3.Expression and antitumor activity of fusion protein RGD-TRAIL in Pichia pastoris.
Acta Pharmaceutica Sinica 2015;50(5):552-559
To compare the activity of RGD-TRAIL in different expression systems, RGD-TRAIL in both Escherichia coli (E.coli) and Pichia pastoris was constructed and expressed. In vitro activity of RGD-TRAIL from Pichia pastoris expression system was also analyzed. Genetic engineering techniques were used to construct recombinant plasmid pET30-rgd-trail and pHBM-rgd-trail. The recombinant protein RGD-TRAIL was purified with Ni ion affinity chromatography after induction. MTT assay, ELISA, scratch wound healing, transwell migration assay and Hoechst 33342 staining were performed to detect the effects of RGD-TRAIL on proliferation, binding activity, migration and apoptosis. The expression of apoptosis-associated proteins was detected by Western blotting. Recombinant protein RGD-TRAIL was successfully expressed in a form of inclusion body in E.coli, while expressed secretorily in Pichia pastoris. It possessed more potent cytotoxicity than RGD-TRAIL in E.coli by MTT assay. The RGD-TRAIL expressed by Pichia pastoris showed powerful binding affinity with cancer cells expressing α(v), DR4, DR5 and highly potent cytotoxicity through inducing apoptosis of cancer cells. Nuclear fragmentation was examined by Hoechst 33342 staining. Cleaved PARP and caspase-3 were also detected after incubation with RGD-TRAIL. Additionally, RGD-TRAIL inhibited migration significantly in A549 and HT1080 cells. The results demonstrate that Pichia pastoris expression system is more suitable for the recombinant protein RGD-TRAIL. Its binding affinity and antitumor activity might make RGD-TRAIL a promising candidate for cancer therapy.
Antineoplastic Agents
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pharmacology
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Apoptosis
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Blotting, Western
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Cell Line, Tumor
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Chromatography, Affinity
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Enzyme-Linked Immunosorbent Assay
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Escherichia coli
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Humans
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Oligopeptides
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biosynthesis
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pharmacology
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Pichia
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metabolism
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Plasmids
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Recombinant Fusion Proteins
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biosynthesis
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pharmacology
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TNF-Related Apoptosis-Inducing Ligand
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biosynthesis
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pharmacology
4.Nasal mucus and nasal inflammation.
Chinese Journal of Otorhinolaryngology Head and Neck Surgery 2007;42(9):707-710
Humans
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Mucus
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Nasal Mucosa
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Rhinitis
5.Meta analysis of efficacy and safety of levofloxacin in treatment of refractory pulmonary tuberculosis hemoptysis
Zhongting FEI ; Zhen CHEN ; Yao TANG
Chinese Journal of Biochemical Pharmaceutics 2014;37(7):164-166
Objective To study efficacy and safety of the levofloxacin in treatment of refractory pulmonary tuberculosis hemoptysis by meta analysis.Methods Pubmed (2000/2014-07),EMbase (2000/2014-07),the Cochrane library (2014,7),the Chinese biomedical literature database (2000/2014-07)were retrieved about the randomized controlled trials of levofloxacin in treatment of refractory pulmonary tuberculosis hemoptysis,and the time was limited from January 2000 to July 2014.After objective evaluation of the retrieved paper quality,Cochrane collaboration with Rev Man 5. 3 software system was applicated to Meta analysis.Results In six reports of the sputum negative rate,levofloxacin group was significantly higher,and the difference was statistically significant (OR=4.21,95%CI:3.08 ~5.75,P<0.0001);adverse reactions in three articles,the difference of adverse reactions incidence had no statistical significance (OR =0.82,95%CI:0.56 ~1.18,P =0.28)between experimental group and control group, respectively.Conclusion The clinical effect of levofloxacin in treatment of refractory pulmonary tuberculosis hemoptysis is significant with a high security, which gives a guidance for clinical treatment and is worth clinical promotion.
6.Laparoscopic hepatectomy for primary liver cancer
Shuying SU ; Lin FEI ; Zuojun ZHEN
Chinese Journal of General Surgery 1997;0(04):-
Objective To evaluate laparoscopic hepatectomy for the treatment of primary liver cancer. Methods Nine patients with primary liver cancers at segment Ⅱ, Ⅲ, Ⅴ, Ⅵ and at the edge of the liver underwent laparoscopic partial hepatectomy with hand-assist devices, harmonic scalpel, and Endo-GIA. Results All operations were successful including resection of tumors involving both Ⅱ and Ⅲ segments, and irregular segmentectomy, and 2 cases with additional laparoscopic splenectomy. Surgery lasted for 80~145 min. Intraoperative bleeding was 150~700 ml, with no postoperative complications. Patients were followed-up for 5~25 months with intrahepatic tumor recurrence on 3rd, 4th and 13rd month in one each respectively. Conclusion Hand-assisted laparoscopic partial hepatectomy is a safe and feasible approach for primary liver cancer in clinically selected patients.
