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OBJECTIVE@#To investigate the regulatory effects of miR-26a-5p on the osteogenic differentiation of adipose-derived mesenchymal stem cells (ADSCs) by regulating cAMP response element binding protein 1 (CREB1).@*METHODS@#The adipose tissues of four 3-4 weeks old female C57BL/6 mice were collected and the cells were isolated and cultured by digestion separation method. After morphological observation and identification by flow cytometry, the 3rd-generation cells were subjected to osteogenic differentiation induction. At 0, 3, 7, and 14 days after osteogenic differentiation induction, the calcium deposition was observed by alizarin red staining, ALP activity was detected, miR- 26a-5p and CREB1 mRNA expressions were examined by real-time fluorescence quantitative PCR, and CREB1 protein and its phosphorylation (phospho-CREB1, p-CREB1) level were measured by Western blot. After the binding sites between miR-26a-5p and CREB1 was predicted by the starBase database, HEK-293T cells were used to conduct a dual-luciferase reporter gene experiment to verify the targeting relationship (represented as luciferase activity after 48 hours of culture). Finally, miR-26a-p inhibitor (experimental group) and the corresponding negative control (control group) were transfected into ADSCs. Alizarin red staining, ALP activity, real-time fluorescent quantitative PCR (miR-26a-5p) and Western blot [CREB1, p-CREB1, Runt-related transcription factor 2 (RUNX2), and osteocalcin (OCN)] were performed at 7 and 14 days after osteogenic induction culture.@*RESULTS@#The cultured cells were identified as ADSCs. With the prolongation of osteogenic induction culture, the number of calcified nodules and ALP activity significantly increased ( P<0.05). The relative expression of miR-26a-5p in the cells gradually decreased, while the relative expressions of CREB1 mRNA and protein, as well as the relative expression of p-CREB1 protein were increased. The differences were significant between 7, 14 days and 0 day ( P<0.05). There was no significant difference in p-CREB1/CREB1 between different time points ( P>0.05). The starBase database predicted that miR-26a-5p and CREB1 had targeted binding sequences, and the dual-luciferase reporter gene experiment revealed that overexpression of miR-26a-5p significantly suppressed CREB1 wild-type luciferase activity ( P<0.05). After 7 and 14 days of osteogenic induction, compared with the control group, the number of calcified nodules, ALP activity, and relative expressions of CREB1, p-CREB1, OCN, and RUNX2 proteins in the experimental group significantly increased ( P<0.05). There was no significant difference in p-CREB1/CREB1 between the two groups ( P>0.05).@*CONCLUSION@#Knocking down miR-26a-5p promoted the osteogenic differentiation of ADSCs by up-regulating CREB1 and its phosphorylation.
Subject(s)
Animals , Female , Mice , Cell Differentiation , Cells, Cultured , Core Binding Factor Alpha 1 Subunit/metabolism , Cyclic AMP Response Element-Binding Protein/metabolism , Mesenchymal Stem Cells , Mice, Inbred C57BL , MicroRNAs/metabolism , Osteocalcin/metabolism , Osteogenesis/genetics , RNA, Messenger/geneticsABSTRACT
Objective:To investigate the effect of transcatheter arterial chemoembolization (TACE) combined with ultrasound-guided radiofrequency ablation (RFA) on the efficacy and immune function in patients with primary liver cancer.Methods:The clinical data of 152 patients with primary liver cancer from February 2019 to February 2021 in the Second Affiliated Hospital of Xi′an Jiaotong University were retrospectively analyzed. Among them, 76 patients were treated with TACE combined with RFA (combined group), and 76 patients were treated with TACE (control group). The efficacy was compared; the α-L fucosidase, T lymphocyte subsets (CD 3, CD 4, CD 8 and CD 4/CD 8), B lymphocyte subsets (CD 19) and tumor markers (alpha-fetoprotein, AFP; carcinoembryonic antigen, CEA; carbohydrate antigen 125, CA125) before treatment and 1 month after treatment were detected. Results:The total clinical effective rate in combined group was significantly higher than that in control group: 81.58% (62/76) vs. 52.63% (40/76), and there was statistical difference ( χ2 = 4.54, P<0.05). There were no statistical difference in all indexes before treatment between 2 groups ( P>0.05); the α-L fucosidase, AFP and CD 8 1 month after treatment in combined group were significantly lower than those in control group: (18.06 ± 5.33) U/L vs. (26.58 ± 7.75) U/L, (87.93 ± 22.55) μg/L vs. (146.83 ± 21.85) μg/L and 0.295 ± 0.052 vs. 0.367 ± 0.064, the CD 3, CD 4 and CD 4/CD 8 were significantly higher than those in control group (0.489 ± 0.054 vs. 0.462 ± 0.063, 0.363 ± 0.059 vs. 0.303 ± 0.075 and 1.43 ± 0.27 vs. 0.89 ± 0.14), and there were statistical differences ( P<0.01 or<0.05); there was no statistical difference in CEA, CA125 and CD 19 1 month after treatment between 2 groups ( P>0.05). Conclusions:TACE combined with RFA in the treatment of primary liver cancer patients can not only improve the total clinical effective rate, but also significantly improve the immune function, and help to reduce level of the liver tumor marker of AFP.
