1.Construction of HPV18E7 recombinant plasmid and exploration of its optimization expression condition in Escherichia coli
Renjian HU ; Jiali CAI ; Li LIU ; Manyu TU ; Tao XU ; Cuirong DU ; Jia LUO ; Sen DING
Chongqing Medicine 2013;(30):3647-3649
Objective To construct recombinant plasmids containing HPV18E7 gene ,and explore the optimization condition of its expression in Escherichia coli .Methods The genomic DNA extracted from HeLa cell line which served as a template to the HPV18 E7 gene was amplified using PCR method ;and the amplified product of HPV18E7 gene was connected to the pET-32a(+ ) vector ,which composed the pET-32a(+ )-HPV18E7 recombinant plasmid ;the positive recombinant plasmids were transformed into BL21-DE3-pLysS competent cells and the optimized expression condition was explored in order to obtain a large amount of HPV18E7 oncogenic protein .Results The fragment length of PCR products of HeLa cell genomic DNA was consistent with that of HPV18 E7 gene .In LB medium ,the expression level of the target protein was not high under such conditions as different concentra-tion of IPTG and lactose ,different temperatures and different induction starting amount .Therefore the ZYM-5052 auto-induction medium was tried in this experiment ,and the expression amount of the fusion protein was much higher than that induced with IPTG and lactose .Conclusion The amount of HPV18E7 fusion protein in ZYM-5052 automatic induction medium is much higher than that induced with IPTG and lactose .
2.Correlation and path analysis on characters related to flower yield per plant of Carthamus tinctorius
Yuxia YANG ; Wei WU ; Youliang ZHENG ; Chunyan HUANG ; Renjian LIU ; Li CHEN
Chinese Traditional and Herbal Drugs 1994;0(01):-
plant height.Multiple regression analysis showed that branch height from base(X_2),number of primary branches(X_4),number of effective cones per plant(X_6),number of ineffective cones per plant(X_7),number of grains per cone(X_9),and diameter of primary head(X_(10)) were the main factors affecting flower yield(Y) per plant.Multiple regression equation of flower yield per plant and six characters was Y=-3.037 0+(0.002 7) X_2+0.045 9 X_4+0.074 5 X_6+0.043 2 X_7+0.023 0 X_9+1.148 2 X_(10)(F=21.84~()).The direct effect of number of effective cones per plant was the strongest,followed by diameter of primary head.There were significant differences within the flower yield per plant and the safflor yellow A content of different species.The correlation between the safflor yellow A content and the flower yield per plant was insignificant.Conclusion High-yield and high-quality are compatible in breeding of safflower which is used as herbal medicine.Number of effective cones per plant and diameter of primary head are focused on the high yield breeding and cultivation of safflower species.The plant type of higher flower yield safflower species should have more effective cone numbers,more number of cones,number of branches,number of primary branches,bigger diameter of primary head,moderate plant height,branch height from base,and ineffective cones per plant.Of all accessions,PI 239226,PI 253540,PI 367833,and Jianyang Honghua are outstanding and optimal for cultivating in Sichuan Province.
3.How massage inhibits scar tissue formation in skeletal muscles after blunt injury
Renjian LIU ; Chenglin TANG ; Min ZHOU ; Jie GAO ; Hui XIE ; Xiaolin CHEN
Chinese Journal of Physical Medicine and Rehabilitation 2012;(10):742-746
Objective To investigate the effect of massage on quadriceps femoris repair after injury by external force and the expression of transforming growth factor β1 (TGF-β1) and collagen-Ⅰ (COL-Ⅰ) mRNA.To explore the molecular mechanisms inhibiting scar tissue formation and promoting muscle repair.Methods Forty New Zealand white rabbits weighing (2.0 ±0.5) kg were randomly divided into a normal control group (A) (n =4),a selfrepair group (B) (n =20,further divided into the 3rd,7th,11th,15th and 19th day time points),and a massage group (C) (n =16,further divided as in group B).In group A the rabbits were not treated,as normal controls.In groups B and C rabbit models of quadriceps femoris injury were prepared using a self-made beater.In group B no massage therapy was given as a natural recovery control; in group C,massage therapy was given after 5 days.Realtime quantitative PCRs were used to detect TGF-β1 and COL-Ⅰ mRNA expression.Resnlts There was no significant difference between groups B and C in the expression of TGF-β1 or COL-Ⅰ mRNA on day 7.At the later time points,expression of both mRNAs in group C was significantly less than in group B.Conclusion Massage can effectively reduce the expression of TGF-β1 and COL-Ⅰ mRNA,inhibit excessive scar formation and promote repair of injured tissue.
