Objective:To investigate the effect of lentivirus-mediated silencing of mitochondrial ribosomal protein L35 (MRPL35) gene on the growth of human esophageal cancer TE-1cells, and to clarify its mechanism.Methods:Three kinds of human esophageal cancer cells, TE-1, ECA109and KYSE150, were selected.The relative expression levels of MRPL35mRNA in three kinds of cells by real-time quantitative PCR.The esophageal cancer TE-1cells were divided into shMRPL35group and shCtrl group, and the cells were infected with si-RNA lentivirus and si-RNA lentivirus;the esophageal cancer cell line stably silenting the MRPL35gene was established.Real-time quantitative PCR and Western blotting methods were used to detect the efficiency of MRPL35gene silencing.The cell growth curves in various groups were detected by CCK-8method, and the apoptotic rates were detected by flow cytometry after AnnexinⅤ-PE/7AAD double staining.Results:Three kinds of esophageal cancer cells expressed MRPL35gene, and the expression levels were not statistically significant between them (P＞0.05) .The results of real-time quantitative PCR and Western blotting methods showed that the mRNA and protein levels of MRPL35in the TE-1cells in shMRPL35group were significantly lower than those in shCtrl group (P＜0.05) .Compared with shCtrl group, the cell growth speed in shMRPL35group was decreased (P＜0.05) , and the apoptotic rate was significantly increased (P＜0.01) .Conclusion:Silencing MRPL35gene can inhibit the proliferation of esophageal cancer TE-1cells and plays a role through the apoptotic pathway.

Objective:To investigate the effect of inspiratory muscle training on the lung function and diaphragm movement of stroke survivors.Methods:Twenty-four stroke survivors were randomly divided into a control group and an experimental group. Both groups were given routine rehabilitation therapy, while the experimental group was additionally provided with 20 minutes of inspiratory muscle training, 5 times per week for 4 weeks. Before and after the treatment, the forced vital capacity (FVC), forced expiratory volume in one second (FEV1) and peak expiratory flow (PEF) expressed as percentages of the predicted values were used to assess ventilation. The maximum inspiratory pressure (MIP) and peak inspiratory flow (PIF) were used to assess inspiratory muscle function. Diaphragm mobility, as well as the diaphragm′s thickness at the end of expiration (DTee) and inspiration (DTei) and the diaphragm thickening fraction (DTF) were measured using ultrasonography. Any pulmonary infection was also recorded.Results:There was no significant difference between the two groups in any of the measurements before treatment. After the intervention the average FVC, FEV1, MIP, PIF, diaphragm mobility, DTei and DTF of the experimental group were all significantly better than before treatment and significantly better than the control group′s averages. However, no significant difference was found in the average PEF or DTee, nor in the rate of pulmonary infection between the two groups.Conclusion:Inspiratory muscle training can effectively improve ventilation, inspiratory muscle function, diaphragm mobility and diaphragm thickness at the end of inspiration among stroke survivors.

Objective To investigate the protective effects of Zhige Jiejiu Baogan Fang(Semen Hoveniae and Radix Puerariae Lobatae hangover-relieving and liver-protecting recipe, ZGJJBGF) on superoxide dismutase (SOD), malondialdehyde (MDA) and glutathione (GSH) rats with alcoholic liver injury. Methods 180 male Wistar rats were randomly divided into 6 groups: blank group, model group, positive control (Compound Methionine and Choline Bitartrate Tablet) group (0.36 g/kg), high-dose, mid-dose and low-dose ZGJJBGF groups (18,9,1.8 g/kg respectively). Except for the blank group, rats in other groups were orally administered with 50% ethanol(diluted with distilled water) for 6 weeks to establish the alcoholic liver injury rat model. Rats in all experimental groups except the blank and model groups received medicinal intervention respectively. The activity of malondialdehyde (MDA), glutathione (GSH),and superoxide dismutase(SOD) in serum and liver were measured at the end of the experiment. Results SOD activity in model group was significantly lower than that in the blank group, and the contents of MDA and GSH in the liver tissues and blood were significantly higher in the blank group (P<0.01). Compared with model group, mid-dose ZGJJBGF significantly increased the serum GSH level (P<0.05) and the activity of SOD (P<0.01) while reducing the content of MDA in liver tissue(P<0.01). Compared with the model group,high-dose ZGJJBGF significantly reduced the content of MDA in liver tissue of rats(P<0.01) and increased the content of GSH(P<0.01). Compared with model group, low-dose ZGJJBGF reduced serum MDA content (P<0.01). Conclusion Zhige Jiejiu Baogan Fang could enhance the antioxidant ability of endogenous antioxidant enzymes such as SOD and GSH. It can also inhibit the damage of liver injury of free radical-induced lipid peroxidation.