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Objective:To develope a self-adjustable automatic planning method of intensity modulated radiotherapy based on predicted dose, in order to enhance the robustness of automatic planning.Methods:After the patients′ dose by 3D U-Res-Net_B network was predicted, the current dose was calculated based on the last iteration result, then the predicted dose was combined to calculate the target dose and optimized. With all iterations completed or exit conditions satisfied, final treatment plannings would be acquired. A total of 30 cases of rectal cancer were tested to verify the effectiveness of the algorithm.Results:The mean value of planning target volumes′ V100% was (95.03±0.91)% for clinical plans, close to (94.67±1.96)% for automatical plans( P>0.05), and better than (92.90±2.13)% for predicted dose with the statisically significant difference ( t=29.0, P<0.05). Automatic planning′s indexes such as V35 of small intestines, V40 of bladders and V20 - V40 of femoral heads were lower than predicted and clinical ones, with the statisically significant difference( t=4.5-118.0, P<0.05). Discrepancy in other indexes of organs at risk was not statistically significantly different( P>0.05). Conclusions:This method made automatic planning processes more robust and more adaptive to difficult clinical situations.
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Objective:To observe the hemodynamic change and inhibitory effect on stress response of oxycodone during induction, maintenance and resuscitation period of general anesthesia in patients undergoing gastric cancer surgery.Methods:60 patients with general anesthesia undergoing elective gastric cancer from March 2018-March 2019 in No.904 Hospital were selected. According to the random number table method, they were randomly divided into two groups ( n=30): Oxycodone group (group O) and Fentanyl group (group F). The anesthesia was induced by intravenous oxycodone 0.3 mg/kg (group O) or fentanyl 3 μg/kg (group F), propofol 2 mg/kg and cisatracurium 0.25 mg/kg. Oxycodone (group O) or fentanyl (group F) with sevoflurane were used for maintenance of general anesthesia. The mean arterial pressure (MAP) and heart rate (HR) were recorded before anesthesia induction (T 0), immediately intubation (T 1), 1 min (T 2) and 5 min (T 3) after intubation, immediately skin incision (T 4), surgery for 1 h (T 5), the end of operation (T 6) and 1 min (T 7) and 5 min (T 8) after extubation. The concentration of glucose (Glu), cortisol (Cor), epinephrine (E) and norepinephrine (NE) were detected in time T 0,T 2, T 4,T 5 and T 7. The time of extubation and the visual analogue scale (VAS) scores after extubation and 30 min after entering postanesthesia care unit (PACU) were recorded. The incidence of cough during induction, dysphoria during extubation and the adverse effects such as nausea, vomit and respiratory depression during PACU were also observed. Results:There were no significant difference between two groups in MAP and HR at the time point from T 0 to T 8 ( P>0.05); there were no significant difference about concentrations of Glu, Cor, E and NE at the time point of T 0,T 2,T 4,T 5 and T 7 ( P>0.05). Extubation time and VAS scores of 30 min after entering PACU in group O were significantly lower than those of group F ( P<0.05). Meanwhile, the incidences of cough during induction, dysphoria during extubation and respiratory depression in group O were significantly lower than in group F ( P<0.05). There was no significant difference in the incidence of nausea and vomiting ( P>0.05). Conclusions:Oxycodone can be safely and effectively used for induction and maintenance of general anesthesia in gastric cancer surgery, inhibit intraoperative stress response, and improve the quality of recovery period.
