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Objective:To observe the effects of overexpression of polypyrimidine tract binding protein-associated splicing factor (PSF) on the endoplasmic reticulum (ER) oxidative stress damage of human retinal microvascular endothelial cells (hRMEC) under high concentration of 4-hydroxynonenal (4-HNE).Methods:The logarithmic growth phase hRMEC cultured in vitro was divided into normal group, simple 4-HNE treatment group (simple 4-HNE group), empty plasmid combined with 4-HNE treatment group (Vec+4-HNE group), and PSF high expression combined with 4-HNE treatment group (PSF+4-HNE group). In 4-HNE group, Vec+4-HNE group, and PSF+4-HNE group cell culture medium, 10 μmol/L 4-HNE was added and stimulated for 12 hours. Subsequently, the Vec+4-HNE group and PSF+4-HNE group were transfected with transfection reagent liposome 2000 into pcDNA empty bodies and pcDNA-PSF eukaryotic expression plasmids, respectively, for 24 hours. Flow cytometry was used to detect the effects of 4-HNE and PSF on cell apoptosis. The effect of PSF overexpression on the expression of reactive oxygen species (ROS) in hRMEC was detected by 2', 7'-dichlorodihydrofluorescein double Acetate probe. Western blot was used to detect ER oxide protein 1 (Ero-1), protein disulfide isomerase (PDI), C/EBP homologous transcription factor (CHOP), glucose regulatory protein (GRP) 78, protein kinase R-like ER kinase (PERK)/phosphorylated PERK (p-PERK), and Eukaryotic initiation factor (eIF) 2α/the relative expression levels of phosphorylated eIF (peIF) and activated transcription factor 4 (ATF4) proteins in hRMEC of normal group, 4-HNE group, Vec+4-HNE group, and PSF+4-HNE group. Single factor analysis of variance was performed for inter group comparison.Results:The apoptosis rates of the simple 4-HNE group, Vec+4-HNE group, and PSF+4-HNE group were (22.50±0.58)%, (26.93±0.55)%, and (11.70±0.17)%, respectively. The intracellular ROS expression levels were 0.23±0.03, 1.60±0.06, and 0.50±0.06, respectively. The difference in cell apoptosis rate among the three groups was statistically significant ( F=24.531, P<0.05). The expression level of ROS in the Vec+4-HNE group was significantly higher than that in the simple 4-HNE group and the PSF+4-HNE group, with a statistically significant difference ( F=37.274, P<0.05). The relative expression levels of ER Ero-1 and PDI proteins in the normal group, simple 4-HNE group, Vec+4-HNE group, and PSF+4-HNE group were 1.25±0.03, 0.45±0.03, 0.63±0.03, 1.13±0.09, and 1.00±0.10, 0.27±0.10, 0.31±0.05, and 0.80±0.06, respectively. The relative expression levels of CHOP and GRP78 proteins were 0.55±0.06, 1.13±0.09, 0.90±0.06, 0.48±0.04 and 0.48±0.04, 1.25±0.03, 1.03±0.09, 0.50±0.06, respectively. The relative expression levels of Ero-1 ( F=43.164), PDI ( F=36.643), CHOP ( F=42.855), and GRP78 ( F=45.275) proteins in four groups were compared, and the differences were statistically significant ( P<0.05). Four groups of cells ER p-pERK/pERK ( F=35.755), peIF2 α/ The relative expression levels of eIF ( F=38.643) and ATF4 ( F=31.275) proteins were compared, and the differences were statistically significant ( P<0.05). Conclusion:PSF can inhibit cell apoptosis and ROS production induced by high concentration of 4-HNE, and its mechanism is closely related to restoring the homeostasis of ER and down-regulating the activation level of PERK/eIF2α/ATF4 pathway.
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Hyperbaric oxygen therapy is a method of breathing pure oxygen or high-concentration oxygen in a highpressure environment to treat hypoxic diseases and related diseases. According to clinical verification, this therapy has an irreplaceable effect on certain diseases and has gradually become a comprehensive clinical treatment. One of the main methods of certain diseases is widely recognized by the medical field at home and abroad. The development history, treatment principles, key technologies, and future development trends of hyperbaric oxygen are discussed in detail, provide a research direction for the development of hyperbaric oxygen therapy in the future, and at the same time, it has also improved physicians' awareness of hyperbaric oxygen therapy, so as to improving Industry influence.
