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Objective The objective of this study was to investigate the proliferation,autophagy and the potential mechanism of Ubiquitin-like with PHD and ring finger domains 1(UHRF1)in lung adenocarcinoma cells. Methods The expression of UHRF1 in lung adenocarcinoma tissues was determined by the bioinformatics website(TCGA). The expression of UHRF1 in lung adenocarci-noma cell lines(PC-9,A549 and H1299)and human bronchial epithelial cells(16HBE)was detected by qRT-PCR and Western blot. After transfection of UHRF1-shRNA,CCK-8,clone formation and ki67 were performed to detect the changes in the prolifera-tive capacity of lung adenocarcinoma A549 cells. Western blot was used to detect the changes of autophagy-associated proteins(LC3-I/II and Beclin-1)and proliferation-related proteins(CDK6,Rb and PCNA). Transmission electron microscopy was used to ob-serve the effect of UHRF1 on autophagosomes in A549 cells. Results The expression of UHRF1 in lung adenocarcinoma tissues was significantly higher than that in adjacent tissues. Compared with normal bronchial epithelial 16HBE cells,the mRNA and protein levels of UHRF1 in lung adenocarcinoma A549 and H1299 cells were significantly increased. In addition,CCK-8 assay and colony forma-tion experiments showed that silencing UHRF1 reduced the growth of A549 cells. Ki-67 immunofluorescence staining showed that the proliferation ability of A549 cells after knocking out UHRF1 was significantly lower than that in the normal control group. Further-more,knockdown of UHRF1 resulted in an increased expression of CKD6 and PCNA proteins in comparison with the control-siRNA group. The expression of Rb protein was down-regulated in the UHRF1-siRNA group. Silencing UHRF1 increased the ratio of LC3-II/LC3-I, induced up -regulation of Beclin -1 expression and promoted the formation of autophagic bodies in A549 cells. Conclusion UHRF1 is highly expressed in lung adenocarcinoma,and silencing UHRF1 can inhibit proliferation. This effect may be regulated by promoting autophagy.
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Objective To explore the clinical significance of interstitial chemotherapy applied in breast cancer.Methods Sixty breast cancer patients with modified radical mastectomy were divided into treatment group (40 cases) and control group (20 cases) according to whether using fluorouracil implants.The control group was given conventional treatment.In treatment group,distilled water washed the wound after conventional treatment,and fluorouracil implants (0.4 g) was placed below local tumor,between pectoralis major and pectoralis minor,below 0.5 cm axillary vessel.The bone marrow suppression,liver and renal function,gastrointestinal reaction,skin flap necrosis,subcutaeous hydrops,local recurrence were observed in the 2 groups.Results There were no significant differences in the rates of bone marrow suppression,liver and renal function abnormal,skin flap necrosis,subcutaeous hydrops and gastrointestinal reaction between treatment group and control group (P ＞ 0.05).The rate of local recurrence in treatment group was significantly lower than that in control group [0 vs.20.0％ (4/20)],there was statistical difference (P ＜0.05).Conclusion Intraoperative application of fluorouracil implants is safe and effective in modified radical mastectomy of breast cancer.
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<p><b>OBJECTIVE</b>To evaluate the therapeutic effects of epidermal growth factor (EGF) combined with plasma cryoprecipitate (CRYO) on the corneal injury induced by paraquat (PQ).</p><p><b>METHODS</b>According to the "Toxicological test methods of pesticides for registration" (GB 15670-1995), the conjunctival sacs of 18 health New Zealand rabbits were exposed to 100 µl 20% PQ, which were randomly divided into EGF, CRYO and EGF plus CRYO groups. The routine treatments (normal saline washing and antibiotic eyedrops) were administrated to the injured eyes of 3 groups, at the same time the left eyes of 3 groups were treated with EGF, CRYO and EGF plus CRYO, respectively. The injury of conjunctival, iris and corneal, fluorescent stranded and pathology changes of corneal were observed. The injury score was calculated and the recovery time of corneal injury was recorded.</p><p><b>RESULTS</b>The recovery time of corneal injury in EGF and EGF plus CRYO groups were 19.50 ± 3.08 and 18.67 ± 2.73 days, respectively which were significantly lower than those (27.33 ± 2.58 and 26.83 ± 3.13 days) in corresponding routine treatment controls (P < 0.05).</p><p><b>CONCLUSION</b>EGF and EGF plus CRYO could be used to treat the corneal injury induced by paraquat.</p>
Subject(s)