ObjectiveTo observe the effect of Hoveniae Semen， Flos Puerariae and their combinations on acute alcoholic liver disease and provide a scientific basis for the drug use in clinical practice and the research on other alcoholic diseases. MethodThe acute alcoholic liver injury model of mice was established by one-time gavage with 56% （V/V） Hongxing Erguotou liquor （12 mL·kg^-1）. One hundred and twenty male ICR mice were randomly assigned into blank group， model group， silybin group， Flos Puerariae group， Hoveniae Semen group， and Flos Puerariae-Hoveniae Semen combination groups （ratios of 1∶1， 1∶2 and 2∶1， respectively）， with 15 mice in each group. Each group was administrated with 10 mL·kg^-1 corresponding preventive drugs for 3 days by gavage. Except the blank group， the other groups were given Erguotou liquor by gavage at 12 mL·kg^-1. The mice were sacrificed 12 h after drinking for the observation of liver function and oxidative stress. The pathological changes of liver were observed via hematoxylin-eosin （HE） staining. Western blot was employed to determine the expression levels of proteins in the Kelch-like Ech-associated protein 1 （Keap1）/nuclear factor E₂-related factor 2 （Nrf2）/antioxidant response element （ARE） signaling pathway. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to detect mRNA levels of related genes. ResultCompared with control group， the modeling elevated the levels of alanine aminotransferase （ALT）， aspartate aminotransferase （AST） and alkaline phosphatase （ALP） in the serum and the content of malondialdehyde （MDA） and reactive oxygen species （ROS） in liver tissue （P<0.01） and decreased the activities of glutathione （GSH） and superoxide dismutase （SOD） （P<0.01）. The mRNA and protein expressions of Keap1 were significantly increased （P<0.05，P<0.01）， mRNA and protein expressions of Nrf2 were significantly decreased （P<0.01）. Compared with model group， Flos Puerariae-Hoveniae Semen 2∶1 lowered the levels of ALT， AST and ALP in the serum （P<0.01） and MDA and ROS in the liver （P<0.01）， and increased the activities of GSH and SOD （P<0.01）. Moreover， it alleviated the hepatic steatosis injury， up-regulated mRNA and protein levels of Nrf2 （P<0.01）， and down-regulated the mRNA and protein levels of Keap1 （P<0.01）. ConclusionFlos Puerariae， Hoveniae Semen and their combinations may exert the pre-protective effect on acute alcoholic liver injury in mice by regulating the Keap1/Nrf2/ARE pathway in the liver and restoring the liver oxidative balance destroyed by ethanol to inhibit the development of alcoholic liver disease .