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Hyperbaric Oxygenation , Oxygen/therapeutic use , Research DesignABSTRACT
The environmental gas concentration affects the storage period and quality of fruits and vegetables. High concentration CO₂ treating for a long time will cause damage to fruits, However, the specific molecular mechanism is unclear. To analyze the mechanism of CO₂ injury in apple, high-throughput sequencing technology of Illumina Hiseq 4000 and non-targeted metabolism technology were used to analyze the transcriptome sequencing and metabolomics analysis of browning flesh tissue of damage fruit and normal pulp tissue of the control group. A total of 6 332 differentially expressed genes were obtained, including 4 187 up-regulated genes and 2 145 down regulated genes. Functional analysis of the differentially expressed genes confirmed that the occurrence of CO₂ injury in apple was related to redox process, lipid metabolism, hormone signal transduction process and energy metabolism process. Twenty candidate browning genes were successfully screened, among which grxcr1 (md14g1137800) and gpx (md06g1081300) participated in the reactive oxygen species scavenging process, and pld1_ 2 (md15g1125000) and plcd (md07g1221900) participated in phospholipid acid synthesis and affected membrane metabolism. mdh1 (md05g1238800) participated in TCA cycle and affected energy metabolism. A total of 77 differential metabolites were obtained by metabolomic analysis, mainly organic acids, lipids, sugars and polyketones, including 35 metabolites related to browning. The metabolism of flavonoids was involved in the browning process of apple. Compared with the control tissue, the content of flavonoids such as catechin and quercetin decreased significantly in the damaged apple tissue, the antioxidant capacity of cells decreased, the redox state was unbalanced, and the cell structure was destroyed, resulting in browning. The results of this study further enrich the theoretical basis of CO₂ damage, and provide reference for the practical application of high concentration CO₂ preservation technology.
Subject(s)
Carbon Dioxide , Fruit , Gene Expression Regulation, Plant , Malus/genetics , Metabolome , TranscriptomeABSTRACT
Objective To prove the feasibility of a Fogging Process in fixing high-concentration radioactive aerosol 131I contamination. Methods High-concentration radioactive aerosol 131I contamination in an 131I-operating glovebox for radiopharmaceutical production was disposed by using fogging and fixing process. The aerosol 131I concentrations were detected and the results were analyzed. Results After a 120 minutes fixing, the 131I contamination in this glovebox reduced from(289 ± 9) DAC and (304 ± 6) DAC to (21.7 ± 2.0) DAC and (26.2 ± 1.8) DAC. After a 180 minutes fixing, the 131I contamination in this glovebox reduced from (259 ± 10) DAC to (1.80 ± 0.18) DAC. These results showed that no aerosol 131I contamination was raised again after 24-hours finishing this task. Conclusion Aerosol 131I concentration in a limited space could be controlled by using a fogging and fixing process, which could reduce the risk of internal exposure of staff. This process could be used by radiopharmaceutical production as an emergency management for dispose high-concentration radioactive aerosol 131I contamination.
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@#AIM:To observe the expression pattern of Cyclin D1 in human lens epithelial cells(HLECs)after traumatic stimulation in high-glucose culture<p>METHODS: The activity of HLECs was detected by MTT method after incubate with differernt concentration glucose for 24h <i>in vitro </i>to determine the optimal glucose concentration. qRT-PCR and Western blot were used to detect the high glucose pretreatment group and the non-high glucose pretreatment group. The expression of Cyclin D1 in HLECs at different time points after traumatic stimulation was detected.<p>RESULTS: The viability of HLECs were increased when treatment with low concentration glucose, but the concentration should not exceed 25.5mmol/L, or it will inhibit the activity of HLECs; The reasult of high glucose pretreatment group reveal that the expression of Cyclin D1 is down-regulated in a time-dependent manner within a certain time range. While the expression of Cyclin D1 was irregular in the non-pretreatment group, it was increased at the time point of 12h and 48h. The score treatment can up-regulate the expression of Cyclin D1 in HLECs in a certain degreen.<p>CONCLUSION: The effects of glucose on HLECs activity and Cyclin D1 experssion are irrugular. Trauma treatment can stimulate the expression of Cyclin D1 in HLECs to some extent.