4.Comparison of volatile oils of cultivated Houttaynia cordata populations with wild.
Lei LIU ; Wei WU ; Zhiping FU ; Chunyan HUANG ; Renjian LIU
China Journal of Chinese Materia Medica 2010;35(7):876-881
OBJECTIVETo provide scientific proofs for the exploitation, utilization, and normalized cultivation of Houttuynia cordata.
METHODThe underground parts of 17 wild H. cordata populations from different valleys and altitudes of mountain. Emei were transplanted to the same growth conditions. After one year's cultivation, volatile oil was obtained by steam distillation from the aerial part of the materials. The chemical constituents were separated and identified by GC-MS, and the relative content of each constituent was determined by area normalization.
RESULTTotally, 31 chemical components were identified, 19 components could be detected in all materials. The t-test results indicated that the contents of alpha-pinene and D-limonene were extremely significantly higher than that in the wild populations, and the contents of camphene and 2-undecanone were also significantly higher than that in the wild populations. And the reverse was found in the content of trans-beta-ocimene. All these 31 components could be divided into 9 chemical compositions, and 7 chemical compositions could be detected in the wild and cultivated. RSD values of 5 chemical compositions in wild populations were higher than that in the cultivated, except for the contents of diterpenyl alcohols and diterpenyl aldehydes. These five chemical compositions accounted for 84.05% and 90.12% of the whole volatile oils in the wild and cultivated conditions, respectively.
CONCLUSIONThe components and contents of the volatile oils between wild and cultivated were different. The volatile oils polymorphism decrease distinctively as all the wild populations of H. cordata were transplanted to the uniform environmental conditions.
Houttuynia ; chemistry ; growth & development ; Oils, Volatile ; analysis ; chemistry ; Organic Chemicals ; analysis
5.Effect of electro-acupuncture on hepatic Toll-like receptor 4 and nuclear factor κB expressions in rats with non-alcoholic fatty liver disease.
Xiaolin CHEN ; Chenglin TANG ; Hui XIE ; Nianzhen TANG ; Jie GAO ; Renjian LIU
Journal of Southern Medical University 2014;34(11):1584-1588
OBJECTIVETo investigate the effect of electro-acupuncture on Toll-like receptor 4 (TLR4) nuclear factor κB (NF-κB) expressions in the liver of rats with non-alcoholic fatty liver disease (NAFLD).
METHODSSD rat models of NAFLD induced by feeding high-fat diet for 5 weeks were subjected to early or late intervention with electro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao for 20 min once daily for 21 consecutive days. The changes in serum ALT, AST, TC, and TG levels were determined in the rats and hepatic expressions of TLR4 and NF-κB detected with quantitative fluorescence PCR and Western blotting after treatments.
RESULTSSerum ALT, AST, TC, and TG levels and TLR4 and NF-κB levels increased significantly after NAFLD onset (P<0.05), and were markedly reduced by interventions with electro-acupuncture (P<0.05).
CONCLUSIONElectro-acupuncture at the acupoints Zusanli, Fenglong and Sanyinjiao can modify blood lipid and hepatic function to produce positive effect on NAFLD possibly by modifying lipid metabolism, lowering expressions of TLR4 and NF-κB in the liver, and reducing the impact of inflammation on NAFLD.
Animals ; Diet, High-Fat ; Disease Models, Animal ; Electroacupuncture ; Inflammation ; NF-kappa B ; metabolism ; Non-alcoholic Fatty Liver Disease ; metabolism ; therapy ; Rats ; Rats, Sprague-Dawley ; Toll-Like Receptor 4 ; metabolism