1.Development and clinical application of a Sandwich ELISA for hK6 detection
Hua ZHANG ; Chengjin HU ; Kuixiang CHEN
International Journal of Laboratory Medicine 2017;38(17):2337-2338,2341
Objective To prepare the mAbs against hK6 for establishing a sandwich enzyme-linked immunosorbent assay(ELISA) of hK6,and exploring its clinical value.Methods The hybridoma technique was used to prepare mAbs against hK6.The mAbs were purified and labelled with horseradish peroxidase for the sandwich ELISA method.The sandwich ELISA method was used to detect the serum hK6 concentrations in patients with malignant gastric neoplasm.Then the best antibody pair was selected from coating antibody and enzyme-linked antibody to establish a sandwich ELISA method through the chessboard titrations.Compared with CEA,we explored the feasibility of hK6 as gastric cancer biomarkers.Results A sandwich ELISA method was established for quantifying hK6 in serum.The results showed that the optimal concentration of coating antibody was 5 μg/mL.The optimal concentration of enzyme-linked antibody was 1:2 000.Serum hK6 in the patients with gastric cancer groups[(5.78±1.66)ng/mL] than healthy individuals groups[(3.35±0.67)ng/mL] and those in gastric ulcer groups[(3.59±1.02)ng/mL],the difference was statistically significant(P<0.05).Furthermore,there was no significant difference in values of serum hK6 between patients with gastric ulcer groups and healthy individuals groups(P>0.05).The hK6 positive rate of gastric cancer was 69.70%,and CEA was 45.46%.In the combined detection,the positive rate was 78.79%.Conclusion A sandwich ELISA is established successfully.As a favorable serum biomarker for gastric cancer,the detection of hK6 together with CEA is helpful in the diagnosis of gastric cancer.
2.Quercetin attenuates cardiomyocyte hypertrophy through proteasome in-hibition
Kuixiang CHEN ; Sujuan LI ; Jiandong LUO ; Yinghua LIU
Chinese Journal of Pathophysiology 2015;(8):1352-1359
AIM:ToinvestigatetheprotectiveeffectofquercetinonangiotensinⅡ(AngⅡ)-inducedcardio-myocyte hypertrophy and its possible mechanism .METHODS: Cardiomyocyte hypertrophy was induced by AngⅡ ( 100 nmol/L) in primary neonatal cardiomyocytes and H 9c2 cells.The cells were treated with different concentration of querce-tin (10 μmol/L, 20 μmol/L and 40 μmol/L) for 48 h and then the cardiomyocyte surface areas were measured by immu-nofluorescence .Proteasome activity was detected by fluorescent peptide substrate .The phosphorylated levels of GSK-3α/βand Akt in H9c2 cells were determined by Western blot .RESULTS:Compared with control group , the cardiomyocyte sur-face areas were both increased in primary cultured neonatal cardiomyocytes and H 9c2 cells, while the surface areas were significantly decreased by quercetin , especially at concentration of 20 μmol/L compared with Ang Ⅱgroup (P<0.05). Compared with control group , the chymotrypsin-like, trypsin-like and caspase-like activities of proteasome were all in-creased in H9c2 cells (P<0.05).The trypsin-like and caspase-like activities of proteasome were inhibited by 20 μmol/L and 40 μmol/L quercetin , while chymotrypsin-like activity was inhibited only at 20 μmol/L of quercetin compared with AngⅡgroup (P<0.05).In addition, phosphorylated levels of GSK-3α-Ser21, GSK-3β-Ser9 and Akt-Ser473 in AngⅡgroup were all increased compared with control group , which were obviously inhibited by in 20 μmol/L and 40 μmol/L quercetin ( P<0.05 ) .CONCLUSION: Quercetin decreases cardiomyocyte hypertrophy through proteasome inhibition , which may be related to the inhibition of Akt and therefore increasing activation of GSK -3α/βin H9c2 cells.
3.Experimental studies of Huoluojiangu capsule on anti-inflammatory efficiency
Jiamao ZHOU ; Fengxiang YAN ; Jianshe QUAN ; Ji ZHOU ; Kuixiang LI ; Xiongxin CHEN ; Ping YU ; Lanxiu NIE ; Shizhu YIN ; Yixi CHEN ; Lan XIE
Journal of Chinese Physician 2001;0(05):-
Objective To observe the effect of Huoluojiangu Capsule(HJC) on anti-inflammatory experiment.Methods The anti-inflammatory experimental models were established in three kinds of rats with pleuritis,the swellen sole of the foot and the PGE_2 content in inflammatory tissues,respectively.Results The swollen reaction in the sole of foot in rats induced by egg white was lessoned with the treatment of low,middle and high dosage of HJC.Compared with the blank control group,the treatment is effective in low dosage group and evidently effective in groups of middle and high dosage after 30 min(P0.05),respectively.Compared with the blank control group,the effect of HJC on the PGE_2 content in the inflammatory tissues of middle and high dosage groups was significant(P