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Objective To establish a bacterial endotoxin test method for high concentration vitamin B6 injection. Method The test was taken according to the bacterial endotoxin test in Chinese pharmacopoeia 2015 edition. Result By diluting the sample concentration to 1.04 mg/ml with the buffer of pH6.5-7.5, and using λ=0.06 EU/ml of TAL reagent, the interference could be effectively avoided. Conclusion The method was useful, which could be used to test the bacterial endotoxin in high concentration vitamin B6 injection. The bacterial endotoxin limit was defined as 0.06 EU/mg.
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Objective To optimize the extraction methods of mitochondrial genome DNA(mtDNA)of Oncomelania hupen-sis. Methods The pyrolysis,protein K variable-temperature digestion and high-concentration potassium acetate purification were applied to optimize the high-concentration-salt precipitation method,and then the optimized method was compared with two common extraction methods,the sucrose density gradient centrifugation method and traditional high-concentration-salt pre-cipitation method. The mtDNA samples were identified by using spectrophotometry,agarose gel electrophoresis and the amplifi-cation products of COX1. The nuclear DNA contamination was tested by the amplification products of ITS. Results The concen-tration and yield of the improved method was significantly higher than those of the traditional method(F=3032.65,10185.00, both P<0.01). The mtDNA samples extracted were essentially free of nuclear DNA and protein,meeting PCR,sequence analy-sis and other molecular biology research requirements. Conclusions The improved high-concentration-salt precipitation meth-od for isolating mtDNA is simple,and it has high yield and low cost. The extracted mtDNA can meet relevant analysis require-ments.
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Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.
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Objective:To investigate the regulation effect of endogenous nicotinamide phosphoribosyl transferase (Nampt) on the Vimentin expression of glomerular cells in high concentration glucose,and to clarify the mechanism of formation of diabetic kidney inflammation fibrosis.Methods:The C57/BL6 diabetic mice were selected and the kidney tissues were collected,and the wild C57/L86 mice were used as control group;the pathological section and tissue fluorescence staining were performed.The expression and location of endogenous Nampt and Vimentin in the glomerular cells were detected by immuno-focused technology.The HBZY-1 cells were randomly divided into 4 groups:low concentration of glucose (LG,0.56 mmol · L-1) control group,high concentration glucose (HG,200 mmol · L-1) group,HG +-FK866 group and HG+nicotinamide mononucleotide (NMN) group.In HG group,the cells were treated with FK866 (10 μmol · L-1) and NMN (1 mmol · L-1) for 24 h after cultured with HG for 5 d.The expression levels of Nampt,Vimentin,nuclear factor-kappa B p65 (NF-κBp65) and sirtuin type 1 (Sirt1) were detected by immunofluorescence and Western blotting methods.The expression levels of Nampt and Vimentin were detected by RT-PCR and Western blotting methods.Results:The shape and size of glomerulus had obvious atrophy of the mice in severe diabetic group compared with normal C57/BL6 mice group.The expression level of Vimentin in glomerular cells was increased with the increasing of endogenous Nampt (P＜0.01).When the HBZY-1 cells were cultured in HG condition,the exprssion levels of Nampt,Vimentin and NF-kB p65 were obviously increased while the Sirt1 expression levels was significantly decreased compared with control group (P＜ 0.01).The expression levels of Nampt,Vimentin and NF-κB p65 in glomerular cells in HG+FK866 HG+ NMN groups were singnificartyly decreased compared with control group (P＜0.01).Conclusion:The endogenous Nampt over-expression in glomerular cells can enhance the expression of Vimentin under high concentration of glucose stress through NF-κBp65 and Sirt1 signal pathway.
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Objective To investigate the necessity of dilution in samples with high concentrations of D-Dimer and optimum dilution multiple.Methods Quality control products and calibration were detected by using Sysmex CS5100 for precision evaluation,including within-batch and between-run precision.Calibration were detected for validation of linear range and clinical reportable.Samples with D-Dimer5 mg/L and FDP>20 μg/mL were also serially diluted and detected to calculated recovery rate.Results Within-batch and between-run coefficients of variation were both less than 3%.Within the scope of 0.207-5.170 mg/L,the linear distribution was fine.The clinical reportable range was 0.207-165.440 mg/L.For samples with D-Dimer5 mg/L and FDP>20 μg/mL,there was obvious antigen excess phenomenon,and gradient dilution was required.Conclusion For samples with D-Dimer>5 mg/L and FDP>20 μg/mL,dilution should be performed to ensure the accuracy of detected results.
