ABSTRACT
Objective To observe the clinical value of indocyanine green retention rate at 15 minutes (ICGR15) in the evaluation of hepatic functional reserve. Methods A total of 185 patients with liver disease, including 45 cases of liver failure, 90 cases of cirrhosis (child A, B and C, respectively), 20 cases of acute hepatitis, 30 cases of chronic hepatitis (mild, moderate). Expression levels of ICGR15 were compared between groups. Values of ICGR15, total bilirubin (TBIL), albumin (ALB), blood coagulation time (PT) were compared before treatment and one month after treatment in hepatic failure group. Levels of alanine aminotransferase (ALT), aspertate aminotransferase (AST), TBIL and ICGR15 were compared before treatment and 1 month after treatment in acute hepatitis group. Results Levels of ICGR15 (%) were 56.3±14.7, 28.9±9.6, 22.4±6.8 and 13.7±2.3 in liver failure group, liver cirrhosis group, acute hepatitis group and chronic hepatitis group, which showed a gradual downward trend (F=125.317, P<0.05). Among them, the levels of ICGR15 (%) were 17.3±5.4, 25.7±7.5 and 34.5±7.3 in Child A, B and C groups of liver cirrhosis group, which showed a gradual upward trend (P<0.05). After one month treatment, levels of TBIL, PT and ICGR15 were significantly lower than T helper 17 cells; intima-media thickness before the treatment in liver failure group. The levels of ALT, AST, TBIL and ICGR15 were significantly lower after treatment than those before treatment in acute hepatitis group (P<0.05). Conclusion ICGR15 can reflect hepatic reserved function, which is not affected by the application of albumin and fresh plasma, and makes up the deficiency of PT and ALB detection.
ABSTRACT
Objective To investigate the effects of silencing of the human cerebral 21 .5 kDa myelin basic protein (MBP) gene on the proliferation and apoptosis of the glioma U251 cells .Methods The 21 .5 kDa MBP sequence‐specific short hair‐pin RNA (shR‐NA) recombinant plasmids pGenesil‐1‐MBP‐3 were transfected into the human glioma cell line(U251) ,the cells of U251 was used as MBP silencing group ,the cells transfected with negative control plasmids used as negative control group ,and the cells transfected with liposomes used as blank control group .Real‐time PCR and Westernblot were used to detect the expression levels of the 21 .5 kDa MBP mRNA and protein in each group ,and the cell proliferation curve was measured by CCK‐8 assay ,the apoptosis rate was a‐nalysised by Flow cytometry .Results Both the mRNA and the protein expression levels of the 21 .5 kDa MBP of MBP silencing group were significantly lower than those in the control groups (P<0 .05);the cellular proliferation activity of the MBP silencing group decreased significantly (P<0 .05)while the cellular apoptotic rate increased significantly (P<0 .05) .Conclusion Silencing of the human cerebral 21 .5 kDa MBP gene may playa dual role in the inhibition of proliferation and the promotion of apoptosis of the glioma U251 cells .
ABSTRACT
Objective To construct luciferase reporter vector containing full-length high-mobility group box 1 ( HMGB1, GenBank NM-010439) promoter for the screening of medicine. Methods The full-length HMGB1 promoter was amplified by polymerase chain reaction ( PCR) , and then was inserted into GV238 vector to construct plasmid GV238-HMGB1-P-Luc. GV238-HMGB1-P-Luc combined with internal reference plasmid pRL was co-transfected into Hela cells ( GV238-HMGB1-P-Luc group, which served as positive control group) . Plasmid pGL3-basic combined with pRL was co-transfected into Hela cells (pGL3-basic group, which served as negative control group) . Additionally, lipopolysaccharides ( LPS, 0.2 μg/mL) was used as the activator for the positive control group (LPS group), and then sodium butyrate (SB, 10 mmol/L) was used as the inhibitor for LPS group ( SB group) . At the end of experiment hour 24, luciferase activity was detected. Results The results of digestion, amplification, sequencing and identification showed that the full length of HMGB1 promoter was 2 140 bp, and the DNA sequence was correct, without mutation. Luciferase activity in GV238-HMGB1-P-Luc group was increased as compared with that of the pGL3-basic group ( P<0.05) . Luciferase activity in the LPS group was increased ( P<0.01, compared with that of GV238-HMGB1-P-Luc group) , and then was decreased after the administration of SB ( P<0.01, compared with that of the LPS group) . Conclusion A model of luciferase reporter vector containing HMGB1 promoter has been successfully constructed. Its activity can be increased by LPS, and then is in hibited by SB. The model can be used for further screening of medicine with the activities of regulating HMGB1 promoter.