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Objective:To investigate the effect of Stigma Maydis Palysaccharide(SMPS)on ATP synthesis in kidney mitochondria of D-galactose-induced aging mice, and to clarify its possible mechanism.Methods:The aging mouse model was established by subcutaneous injection of D-galactose solution in the back of the neck.The 48 SPF male mice were randomly divided into normal control group(control group), D-galactose model group(D-Gal group), SMPS low-dose group and SMPS high-dose group(n=12 for each). The control group was subcutaneously injected with 150 mg/kg normal saline on the back of the neck every day, while the other three groups were subcutaneously injected with 150 mg/kg of D-gal solution on the back of the neck every day.SMPS-L and-H dose groups were given 30 mg/kg and 60 mg/kg of SMPS solution by gavage at the same day of D-Gal injection.The control group and D-GAL group were given the same volume of normal saline daily by gavage for 42 days.Blood samples were collected from the eyeball under general anesthesia after 42 days of intervention for the detection of serum levels of superoxide dismutase(SOD), glutathione peroxidase(GSH-Px)and MDA.After harvesting the kidney tissue, various tests were used to detect ATP content, the mRNA expression levels and protein expression levels in kidney.Luciferase assay was used to detect ATP content in renal tissue.Real-time fluorescent quantitative PCR was used to detect the mRNA expression levels of succinate dehydrogenase(SDH)of complex Ⅱ, cytochrome C reductase(Cycs)of complex Ⅲ, complex Ⅳ(COXⅣ)and ATP5b in ATP synthase in mitochondrial oxidative respiratory chain.Western blot was used to detect the expression levels of mitochondrial fusion protein 2(MFN2), dynamin-related protein1(DRP1)and mitochondrial autophagy related protein P62 in renal tissues of each group.Results:Compared with control group, the activities of serum of SOD(116.53±10.01)U/mg and GSH-Px(127.58±8.74)μmol/L were significantly decreased in D-GAL group(both P< 0.01), and serum MDA content(15.42±0.91)μmol/L increased significantly in D-GAL group( P<0.01). Compared with D-GAL group, the activities of SOD(152.80±9.29)U/mg and GSH-Px(274.07±10.73)μmol/L were significantly increased in SMPS intervention group( P< 0.01), while the MDA content(8.10±0.66)μmol/L decreased significantly in SMPS intervention group( P< 0.01). Compared with control group, the content of ATP(178±4)10 -4 μmol in D-gal group was significantly decreased( P<0.01), the mRNA expression levels of SDH, Cycs and COXⅣ were not significantly changed in D-gal group, and the mRNA expression level of ATP5b(0.67±0.01)was down-regulated in D-gal group( P<0.01), the expression of MFN2 protein(0.29±0.02)was significantly decreased in D-gal group( P<0.01), and the expression of DRP1 and P62 protein(0.31±0.02 and 0.21±0.01)was significantly increased in D-gal group(both P<0.01). Compared with the D-gal group, the ATP content(193±1)10 -4 μmol in the kidney tissue of the mice was significantly increased in SMPS intervention group( P< 0.01), and the ATP5b mRNA expression and MFN2 protein expression(0.87±0.05 and 0.71±0.08)were significantly increased in SMPS intervention group(both P< 0.01), DRP1 and P62 protein expressions(0.20±0.01 and 0.10±0.01)were significantly down-regulated in in SMPS intervention group(both P< 0.01). Conclusions:SMPS can improve the mitochondrial dynamic homeostasis disorder in aging mice by increasing the activity of antioxidant enzymes, up-regulating the expression of ATP5b mRNA and MFN2 protein, down-regulating the expression of DRP1 and P62 protein, and promoting the generation of mitochondrial ATP in D-gal-induced aging mice kidney tissue.