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Objective To investigate the metabolism of strychnine (STN) and the metabolic interaction between STN and glycyrthetic acid (GA) in vitro.MethodsHuman liver microsomes (HLM) and human recombinant cytochrome P450 (CYP) isoforms were employed to study the metabolism of STN and the metabolic interaction of STN with GA in vitro.ResultsIn HLM,the Km,Vmax,and clearance of STN were 88.50 μmol/L,0.88 nmol/(mg·min),and 9.93 mL/(mg·min),respectively.STN was metabolized mainly by CYP3A4.However,STN noncompetitively inhibited CYP3A4-catalyzed testosterone 6β-hydroxylation with IC50 value of 5.9 μtmol/L and Ki value of 5.5μmol/L.Moreover,GA competitively inhibited STN metabolism with IC5o value of 10.6 μmol/L and Ki value of 17.7 μmol/L.ConclusionAlthough STN is mainly metabolized by CYP3A4 in vitro,STN has noncompetitive inhibition on CYP3A4-catalyzed testosterone 6β-hydroxylation.Moreover,GA could competitively inhibit STN metabolism.The present work is helpful to elucidate the metabolic interaction between STN and GA.
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Abstract: The activities of four CYP450 enzymes (CYP3A, 1A2, 2El and 2C) and the mRNA expression levels of CYP1A2, 2El, 2Cll and 3A1 in rat liver were determined after Wistar rats were orally administered with brucine (BR) at three dosage levels (3, 15 and 60 mg.kg-1 per day) and the high dose of BR combined with glycyrrhetinic acid (GA, 25 mg.kg-1 per day) or liquiritin (LQ, 20 mg.kg-1 per day) for 7 consecutive days. Compared with the control, brucine caused 24.5% and 34.6% decrease of CYP3A-associated testosterone 6beta-hydroxylation (6betaTesto-OH) and CYP2C-associated tolbutamide hydroxylation (Tol-OH), respectively, and 146.1% increase of CYP2El-associated para-nitrophenol hydroxylation (PNP-OH) at the high dose level. On the other hand, (BR+GA) caused 51.4% and 33.5% decrease, respectively, of CYP2El-associated PNP-OH and CYP1A2-associated ethoxyresorufin-O-de-ethylation (EROD) as compared with the high dose of BR group. Meanwhile, (BR+LQ) caused 41.1% decrease of CYP2El-associated PNP-OH and 37.7% increase of CYP2C-associated Tol-OH. The results indicated that the co-administration of BR with GA or LQ had effect on mRNA expression and activities of the CYP450 enzymes mentioned above to some extent, and the in vivo antagonism of LQ on BR-induced CYPs adverse effects and the in vivo inhibitory action of GA on CYP2E1 and 1A2 might play an important role in the detoxification of Radix Glycyrrhizae against Strychnos nux-vomica L.
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Jaundice is frequent clinic symptoms with high incidence especially among the critical patients. The sepsis complicated with jaundice has drawn more and more consideration. However, the pathogenesis about sepsis complicated with jaundice is not confirmed and it is sometimes identified incorrectly as cholestatic jaundice. This article summarises the pathogenesis and elucidate the management about sepsis complicated with jaundice.
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Objective:To compare the effects of different methods through applying five teaching methods in kidney disease teaching.Method:29 students of seven-year-training project of Grade 2001 of Nanjing Medical University are teaching objects.Results: Clinical thinking training and case report teaching get better general evaluations.Conclusion:Using different methods of teaching in different courses can get better effects.