2.Significance of siRNA-mediated TGFBR2 gene silencing on HepG2 cell proliferation
Lin CHENG ; Wujian DENG ; Xiaofeng JIANG ; Kun YU ; De CHEN
The Journal of Practical Medicine 2014;(14):2200-2203
Objective To value the significance of TGFBR2 gene in mediating HepG2 cell proliferation by RNA interference technology. Methods Three kinds of siRNAs targeting TGFBR2 gene were designed, synthesized and transfected into HepG2 cells via lipofetamine2000. Among three kinds of siRNAs, only the one with the most interference efficacy was selected and the correspondent DNA sequence was inserted into plasmid pEGFP-N3. Then the recombinant plasmid of siRNA-pEGFP-N3 was transfected into HepG2 cell and western blot was used to detect the protein level of TGFBR2. Then, TGF-β1 was used to stimulate HepG2 cells with or without siRNA interference and proliferation of HepG2 cells was observed. Results Among these three siRNAs, siRN-1 appeared to be the most effective. After stimulated by 5ng/mL TGF-β1, proliferation of HepG2 cells showed a marked increase in siRNA-1 group compared with blank and siRNA-NC groups (P<0.05). For all that, the proliferation rate was still lower than that in normal HepG2 cell group without TGF-β1 stimulation. Conclusion By silencing TGFBR2 gene, inhibition of TGF-β1 signaling pathway to HepG2 cells could be decreased, thereby enhancing the cell proliferation.
3.STUDIES ON BLEACHING OF PULP BY WHITE-ROT FUNGUS
De-Qing ZHAO ; Lu LIN ; Li-Ping JIANG ;
Microbiology 1992;0(06):-
This article summarizes the bleaching of pulp by white-rot fungus, including the type of white-rot fungus, the type of pulp, the equipment used to bleach by white-rot fungus, and some problems and prospect in practical application by white-rot fungus.
4.Progress in the development of crystallized proteins as drug delivery system.
Kai SHI ; Yan-Bo JIANG ; Lin-Lin YANG ; Fu-de CUI
Acta Pharmaceutica Sinica 2009;44(8):845-851
Crystallization has been widely applied in pharmaceutical formulations as an effective approach to improve the stability and efficacy of small agents. However protein crystals are suffered from limitation in the drug delivery system due to their complex crystallization behaviors. With development of crystallization technologies and their industrial application, protein crystals are receiving more and more attentions as a novel delivery system for biomacromolecules. Crystals with thermodynamic stable structure can improve the physical and chemical stability of protein drugs and present a sustained release behavior. On the basis of pertinent literatures, this review introduces the recent research situation and development process of protein crystals as drug delivery system. Moreover, the crystallization process of proteins, as well as the preparation and potential application are discussed systematically.
Crystallization
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Drug Delivery Systems
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Pharmaceutical Preparations
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administration & dosage
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Proteins
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chemistry
5.Inhibition of tropical injected bevacizumab on haze formation after off-flap epipolis laser in situkeratomileusis
Jing, LI ; Zheng-wei, SHEN ; De-zhong, LI ; Ya, YE ; Li, JIANG ; He, YIN ; Lin-ping, XUE
Chinese Journal of Experimental Ophthalmology 2013;(6):529-534
Background Haze formation is a key factor of vision reduce following corneal refractive surgery.Transforming growth factor-β1 (TGF-β1) and α-smooth muscle actin (α-SMA) are documented to participate in haze formation.Laboratory study showed that bevacizumab can not only inhibit corneal neovascularization,but also promote the healing of corneal epithelial basement membrane.However,the impact of bevacizumab on corneal healing after Off-flap epipolis laser in situkeratomileusis (Off-flap Epi-LASIK) is unclear.Objective The present study was to investigate the inhibition effect of bevacizumab on corneal haze after off-flap Epi-LASIK and its active mechanism.Methods Off-flap Epi-LASIK was performed in 24 adult pigmented rabbits and these rabbits were randomized into three groups.Bevacizumab of 0.1 ml (2.5 mg) was subconjunctivally injected 10 minutes after surgery in 16 rabbits and the same amount of bevacizumab was repeatedly injected 4 days after the initial injection in 8 eyes of 16 eyes.In addition,equivalent amount of normal saline solution was used in the same way in the other 8 rabbits.Another 2 health rabbits were used as the blank controls.Operative eyes were examined by slit lamp biomicroscope daily after surgery and haze was scored based on SundarRayde criteria.Corneas were obtained 4 weeks after operation for hematoxylin & eosin and periodic acid Schiff staining.Expressions of TGF-β1 and α-SMA in corneal tissue were detected by immunochemistry.Results Corneal epithelium healed completely in all eyes 4-5 days after