[Effects of adenovirus-mediated shRNA down-regulates PTEN expression on fibril-binding proteins vinculin, filamin A and cortactin in activated hepatic stellate cells].

Objective: To investigate the effect of adenovirus-mediated shRNA down-regulating phosphatase and tensin homolog deleted on chromosome 10 (PTEN) expression on vinculin, filamin A, and cortactin in activated hepatic stellate cells (HSCs). Methods: Activated rats hepatic stellate cell line (HSC-T6) was cultured in vitro. Recombinant adenovirus Ad-shRNA/PTEN carrying PTEN targeted RNA interference sequence [short hairpin RNA (shRNA)] and empty control virus Ad-GFP were transfected into HSCs. The PTEN mRNA and protein expression of HSCs in each group were detected by real-time fluorescence quantitative PCR and Western blot. The expressional change of vinculin, filamin A and cortactin in HSCs of each group were detected by confocal laser scanning immunofluorescence microscope. Image-pro plus 6.0 software was used for image analysis and processing. The integrated optical density (IOD) of the fluorescence protein expression was measured. The experiment was divided into three groups: control group (DMEM instead of adenovirus solution in the adenovirus transfection step), Ad-GFP group (transfected with empty virus Ad-GFP only expressing green fluorescent protein), and Ad-shRNA/PTEN group (recombinant adenovirus Ad-shRNA/PTEN carrying shRNA targeting PTEN and expressing green fluorescent protein). One-way analysis of variance was used for comparison of mean value among the three groups, and LSD-test was used for comparison between the groups. Results: shRNA targeted PTEN was successfully transfected and the expression of PTEN mRNA and protein in HSC (P < 0.05) was significantly down-regulated. HSCs vinculin was mainly expressed in the cytoplasm. HSCs vinculin fluorescence IOD in the Ad-shRNA/PTEN group (19 758.83 ± 1 520.60) was higher than control (7 737.16 ± 279.93) and Ad-GFP group (7 725.50 ± 373.03) (P < 0.05), but there was no statistically significant difference between control group and Ad-GFP group (P > 0.05). There was no statistically significant difference in the fluorescence IOD of Filamin A among the three groups (P > 0.05), but the subcellular distribution of Filamin A among the three groups were changed. Filamin A in the Ad-shrNA /PTEN HSC group was mainly distributed in the cytoplasm. Filamin A HSC was mainly located in the nucleus.The filamin A HSC in the control group and Ad-GFP group was mainly located in the nucleus. The nucleocytoplasmic ratio of Filamin A in the AD-shrNA /PTEN group (0.60 ± 0.15) was significantly lower than control group (1.20 ± 0.15) and Ad-GFP group (1.08 ± 0.23), P < 0.05. but there was no statistically significant difference in filamin A nucleocytoplasmic ratio of HSC between the control group and the Ad-GFP group (P > 0.05). Cortactin HSCs in the three groups was mainly distributed in the cytoplasm. The cortactin fluorescence IOD of HSCs in the Ad-shRNA/PTEN group was significantly higher than control group (22 959.94 ± 1 710.42) and the Ad-GFP group (22 547.11 ± 1 588.72 ) (P < 0.05), while there was no statistically significant difference in the IOD of cortactin fluorescence in HSCs between the control group and the Ad-GFP group (P > 0.05). Conclusion: The down-regulation of PTEN expression raises the expression of microfilament-binding protein vinculin and cortactin, and changes the subcellular distribution of another microfilament binding protein filamin A, that is, translocation from nucleus to the cytoplasm in activated HSC in vitro.

[Efficacy of minimally invasive pulmonary surfactant administration in preterm infants with neonatal respiratory distress syndrome: a multicenter clinical trial].