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Objective:To investigate the effects of pressure gradient controlled carbon dioxide (CO 2) pneumoperitoneum establishment in patients with gynecological laparoscopic surgery on early circulatory and respiratory function. Methods:From November 1, 2018 to March 31,2019,100 case of gynecological laparoscopic surgery who were scheduled to undergo elective surgery in Jiangsu Hospital of Traditional Chinese Medicine were enrolled and divided into experimental group(50 cases) and control group(50 cases) by random number table method. The experimental group used pressure gradient control method to establish CO 2 pneumoperitoneum, that is, the pneumoperitoneum pressure was set to 5, 9, 12 mmHg(1 mmHg=0.133 kPa) gradually rising three gradients, and after reaching the corresponding gradient, they were maintained for 1 minute to 12 mmHg; the control group used conventional method, that was, the pneumoperitoneum pressure was set directly to 12 mmHg, and began to inflate until reaching the preset pressure. Systolic blood pressure (SBP), diastolic blood pressure (DBP), mean arterial pressure (MAP), heart rate (HR), end-of-breath partial pressure of CO 2 (P ETCO 2), peak airway pressure (P peak), arterial blood partial pressure of CO 2 (PaCO 2) and the intervention of cirulation and respiration were compared between the two groups before and within 15 minutes after pneumoperitoneum. Results:The max values of SBP, DBP, MAP, HR, P ETCO 2, Ppeak and PaCO 2 within 15 minutes after pneumoperitoneum in the experimental group were (117.08±13.07) mmHg, (77.08±9.43) mmHg, (90.06±10.33) mmHg, (69.04±9.10) times/min, (36.00±3.37) mmHg, (20.18±2.74) cmH 2O(1 cmH 2O=0.098 kPa), (40.65±3.31) mmHg, higher than that of the control group (140.63±18.34) mmHg, (91.90±11.79) mmHg, (107.25±12.85) mmHg, (77.67±13.57) times/min, (38.31±4.31) mmHg, (24.81±4.26) cmH 2O, (45.19±4.49) mmHg, the differences were statistically significant( t values were-7.269--2.945, all P<0.01). The amplitudes of SBP, DBP, MAP, HR, P ETCO 2, Ppeak and PaCO 2 fluctuations before and after pneumoperitoneum in the experimental group were (10.14±6.34) mmHg, (8.98±5.88) mmHg, (9.14±5.44) mmHg, (5.80±2.48) times/min, (3.27±1.43) mmHg, (2.65±1.54) cmH2O, (4.08±1.74) mmHg, while the control group were (33.65±14.87) mmHg, (26.17±9.73) mmHg, (28.04±9.97) mmHg, (17.63±9.77) times/min, (6.98±2.89) mmHg, (7.44±2.35) cmH 2O, (9.52±3.92) mmHg, the differences were statistically significant( t values were -11.841--8.048, all P<0.01). Within 15 minutes after pneumoperitoneum, circulatory intervention was 4.08% (2/49) in the experimental group, lower than that in the control group 22.92% (11/48), the difference was statistically significant( χ2=7.412, P<0.01). Respiratory intervention in the experimental group was 0 (0/49), lower than that in the control group 10.42%(5/48), the difference was statistically significant(Fisher test, P<0.05). Conclusions:In gynecological laparoscopic surgery, using pressure gradient control method to establish CO 2 pneumoperitoneum is conducive to reducing the effect of early pneumoperitoneum on circulatory and respiratory function, maintaining the relative stability of circulatory and respiratory function, effectively reducing anesthesia-related interventions after circulatory and respiratory fluctuations, and is conducive to the safety of patients.