operation.The haze scores were lower in the bevacizumab single injection group and repeat injection group than those in the normal saline solution group (P<0.05) in 1 week and 4 weeks after operation.However,no significant difference was seen in the haze scores between the bevacizumab single injection group and repeat injection group (P > 0.05).The hostopathological examination showed that the fibrosis response of cornea tissue was slight in the bevacizumab single injection group and repeat injection group comparison with the normal saline solution group.At 1 week after operation,the expression levels of TGF-β1 were (49.8 ± 2.1) PU and (38.6 ±4.4) PU in the bevacizumab single injection group and repeat injection group,and those of 4 weeks were (37.7 ±4.8) PU and (28.3 ± 3.5) PU,indicating significant decrease in the TGF-β1 expression compared with (65.1 ±5.3) PU and (51.6±2.2) PU of the normal saline solution group in both 1 week and 4 weeks (P<0.01).The expression levels of α-SMA in corneas were (67.2±10.0) PU and (32.7±3.1) PU at 1 week,and (34.2±5.7) PU and (22.8±3.0) PU at4 weeks after operation in the bevacizumab single injection group and repeat injection group,which were significantly lower than (87.8±7.7) PU and (59.4±5.6) PU in the normal saline solution group in both 1 week and 4 weeks (P<0.05).Meanwhile,the expression levels of TGF-β1 and α-SMA were declined in the bevacizumab repeat injection group compared with single injection group (P<0.01).Periodic acid Schiff staining exhibited that the basement membrane of cornea was intact and continued in bevacizumab injection group.But corneal basement membrane was discontinuous in the normal saline solution group.Conclusions Subconjunctival injection of bevacizumab downregulates the expressions of TGF-β1and α-SMA in cornea after Off-flap Epi-LASIK and thus prevents haze formation.
6.The significance of multi-slice helical CT multiplanar reconstruction in the diagnoses of laryngeal carcinoma
Lin LI ; De-Hong LUO ; Chun-Wu ZHOU ; Xin-Ming ZHAO ; Li-Ming JIANG ; Yao HUANG ; Ling-Xia JIANG ; Jing LI ; Ning WU ;
Chinese Journal of Radiology 1999;0(10):-
Objective To evaluate the significance of multi-slice helical CT with multiplanar reconstruction in laryngeal carcinoma.Methods Thirty-five patients with laryngeal carcinoma were studied by helical CT,MPR were subsequently done.The lesion extent of the axial image findings,MPR findings and the combined image findings were compared with the pathological results respectively.The data were statistically analyzed.Results In the evaluation of the anterior commissure,the axial image findings,MPR findings and the combined image findings were 82.9%,68.6% and 91.4% in accuracy respectively,the results were statistically different(P0.05).The combined images were superior to the axial images and the MPR images in sensitivity,specificity and accuracy of the lesion extent.Conclusion The axial images could show the shape,size,extension of the tumor and the lymphadenopathy,MPR images displayed the shape,size and extension roundly and directly,they were the supplement for the axial images.Axial images combined with MPR could improve the accuracy in the diagnoses of laryngeal carcinoma.
7.Clinical comparative study of minimally invasive esophagectomy versus open esophagectomy for esophageal carcinoma.
Bao-fu CHEN ; Cheng-chu ZHU ; Chun-guo WANG ; De-hua MA ; Jiang LIN ; Bo ZHANG ; Min KONG
Chinese Journal of Surgery 2010;48(16):1206-1209
OBJECTIVETo explore and evaluate the feasibility, safety, radicality and the short-term outcome of minimally invasive esophagectomy versus open esophagectomy for esophageal cancer.
METHODSFrom July 2007 to October 2009, 67 patients with esophageal cancer received minimally invasive esophagectomy (MIE group), while 38 patients underwent conventional open esophagectomy (OE group: via right thorax, abdomen, left neck). The operative procedures, clinicopathological data and short-term outcome were collected and compared between the two groups.
RESULTSThe clinical data of the two groups were comparable. No significant differences was found in demographics between the two groups. Median blood loss in MIE group was less than that in OE group (chest: 112.3 ml vs. 175.3 ml, P = 0.035, abdominal: 31.4 ml vs. 100.5 ml, P = 0.026). More patients in OE group were transferred to ICU (P = 0.042) and more obvious pain (P = 0.005). The rate of pulmonary infection and intestinal obstruction in OE group were higher than MIE group (P = 0.046 and 0.045). There were no differences in the number of lymph node dissection for two groups, the average was 20.9 and lymph node metastasis rate was 26.9% in MIE group. Mean follow up was (14.0 ± 2.2) months (range, 2 to 29 months). Recurrence rate and survival rate were no differences.
CONCLUSIONThe Minimally invasive esophagectomy for esophageal cancer is feasible, safe, and reliable short-term effect, and can achieve radical tumor resection, which may lead to better future of surgical treatment for esophageal carcinoma.
