3.Biochemical process engineerinmg and modernization of Chinese materia medica (Ⅰ)
Chinese Traditional and Herbal Drugs 1994;0(08):-
Some studies corresponding modernization of Chinese materia m edica(CMM)were performed in the institute for the new technologies,materials,and facilities based on the achievements and experiences gotten in the past two dec ades.The technologies of large-scale propagation of plant cell,tissue,and organ were applied to the products of cell metabolism,biotransformation,and artificial breeding and rooting.Many new types of large-scale bioreactors were designed an d used not only for laboratory but also for manufactory.Reaction/separation inte gration technology,microwave-assisted extraction,and the other new extraction te chnologies were studied to realize high efficiency and low energy.Besides tradit ional separation and purification methods,new technologies,including extraction in reversed micelles,one step three-phase extraction on Penicillin,foam fraction ation,membrane separation,and high-speed counter-current chromatography(HSCC),we re developed.HSCCC is a kind of liquid-liquid partition chromatography without a ny solid matrix,which eliminates irreversible adsorption of samples on solid sup port in the conventional chromatographic column.It has been successfully applied to the analysis and separation of various natural products.At the same time,man y types of microsphere and microencapsules for controlled release and separation media were prepared and lots of attention was paid on drug modifying and embedd ing techniques.HSCCC was also applied as a new method to the study of CMM finger print and showed good precision and repeatability.High performance liquid chroma tography(HPLC),gas chromatography(GC),high performance capillary electrophoresis (HPCE),and mass spectrometer(MS)played a very important role in fingerprinting.C ontinuous propagation-continuous collection technological process was set for la rge-scale propagation of micro-algae in the study of marine drugs.Transgenic alg ae were cultured to prduce genetically engineered products.
4.Research progress of cervical carcinoma risk factors
Practical Oncology Journal 2014;(4):367-370
Cervical carcinoma is a serious threat to the health of women around the world ,and its inci-dence ranks at the second position after breast cancer in female reproductive system .In addition to oncogene acti-vation and inactivation of tumor suppressor gene ,endogenous and exogenous factors also affect the development of cervical cancer .
5.The roles of motility in colonization of Helicobacter pylori in human stomach
Chinese Journal of Laboratory Medicine 2008;31(7):733-736
Helicobacter pylori is able to colonize the human stomach and dwell in the human stomach for decades or for whole lifetime. A number of potential virulence factors contribute to Helicobacter pylori to colonize this unusual niche. Motility is an essential colonization factor based on the fact that nonmotile variants of Helicobacter pylori can't infect gnotobiotic piglets. Motility is not a colonization factor based on rapid loss of motility of Helicobacter pylori in the gastric lumen in vivo. The exact roles of Helicobacter pylori motility are not yet known. The aim of this article is to discuss correlation between colonization and motility of Helicobacter pylori.
7.Construction of shRNA eukaryotic expression vector targeting OPTN gene and detection of interference effect
Yu GU ; Libin FAN ; Xiaoyin LIU
Acta Universitatis Medicinalis Anhui 2014;(1):1-4
Objective To construct shRNA against optineurin ( OPTN ) clone and transfect the construction into HEK293FT cells for inhibiting the expression of OPTN, which is laid the foundation for further research of molecu-lar mechanism OPTN protein. Methods The insert of this clone was a double-strained DNA sequence against OPTN which would be transcripted into shRNA and it was synthesized according to the sequence in Origene. Ex-pression vector pRFP-C-RS was employed to fuse the insert to get the construction and finally confirmed by sequen-cing. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN and RFP signals could be detected using fluorescence microscope. Western blot was further employed to check the protein level of OPTN. HEK293FT cells were transfected with the very clone pRFP-C-RS-shOPTN was then obtained to investigate the functional study of OPTN which was measured by Salmonella infection experiment. Results shRNA eukaryotic expression vector targeting OPTN gene was successfully constructed. HEK293FT cells were transfected with pRFP-C-RS-shOPTN and 72 hours later we observed strong RFP signals showing that shOPTN expressed. Western blot was further employed to check the protein level of OPTN which showed that OPTN expression was indeed interfered. Salmonella infection assays was then performed and showed that OPTN could inhibit the proliferation of Salmonella that has invaded HEK293FT cells. Conclusion HEK293FT cell line which expresses shRNA against OPTN show a sharp inhibition of OPTN protein expression by 80%. So this cell line can be further used to investigate how OPTN regulates auto-phagy as an autophagy receptor. Meanwhile we find that OPTN can be as a autophagy regulator and strongly sup-press the proliferation of invasive Salmonella in vivo.
9.Research progress on root canal biofilm
Ying ZHENG ; Lijie FAN ; Zhiyuan GU
Journal of Practical Stomatology 2010;26(2):252-254
Root canal biofilm is frequently detected in the canal wall of infected root canal and the root canal with failed root canal therapy. Due to its special structure and diverse composition, root canal biofilm has the ability of the drug tolerance and antiimmunity, which lead to apical periodontitis. This review summarizes the features of the root canal biofilm and latest clinical methods to remove it.
10.Down-regulation of midkine by small interfering RNA increase chemosensitivity in bladder cancer cells
Yu FAN ; Xiao GU ; Xiaoda TANG
Chinese Journal of Urology 2010;31(9):604-607
Objective To study the effects of knock down midkine(MK)by siRNA on chemosinsitivity in bladder cancer cells. Methods Three MK siRNAs were designed and constructed. After transfected with MK siRNA or scrambled siRNA for different time, cultured cells were harvested to carry on the next experiments. Expression of MK was determined by real-time RT-PCR and western blot, and apoptosis were evaluated by caspase-3 activity and TUNEL assay. MTT was performed to examine the inhibition effect of Paclitaxel (PTX) on cells. Results MK siRNA could down-regulate the MK expression in a dose- and time-dependent manner. The MTT results showed that the inhibit rates were (18.21±0.36)%, (18.19±0.29)%, (17.89±0.33)%, (1.86±0.52)%, (32.56±0.53) %, (53. 83±0.38) % and (78. 95±0.55) % in different groups PTX alone(0.2μmol/L), ConA +PTX(0.2 μmol/L), Con-B +PTX(0.2 μmol/L), siRNA alone(12. 50 nmol/L), siRNA(3. 125nmol/L) + PTX (0. 2 μmol/L), siRNA (6. 25 nmol/L) + PTX (0. 2 μmol/L) and siRNA (12. 50nmol/L)+PTX(0.2 μmol/L), respectively. The TUNEL results showed that apoptosis index was (1.81 ±0. 36)%, (1. 89±0. 38)%, (5. 56±0. 58)%, (9. 68±0.55)% and (15. 25±0.56)% in different groups (Con-A, Con-B, siRNA (3. 125 nmol/L), siRNA (6. 25 nmol/L) and siRNA ( 12. 5nmol/L), respectively. The activity of caspase-3 increased significantly in transefected cells with a dose-and time-dependent manner. Conclusion MK siRNA could sensitize human bladder cancer cells to chemotherapy which might be through the apotosis.