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OBJECTIVE:To prepare Venlafaxine hydrochloride sustained-release tablets,and to investigate the characteristics of drug release. METHODS:Using glyceryl behenate as lipidic matrix material and HPMC as hydrophilic matrix material,the kind and dosage of excipients were screened by single factor experiment. Using 4,8,24 h accumulative release rate and the deviation summation of ideal values as index,the viscosity of HPMC,the amounts of HPMC K15M and glyceryl behenate were optimized by orthogonal test. The dissolution curves were fitted by different equations. RESULTS:The optimal formulation was as follows as venlafaxine hydrochloride 8.5 kg,HPMC K15M 15 kg,glyceryl behenate 26 kg,magnesium stearate 0.5 kg. 4,8,24 h accumula-tive release rates of prepared matrix sustained-release tablets were 34.3%,63.9% and 99.2%,respectively. The releases profiles of hydrophilic and lipidic matrix sustained release tablets followed first-order equation in vitro mainly through matrix erosion. CON-CLUSIONS:Venlafaxine hydrochloride hydrophilic and lipidic matrix sustained-release tablets with good sustained-release effect have been prepared successfully.
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Objective:To investigate the protective effect of schisandrin B (SchB)on the cerebral ischemia reperfusion injury of the rats and the influence in HSPA12B/PI3K/Akt signaling pathway,and to explore the mechanisms.Methods:130 SD rats were divided into sham group,cerebral ischemia reperfusion injury model group (model group),low dose of SchB group (SchB 3 mg· kg-1 ,SchB1 group),middle dose of SchB group (SchB 10 mg·kg-1 ,SchB2 group)and high dose of SchB group (SchB 30 mg·kg-1 ,SchB3 group)(n=26).The rats in sham group didn’t plug lines;the rats in model were used to establish ischemia reperfusion models;the rats in SchB1 ,SchB2 and SchB3 groups were firstly pretreated with different doses of SchB for 7 d,and then they were used to build cerebral ischemia reperfusion injury models.The nerve dysfunction of rats was evaluated by neurologic deficit score.The cerebral edema was detected by measuring the content of water in brain tissue.The morphological changes of brain tissue were observed by toluidine blue staining.The levels of nuclear factor kappa B (NF-κB), tumor necrosis factor-α(TNF-α),interleukin-1 (IL-1)and interleukin-6 (IL-6)in the brain tissue were detected by ELISA.Western blotting method was used to detect the protein expression levels of heat shock protein A12B (HSPA12B ), serine-threonine kinase (Akt ) and phosphorylation serine-threonine kinase (p-AKT ). Results:Compared with sham group,the neurologic deficit score of rats in model group was significantly increased (P <0.01),and the content of water in brain tissue was increased (P < 0.01 );the brain tissue structure was loosened,and the mesenchyme appeared edema;the NF-κB,TNF-α,IL-1,and IL-6 levels were increased (P <0.01),and the expression levels of HSPA12B and p-Akt proteins were decreased (P <0.01).Compared with model group,the neurologic deficit scores of the rats in SchB1 ,SchB2 ,and SchB3 groups were decreased (P <0.01),and the contents of water in brain tissue of the rats in SchB2 and SchB3 groups were decreased (P <0.05);the edema of nerve cells was alleviated,and the cavities were reduced;the NF-κB,TNF-α,IL-1,and IL-6 levels were decreased (P <0.05 or P <0.01),the expression levels of the HSPA12B protein in SchB2 ,and SchB3 groups were increased (P <0.05),and the p-Akt protein expression levels of the rats in SchB1 ,SchB2 ,and SchB3 groups were increased (P <0.01).Conclusion:SchB could protect the cerebral ischemia reperfusion injury of rats,its mechanism may be related to regulating the HSPA12B/PI3K/Akt signaling pathway and inhibiting the inflammatory reaction damage to the nerve cells of reperfusion.