ObjectiveTo explore the relationship between Kansui Radix and Glycyrrhizae Radix et Rhizoma in the Gansui Banxiatang based on cytochrome P450 （CYP450） enzyme and ascites rats. MethodA total of 150 Wistar rats were randomized into blank group （distilled water， ig）， model group （distilled water， ig）， Gansui Banxiatang group （5.68 g·kg^-1·d^-1， ig）， Gansui Banxiatang without Kansui Radix group （5.57 g·kg^-1·d^-1， ig）， Gansui Banxiatang without Glycyrrhizae Radix et Rhizoma group （4.01 g·kg^-1·d^-1， ig）， and Gansui Banxiatang without Kansui Radix And Glycyrrhizae Radix et Rhizoma group （3.90 g·kg^-1·d^-1， ig）， with 25 rats in each group. The Gansui Banxiatang was composed of 1.1 g Kansui Radix， 9 g Pinelliae Rhizoma Praeparatum， 15 g Paeoniae Radix Alba， 16.7 g Glycyrrhizae Radix et Rhizoma praeparata cum melle， and 15 g honey. The rats， except for the blank group， were injected （ip） with Walker-256 cells to induce cancerous ascites. The administration lasted 7 days for each group. The concentration of probe drugs was detected by high performance liquid chromatography （HPLC） and based on the metabolic rates of the drugs， the CYP450 enzyme activity was yielded. The CYP450 enzyme gene expression was determined by Real-time PCR， and CYP450 enzyme protein expression by Western blot. ResultCompared with the blank group， the model group showed high activity of CYP1A2 and CYP2C9， low activity of CYP2E1， CYP2C19， and CYP3A4， low mRNA expression of CYP2E1， CYP2C19， CYP2C9， and CYP3A4， and low protein expression of CYP2E1 and CYP3A4 （P<0.05）. Compared with the model group， Gansui Banxiatang group displayed low activity of CYP2C9 and high expression of CYP3A4 mRNA （P<0.01）， and Gansui Banxiatang without Kansui Radix group demonstrated low activity of CYP1A2， high activity of CYP2E1 and CYP3A4， high mRNA expression of CYP1A2， CYP2E1， CYP2C19， and CYP3A4， and high protein expression of CYP2E1 and CYP3A4 （P<0.05）. In comparison with the model group， the Gansui Banxiatang without Glycyrrhizae Radix Et Rhizoma group showed low activity of CYP2C9， high activity of CYP3A4， and high mRNA expression of CYP2E1 and CYP2C19 （P<0.05）， and Gansui Banxiatang without Kansui Radix and Glycyrrhizae Radix Et Rhizoma group had low activity of CYP2C9， high activity of CYP3A4， and high CYP3A4 mRNA expression （P<0.01）. The activity of CYP2E1， CYP2C9， and CYP3A4 was higher， and the mRNA expression of CYP1A2， CYP2C19， and CYP3A4 and CYP3A4 protein expression were higher in the Gansui Banxiatang without Kansui Radix group than in the Gansui Banxiatang group （P<0.05）. The activity of CYP2C9 and CYP3A4， and CYP2C19 mRNA were higher and mRNA expression of CYP3A4 was lower in the Gansui Banxiatang without Glycyrrhizae Radix Et Rhizoma group than in the Gansui Banxiatang group （P<0.05）. The activity of CYP2C9 and CYP3A4 in the Gansui Banxiatang without Kansui Radix and Glycyrrhizae Radix et Rhizoma group was higher than that in the Gansui Banxiatang group （P<0.01）. ConclusionThe four prescriptions all showed certain efficacy in the perspective of CYP450 enzyme， among which Gansui Banxiatang without Kansui Radix had the best effect and was better than Gansui Banxiatang. The Kansui Radix and Glycyrrhizae Radix et Rhizoma did not show antagonism in Gansui Banxiatang， but reduced the effect. Thus， in this study， the medicinal pair may show mutual inhibition.