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AIM:To study the expression of glycine receptorα1 subunit in neonatal rat myocardial cells and to investigate the effect of lipopolysaccharide (LPS), hypoxia/reoxygenation, isoproterenol (ISO) and high concentration of glucose (HG) on the expression of glycine receptorα1 subunit in the neonatal rat myocardial cells.METHODS:Neonatal rat myocardial cells were cultured in vitro.The expression of glycine receptorα1 subunit was detected by Western blotting. The neonatal rat myocardial cells were treated with LPS (20 mg/L), ISO (100 μmol/L) or high concentration of glucose (25 mmol/L) for 24 h, or were exposed to hypoxia for 3 h followed by reoxygenation for 3 h.Subsequently, the cell viabil-ity was measured by CCK-8 assay, and the expression of glycine receptorα1 subunit was determined by Western blotting. RESULTS:The expression of glycine receptor α1 subunit in the neonatal rat myocardial cells was positively detectable by Western blotting.Compared with control group, no significant difference of the cell viability ( P>0.05) in LPS group, ISO group, hypoxia/reoxygenation group and HG group was observed.The expression of glycine receptor α1 subunit was in-creased (P<0.01) in LPS group, ISO group and hypoxia/reoxygenatio group, but decreased (P<0.01) in HG group. CONCLUSION:Glycine receptorα1 subunit exists in the neonatal rat myocardial cells.A certain concentration of LPS or ISO, or hypoxia/reoxygenation for a certain period upregulate the expression of glycine receptorα1 subunit, but HG down-regulates the expression of glycine receptor α1 subunit in cultured neonatal rat myocardial cells.
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Objectives: Recently, it has been reported that the effects of artificial high concentration carbon-dioxide (CO2) on core temperature, cutaneous blood flow, thermal sensation. However, the effect of artificial high concentration CO2 water foot baths for spasticity, lower extremity motor function and walking ability was not identified. The purpose of this study was to investigate whether the newly artificial high concentration CO2 water foot bath inhibits spasticity and improves lower extremity motor function and gait speed in spastic paraplegia patient. Case Presentation: The patient was a 37 years old man with spastic paraplegia of human immunodeficiency virus encephalopathy, without signs of cognitive impairment. The patient was able to walk without assistance using a T-cane or an ankle-foot orthosis. He had no medical condition that limited footbath usage (such as uncontrolled cardiopulmonary disease, severe joint disability and severe sensory disturbance), severe aphasia that made it impossible to follow verbal instructions, and cognitive dysfunction that interfered with outcome assessments. Informed consent was obtained from him according to the ethical guidelines of the hospital, after he fully understood the purpose and methodology of the study. This work was carried out with permission from the Ethical Committee of Kagoshima University. Methods: This case study was before and after intervention trial. Six outcome instruments were used at baseline and after the artificial high concentration CO2 water foot bath: the modified Ashworth scale (MAS) score for the gastrocnemius muscles as a measure of spasticity, ankle clonus, muscle stiffness at triceps muscle of calf, deep body and surface skin temperature as a monitor for physical condition, the active range of motion as an assessment tool for motor function, and the 10-m walk test as a measure of walking ability. Lower-extremity movement acceleration was also measured using an accelerometer. The subject rested in a chair for 10 min and the above-noted physiological reactions during the last 5 min of the resting period were recorded as baseline values. Next, the subject received a 20-min foot bath in water at 38 °C, with a 10-min recovery period. The artificial high concentration CO2 water foot bath improved the acceleration of the spastic lower extremities and this improvement in acceleration lasted for 10 min after the footbath usage. Results: The subject experienced no discomfort before, during or after the intervention, and all assessments were completed safely. The deep body temperature and skin temperature increased immediately after and 10 minutes after the artificial high concentration CO2 water foot baths. The MAS score, ankle clonus and the muscle stiffness for the triceps muscle of calf were decreased. The active range of motion for ankle dorsiflexion and gait speed improved after the 20-min intervention. Conclusion: These findings suggest that artificial high concentration CO2 water foot bath is an effective method for controlling spasticity, and improves motor function and walking ability in spastic paraplegia patients.