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[ ABSTRACT] AIM:To investigate the changes of migration of lung adenocarcinoma cells promoted by IL-8 and the inner and outer mitochondrial membrane dynamic changes during this process.METHODS:Human lung adenocarci-noma cell line A549 was divided into control group and IL-8 group.Cell migration was analyzed by scratch detection and Transwell assay.The secretion of endogenous IL-8 was detected by ELISA.The protein levels of mitochondrial cytochrome C ( Cyt C) and mitochondrial outer membrane protein Tom20 was detected by Western blot.The mRNA expression of mito-chondrial fusion genes Mfn1, Mfn2 and OPA1 and fission genes Fis1, Drp1 and MTP18 was detected by RT-PCR.The morphological changes of mitochondria were observed by MitoTracker Red CMXRos dye staining and confocal microscopy. RESULTS:The migratory rate of A549 cells and endogenous secretion of IL-8 in A549 cells were higher than those in SPC-A-1 cells.The migratory rate of A549 cells was improved by IL-8 in a time-dependent manner.Compared with control group, the Tom20 protein expression was increased ( P0.05).Under confocal microscope, the punctate aggregates in the mitochondria of the A549 cells treated with IL-8 were observed.CONCLU-SION:The migratory rate of A549 cells is increased by IL-8, which is related to the changes of mitochondrial fusion genes and the fission genes.
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Objective To explore the mechanism of protective effect of Rhodioloside in cerebral ischemia-reperfusion rats and its relevance to phosphatidylinositol 3-kinases ( PI3-K)/protein serine-threonine kinases ( AKT) signaling pathway .Methods Forty eight Sprague-Dawley rats were randomly divided into four groups: sham-operation group , ischemia-reperfusion group , and Rhodiolo-side treatment groups (5 and 10 mg/kg).The model of right middle cerebral artery occlusion was established with thread ligation meth -od.The score of the neurological deficit was estimated 2 h followed by 24 h reperfusion.Histopathological changes were observed by hematoxylin-eosin(HE) staining.The infarct volume was measured with triphenyltetrazolium chloride (TTC) staining.Apoptotic cells were assessed with terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) method.The expressions of PI3-K and p-AKT were evaluated with immunohistochemistry .Results The score of the neurological deficit was decreased more ob-viously, the number of apoptotic were decreased more significantly , the expressions of PI3-K and p-AKT were increased more signifi-cantly in the Rhodioloside treatment groups (5 and 10 mg/kg) than in the ischemia-reperfusion group ( P <0.05).The score of the neurological deficit was decreased , the number of apoptotic was decreased , and the expressions of PI 3-K and p-AKT were increased in the Rhodioloside treatment group (10 mg/kg) than the Rhodioloside treatment group (5 mg/kg) ( P <0.05).Conclusions The protective mechanism of Rhodioloside therapy against cerebral ischemia r-eperfusion injury might be associated with activating the PI 3-K/AKT signaling pathway and then inhibiting neuronal apoptosis .
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Objective To copy the mouse aging model with D-galactose,and to investigate the role of Schisandra total lignin (SCL)in the mouse brain tissue aging and its mechanism.Methods 50 mice were radomly divided into control group,model group (100 mg·kg-1 ·d-1),low dose (35 mg·kg-1 ·d-1)of SCL group (SCL-L), middle dose (70 mg· kg-1 · d-1 )of SCL group (SCL-M)and high dose (140 mg· kg-1 · d-1 )of SCL group (SCL-H)(n=10).D-galactose (100 mg·kg-1 ·d-1 )was injected into the mice hypodermically for 10 weeks to induce aging models in all the groups except control group,and 35,70,and 140 mg· kg-1 · d-1 SCL were administered for 10 weeks in SCL groups.The learning and memory abilities were measured by the Water Maze test.The expression levels of Bax,Bcl-2,ubiquitin (Ub),microtubule-associated protein light chain 3 (LC3)in the brain tissue of the mice in various groups were observed by Western blotting method. The LC3 protein expressions in mouse brain cortex and hippocampus were observed by immunohistochemistry.Results In learning and memory test,compared with control group,the swimming time of the mice in model group was increased (P<0.05),and the number of errors was increased (P<0.05);compared with model group,the swimming time in SCL-L,SCL-M and SCL-H groups was decreased (P<0.05)and the number of errors was also decreased (P<0.05). Compared with control group,the expression level of Bax was increased (P<0.05),the expression level of Bcl-2 was decreased (P<0.05),the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were increased (P<0.05)in model group;compared with model group,the expression level of Bax was decreased (P<0.05),the expression level of Bcl-2 was incerased (P<0.05),and the expression levels of Ub and LC3-Ⅱ/LC3-Ⅰ proteins were decreased (P<0.05)in SCL-L,SCL-M and SCL-H groups.In control group,the neuronal morphology was normal,and none of brown granules were visible in the cytoplasm of mouse brain cortex and hippocampus and the expression of LC3 protein was negative.In model group,the neurons were degeneration,and the number of LC3 protein positive cells in the cerebral cortex and hippocamptal tissue was increased (P<0.05).In SCL-L,SCL-M and SCL-H groups,the number of degenerative neurons was decreased,and the number of LC3 protein positive cells was decreased (P<0.05).Conclusion SCL can inhibit the D-galactose-induced brain tissue aging in the mice, and the mechanism is related to regulating autophagy and inhibiting apoptosis.
