1.Analysis of the Genetic Diversity of Germplasm Resources of Duchesnea indica from Hunan by Random Amplified Polymorphic DNA
Xiangdan LIU ; Yu GAO ; Yali ZHANG ; Jialuo CAI ; Qiaozhen TONG ; Congzi ZHANG
China Pharmacy 2015;(19):2628-2631
OBJECTIVE:To study the genetic diversity of germplasm resources of Duchesnea indica from Hunan. METHODS:The random amplified polymorphic DNA(RAPD)reaction system of germplasm resources was established and RAPD-PCR was ad-opted to detect the expressions of 24 groups of D. indica from Hunan(including 21 wild and 3 cultivated varietos in different geo-graphical distribution). Agarose gel electrophoresis was used to analyze the results and calculate allele number (Na),effective al-lele number (Ne) the polymorphism points percentage (PPB),Nei’s gene diversity index (H) and Shannon’s information index (I). RESULTS:Totally 14 polymorphic primers were screened and 90 electrophoresis bands were amplified,including 81 polymor-phic bands with Na of 1.900 0,Ne of 1.540 1,PPB of 90.0%,H of 0.312 8 and I of 0.467 5. CONCLUSIONS:The genetic diver-sity of D. indica from Hunan is rich,and there were differences in genetic background between the cultivated species and wild spe-cies;clustering results show significant correlation with geographic distribution.
2.Mechanism of inflammatory cancer-associated fibroblast-mediated drug resistance in colorectal cancer cells
Jialuo CAI ; Ruiqiu ZHU ; Sen LI ; Yijun CAO ; Fang HUANG
China Oncology 2023;33(12):1065-1072
Background and purpose:Colorectal cancer(CRC)is one of the common malignancies,but the mechanism by which it develops resistance to drug remains unclear.The tumor microenvironment(TME),especially cancer-associated fibroblast(CAF),plays an important role in the occurrence,development and drug resistance of tumors.This study aimed to investigate the effect of inflammatory cancer-associated fibroblasts(iCAF)on drug resistance in CRC cells and its possible mechanism.Methods:The primary CAFs were collected from CRC patients underwent surgery in Putuo Hospital,Shanghai University of Traditional Chinese Medicine from Aug.2022 to Sep.2022,and the primary cells were sorted according to the surface marker of CAF[approved by the Ethics Committee of Putuo Hospital Affiliated to Shanghai University of Traditional Chinese Medicine:PTEC-A-2023-5(S)-1],platelet derived growth factor receptor alpha(PDGFRA),to screen iCAF.Human intestinal fibroblast(HIF)and iCAF cells were cultured using serum-free medium to obtain conditioned medium.According to the treatment method,colon cancer cells were divided into control group(no treatment),experimental group 1(treated with HIF-CM)and experimental group 2(treated with iCAF-CM).We observed the changes in the survival rate and apoptotic rate of CRC cells,the changes in protein and mRNA levels and the effect on the Wnt/β-catenin signaling pathway after stimulation with HIF-CM or iCAF-CM.Results:After iCAFs stimulation,the half inhibition concentration(IC50)of CRC cells was higher compared with the control group and HIF-CM group(P<0.05).Compared with the control group and HIF-CM group,the apoptotic rate of tumor cells in iCAF-CM group decreased significantly,the expression of caspase-3 was decreased,and the expressions of Bcl-2,Bcl-xL and survivin were increased(P<0.05).The Wnt/β-catenin signaling pathway was activated in the iCAF-CM group.Conclusion:iCAFs can mediate drug resistance in CRC cells,and the mechanism is related to the activation of Wnt/β-catenin signaling pathway.
3.Intervention effect and mechanism of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in RA model rats
Yuanshan HAN ; Gangqiang YI ; Xin LI ; Lin OUYANG ; Jialuo CAI ; Manshu ZOU
China Pharmacy 2022;33(16):1974-1979
OBJECTIVE To evaluate the intervention effect of Caulis sinomenii compatible with prepared Aconiti Lateralis on bone destruction in rheumatoid arthritis (RA)model rats ,and to investigate its mechanism. METHODS Totally 40 SD rats were randomly divided into blank group ,model group ,positive control group (indomethacin 0.013 5 g/kg)and C. sinomenii compatible with prepared Aconiti Lateralis group (C. sinomenii 1.08 g/kg+prepared Aconiti Lateralis 1.35 g/kg)according to body mass ,with 10 rats in each group. Except for the blank group ,all the other groups made RA rat models by injecting type Ⅱ bovine collagen. Rats in each group were given corresponding drugs or distilled water intragastrically. The general information ,body weight ,foot swelling and arthritis index (AI)scores of rats in each group were recorded. After the 30th day of administration ,the changes of ankle bone in rats were detected by small animal CT machine. The levels of inflammatory factors [interleukin- 31(IL-31),IL-25 and IL- 3] and chemokines [receptor activator of nuclear factor κB ligand(RANKL),receptor activator of nuclear factor κB (RANK)and osteoprotegerin (OPG)] in serum were detected by enzyme-linked immunosorbent assay. Pathological indexes of rat ankle joint were observed by HE staining. Immunohistochemical method was used to detect the expression of RANKL ,RANK and OPG in synovial tissue of rat ankle joint. RESULTS Compared with blank group ,the mental state of the model group was weak , the activity decreased significantly ,the hair lost luster ,and the body weight decreased significantly on the 12th to 30th days (P< 0.05 or P<0.01);the swelling degree of the foot was significantly increased and the AI score was significantly increased on the 12th to 30th days(P<0.01);the ankle joint in model group had rough surface ,obvious tissue damage and serious bone erosion ; serum levels of IL- 31,IL-25,IL-3,RANKL and RANK were increased significantly ,while the level of OPG was decreased significantly (P<0.01); the expression of RANKL and RANK in synovium of ankle joint increased significantly , while the expression of OPG decreased significantly (P<0.01). Compared with model group ,the above indexes of administration groups were improved to varying degrees ,and most of the differences were statistically significant (P<0.05 or P<0.01). CONCLUSIONS By inhibiting the RANKL/RANK/OPG signaling pathway ,C. sinomenii compatible with prepared Aconiti Lateralis can inhibit the excessive proliferation of osteoclasts and restore the balance of bone metabolism so as to play a role in protecting bone joints and treating RA.