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Objectives: The warm-water foot bathing is widely used as a clinical method for hemiplegic limb. Recent research have reported that the artificial high concentration carbon-dioxide (CO2) water foot bathing have a potent vasodilative action. However, the definite effects of the artificial high concentration CO2 water foot bath for hemiplegic limbs remain uncertain. We examined that the effects of the artificial high concentration CO2 water foot bath for patients after stroke. Patients: Three inpatients after stroke were recruited for this study. The age and duration after onset were 58.3 ± 21.4 years and 63.0 ± 38.9 months, respectively. Of the three patients (two males and one female), two were diagnosed with cerebral hemorrhage, one with cerebral infarction. Methods: The artificial high concentration CO2 water foot bath and tap water foot bath were prepared. The concentration of CO2 water foot bath was approximately 1000-1,200 ppm, and both lower limbs (under the knee joint) were immersed in 38 °C water for 20 minutes. Foot bathing in tap water was also carried out under the same conditions in the another day. The following physiological data were measured before foot bathing and after the end of foot bathing. Not only the deep body temperature at axillary, the surface skin temperature at the front of femur, the calf of the leg and the dorsal foot, but also the muscle stiffness at triceps muscle of calf were evaluated. Results: None of the subjects experienced discomfort before and after both the high concentration CO2 water and the tap water foot bath. The physiological examination was completed safely in all subjects. The results were as follows: The deep body temperature and the surface skin temperature had increased, and the muscle stiffness had been relieved in the high concentration CO2 water foot bath compared with the tap water bathing. The deep body temperature of the high concentration CO2 water foot bath have risen from 36.4 °C to 36.9 °C, the surface-skin temperature of the front of femur (from 26.7 °C to 28.1 °C), the calf of the leg (from 29.5 °C to 31.9 °C) and the dorsal foot (from 29.9 °C to 32.3 °C) have risen, respectively. The muscle stiffness have been relieved from 55.3 to 51.8 before and after. There was no change that the tap water had increased in the deep body temperature and the surface-skin temperature, and the muscle stiffness had been relieved before and after. Conclusion: These results suggested that the use of the high concentration CO2 water foot bath was more effective in hyperthermia compared with the tap water. Furthermore, we considered that carbon dioxide had promoted to increase the skin and the muscle blood flow by vasodilative action to the arteriole, and use of the high concentration CO2 water foot bath contribute to improve the circulatory dynamics for the hemiplegic limb. These findings may suggest that the use of the high concentration CO2 water foot bath is an effective physiotherapy for circulatory dynamics treatment that might facilitate stroke rehabilitation
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Objectives: Spasticity is defined as a pathological increase in muscle tonus, and increased muscle tonus of lower limbs is a major obstacle to the stroke rehabilitation. Foot baths are considered to provide beneficial thermal therapy for post-stroke patients with spasticity, but their anti-spastic effects have not been investigated comprehensively. The present study aimed to evaluate alterations in spasticity and motor function using foot baths in post-stroke patients with spastic hemiplegia. Methods: We underwent two separate experiments each consisting of immersion in warm water up to the knee joint level, and measuring spasticity, physiological examination and motor function. Experiment 1; Fourteen post-stroke patients with lower limb spasticity were enrolled in this study (nine males and five females; mean age 50.4±12.9 years; range, 28-65 years). The subjects’ legs from below the knee joint were immersed in water at 41°C for 15 min. Measurements of F-waves and a physiological examination were carried out immediately (within 5 min) after the foot-bath session, and again 30 min later, while the subject remained wrapped in blankets on the lift-bath stretcher. Experiment 2; Six post-stroke patients with lower limb spasticity were enrolled in this study (five males and one female; mean age 55.2±14.6 years; range, 39-68 years). The subjects’ legs from below the knee joint were immersed in the artificial high concentration carbon-dioxide (CO2) water or tap water foot bath at 38°C for 30 min. Measurements of muscle stiffness, motor function (active range of motion: A-ROM) and a physiological examination were carried out immediately (within 5 min) after the foot-bath session, and again 10 min later, while the subject remained wrapped in blankets. Results: None of the subjects experienced discomfort before, during or after the foot-bath treatment. The physiological examination was completed safely in all subjects. Experiment 1; The mean values of F-wave parameters were significantly reduced after foot-bath treatment (P<0.01). The anti-spastic effects of foot-bath treatment were indicated by decreased F-wave parameters, in parallel with decreases in modified Ashworth scale (MAS) score. The body temperature was significantly increased both immediately after, and 30 min following foot-bath treatment. Experiment 2; The changes both in the body and surface skin temperature were higher in the artificial high concentration CO2 water foot bath compared with the tap water foot bath. The changes in the MAS score, muscle stiffness and A-ROM were also higher in the high concentration CO2 water foot bath than in the tap water foot bath. Conclusion: These findings demonstrate that the use of foot baths is an effective non-pharmacological anti-spastic treatment that might facilitate stroke rehabilitation. In addition, the high concentration CO2 water foot baths appeared to play an important role in decreased spasticity.
