1.The role of pituitary adenylate cyclase-activating polypeptide in the growth modulation of human pancreas carcinoma
Min ZHAO ; Ruolan JIANG ; Lumei SUN
Chinese Journal of Digestion 2001;21(2):86-89
Objective To investigate the role pituitary adenylate cyclase-activating polypeptide (PACAP) in the growth modulation of PACAP of human pancreas carcinoma cells and determine whether sphingomyolin (SM) may act as a second messenger involved in the postreceptor signal transduction. Methods Human pancreas carcinoma cell strains, JF305, HS766T and ASPC-1 cells were cultivated, reproduced and then treated with PACAP1-38 (10- 12 - 10- 6 M). The amounts of proliferated carcinoma cells were estiimated with Mosmann's method (MTT). The concentrations of intracellular SM in cells were determined with thin layer chromotograph. Intracellular adenosine monophosphate and Ca2 + levels were detected by radioimmunoassay and Fura-2/AM respectively. ResultsIt was found that three kind of human pancreatic cancer cells were proliferated and the intracellular levels of SM, cAMP and cytosolic Ca2+ were increased by treating PACAP1-38. The effect of PACAP1-38 in JF305, HS766T and ASPC-1 could be inhibited by Somatostatin.ConclusionPACAP1-38 may play a role in the proliferation of human pancreatic cancer cells. The postreceptorsignal transduction of PACAP may be mediated by both adenosine cyclinase and Calcium-calmodin pathways. SM may be a second messenger involved in this process.
2.ERS response mediates glucolipotoxicity-induced β-cell apoptosis
Yanli YANG ; Ruolan XIANG ; Qi SUN
Basic & Clinical Medicine 2009;29(12):1337-1340
As one of the most sensitive cells of endoplasmic retieulum stress (ERS), pancreatic β-cells have an a-bundance of endoplasmic reticulum. Fatty acids cause apoptosis of β-cells and might contribute to β-cell loss in type 2 diabetes mellitus via the induction of ERS. Glucose is an amplifier of the ERS response to fatty acid, leading to increased β-cell apoptosis. ERS response mediates glucolipotoxicity-induced β-cell apoptosis.
3.The role of pituitary adenylate cyclase activating polypeptide in the growth modulation of human pancreas carcinoma
Min ZHAO ; Lumei SUN ; Ruolan JIANG
Chinese Journal of Digestion 2001;0(02):-
Objective To investigate the role pituitary adenylate cyclase activating polypeptide (PACAP) in the growth modulation of PACAP of human pancreas carcinoma cells and determine whether sphingomyolin (SM) may act as a second messenger involved in the postreceptor signal transduction. Methods Human pancreas carcinoma cell strains, JF305, HS766T and ASPC 1 cells were cultivated, reproduced and then treated with PACAP 1 38 (10 -12 -10 -6 M). The amounts of proliferated carcinoma cells were estiimated with Mosmann's method (MTT). The concentrations of intracellular SM in cells were determined with thin layer chromotograph. Intracellular adenosine monophosphate and Ca 2+ levels were detected by radioimmunoassay and Fura 2/AM respectively. Results It was found that three kind of human pancreatic cancer cells were proliferated and the intracellular levels of SM, cAMP and cytosolic Ca 2+ were increased by treating PACAP 1 38 . The effect of PACAP 1 38 in JF305, HS766T and ASPC 1 could be inhibited by Somatostatin. Conclusion PACAP 1 38 may play a role in the proliferation of human pancreatic cancer cells. The postreceptorsignal transduction of PACAP may be mediated by both adenosine cyclinase and Calcium calmodin pathways. SM may be a second messenger involved in this process.
