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Objective To investigate the effects of harmine(HM)on the expression level of mitochondrion fusion related proteins and mitochondrial function injury in PC 12 cells.Methods PC 12 cells were divided into cell control group,HM group,mitochondrion mitosis inhibitor Mdivi-1 group,HM+Mdivi-1 group,mitochondrion fission agonist WY14643 group,HM+WY14643 group,with drug concentrations of 1,10,25,50,100 μmol·L-1.After 24 h treatment,the MTT method was used to detect the cell survival rate,and a microscope was used to observe the cell morphology,MitoTracker Red probe staining was used to observe the mitochondrial morphology and the length ratio of vertical and horizontal axes,JC-1 staining was used to detect the mitochondrial membrane potential,and a kit was used to detect ATP level and lactate dehydrogenase(LDH)activity.Immunofluorescence staining and Western blotting were used to assess the expression levels of caspase-3,apoptosis-promoting protein(Bax)cytochrome C(cyt-c),mitochondrial fusion protein(Mfn2)and mitochondrial mitotic protein(Drp-1).The interference sequence of Drp1 was transfected by electroporation,and the siRNA sequence with good transfection effect was screened.The related indicators were detected by fluorescence method,MTT method,and immunoblotting method in cooperation with drug intervention.Results MTT results showed that compared with the cell control group,the survival rate of HM group,Mdivi-1 group,HM+Mdivi-1 group,WY14643 group and HM+WY14643 group decreased significantly(P＜0.01),and the EC50 were(11.48±2.32),(12.35±1.67),(14.88±2.07),(39.14±3.25),(20.09±1.97),respectively.According to this,subsequent experiments selected 20 μmol·L-1for HM,WY 14643 and HM+WY14643 as working concentrations to construct PC 12 cell model.Microscopic observation and MitoTracker Red probe staining showed that the cell density in the drug group decreased in varying degrees,and a transition from branched to round morphology in the drug-treated groups was observed.The morphology of mitochondria tended to be round,and the ratio of the length of the longitudinal axis to transverse axis was(3.33±0.72)in the cell control group,(2.19±0.58)in the HM group,(2.45±0.44)in Mdivi-1 group,and(1.43±0.62)in HM+Mdivi-1 group,respectively.The results of JC-1 staining showed that compared with the cell control group,the mitochondrial mode potential of the HM group significantly decreased(P＜0.01).ROS significantly increased(P＜0.01)and ATP levels decreased(P＜0.01),and LDH enzyme activity increased(P＜0.01).Immunofluorescence staining and Western blotting results showed that compared with the cell control group,the expression levels of proapoptotic proteins Bax,cytochrome C,and caspase-3 in the HM group were significantly increased(all P＜0.01).Compared with the cell control group,the expression level of mitochondrial fission related protein Drp1 in HM group was significantly higher(P＜0.01).The expression level of mitochondrial fusion related protein Mfn2 significantly decreased(P＜0.01).After specific interference with Drp1 and synergistic intervention with HM,the survival rate of PC 12 cells in each interference group decreased compared to each drug intervention group.The expression of Drp1 and Mfn2 was downregulated,and the differences were statistically significant(P＜0.05 or P＜0.01).Conclusion HM can reduce the mitochoudrial membrane potential and ATP levels by accumulating ROS,there by activating the caspase-3 apoptosis pathway and promoting cell apoptosis.Mitochondrial fusion division may be involved in the damage of PC12 cells caused by HM,initiating apoptosis through the mitochondrial pathway.