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AIM: To observe the efficacy of high concentrations of sodium hyaluronate ( 3g/L SH ) for moderate to severe dry eye. METHODS: Forty moderate to severe dry eye patients were included in the study according to the diagnosis criteria and randomized into two groups. The patients of the trial group received topical administration of high concentration sodium hyaluronate (3g/L), and those of the control group received sodium hyaluronate ( 1g/L ) plus recombinant human epidermal growth factor. The dry eye symptom scores, ocular surface disease index ( OSDI) scores, tear film break-up time ( BUT) , SchirmerⅠ test and corneal fluorescein staining score were evaluated. All the indexes were compared between the two groups 2wk before and after treatment. RESULTS: There were no significant differences of the indicators between the two groups before treatment. After 2wk treatment, the differences were statistically significant compared to former except for the SchirmerⅠtest. Compared with the control group, the symptom scores and the OSDI scores were lowered. No significant differences were found in the other indicators between these two groups. CONCLUSION: Topical usage of highconcentrations of sodium hyaluronate (3g/L) is beneficial for remitting the ocular symptoms in moderate to severe dry eyes, and also improve the quality of life of patients.
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<b>Objectives: </b>Recently, it has been reported that the effects of artificial high concentration carbon-dioxide (CO<sub>2</sub>) on core temperature, cutaneous blood flow, thermal sensation. However, the effect of artificial high concentration CO<sub>2</sub> water foot baths for spasticity, lower extremity motor function and walking ability was not identified. The purpose of this study was to investigate whether the newly artificial high concentration CO<sub>2</sub> water foot bath inhibits spasticity and improves lower extremity motor function and gait speed in spastic paraplegia patient.<BR><b>Case Presentation: </b>The patient was a 37 years old man with spastic paraplegia of human immunodeficiency virus encephalopathy, without signs of cognitive impairment. The patient was able to walk without assistance using a T-cane or an ankle-foot orthosis. He had no medical condition that limited footbath usage (such as uncontrolled cardiopulmonary disease, severe joint disability and severe sensory disturbance), severe aphasia that made it impossible to follow verbal instructions, and cognitive dysfunction that interfered with outcome assessments. Informed consent was obtained from him according to the ethical guidelines of the hospital, after he fully understood the purpose and methodology of the study. This work was carried out with permission from the Ethical Committee of Kagoshima University.<BR><b>Methods: </b>This case study was before and after intervention trial. Six outcome instruments were used at baseline and after the artificial high concentration CO<sub>2</sub> water foot bath: the modified Ashworth scale (MAS) score for the gastrocnemius muscles as a measure of spasticity, ankle clonus, muscle stiffness at triceps muscle of calf, deep body and surface skin temperature as a monitor for physical condition, the active range of motion as an assessment tool for motor function, and the 10-m walk test as a measure of walking ability. Lower-extremity movement acceleration was also measured using an accelerometer. The subject rested in a chair for 10 min and the above-noted physiological reactions during the last 5 min of the resting period were recorded as baseline values. Next, the subject received a 20-min foot bath in water at 38 °C, with a 10-min recovery period. The artificial high concentration CO<sub>2</sub> water foot bath improved the acceleration of the spastic lower extremities and this improvement in acceleration lasted for 10 min after the footbath usage.<BR><b>Results: </b>The subject experienced no discomfort before, during or after the intervention, and all assessments were completed safely. The deep body temperature and skin temperature increased immediately after and 10 minutes after the artificial high concentration CO<sub>2</sub> water foot baths. The MAS score, ankle clonus and the muscle stiffness for the triceps muscle of calf were decreased. The active range of motion for ankle dorsiflexion and gait speed improved after the 20-min intervention.<BR><b>Conclusion: </b>These findings suggest that artificial high concentration CO<sub>2</sub> water foot bath is an effective method for controlling spasticity, and improves motor function and walking ability in spastic paraplegia patients.
