1.17, 95%CI was 0.99-1.37, P = 0.053). Kaplan-Meier survival curve showed that the 28-day survival rate of elderly septic patients in 100 g/L ≤ Hb < 130 g/L group was significantly higher than that in Hb < 100 g/L, 130 g/L ≤ Hb < 150 g/L and Hb ≥ 150 g/L groups (85.26% vs. 77.33%, 79.81%, 74.33%; Log-Rank test: χ² = 71.850, P < 0.001). CONCLUSIONS Elderly patients with sepsis exhibited low mortality risk if their 100 g/L ≤ Hb < 130 g/L within 24 hours after admission to ICU, and both higher and lower Hb levels led to increased mortality risks.
Humans ; Male ; Female ; Aged ; Retrospective Studies ; Sepsis/diagnosis* ; Critical Care ; Intensive Care Units ; Prognosis ; Hemoglobins ; ROC Curve