ObjectiveTo explore the improvement effect of Flos Puerariae， Hoveniae Semen， and their compatibility on acute alcoholic gastric mucosal injury， and lay a foundation for further development of Flos Puerariae， Hoveniae Semen， and their compatibility in the prevention and treatment of alcohol-induced multiple organ injury. MethodThe acute alcohol-induced gastric mucosal injury model of mice was established by multiple intragastric administration of 56% Hongxing Erguotou liquor （15 mL·kg^-1）. A total of 120 male ICR mice were randomly divided into 8 groups， namely， the blank group， model group， omeprazole group （0.026 g·kg^-1）， Flos Puerariae-Hoveniae Semen （compatibility） high， medium， and low-dose groups （29.2，14.6， 7.3 g·kg^-1）， Flos Puerariae group （19.5 g·kg^-1）， and Hoveniae Semen group （19.5 g·kg^-1）， with 15 mice in each group. After one week of adaptive feeding， the animals were pre-administrated with the corresponding drug at the rate of 10 mL·kg^-1 for 3 d. From the 4^th day， after 1 h of administration， Erguotou liquid was administrated at the rate of 15 mL·kg^-1 and the blank group was administrated with the same volume of deionized water to record the drunkenness and sober up time. The administration was lasted for 3 d. One hour after the last administration， the eyeballs were removed and the mice were sacrificed. The concentration of ethanol in serum was determined by gas chromatograph， and the activity of ethanol dehydrogenase （ADH） in gastric mucosa was determined by ultraviolet-vis spectrophotometer. Hematoxylin-eosin （HE） staining was used to observe the pathological changes in gastric mucosa. Serum inflammatory factors were determined by enzyme-linked immunosorbent assay （ELISA）. The mRNA expression of nuclear transcription factor-κB （NF-κB） p65 and NF-κB inhibitory protein α （IκBα） were detected by real-time polymerase chain reaction （Real-time PCR）. ResultAs compared with the normal group， the content of interleukin-6 （IL-6）， interleukin-1β （IL-1β）， and tumor necrosis factor-α （TNF-α） in serum of mice in the model group was increased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues was increased （P<0.01）， and the mRNA expression of IκBα was decreased （P<0.01）. As compared with the model group， the drunkenness time of the omeprazole group， high and medium-dose compatibility groups， and Flos Puerariae group was prolonged （P<0.05）， the sober up time of the high and medium-dose compatibility groups was shortened （P<0.05）， the ethanol concentration in the serum of the high-dose compatibility group was decreased （P<0.05）， the ADH activity in the gastric mucosa of the omeprazole group and high and medium-dose compatibility groups was increased （P<0.05）， the macroscopic injury score of the high， medium， and low-dose compatibility groups and Flos Puerariae group was decreased （P<0.05）， the score of pathological injury in the omeprazole group， high， medium， and low-dose compatibility groups， and Flos Puerariae group was decreased （P<0.01）， the expression of IL-6 in serum of all drug groups was decreased （P<0.05）， the expression of IL-1β in serum of the omeprazole group， high， medium， and low-dose Flos Puerariae groups， and Hoveniae Semen group was decreased （P<0.05）， the expression of TNF-α in serum of high and medium-dose groups was decreased （P<0.05）， the mRNA expression of NF-κB p65 in gastric mucosa tissues of all drug groups was decreased （P<0.05）， and the mRNA expression of IκBα in gastric mucosa tissues of the omeprazole group and high， medium， and low-dose compatibility groups was increased （P<0.05）. As compared with the high-dose compatibility group， the drunkenness time in the low-dose compatibility group and Hoveniae Semen group was shortened （P<0.01）， the sober up time in the Flos Puerariae and Hoveniae Semen groups was prolonged （P<0.01）， the concentration of ethanol in the serum of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group increased （P<0.05）， the macroscopic injury score of the medium and low-dose compatibility groups and Hoveniae Semen group was increased （P<0.05）， the pathological injury score of the medium and low-dose compatibility groups， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， the content of IL-1β in serum of low-dose compatibility group， Flos Puerariae group， and Hoveniae Semen group was increased （P<0.01）， and the mRNA expression of IκBα in gastric mucosa of the Flos Puerariae group and Hoveniae Semen group was decreased （P<0.05）. As compared with the medium-dose compatibility group， the drunkenness time in the Hoveniae Semen group was shortened （P<0.05）， the sober up time in the Flos Puerariae group was prolonged （P<0.05）， the pathological injury score in the Flos Puerariae group and Hoveniae Semen group was increased （P<0.01）， and the content of IL-1β in serum of the low-dose compatibility group， the Flos Puerariae group， and Hoveniae Semen group was increased （P<0.05）. As compared with the low-dose compatibility group， the pathological injury score of the Hoveniae Semen group was increased （P<0.05）. ConclusionFlos Puerariae， Hoveniae Semen， and their compatibility play a role in preventing and treating acute alcoholic gastric mucosal injury in mice， which may be related to the inhibition of the expression of NF-κB signal pathway in gastric mucosa， and the high-dose compatibility group has the optimal effect.