6.Construction and identification of eukaryotic expression vector expressing Islet-brain 1 gene
Qi SUN ; Ruolan XIANG ; Yuxiu LI ; Heng WANG
Basic & Clinical Medicine 2006;0(07):-
Objective To construct and to identify eukaryotic expression vector expressing Islet-brain 1(IB1) gene.Methods Total RNA was extracted from human insulinoma.IB1 gene was amplified by PCR from human IB1cDNA library.The eukaryotic expression vector encoding IB1 was constructed by inserting the IB1 cDNA into EcoR I/Kpn I sites of the pEGFP-N1 vector with the green fluorescent.The construct was transfected into RINm5F cell line,screened by G418.The phase contrast fluorescence microscope,flow cytometer,and Western blot were used to identify the recombinant plasmid and transfeced cell line.Results The RT-PCR products for IB1(AA1-280)generated from human insulinoma was 840 bp.Sequence analysis proved the same sequence as published in Gen-Bank.Two bands showed that pEGFP-N1 vector encoding IB1 digested by EcoR I or Kpn I.Western blot showed IB1 gene was expressed in RINm5F cells.Conclusion The recombinant prokaryotic expression plasmid pEGFP-N1-IB1 has been successfully constructed.
7.The Growth Regulation and Postreceptor Signal Transduction of Somatostatin on Cell Line of Human Colon Cancer
Mingjun SUN ; Ying PANG ; Ruolan JIANG ; Baoyu FU ; Yingjei LIU
Journal of China Medical University 2001;30(1):33-35
Objective:Our purpose was to investigate the growth regulation and postreceptor signal transduction of somatostatin (SS) on human colon cancer cell line. Methods:Low differentiated clone A cells of human carcinoma were treated with different concentrations of SS and the growth status was observed. Intracellular 1,4,5-trisphosphate (IP3) and cyclic adenosine monophosphate (cAMP) were extracted, then the concentrations of them were measured by liquid scintillation technique and gamma scintillation counter.Intracellular free calcium concentration was detected by loading Fura-2 and fluorescental technique. Protein kinase C (PKC) was extracted from cytosol and membrane, then the activity of both parts was determined with TaKai method. Results:The clone A cell growth was inhibited greatly by different concentrations of SS (10-10 to 10-5 mol/L) and it is related to the doses of SS.Somatortatin could largely inhibit the production of intracellular IP3, [Ca2+]i, and cAMP, and decrease the activity of PKC. Conclusion:The growth of Clone A cell can be inhibited by SS. The inhibition may be mediated by phosphate inositol pathway, so intracellular IP3, [Ca2+]i decreased, or inhibited the cell growth by inhibiting the activity of PKC.On the other hand, the cell proliferation may be inhibited by adenosine cyclase pathway, that is decreasing intracellular cAMP ,inhibiting cAMP-depending protein kinase.
8.STUDY ON THE PROTECTIVE EFFECT OF ENTERAL NUTRITION WITH SALVIA MILTIORRHIZA AGAINST MULTIPLE ORGANS INJURIES IN RABBITS
Ruolan DOU ; Jie XU ; Yugang JIANG ; Zhihui SUN ; Wei PANG ; Xinhua XIA
Acta Nutrimenta Sinica 2004;0(05):-
Objective: To study the protective effect of enteral nutrition with Salvia miltiorrhiza on injuries multiple organs in rabbits. Method: Forty-four rabbits were randomly divided into four groups: Normal control group (N) 8 rabbits, Endotoxin group (LPS), Enteral nutrition group (LPS+EN) and Enteral nutrition with Salvia miltiorrhiza group (LPS+DS),each 12 rabbits. Three days before given endotoxin, LPS+DS group was fed enteral nutrition fluid with Sallvia miltiorrhiza 80 ml/kg bw (15g salvia per 100 ml fluid) and LPS+ EN group was fed enteral nutrition only.N group was fed the same quantity of normal saline. At the end of 10 d, the rabbits were killed and serum SOD, MDA, endotoxin and intestinal mucus SIgA contents were determined. Small intestine, liver, lung, kidney and heart were taken for pathological examination. Results: Serum MDA of LPS+DS group was much lower than LPS group, endotoxin level also lower , but intestinal SIgA content higher LPS group . The pathological examination showed that the intestinal villi were shortend, injured , and necrotic in LPS group; liver was hemorrhagic at portal area with cytoplasmic vacuolization, lung and kidney were also injuried pathologically. These pathological features were comparatively mild in LPS+EN group and much better in LPS+ DS group. Conclusion: Enteral nutrition with Salvia miltiorrhiza may reduce the oxidative damages of multiple organs induced by endotoxin as shown by increased SIgA of small intestine, maintenance of intestinal mucosa integrity and alleviation of pathological changes of multiply organs in rabbits.