Animals , Rabbits , Blood Transfusion, Autologous , Cornea , Corneal Injuries , Epidermal Growth Factor , Therapeutic Uses , Eye Injuries , Drug Therapy , Factor VIII , Therapeutic Uses , Fibrinogen , Therapeutic Uses , Ophthalmic Solutions , Paraquat , Plasma , Treatment Outcome , Wound HealingABSTRACT
ObjectiveTo investigate the effect of dexmedetomidine on tracheal extubation response in patients undergoing thyroid surgery after general anesthesia.Methods Forty-two patients who ASA classification Ⅰ - Ⅱ,were performed thyroid surgery under total intravenous anesthesia.They were divided into dexmedetomidine group and control group by random number table with 21 cases each.Both groups patients anesthesia induction and maintenance with the same methods,dexmedetomidine group patients at 15 min before anesthesia induction with venous infusion dexmedetomidine 0.6 μ g/kg(finished 10 min infusion),followed by 0.4 μ g/(kg · h) continuous infusion,control group patients with pump equivalent in 0.9％ sodium chloride injection in the same way.Systolic blood pressure (SBP),diastolic blood pressure (DBP) and heart rate (HR),surgery time,recovery time,extubation time,orientation recovery time,modified Aldrete score ≥9 scores time,rate of recovery from restlessness,tolerance score and recovery from any adverse effects were recorded.ResultsThere were no significant differences between two groups in surgery time,recovery time,extubation time,orientation recovery time,modified Aldrete score ≥ 9 scores time (P ＞0.05).SBP,DBP,HR at immediately and after extubation with 1,3,5 min of dexmedetomidine group were lower than those of control group(P ＜ 0.05 or ＜ 0.01 ).Tolerance score excellent rate of dexmedetomidine group was higher than that of control group[95.2％ (20/21) vs.28.6％ (6/21)] (P ＜ 0.05),the rate of recovery from restlessness of dexmedetomidine group was lower than that of control group[0 vs.28.6％(6/21 ) ] (P ＜ 0.05).ConclusionBefore anesthesia induction in patients with venous infusion dexmedetomidine 0.6 p g/kg(finished 10 min infusion),followed by 0.4 p g/(kg·h) continuous infusion can effectively reduce the patients' cardiovascular response to extubation,strengthen the patients' tolerance to endotracheal tube,reduce the patients' agitation and recover consciousness comfortable.
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Radioactive 125I interstitial brachytherapy is easy to cause the intestinal tract tissue damage.Its mechanism possibly correlates to the quick renewal speed of the intestinal tract tissue and a majority of ceils in M, G2 and late S phases. The tolerant dose of intestinal tract tissue is approximately 160 Gy. If the absorbed dose is higher than the tolerant one, the intestinal tract tissue will be damaged. In the clinical practice of radio-active 125I interstitial brachytherapy, through the strict plan of the TPS system before the operation, by using ul-trasound, CT, MR/and other imaging technologies in the operating process and establishing the suitable barrier protection between the seeds and intestinal tract tissue, the radioactive damage of the intestinal tract tissue my be reduced.
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We developed a novel method for constructing nearly random peptide library. Genomic DNAs extracted from tissue or cells of large genome species were digested with frequent cutter to produce short DNA fragments. These short fragments can be considered nearly random. Nearly random peptide libraries can be constructed by cloning the short fragments into appropriate expression vectors and transformation into host cells. Genomic DNA from one species can be digested with different restriction enzymes and ligated to different reading frames to produce several different libraries. In this study, we digested tobacco genomic DNA with two enzymes and cloned into three different reading frames to make totally six nearly random peptide libraries.
Subject(s)
DNA, Plant , Genetics , Genome, Plant , Genetics , Peptide Library , Nicotiana , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To deduce all potential ligands undiscovered experimentally by searching all the proteins containing same C-termini, which can bind a certain PDZ domain.</p><p><b>METHODS</b>We developed a JAVA program for searching short exact sequence matches at C-terminus. According to the known C-termini, which PDZ domains recognized experimentally, Swissprot database has been searched by this program for all potential ligands.</p><p><b>RESULTS</b>Some PDZ domains may have more potential ligand proteins, which are undiscovered yet experimentally. These bioinformatic results also provide clues for studying functions of hypothetical proteins and PDZ domains' protein interactions in many different organisms.</p><p><b>CONCLUSION</b>The results may provide useful clues for discovering potential functions of hypothetical proteins and new functions of known proteins.</p>