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<b>Objectives:</b> The warm-water foot bathing is widely used as a clinical method for hemiplegic limb. Recent research have reported that the artificial high concentration carbon-dioxide (CO<sub>2</sub>) water foot bathing have a potent vasodilative action. However, the definite effects of the artificial high concentration CO<sub>2</sub> water foot bath for hemiplegic limbs remain uncertain. We examined that the effects of the artificial high concentration CO<sub>2</sub> water foot bath for patients after stroke. <BR><b>Patients: </b>Three inpatients after stroke were recruited for this study. The age and duration after onset were 58.3 ± 21.4 years and 63.0 ± 38.9 months, respectively. Of the three patients (two males and one female), two were diagnosed with cerebral hemorrhage, one with cerebral infarction. <BR><b>Methods: </b>The artificial high concentration CO<sub>2</sub> water foot bath and tap water foot bath were prepared. The concentration of CO<sub>2</sub> water foot bath was approximately 1000-1,200 ppm, and both lower limbs (under the knee joint) were immersed in 38 °C water for 20 minutes. Foot bathing in tap water was also carried out under the same conditions in the another day. The following physiological data were measured before foot bathing and after the end of foot bathing. Not only the deep body temperature at axillary, the surface skin temperature at the front of femur, the calf of the leg and the dorsal foot, but also the muscle stiffness at triceps muscle of calf were evaluated. <BR><b>Results: </b>None of the subjects experienced discomfort before and after both the high concentration CO<sub>2</sub> water and the tap water foot bath. The physiological examination was completed safely in all subjects. The results were as follows: The deep body temperature and the surface skin temperature had increased, and the muscle stiffness had been relieved in the high concentration CO<sub>2</sub> water foot bath compared with the tap water bathing. The deep body temperature of the high concentration CO<sub>2</sub> water foot bath have risen from 36.4 °C to 36.9 °C, the surface-skin temperature of the front of femur (from 26.7 °C to 28.1 °C), the calf of the leg (from 29.5 °C to 31.9 °C) and the dorsal foot (from 29.9 °C to 32.3 °C) have risen, respectively. The muscle stiffness have been relieved from 55.3 to 51.8 before and after. There was no change that the tap water had increased in the deep body temperature and the surface-skin temperature, and the muscle stiffness had been relieved before and after. <BR><b>Conclusion:</b> These results suggested that the use of the high concentration CO<sub>2</sub> water foot bath was more effective in hyperthermia compared with the tap water. Furthermore, we considered that carbon dioxide had promoted to increase the skin and the muscle blood flow by vasodilative action to the arteriole, and use of the high concentration CO<sub>2</sub> water foot bath contribute to improve the circulatory dynamics for the hemiplegic limb. These findings may suggest that the use of the high concentration CO<sub>2</sub> water foot bath is an effective physiotherapy for circulatory dynamics treatment that might facilitate stroke rehabilitation
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<b>Objectives:</b> Spasticity is defined as a pathological increase in muscle tonus, and increased muscle tonus of lower limbs is a major obstacle to the stroke rehabilitation. Foot baths are considered to provide beneficial thermal therapy for post-stroke patients with spasticity, but their anti-spastic effects have not been investigated comprehensively. The present study aimed to evaluate alterations in spasticity and motor function using foot baths in post-stroke patients with spastic hemiplegia. <BR><b>Methods:</b> We underwent two separate experiments each consisting of immersion in warm water up to the knee joint level, and measuring spasticity, physiological examination and motor function. <BR><b>Experiment 1;</b> Fourteen post-stroke patients with lower limb spasticity were enrolled in this study (nine males and five females; mean age 50.4±12.9 years; range, 28-65 years). The subjects’ legs from below the knee joint were immersed in water at 41°C for 15 min. Measurements of F-waves and a physiological examination were carried out immediately (within 5 min) after the foot-bath session, and again 30 min later, while the subject remained wrapped in blankets on the lift-bath stretcher. <BR><b>Experiment 