Objective: To explore the feasibility and safety of minimally invasive surfactant administration (MISA) in preterm neonates with respiratory distress syndrome (NRDS). Methods: In this multicenter prospective randomized controlled trial, 92 preterm infants with gestation age ≤30 weeks and diagnosed with NRDS were enrolled in 8 level Ⅲ neonatal intensive care units (NICU) in Beijing-Tianjin-Hebei Region from 1(st) July 2017 to 31(st) December 2018. They were randomly assigned to minimally invasive surfactant administration (MISA) group or endotracheal intubation surfactant administration (EISA) group according to random number generated by computer. Infants in both groups received calf pulmonary surfactant preparation at a dose of 70-100 mg/kg. The data of demography, perinatal situation, medication administration, complications, clinical outcomes in the two groups were compared with Chi-square test, Student's t-test, Mann-Whitney U test or Fisher's exact test. Results: Among the 92 preterm infants, 53 were males, 39 were females; 47 were in the MISA group (25 males), and 45 were in the EISA group (28 males). The gestational age and birth weight were (29.5±1.2) weeks and (1 271±242) g in all patients, (29.5±1.4) weeks and (1 285±256) g in the MISA group, and (29.6±0.9) weeks and (1 255±227) g in the EISA group. The duration of surfactant infusion and the length of whole procedure in the MISA group were significantly longer than that in the EISA group (60 (18, 270) s vs. 50 (30, 60) s, Z=3.009, P=0.003; 90 (60, 300) s vs. 60 (44, 270) s, Z=3.365, P=0.001). For the outcomes, the incidence of hemodynamically significant patent ductus arteriosus (hsPDA) and bronchopulmonary dysplasia (BPD) were lower in the MISA group than in the EISA group (36% (17/47) vs. 67% (30/45), χ(2)=8.556, P=0.003; 26% (12/47) vs. 47% (21/45), χ(2)=4.464, P=0.035). Conclusions: Minimally invasive surfactant administration is applicable in preterm infants ≤30 weeks gestational age with NRDS. Although the length of whole procedure is longer than route endotracheal administration, the benefit of decreasing the incidences of hsPDA and BPD outweighs this demerit.

Analysis of Driver-Passenger Relationship and Restoration of Accident Process Based on 3D Laser Scanning Technology.

ABSTRACT: Objective To discuss the application of 3D laser scanner and computer technology in restoration of the accident scene and reconstruction of the accident process, as well as identification of the driver-passenger relationship. Methods The scene of a traffic accident, the accident vehicle and the vehicle of the same type as accident vehicle were scanned using 3D laser scanner. The accident scene, traces and accident vehicle were integrated using computer technology to restore the accident scene, and the accident process was reconstructed and analyzed by combining the characteristics of the body injuries. Results By restoring the accident scene and reconstructing the accident process with 3D laser scanner, it was determined that Wu was in the driving seat at the time of the accident. Conclusion It is more objective and scientific to use 3D laser scanning technology to restore the accident scene, reconstruct the accident process and analyze the moving track of the driver and passengers in the vehicle. It will help to improve the accuracy of forensic identification of road traffic accidents.

Comparison of a central location test versus a home usage test for consumer perception of ready-to-mix protein beverages.

Ready-to-mix (RTM) whey protein beverages are an expanding product category, and sensory properties strongly affect consumer acceptance and purchase intent. Because consumers themselves prepare RTM whey protein beverages, understanding possible gaps between central location test (CLT) and home usage test (HUT) results is critical. The objectives of this study were to compare results obtained from a CLT and a HUT and to identify the drivers of liking and disliking vanilla-flavored RTM whey protein beverages. Fourteen commercial vanilla-flavored RTM whey protein beverages were rehydrated with spring water at 15% solids (wt/vol) and evaluated by a trained panel (n = 8). Ten representative products were selected for consumer testing. Rehydrated beverages were subsequently evaluated by protein beverage consumers (n = 160) in a CLT. Nine representative products were selected for the HUT. Consumers prepared and evaluated individual beverages over 3 consecutive weeks, trying 3 samples each week. Overall liking and other attributes were scored by consumers in both tests. Data were evaluated by univariate and multivariate statistical analyses. Overall liking scores from the HUT were higher than scores from the CLT. The products with the highest and lowest overall liking scores were consistent between the CLT and HUT. More differences were observed among beverages by CLT compared with HUT when liking was averaged across all consumers. Both methods identified 2 distinct consumer clusters. Fruity flavor and sweet taste were drivers of liking, whereas cardboard flavor and bitter taste were drivers of disliking in both methods. The HUT exclusively identified thickness (viscosity) as a driver of liking and astringency as a driver of disliking. These results suggest that a CLT can be used to differentiate consumer acceptance among vanilla-flavored RTM whey protein beverages. A HUT should be used to provide more intensive insights for mouthfeel and mixing experience-related attributes.

[Effect of downregulation of phosphatase and tensin homolog gene expression on p130crk- related substrate protein and paxillin signal transduction in activated hepatic stellate cells 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.