Aged ; Esophageal Neoplasms ; surgery ; Esophagectomy ; methods ; Female ; Follow-Up Studies ; Humans ; Laparoscopy ; Male ; Middle Aged ; Retrospective Studies ; Thoracoscopy ; Treatment Outcome
8.Expression of porcine beta-defensin 1 gene in Pichia pastoris.
Li-Hua JIANG ; Hai-Rong LU ; De-Xin HUANG ; Jun-Bo YI ; Ling-Yun LI ; Feng LIN
Chinese Journal of Biotechnology 2006;22(6):1036-1039
PBD-1 is an antibacterial peptide that plays an important role in defence system of porcine. To produce PBD-1 with bioactivity in Pichia pastoris, according to published amino acid sequence of porcine beta-defensin 1(PBD-1) and the partiality codon of yeast, the PBD-1 gene was synthesized by PCR and cloned into pPIC9K to construct the recombinant expression vector pPIC9K-PBD-1, the obtained recombinant plasmid was linearized by Sal I, and then transformed into SMD1168 by electroporation. Under the control of the promoter AOX1, an approximately 4.5 kD PBD-1 peptide was expressed. Antibacterial activity assay shows that the PBD-1 has the antibacterial activity on Staphylococcus aureus. This is the first secreted expression of porcine beta-defensin 1 gene in Pichia pastoris.
Animals
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Anti-Bacterial Agents
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biosynthesis
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isolation & purification
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pharmacology
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Cloning, Molecular
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Gene Expression
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Genetic Vectors
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genetics
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Pichia
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genetics
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Protein Engineering
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Staphylococcus aureus
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drug effects
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Swine
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genetics
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beta-Defensins
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biosynthesis
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genetics
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isolation & purification
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pharmacology
9.Effect of Siwu decoction on function and expression of P-glycoprotein in Caco-2 cells.
Yi JIANG ; Zeng-chun MA ; Xian-ju HUANG ; Qing YOU ; Hong-ling TAN ; Yu-guang WANG ; Qian-de LIANG ; Xiang-lin TANG ; Cheng-rong XIAO ; Yue GAO
China Journal of Chinese Materia Medica 2015;40(5):933-937
To study the effect of Siwu decoction on the function and expression of P-glycoprotein (P-gp) in Caco-2 cells. The Real-time quantitative poly-merase chain reaction (Q-PCR) was used to analyze the mRNA expression of MDR1 gene in Caco-2 cells. Flow cytometer was used to study the effect of Siwu decoction on the uptake of Rhodamine 123 in Caco-2 cells, in order to evaluate the efflux function of P-gp. Western blotting method was used to detect the effect of Siwu decoction on the P-gp protein expression of Caco-2 cells. Compared with the blank control group, after Caco-2 incubation with Siwu decoction at concentrations of 3.3, 5.0, 10.0 g x L(-1) for 24, 48, 72 h, the mRNA expression of MDR1 was up-regulated, suggesting the effect of Siwu decoction in inducing the expression of MDR1. After the administration with Siwu decoction in Caco-2 cells for 48 h, the uptake of Rhodamine 123 in Caco-2 cells decreased by respectively 16.6%, 22.1% (P < 0.05) and 45.4% (P < 0.01), indicating that the long-term administration of Siwu decoction can enhance the P-gp efflux function of Caco-2 cells. After the incubation of Caco-2 cells with Siwu decoction for 48 h, the P-gp protein expression on Caco-2 cell emebranes, demonstrating the effect of Siwu decoction in inducing the protein expression of P-gp.
ATP Binding Cassette Transporter, Sub-Family B
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genetics
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metabolism
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ATP-Binding Cassette, Sub-Family B, Member 1
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genetics
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metabolism
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Caco-2 Cells
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Drugs, Chinese Herbal
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pharmacology
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Humans
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Up-Regulation
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drug effects
10.Evidence grading system and its usage in clinical decision-making analysis.
De-ying KANG ; Jia-liang WANG ; Qi HONG ; Jia-lin JIANG
Chinese Journal of Epidemiology 2004;25(4):351-354
OBJECTIVETo explore feasibility of applying evidence grading system in clinical decision-making analysis.
METHODSUsing grading system as well as a quantitative approach to generate the threshold NNT((T)), and conducting clinical decision-making analysis through one study on Asymptomatic carotid atherosclerosis.
RESULTSNNT((T)) was 12, comparing with no-intervention in the ACA study that NNT = 6. NNT < NNT((T)) indicated the benefits of intervention outweigh the risks and costs.
CONCLUSIONTo obtain the rate of adverse event and the specifying approximate cost were vital in the process of applying grading system in clinical decision-making.
Advance Care Planning ; Algorithms ; Carotid Artery Diseases ; economics ; therapy ; Decision Making, Organizational ; Evidence-Based Medicine ; methods ; Health Care Costs ; Humans ; Models, Theoretical