7.EXPRESSION OF SV40 LARGE TUMOR ANTIGEN AND FORMATION OF COMPLEXES WITH P53 AND PRb IN HUMAN BRAIN TUMORS
Xiang ZHANG ; Haining ZHEN ; Zho FEI
Medical Journal of Chinese People's Liberation Army 1981;0(06):-
SV40 large tumor antigen(Tag) expression was investigated by immunoprecipitation followed by silver staining and Western blot in 65 cases of human brain tumors and 8 cases of normal brain tissues, Tag P53 and Tag PRb complexes were screened respectively in 18 and 15 Tag positive tumor tissues. SV40 Tag was found in ependymomas, choroid plexus papillomas, pituitary adenomas, astrocytomas, meningiomas, glioblastomas multiforme and medulloblastomas, whereas 8 normal brain tissues were all negative for Tag. Tag P53 and Tag PRb complexs were detected respectively in Tag positive tumors. The results indicated that SV40 is associated with human brain tumori genesis, and the in activation of P53 and PRb due to the formation of Tag P53 and Tag PRb complexes is possibly an important mechanism in the etiopathogenesis of human brain tumors.
9.Thermal comfort and thermoregulation in manned space flight.
Zhen-Zhong YANG ; Jin-Xue FEI ; Xue-Jun YU
Chinese Journal of Applied Physiology 2013;29(6):518-524
Exposure to thermal environment is one of the main concerns for manned space exploration. By focusing on the works performed on thermoregulation at microgravity or simulated microgravity, we endeavored to review the investigation on space thermal environmental physiology. First of all, the application of medical requirements for the crew module design from normal thermal comfort to accidental thermal emergencies in a space craft will be addressed. Then, alterations in the autonomic and behavioral temperature regulation caused by the effect of weightlessness both in space flight and its simulation on the ground are also discussed. Furthermore, countermeasures like exercise training, simulated natural ventilation, encouraged drink, etc., in the protection of thermoregulation during space flight is presented. Finally, the challenge of space thermal environment physiology faced in the future is figured out.
Aerospace Medicine
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Body Temperature Regulation
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Environment
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Exercise
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Humans
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Space Flight
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Weightlessness
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Weightlessness Simulation
10.Comparison of a fractional micro-plasma radio-frequency technology versus fractional ultrapulsed CO2 laser for the treatment of atrophic acne scars
Wenli LU ; Zhen ZHANG ; Ye FEI ; Xiangdong CHEN
Chinese Journal of Dermatology 2012;45(3):165-168
Objective To compare the efficacy of and adverse reactions to a fractional micro-plasma radio-frequency technology versus fractional ultrapulsed CO2 laser in the treatment of atrophic acne scars.Methods Twenty-one patients with atrophic acne scars were enrolled in this study.Half of each subject's face was treated with micro-plasma,and the other half with fractional ultrapulsed CO2 laser,for one session.Treatments were randomly administered in a split-face manner.The efficacy and adverse effects were evaluated 6 months after the treatment by using the following outcome parameters:ECCA grading scale (échelle d' évaluation clinique des cicatrices d'acné),degrees of post-treatment pain and edema,time taken for crusting and durations of inflammatory erythema,patients' subjective rating of improvement in scar by using a quartile grading scale.A non-parametric rank test was conducted to compare the efficacy and adverse effects between the two treatments.A scar model was established on both ears of a New Zealand big ear rabbit,and treated with the CO2 laser and micro-plasma respectively,immediately followed by the resection of scar tissue for the observation of histological changes.Results A decrease of ECCAscore was observed in 90.5% (19/21) of the patients after micro-plasma treatment,and in 86.7% (18/21) after CO2 laser treatment; no significant difference was observed in the response rate (P > 0.05).In detail,ECCA score decreased by 31.5% (from 50.71 to 34.76 ) after micro-plasma treatment,compared to 29.9% (from 53.57 to 38.10) after CO2 laser treatmet.Mild edema was observed after micro-plasma treatment,with the duration of erythema and crusting being 13.95 days and 6.95 days,respectively,compared to 45.81 days and 10.10 days respectively,after CO2 laser treatment.Pigmentation occurred in none of the patients after micro-plasma treatment,but in 19% after CO2 laser treatment.Animal experiment showed that microscopic ablation zones were broad and shallow after micro-plasma treatment,but narrow and deep after CO2 laser treatment.Conclusions Fractional micro-plasma and CO2 laser are both effective for the treatment of atrophic acne scars,but the former seems to have less side effects with a lower incidence of pigmentation.