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Objective To evaluate the effect of global end diastolic volume index (GEDVI) on fluid resuscitation in elderly patients with septic shock. Methods A prospective randomized controlled trial (RCT) was conducted. Septic shock patients over 65 years admitted to intensive care unit (ICU) of Shandong Province, Zibo Central Hospital from January 2013 to December 2015 were enrolled. The patients were randomly divided into control group and observation group, 20 cases in each group. In accordance with the guidelines for the treatment of septic shock, early goal-directed therapy (EGDT), rehydration in the control group was treated with the guide of central venous pressure (CVP); observation group was received pulse indicator continuous cardiac output (PiCCO) monitoring, and rehydration was treated according to the GEDVI and extravascular lung water index (EVLWI), i.e. GEDVI was maintained in 650-800 mL/m2, EVLWI was not obviously increased compared with the basic value and without the emphasis of CVP. Initial acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ ) score, sequential organ failure score (SOFA), procalcitonin (PCT), mean arterial pressure (MAP), lactic acid (Lac) and oxygenation index (PaO2/FiO2); serial Lac, central venous to arterial carbon dioxide pressure (Pcv-aCO2), fluid balance, the amount of noradrenaline accumulation, PaO2/FiO2 after 6, 24 and 48 hours resuscitation; mechanical ventilation time, incidence of acute heart failure, ICU hospitalization time and 28-day mortality were recorded. Results There was no significant difference in gender,age, APACHE Ⅱ score, SOFA score, primary disease, infection site and basal PCT, MAP, Lac, PaO2/FiO2 between the two groups. Compared with the control group, 6 hours Lac, Pcv-aCO2 recovery, positive fluid balance, noradrenaline accumulation and PaO2/FiO2 of the observation group showed no significant difference; positive fluid balance 24 hours in the recovery was significantly reduced (mL: 2919.80±986.44 vs. 3991.40±933.53), Pcv-aCO2 significantly decreased [mmHg (1 mmHg = 0.133 kPa): 5.55±1.43 vs. 7.10±2.38], PaO2/FiO2 significantly improved (mmHg: 194.80±28.57 vs. 177.65±23.46), and noradrenaline accumulation was increased significantly (mg: 40.99±20.69 vs. 27.31±19.34) with statistically significant difference (all P < 0.05); the blood level of Lac 48 hours in the recovery was significantly decreased (mmol/L: 1.16±0.89 vs. 1.85±1.01), Pcv-aCO2 (mmHg: 5.35±1.18 vs. 6.70±2.34), and PaO2/FiO2 (mmHg: 215.75±33.84 vs. 190.60±32.89) were further improved, the positive fluid balance was significantly reduced (mL: 3141.55±1245.69 vs. 4533.85±1416.67, all P < 0.05). Compared with the control group, mechanical ventilation time (days: 3.65±1.31 vs. 4.50±1.19), ICU hospitalization time (days: 5.80±1.67 vs. 7.15±2.30) was significantly shorter in the observation group (both P < 0.05), acute heart failure rate was decreased significantly (5.0% vs. 30.0%, P < 0.05), but the 28-day mortality showed no statistical significance (25.0% vs. 40.0%, P = 0.311). Conclusions Compared to the conventional EGDT methods, fluid resuscitation under the guidance of GEDVI in elderly patients with septic shock with less liquid loading, can achieve better oxygenation and reduce heart failure, shorten the duration of mechanical ventilation and ICU stay, and play an important significant guidance for elderly patients' fluid resuscitation with septic shock.
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Objective To propose an accurate method of noninvasive determination of central venous pressure(CVP ) by locating the central point of right atrium (RA ) using echocardiography .Methods Through the 3D reconstruction ,the accurate positions of RA of 30 patients who had been examined by multislice 3‐dimensional computed tomography for chest imaging were recorded .Based on solid geometric principles ,the central point in RA was located by echocardiography and then compared with CT‐location point .The accuracy and feasibility were assessed by absolute distance (Da) ,vertical distance (Dv) and the whole time of location (T) between the two points .Results Mean Da ,Dv and T of the whole subjects were 07.6cm(95% CI:06.2to08.1cm),01.6cm(95% CI:-00.2to03.4cm),and438.0s(95% CI:400.1to 47 4.0 s) ,respectively .Conclusions The echocardiographic method on the basis of solid geometry proposed in this study could be used to locate the central point in RA accurately and simply .Thus it would be helpful to improve the accuracy of noninvasive determination of central venous pressure .
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Objective To determine the central venous pressure (CVP) noninvasively based on hemodynamics principles using ultrasound location of the collapse point of the internal jugular vein.Methods Forty patients were enrolled in this study.The collapse point of the internal jugular vein was located and marked by a linear transducer,the body mark of right atrium was marked on the right lateral wall of the chest.The noninvasive CVP was calculated according to the vertical distance between those two points.The invasive CVP determination by central venous catheter was also carried out on all the patients.Correlation analysis was used to compare the invasive and noninvasive methods.With invasive determination of CVP as the gold standard,the ROC curve of the noninvasive ultrasound method was sketched to explore the optimal cut-off points.Results The correlation analysis reveal high positive correlation between CVPs determined by ultrasound imaging and central venous catheter (r =0.906,P ＜0.01).By the ROC curve test,fluid column height of 10.75 cm by ultrasound method was determined as the cut-off point,with the sensitivity and specificity of diagnosing elevation of CVP being 88.9％ and 93.5 ％ respectively.The corresponding area under the curve was 0.971.Conclusions Ultrasound imaging could be used to determine CVP noninvasively,which would be helpful in diagnosis of the circulating load of patients.
