1.Effect of all-trans retinoic acid on expression of gap junction genes of hepatocellular carcinoma cell lines
Bin XU ; Jianwei WANG ; Heqing FANG ; Yingbin LIU ; Yiping MU ; Xiujun CAI ; Jiangtao LI ; Shuyou PENG
Chinese Journal of General Surgery 1993;0(03):-
Objective To investigate the effect of all-trans retinoic acid (ATRA) on expression of gap junction genes 26, 32 and 43 in two human hepatocellular carcinoma cell lines. Method Human hepatocellular carcinoma cell lines SMMC-7721 and BEL-7404 were cultured and inoculated in culture plate. Cells were divided into 2 groups and cultured respectively with normal medium and medium contains ATRA at a concentration of 10-mol/L. After 24, 48 and 72 hours, cells were collected and total mRNA were extracted. RT-PCR was used to detect the transcription of gap junction genes 26, 32 and 43. Result SMMC-7721 and BEL-7404 cultured under condition of normal medium do not express CX26 mRNA and CX32 mRNA. With the effect of ATRA for 48 and 72 hours, SMMC-7721 cell expressed CX26 mRNA and CX32 mRNA. BEL-7404 expressed CX26 mRNA and CX32 mRNA while cultured with ATRA for 72 hours. CX43 mRNA expression of the two cell lines was not altered by ATRA. Conclusion ATRA affects the expression of CX26 and CX32 in HCC cell lines SMMC-7721 and BEL-7404 at the transcription level.
2.Raman spectroscopic study on effect of danshen injection on human erythrocyte membranes and its mechanism.
Heqing CAI ; Yanping ZHAO ; Welling ZHU ; Chao YU ; Shu GE ; Songhao LIU
China Journal of Chinese Materia Medica 2012;37(10):1467-1471
OBJECTIVEDanshen, as a traditional Chinese medicine for promoting blood circulation and removing blood stasis, is widely applied in improving human erythrocyte deformability in clinics. But its direct effect on erythrocyte membranes is still unclear.
METHODIn this essay, the confocal Raman technique was adopted to measure the changes in Raman spectra of human erythrocytes before and after the administration of Danshen injection.
RESULTThe results showed slight changes in group conformations corresponding to erythrocyte membranes after the administration of danshen injection. Specifically, 1064, 1126 cm(-1) spectral lines attributed to phospholipid molecule acyl C-C skeleton anti-conformation were obviously weakened, whereas 1091 cm(-1) spectral line attributed to phospholipid molecule acyl C-C skeleton guache conformation notably intensified. Besides, the longitudinal order-parameter in chains (Strans) of phospholipids was reduced significantly.
CONCLUSIONDanshen injection can transfer erythrocyte membrane phospholipid molecule acyl C-C skeleton anti-conformation to guache conformation, indicating the increase in liquidity of erythrocyte membrane phospholipids and erythrocytes. Danshen injection's effect in improving erythrocyte membrane structure and function may be the intrinsic mechanism for its pharmacological effect as a traditional Chinese medicine.
Adult ; Blood Proteins ; chemistry ; Erythrocyte Membrane ; chemistry ; drug effects ; Female ; Humans ; Injections ; Male ; Molecular Conformation ; Phospholipids ; chemistry ; Salvia miltiorrhiza ; Spectrum Analysis, Raman ; methods ; Young Adult
3.The clinical features and surgical complications of elderly gastric carcinoma cases
Yingbin LIU ; Weilong CAI ; Yong WANG ; Xuan WANG ; Xiaoming MA ; Ying KONG ; Yan CHEN ; Kr DEVKOTA ; Jianwei WANG ; Jiangtao LI ; Zhe TANG ; Heqing FANG ; Shuyou PENG
Chinese Journal of General Surgery 2008;23(10):777-780
Objective To study the clinical features and postoperative complications for gastric carcinoma in the patients older than 65 years. Methods From January 2005 to January 2007, 65 elderly gastric carcinoma cases underwent surgical resection, the age related preoperative and postoperative complications were analyzed. Result The incidence of preoperative complications in elderly gastriccarc inoma patients is 83%, compared with younger group of 59%; Preoperatively, 52% elderly cases presented with two or more complications. The incidence of hypertension was the highest (40%), the radical curative rate for the elderly group was 86% as compared with younger group of 93%. The postoperative complications in elderly group was 37%. In elderly cases, postoperative complications were common in those suffering from preoperative complications such as hypertension, diabetes, respiratory diseases, anaemia, hypoalbuminemia. Conclusion The age itself is not a single influential factors indicating postoperative complications. There was no significant difference in the morbidity and mortality between elderly and younger gastric carcinoma cases undergoing a surgery.
4.Propagation and phenotypic analysis of mutant rabbits with MSTN homozygous mutation.
Liqing SHANG ; Shaozheng SONG ; Ting ZHANG ; Kunning YAN ; Heqing CAI ; Yuguo YUAN ; Yong CHENG
Chinese Journal of Biotechnology 2022;38(5):1847-1858
Myostatin gene (MSTN) encodes a negative regulator for controlling skeletal muscle growth in animals. In this study, MSTN-/- homozygous mutants with "double muscle" phenotypic traits and stable inheritance were bred on the basis of MSTN gene editing rabbits, with the aim to establish a method for breeding homozygous progeny from primary MSTN biallelic mutant rabbits. MSTN-/- primary mutant rabbits were generated by CRISPR/Cas9 gene editing technology. The primary mutant rabbits were mated with wild type rabbits to produce F1 rabbits, whereas the F2 generation homozygous rabbits were bred by half-sibling mating or backcrossing with F1 generation rabbits of the same mutant strain. Sequence analysis of PCR products and its T vector cloning were used to screen homozygous rabbits. The MSTN mutant rabbits with 14-19 week-old were weighed and the difference of gluteus maximus tissue sections and muscle fiber cross-sectional area were calculated and analyzed. Five primary rabbits with MSTN gene mutation were obtained, among which three were used for homozygous breeding. A total of 15 homozygous rabbits (5 types of mutants) were obtained (M2-a: 3; M2-b: 2; M3-a: 2; M7-a: 6; M7-b: 2). The body weight of MSTN-/- homozygous mutant rabbits aged 14-19 weeks were significantly higher than that of MSTN+/+ wild-type rabbits of the same age ((2 718±120) g vs. (1 969±53) g, P < 0.01, a 38.0% increase). The mean cross sections of gluteus maximus muscle fiber in homozygous mutant rabbits were not only significantly higher than that of wild type rabbits ((3 512.2±439.2) μm2 vs. (1 274.8±327.3) μm2, P < 0.01), but also significantly higher than that of MSTN+/- hemizygous rabbits ((3 512.2±439.2) μm2 vs. (2 610.4±604.4) μm2, P < 0.05). In summary, five homozygous mutants rabbits of MSTN-/- gene were successfully bred, which showed a clear lean phenotype. The results showed that the primary breeds were non-chimeric mutant rabbits, and the mutant traits could be inherited from the offspring. MSTN-/- homozygous mutant rabbits of F2 generation could be obtained from F1 hemizygous rabbits by inbreeding or backcrossing. The progenies of the primary biallelic mutant rabbits were separated into two single-allelic mutants, both of which showed a "double-muscle" phenotype. Thus, this study has made progress in breeding high-quality livestock breeds with gene editing technology.
Animals
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CRISPR-Cas Systems/genetics*
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Gene Editing
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Muscle, Skeletal/metabolism*
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Mutation
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Myostatin/metabolism*
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Phenotype
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Rabbits