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Objective:To optimize the alcohol precipitation process of Quhan Zhufeng Granules.Methods:Taking the volume fraction of ethanol, the relative density of the concentrated solution and the standing time as the investigation factors, the Z value of the total evaluation of gentiopicrin content, oleanolic acid content and dry paste yield of gentiana macrophylla as the evaluation indexes, the star-point design-response surface method was used to optimize the alcohol precipitation process.Results:The optimal the alcohol precipitation process of Quhan Zhufeng Granules: concentration relative density 1.08 g/ml (90-95 ℃), alcohol precipitation at the end of volume fraction of 62% ethanol, standing for 16 h.Conclusion:The alcohol precipitation process using overall desirability and central composite design is stable and feasible, and has good predictability, which can provide experimental basis for further scale production.
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Objective:To observe the effect of Yunpi-Xiefei-Huatan Decoction on airway mucus hypersecretion of asthmatic rats and its regulation on epidermal growth factor receptor (EGFR) / mucin 5AC (MUC5AC) signal pathway. Methods:Seventy SD rats were randomly divided into normal group, model group, high dose group, medium dose group, low dose group, western medicine group and combined group, with 10 rats in each group. Except the normal group, the other groups were sensitized with 1 ml ovalbumin and aluminum hydroxide mixture to establish the asthma rat model. On the 16th day of the experiment, the high, medium and low dose groups were given Yunpi-Xiefei-Huatan Decoction of 40, 20, 10 g/kg, respectively, the western medicine group was given carboxymethylstein tablets of 150 mg/kg, and the combined group was given Yunpi-Xiefei-Huatan Decoction of 20 g/kg and carboxymethylstein tablets of 150 mg/kg, once a day, for 4 weeks. The levels of tumor necrosis factor (TNF-α) and interleukin-13 (IL-13) in serum of rats were detected by Enzyme-linked Immunosorbent Assay (ELISA), the total number and classification of leukocytes in BALF were observed by Wright Giemsa staining, the pathological changes of lung tissue were observed by glycogen staining (PAS). The protein expression of epidermal growth factor receptor (EGFR) and MUC5AC (MUC5AC) were detected by Western blotting, and the mRNA expression of EGFR and MUC5AC was detected by Reverse Transcriptase Polymerase Chain Reaction (RT-PCR). Results:Compared with the model group, the level of IL-13 and TNF-α in the high, medium and low dose groups of traditional Chinese medicine, western medicine group and combined group was significantly decreased ( P<0.05). The levels of WBC, eosinophils and neutrophils in rat alveolar lavage fluid were significantly decreased ( P<0.05). The expression of EGFR (0.466 ± 0.023, 0.354 ± 0.047, 0.667 ± 0.066, 0.553 ± 0.065, 0.290 ± 0.033 vs. 0.782 ± 0.047) and MUC5AC (0.424 ± 0.022, 0.313 ± 0.033, 0.603 ± 0.051, 0.495 ± 0.041, 0.243 ± 0.024 vs. 0.806 ± 0.090) significantly decreased ( P<0.05), the m RNA expression of EGFR (2.302 ± 0.321, 2.549 ± 0.623, 3.084 ± 0.453, 2.585 ± 0.314, 1.810 ± 0.379 vs. 4.101 ± 0.567), MUC5AC (3.243 ± 0.742, 3.283 ± 1.064, 4.419 ± 0.572, 3.817 ± 0.637, 2.469 ± 0.424 vs. 5.840 ± 0.661) in the high, medium and low dose groups, western medicine group and combined group was significantly decreased ( P<0.05). Conclusion:Yunpi-Xiefei-Huatan Decoction ccould inhibit asthma, and its mechanism mightbe related to the EGFR/MUC5AC signaling pathway.