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Objective To investigate the incidence of nosocomial infection of placing peripherally inserted central catheter (PICC) and venous port acess(VPA) in children with leukemia. Methods 60 cases with leukemia were treated in our hospital from January 2005 to September 2009.35 cases were mail, 25 cases were femail.Their ages ranged from 2.0 to 16.5 years old.42 cases fell to the PICC group, hospital-ization for 202 times, 18 cases belonged to the VPA group, hospitalization for 109 times. The incidence of infection in hospital, infection site, common pathogens and prognosis of two groups were analyzed retro-spectively.Results The incidence of fever and increasing of C-reactive protein of the PICC group were significantly lower than those of the VPA group. There was no difference of the incidence of white blood cells decreasing between two groups. The incidence of infection in hospital in the PICC group was 38.61%, higher than 28.44% of the VPA group. Their difference was significant. There was significant difference of the distribution of infection between two groups. There was no significant difference of the major pathogens of sputum culture and blood culture between two groups. Conclusions Using venous port acess(VPA) could reduce the incidence of infection in hospital compared using peripherally insertied central catheters in children with leukemia, so venous port acess( VPA ) may be safer.
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Objective:To setup a stable cell line which can overexpress human p100 protein,and then st udy the physiologic functions of p100 in regulation of STAT6-mediated gene tran scription.Methods:Using coimmunoprecipitation method to identify protein-protein interactions.STA T6-mediated gene transcriptional activation was detected by luciferase assay.Results:Human p100 protein interacted with both STAT6 and RNA polymerase Ⅱ,resulting in the enhancement of STAT6-mediated gene transcriptional activation.Conclusion:Human p100 protein acts as a bridging factor between STAT6 and basal transcripti onal machenary. [
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Blood glucose levels of 220 days were recorded in 85 type 2 diabetic patients for monitoring glycemia for 48-72 h each one by the continuous glucose monitoring system (CGMS). Frequencies of nocturnal hypoglycemia (blood glucose≤2. 8mmol/L) were estimated when bedtime blood glucose levels were≤4.4mmol/ L,≤5.0mmol/L,≤5. 6mmol/L,≤6.1mmol/L,≤6. 7mmol/L, or≤7. 2mmol/L respectively. Nocturnal hypoglycemia occurred in 39 nights of 220 days (17.7%) of 26 patients and their bedtime blood glucose level was (4.4?2.6)mmol/L with a range from 2. 2 to 12. 6 mmol/L. Using a cut-off of bedtime blood glucose at≤6. 7 mmol/L, area under the ROC curve was reached maximal value (0. 359?0. 046). The positive predictive value was 25% and the negative predictive value was 91. 7%. CCMS is a useful tool to detect unaware nocturnal hypoglycemia.