[A pilot study to explore a registry method for acute cardiovascular events].

Objective: To explore a registry method for acute cardiovascular events in large-scale fields across China. Methods: This study was based on a community-based Kazakh cohort, including 1 668 persons with complete baseline data, conducted in Altay, Xinjiang. Acute cardiovascular events included acute myocardial infarction(ICD-10: I21-I22), sudden cardiac death (I46.1), and incident stroke (I60-I64) that occurred during the period from Oct 1, 2012 to Jun 30, 2016. There were three stages, a hospital-based search for checking medical records of inpatients in all four local hospitals, a community-based supplementary registry, and an extra survey for reducing false negatives, to complete this registry. The incidence referred to all events within one year per 100 000 people and mortality rate was the number of fatal events within 28 days per 100 000 people. The incidence and mortality was standardized with the weights from distribution of age and gender in the Sixth National Census of China. Results: Forty two cases with 46 acute cardiovascular events including 7 acute myocardial infarctions, 3 sudden cardiac deaths, and 36 incident strokes were found. The ratio of events was 3.6∶1 for stroke and heart attacks and this ratio was 1.6∶1 for ischemic or hemorrhagic strokes. A total of 28(60.9%) events and 18(39.1%) events were collected in the hospitals and by community-based supplementary registry physicians, respectively. False negatives were not found at the last stage. The crude and standardized incidence rates were 742 and 926 per 100 000 people and the related mortality rates were 194 and 272 per 100 000 people, respectively. Conclusion: Primary Care physicians are"net bottom" of a monitoring system which needs to combine preventive and clinical medicine with all kinds of governmental resources including health and social benefit.

[Clinical and experimental studies on effects of chronic hepatitis B treated with astragali composita].

Eighty-seven cases of chronic persistent hepatitis (CPH) and 29 cases of chronic active hepatitis (CAH) were treated with Astragali Composita (AC), the clinical markedly effective rates were 69.0% and 55.2%, the total effective rates were 90.8% and 89.7% respectively; the sero-negative conversion rates of HBeAg and HBV-DNA were 27.7% and 28.0% respectively, they were higher than that of control group (P < 0.05-0.01). Eight ducks infected hepatitis B virus were treated with AC, sero-negative conversion of DHBV-DNA was seen in 3 ducks, DHBV-DNA levels of duck sera were lower after treatment than that before treatment (P < 0.05); mild inflammatory changes in liver tissues were seen in 3 ducks, its pathologic changes were milder than that of control groups; DHBV-DNA in liver tissues of 2 ducks was negative in situhybridization. The results indicated that AC had obvious effects of protecting liver tissues and some effects of inhibiting sera HBV and HBV replication.

Five-year follow-up of immune response to hepatitis B vaccine in juveniles.

The persistence of anti-HBs was investigated in 95 juveniles who received plasma-derived HBV vaccine (vaccine group) and 63 counterparts who got anti-HBs from natural HBV infection (infection group) for a period of five years. The positive rates of anti-HBs from the first to fifth year in the vaccine group are 97.89%, 98.95%, 81.05%, 78.95% and 72.63% respectively with one recipient remaining anti-HBs negative after being immunized with three dosages of 10 micrograms plasma-derived HBV vaccine in 0, 1st, 2nd month and the mean S/N values (GMV) are 30.94, 22.18, 13.61, 12.02 and 9.18 respectively. There are 26 recipients whose anti-HBs turned from positive to negative at the end of the study with a negative rate of 27.37%. The S/N GMVs in the infection group are 36.37, 27.33, 24.08 at the first, third and fifth year of the study, respectively. Both the S/N GMV and negative rate are lower than that of the vaccine group (P < 0.01). No one was found to have positive HBsAg or elevated ALT in both groups. Questions such as immune dosage, immune program and booster immunization in juvenile population are discussed.

[The immune response and its dynamic changes of anti-HBs in 98 juveniles vaccinated with native hepatitis B vaccine].

The immune response and its dynamic changes of anti-HBs in 98 Juveniles vaccinated with three doses of native plasma derived HB vaccine (have been were studied and) followed up for three years. The results showed that the cumulative positive rate and its mean S/N values of anti-HBs at T1 were 70.41% and 19.89 respectively, at T3, T6, T12, T24 all 98% and 65.54, 41.75, 53.95, 23.79 respectively. However out of 98 mentioned recipients 18 subjects become negative for anti-HBs at T36.