ObjectiveTo observe the effects of Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the Haizao Yuhutang （HYT） on oxidative stress in the liver of goiter rats under the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020. MethodA total of 128 male Wistar rats were randomly divided into a blank group， a model group， a euthyrox group （20 μg·kg^-1）， a HYT group （12.06 g·kg^-1）， a HYT without Sargassum （HYT-H） group （9.90 g·kg^-1）， a HYT without Glycyrrhizae Radix et Rhizoma （HYT-G） group （10.26 g·kg^-1）， a HYT without Sargassum and Glycyrrhizae Radix et Rhizoma （HYT-HG） group （8.10 g·kg^-1）， and a Sargassum and Glycyrrhizae Radix et Rhizoma （HG） group （3.96 g·kg^-1）. The blank group was given deionized water by gavage， and the others were given propylthiouracil （PTU） to replicate the goiter pathological model. Euthyrox was taken as a positive control drug， and the rest of the Chinese medicine groups were given the corresponding decoction by gavage， the material was collected 12 hours after the last dose. The serum levels of aspartate aminotransferase （AST）， alanine aminotransferase （ALT）， alkaline phosphatase （ALP）， and the contents of superoxide dismutase （SOD）， glutathione peroxidase （GSH-Px）， malondialdehyde （MDA）， reactive oxygen species （ROS） in liver tissue were detected in each group. The pathological changes in the liver were observed via hematoxylin-eosin （HE） staining. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was utilized to detect the mRNA expressions of Kelch-like Ech-associated protein 1 （Keap1）， nuclear factor E₂-related factor 2 （Nrf2）， heme oxygenase-1 （HO-1）， p53 and Caspase-3 in liver tissues. Western blot was adopted to detect the protein expressions of Nrf2 and HO-1 in liver tissues in oxidative stress-related signaling pathways. ResultCompared with control group， the model group showed significantly increased serum ALT level and contents of MDA and ROS in liver tissues （P<0.05， P<0.01）， significantly reduced activities of SOD and GSH-Px in the liver （P<0.01）， significantly increased mRNA expression of Keap1 （P<0.01）， and significantly decreased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. Compared with the model group， the HYT group manifested significantly reduced serum levels of AST， ALT， and ALP （P<0.05， P<0.01）， significantly reduced contents of MDA and ROS in liver tissue （P<0.01）， significantly increased the activities of SOD and GSH-Px （P<0.01）， significantly decreased mRNA expressions of Keap1， p53， and Caspase-3 （P<0.01）， and significantly increased mRNA and protein expressions of Nrf2 and HO-1 （P<0.05， P<0.01）. ConclusionUnder the condition of 2 times the dose limit of the Pharmacopoeia of the People's Republic of China 2020， Sargassum and Glycyrrhizae Radix et Rhizoma incompatible pair with the HYT on oxidative stress in the liver of goiter rats had different effects. The HYT that contains Sargassum and Glycyrrhizae Radix et Rhizoma has a protective effect on the liver of goiter rats， and the effect is better than that of the HG group， the euthyrox group， and the incomplete groups. Its mechanism may be related to activating the Nrf2/HO-1 signaling pathway to alleviate liver oxidative stress and inhibiting the p53/Caspase-3 signaling pathway to reduce hepatocyte apoptosis.