2;</b> Six post-stroke patients with lower limb spasticity were enrolled in this study (five males and one female; mean age 55.2±14.6 years; range, 39-68 years). The subjects’ legs from below the knee joint were immersed in the artificial high concentration carbon-dioxide (CO<sub>2</sub>) water or tap water foot bath at 38°C for 30 min. Measurements of muscle stiffness, motor function (active range of motion: A-ROM) and a physiological examination were carried out immediately (within 5 min) after the foot-bath session, and again 10 min later, while the subject remained wrapped in blankets. <BR><b>Results: </b>None of the subjects experienced discomfort before, during or after the foot-bath treatment. The physiological examination was completed safely in all subjects. <BR><b>Experiment 1; </b>The mean values of F-wave parameters were significantly reduced after foot-bath treatment (P<0.01). The anti-spastic effects of foot-bath treatment were indicated by decreased F-wave parameters, in parallel with decreases in modified Ashworth scale (MAS) score. The body temperature was significantly increased both immediately after, and 30 min following foot-bath treatment. <BR><b>Experiment 2;</b> The changes both in the body and surface skin temperature were higher in the artificial high concentration CO<sub>2</sub> water foot bath compared with the tap water foot bath. The changes in the MAS score, muscle stiffness and A-ROM were also higher in the high concentration CO<sub>2</sub> water foot bath than in the tap water foot bath. <BR><b>Conclusion:</b> These findings demonstrate that the use of foot baths is an effective non-pharmacological anti-spastic treatment that might facilitate stroke rehabilitation. In addition, the high concentration CO<sub>2</sub> water foot baths appeared to play an important role in decreased spasticity.
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Objective To observe insulin synthesis and secretion in INS-1 under high glucose, and to clarify the effect of PTH1R. Methods After successful construction of recombinant PTH1R-siRNA vectors in INS-1 cell, insulin secretion and intracellular insulin content of control group, siPTH1R-Negative control group, PTHrP group, and siPTH1R group under 25 mmol/L glucose were measured by radioimmunoassay in INS-1 cell. Intracellular calcium were detected by Fluo-3/AM and the capability of glucose transport was calculated by assaying the uptake of [3H]-2-deoxy-D-glucose in cells.Results Compared with control group, and siPTH1R-NC group, PTHrP group showed increased capability of insulin secretion; PTHrP group had higher intracellular insulin levels than others; PTHrP group showed increased intracellular calcium; the uptake of [3H]-2-deoxy-D-glucose under high glucose after 48h of PTHrP group was increased(all P＜0.01). Conclusion There is a close relationship between PTH1R activation and insulin secretion and synthesis, PTH1R activation may be one of the protective mechanisms in maintaining function of β-cell under high glucose.
ABSTRACT
AIM: To explore the effect of Snail1 siRNA on high-glucose induced tubular epithelial-to-mesenchymal transition (TEMT). METHODS: Subconfluent renal tubular epithelial cells were incubated in serum-free DMEM for 24 h to arrest and synchronize the cell growth. Then cells were treated with normal glucose (5.5 mmol/L D-glucose) or high glucose (25 mmol/L D-glucose) for 72 h. Meanwhile 19.5 mmol/L D-manntiol was used as high osmotic control. Snail1 siRNA was transfected into tubular epithelial cells. In parallel, cells were transfected with non-specific siRNA which served as the control data sets. Cells were then treated with 25 mmol/L D-glucose for 72 h. RNA and cell lysates were collected to determine the protein and mRNA levels of Snail1, TGF-β_1, α-SMA, vimentin and E-cadherin. RESULTS: Transfection caused the decreases in Snail1 at mRNA and protein levels by 62% and 68% respectively as compared to those in untransfected cells cultured in high glucose medium. Western blotting exhibited that Snail1 siRNA transfection restored E-cadherin protein expression by 61% compared to that in high-glucose-treatment cells, whereas it inhibited high-glucose-induced induction of α-SMA protein by 58%. Similarly, RT-PCR revealed that Snail1 siRNA transfection dramatically suppressed the high-glucose-induced mRNA expressions of α-SMA and vimentin by 72% and 61%, respectively, while E-cadherin mRNA increased by 53%. CONCLUSION: Our study provides direct evidence that Snail1 is able to control TEMT.