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@#Objective To explore the anti-epileptic effect and mechanism of Shenpu Decoction.MethodsThe pilocarpine-induced epilepsy model was adopted. 50 rats were randomly divided into the Shenpu Decoction control group (n=8), blank control group (n=8), Shenpu Decoction plus pilocarpine group (n=8) and pilocarpine-induced epilepsy model group (n=26), the last group was re-divided into the Shenpu Decoction treatment group and model control group. The incidence of convulsion and the content of free amino acids in brain tissues were observed.ResultsThe incidence of convulsion was 0 in the Shenpu Decoction plus pilocarpine group and 88.5% in the pilocarpine-induced epilepsy model group, there was a significant difference between two groups ( P<0.05). The content of glutamic acid in brain tissue of the Shenpu Decoction plus pilocarpine group and pilocarpine-induced epilepsy model group higher than that of the blank control group ( P<0.05～0.01). The content of gamma-aminobutyric acid in brain tissue of the Shenpu Decoction plus pilocarpine group and Shenpu Decoction treatment group was higher than that of the model control group ( P<0.05).ConclusionShenpu Decoction can effectively prevent and terminate the episodes of convulsion.
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AIM: To compare the bioequivalence of glimepiride tables and capsules with a single dose. METHODS: Twenty Chinese healthy male volunteers were enrolled in a randomized crossover study with a single oral dose 3 mg of two formulations respectively. The blood drug concentration in serum was measured by HPLC, and the pharmacokinetic parameters were calculated by 3p97 software and compared by two one side t test. RESULTS: The parameters of the tables and the capsules were( 1.23 ? 0.19 ) and ( 1.31 ? 0.22 ) mg?h?L -1 at AUC (0-t) ,( 1.32 ? 0.20 ) and ( 1.45 ? 0.24 ) mg?h?L -1 at AUC (0-inf) ,( 0.30 ? 0.05 ) and ( 0.30 ? 0.06 ) mg?L -1 at C max ,( 6.6 ? 2.5 ) and ( 9.3 ? 7.9 ) h the peak time (T peak ), respectively. There were no significant differences between the two formulations. F was 96.4 ? 21.1 calculated by AUC (0-t) and 96.4 ? 21.1 by AUC (0-inf) . CONCLUSION: Glimepiride tables and capsules are of bioequivalence.
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Aim To compare the relative bioavailabilities of domestic acyclovir chewabletablets and normal tablets. Methods A single oral dose of 800 mg acyclovir chewable tablets or normal tablets was given to 8 volunteers respectively in a randomized, cross-over study, acyclovir serum concentration was determined by high performance liquid chromatography. Results The AUC of chewable and normal tablets was respectively 0.42 and 0.40 μg · ml · h-1, and the relative bioavailability of a cyclovir chewable tablets was (105.8 + 13.1) %. Conclusion The chewable tablets and reference tablets is bioequivalent in AUC.
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AIM: To verify the bioequivalence between sustained released tablet of nepopam and normal one. METHODS: 18 volunteers were randomly devided into two groups. Double periodical crossed design was used, and poly dose of nefopam was administered to 18 volunteers following single dose after one week interval. The concentration of nefopam hydrochloride in serum was determinated by HPLC, and the related parameters came out through 3p97 programme. RESULTS: In the single dose test the drug concentration of sustained released tablet maitained 2040 mg?L -1 for 10 h ,c max was ( 45.8 ?15.7) mg?L -1 ,t peak was ( 3.4 ? 0.8) h , and the corresponding parameters of normal tablet were over 20 mg?L -1 for 7.5 h ,( 72.7 ?26.0) mg?L -1 ,and ( 1.6 ? 0.6) h . The AUC was ( 363.4 ? 107.1 ) and ( 374.8 ?125.7) mg?h?L -1 respectively, and F was ( 1.02 ? 0.25 ). In the poly dose test the c max of sustained released and normal one was ( 31.50 ? 12.65 ) and ( 33.68 ?10.51) mg?L -1 ,c min was ( 13.4 ? 4.4 ) and ( 10.9 ?5.4) mg?L -1 , t peak was ( 2.6 ? 0.6 ) and ( 1.22 ? 0.46) h , and FI was ( 0.77 ? 0.26 ) and ( 1.04 ? 0.18 ) respectively. CONCLUSION: The sustained released tablet is credible and the two types of tablet are equieffective in AUC.