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This study firstly introduced the mechanism, benefits and applications of irreversible electroporation(IRE) for tumor ablation. In addition, this study also introduced the most advanced IRE systems cleared by FDA or CFDA and IRE research equipment. The clinically licensed IRE systems include the Nanoknife 3.0 of Angiodynamics, the Dophi
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Humanos , Eletricidade , Eletroporação , Frequência Cardíaca , Neoplasias/terapiaRESUMO
OBJECTIVE: To establish the method for content determination of ligustilide and to optimize the extraction technology of volatile oil and inclusion technology in Quhan zhufeng granules. METHODS: HPLC method was adopted. The determination was performed on Waters C18 column with mobile phase consisted of methanol-water (70 ∶ 30, V/V) at the flow rate of 1 mL/min. The detection wavelength was set at 327 nm, and the column temperature was 30 ℃. The sample size was 10 μL. Using yield of volatile oil and the content of ligustilide as index, with soaking time, the amount of adding water and extraction time as factors, the extraction technology was optimized by orthogonal test. Using inclusion rate, the yield of inclusion compound and yield of volatile oil as index, with ratio of volatile oil to β-cyclodextrin, inclusion temperature and inclusion time as factors, the inclusion technology of volatile oil was optimized by orthogonal test. RESULTS: The linear range of ligustilide was 0.4-4 μg(r=0.999 9); RSDs of precision, stability and reproducibility tests were all lower than 2% (n=6). The recoveries were 96.75%-102.03%(RSD=2.06%,n=6). The optimal extraction technology of volatile oil included 10-fold water (mL/g), soaking for 15 min, extracting for 8 h. Average yield of volatile oil was 0.310 7%, and average content of ligustilide was 0.418 0 mg/g. The optimal inclusion technology of volatile oil included ratio of β-cyclodextrin and volatile oil was 1 ∶ 8 (mL/g); inclusion temperature was 50 ℃; inclusion time was 3 h. Average inclusion rate was 69.43%, and the yield of inclusion compound was 58.89%; the yield of volatile oil was 14.15%. CONCLUSIONS: Established determination method is simple, accurate and stable. The optimal extraction technology of volatile oil and inclusion technology are stable and feasible.
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Objective To investigate the effect of MMP-8 on cornea. Methods Fifteen C57BL/6J healthy mice were selected. The right eyes corneal stroma was injected by 10μL MMP-8 as the experimental group and the left eyes were injected by same amount of normal saline as the control group. At 0,4,8 h, the two-photon microscope second harmonic generation imaging technology was used to scan mice corneal stroma layer by layer in vivo. The obtained images were performed the 3D reconstruction by Imaris software and the signal intensity of the images were calculated. At 4,8 h, the corneal opacity degree was evaluated under slit lamp. At 8 h,mice were killed and corneas were collected to determine the hydroxyproline concentration. Results The cornea stromal fiber signal strengthes at 0 h in the experimental group and control group were (89.7±11.2) and (85.3±7.0),which at 4 h were (14.5±3.4) and (46.6±14. 0) respectively,which at 8 h were (11.0±4.6) and (34.6±12.5) respectively. The cornea stromal signal strength at 4,8 h in the experiemental group was significantly decreased compared with that in the control group (P<0.05) ;the cornea at 4 ,8 h in the experimental group was significantly turbid than that in the control group (P<0.05);the cornea hydroxyproline concentrations detected at 8h in the experiemental group and control group were (0.433±0. 090) μg/mg and (0. 590±0. 133) μg/mg respectively,the experimental group was significantly lower than the control group (F=7. 193,P=0. 014). Conclusion MMP-8 has obvious degradation and destroy effect on mice corneal stroma collagen,which leads to the decrease of corneal opacity.
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OBJECTIVE@#To investigate the effects of LCL161, a Smac mimetic, on the proliferation and apoptosis in hepatocellular carcinoma cells and the underlying mechanisms. @*METHODS@#The effect of LCL161 on the cell viability of HepG2 and SMMC7721 cells was measured by MTT assay. The effect of LCL161 at lower concentrations on the proliferation in hepatocellular carcinoma (HCC) cells was detected by colony formation assay. Apoptosis was assessed by flow cytometry with PI staining. The mitochondrial membrane potential was measured by JC-1 staining. The expression of PARP, p-Akt, cIAP1 and XIAP protein was analyzed by Western blot. @*RESULTS@#LCL161 displayed notable antiproliferative activity on HCC cells at the concentrations of 1-16 μmol/L (P<0.01), with IC50 values of 4.3 and 4.9 μmol/L for HepG2 and SMMC7721 cells, respectively, after treatment for 48 h. LCL161 at lower concentrations obviously inhibited the colony formation of HCC cells. LCL161 induced significant apoptosis in HCC cells (P<0.01), and resulted in the apoptotic rate at (1.5±0.8)% or (1.8±0.6)% , (15.2±2.8)% or (12.2±2.4)%, (28.7±3.0)% or (22.4±2.7)%, (34.6±2.3)% or (30.2±2.4)% for HepG2 cells or SMMC7721 cells at the concentration of 0, 2, 4 or 8 μmol/L, respectively. The result of JC-1 staining indicated that the mitochondrial membrane potential of HCC cells was reduced by LCL161. In addition, LCL161 promoted the cleavage of PARP, down-regulated the protein expression of p-Akt, and degraded cIAP1. @*CONCLUSION@#LCL161 possesses significant anti-proliferative activity and pro-apoptotic action in HepG2 and SMMC7721 cells, which might be correlated with reduction in mitochondrial membrane potential, down-regulation of p-Akt and degradation of cIAP1.