9.Application of mixed reality technology in clinical teaching of airway anatomy under bronchoscopy
Ruolan WU ; Runchang LI ; Lin YE ; Xiaoxuan ZHENG ; Fangfang XIE ; Jiayuan SUN
Chinese Journal of Medical Education Research 2022;21(8):1038-1041
Objective:To explore the effect of introducing mixed reality technology into traditional atlas teaching to teach airway anatomy under bronchoscopy.Methods:A total of 30 Batch 2017 fifth-year clinical medicine students from Shanghai Jiao Tong University School of Medicine were randomly divided into control group and test group by RAND function in Excel, with 15 students in each group. The control group was taught with the traditional bronchoscopic atlas teaching, and the test group was combined with mixed reality technology. The two groups had the same class time. After teaching, the teaching effect was evaluated by examination and evaluation questionnaire. SPSS 25.0 software was conducted for t test and Mann-Whitney U test. Results:The average score after teaching of test group was (61.67±20.15), and that of control group was (36.67±13.32), with statistically significant differences ( t=4.01, P<0.001). According to the questionnaire results, the scores of the test group on course understanding, course concentration, participation, mastery and satisfaction were better than those of the control group, and the differences were statistically significant ( P<0.05). Conclusion:Using mixed reality technology to assist the clinical teaching of airway anatomy under bronchoscopy can improve the quality of students' study and enhance their understanding of the teaching content and students' participation passion, achieving better teaching effect.
10.Exploring the Mechanism of Action of Qizhu Kang'ai Formula (芪术抗癌方) for the Treatment of Colorectal Cancer based on Network Pharmacology and Experimental Validation
Ruolan SUN ; Yan LIANG ; Fan ZHAO ; Shijiao ZHU ; Linlu WAN ; Xu WANG ; Qihang YIN ; Gang YIN ; Decai TANG
Journal of Traditional Chinese Medicine 2024;65(4):404-413
ObjectiveTo explore the potential molecular mechanism of Qizhu Kang'ai Formula (芪术抗癌方, QZKAF) for the treatment of colorectal cancer (CRC). MethodsNetwork pharmacology was used to analyze the active ingredients and targets of QZKAF for CRC, and analyze the key targets of QZKAF for the treatment of CRC by gene function annotation (GO) and Kyoto Encyclopedia of Genomes (KEGG) pathway enrichment analysis. Molecular docking was applied to predict the binding activity of the core active ingredients to the key targets. A orthotopic transplantation tumor mice model of CRC was established to validate the key targets of QZKAF for CRC obtained from network pharmacology analysis. Forty-eight mice were randomly divided into the sham operation group, the model group, the 5-fluorouracil (5-Fu) group, and the QZKAF low-, medium-, and high-dose groups, with 8 mice in each group. Except for the sham operation group, the remaining groups underwent colon cancer orthotopic transplantation tumor modeling. The 5-Fu group was given 30 mg/kg of 5-Fu by intraperitoneal injection once every 3 days on the alternate day after modeling, while the QZKAF low-, medium-, and high-dose groups were given 2.925, 5.85, and 11.7 g/(kg·d) of QZKAF by gastric gavage, respectively, and the sham-operation group and the model group were gavaged with 0.1 ml/10 g of normal saline every day, all for 21 days. The in situ tumors mass and the number of liver metastases were compared between the groups. The pathological changes of colon tumor tissues were observed by HE staining, and the protein expression of protein tyrosine phosphatase nonreceptor type 1 (PTPN1), vinculin, integrin subunit αν, integrin subunit β3, and E-cadherin were detected in colon tumor tissues by Western blot. ResultsNetwork pharmacology screening yielded that the top six core active ingredients of QZKAF intervening in CRC were quercetin, kaempferol, apigenin, luteolin, baicalein and ursolic acid. There were 212 targets of action, and the ranked top three were prostaglandin endoperoxide synthase 1 (PTGS1), prostaglandin endoperoxide synthase 2 (PTGS2), and PTPN1, which may be the key targets of QZKAF in the treatment of CRC. These key targets were significantly enriched mainly in phosphatidylinositol 3-kinase/protein kinase B (PI3K-Akt) signaling pathway, focal adhesion and adhesion junction. Molecular docking results: except for PTGS1 with better binding activity to quercetin, kaempferol, and apigenin (binding energy ≥