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Humanos , Apoptose , Carcinoma Hepatocelular , Tratamento Farmacológico , Genética , Patologia , Linhagem Celular Tumoral , Proliferação de Células , Sobrevivência Celular , Regulação para Baixo , Células Hep G2 , Proteínas Inibidoras de Apoptose , Metabolismo , Neoplasias Hepáticas , Potencial da Membrana Mitocondrial , Proteínas Proto-Oncogênicas c-akt , Genética , Tiazóis , Farmacologia , Ubiquitina-Proteína Ligases , Metabolismo , Proteínas Inibidoras de Apoptose Ligadas ao Cromossomo XRESUMO
Objective To investigate the changes and clinical significance of plasma N‐terminal pro‐brain natriuretic peptide (Nt‐proBNP) in patients with asphyxia of newborns combined with myocardial injury .Methods Fifty‐eight term neonates with neonatal asphyxia born within 24 h were included in our study ,18 of the 25 severe asphyxia cases were combined with myocardial injury ,15 of 33 mild asphyxia cases were with myocardial injury ,there were 25 subjects were not cardiac injured after asphyxia to‐tally .Patients of the same age without asphyxia or any cardiovascular disease were recruited into the control group .Levels of plasma Nt‐proBNP and creatine kinase isoenzyme (CK‐MB) were detected on the first ,third and the fifth days after birth for statistical a‐nalysis .Results On the first and third days after birth ,levels of plasma Nt‐proBNP in severe and mild asphyxia combined with my‐ocardial injury group were significantly higher than the groups without myocardial injury and the control (P0 .05) .On the first day after birth ,levels of Nt‐proBNP ,CK‐MB were positively corre‐lated in the mild asphyxia group (r=0 .58 ,P<0 .05) and severe asphyxia group (r=0 .47 ,P<0 .05) .On the first day after birth , the cutoff value for myocardial injury was 695 .87 fmol/mL ,the area under the ROC curve was 0 .763 ,with the sensitivity of 71 .9%and specificity of 65 .4% ,the positive predictive value was 73 .3% and negative predictive value was 63 .8% .Conclusion The Nt‐proBNP level has important clinical value for early evaluation of the severity of myocardial injury after asphyxia .
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Objective To analyze the morbidity,mortality,and hospitalization of complications in late preterm infants.Meth-ods A total of 389 late preterm infants were retrospectively analyzed.The influence of delivery mode on respiratory complications was compared.And the influence of respiratory complications on length of hospital stay,hospital expenses and mortality were ana-lyzed and compared.Results Our data showed that main complications were diagnosed as respiratory complications(23.65%).Oth-er complications were diagnosed as hypoglycemia (20.57%),feeding intolerance (1 7.48%),hyperbilirubinemia (14.91%),and in-tracranial hemorrhage (8.23%).Respiratory morbidity was significantly higher in preterm infants by caesarean section than by va-ginal delivery (32.34% vs .10.39%,P <0.05).The time and cost of hospitalization and the mortality were higher in late preterm infants accompanied by respiratory complications than those in infants without respiratory complications(P <0.05).Conclusion There were very prone to a variety of complications in late preterm infants.In order to reduce the morbidity of respiratory complica-tions in late preterm infant,indications for caesarean section must be strictly mastered.We should pay more attention to respiratory complications,which might be helpful to short the length of hospital stay,save hospitalization expenses and reduce the mortality.
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Objective:To investigate the influence of hepatocyte growth factor(HGF)on the expression of vascular endothelial growth factor C(VEGF-C)and the mechanism of HGF-induced VEGF-C expression in tongue squamous cell carcinoma Tca8113 cells.Methods:Tca8113 cells were cultured and exposed to HGF with various concentrations.The expression level of VEGF-C was assessed by ELISA.Signaling transduction inhibitors LY294002,U0126,SP600125,SB203580 was used to block PI3K/Akt,P44 /P22MAPK,JNK,P38MAPK signaling pathways,respectively.Then,the expression level of VEGF-C was detected by ELISA.Re-sults:The VEGF-C expression of Tca8113 cells increased at the beginning and decreased later with the increase of HGF concentra-tion.When the concentration of HGF was 40 ng/ml,VEGF-C expression level was the highest.Inhibitor LY294002 of PI3K/Akt and Inhibitor U0126 of P44 /P22MAPK significantly blocked the effects on HGF-induced VEGF-C up-regulation(P <0.01 ).Inhibitor SP600125 of JNK and inhibitor SB203580 of P38MAPK didn't interfere HGF-induced VEGF-C expression(P >0.05).Conclusion:HGF contributed to the expression of VEGF-C,PI3K/Akt and P44 /P22MAPK signaling pathways may be involved in HGF-induced VEGF-C up-regulation,and may play potential roles in lymphatic metastasis of oral squamous cell carcinoma.