ObjectiveTo investigate the effect of mitochondrial calcium uniporter （MCU） on the cytoskeleton of pancreatic ductal epithelial cells in a mouse model of acute pancreatitis （AP） induced by caerulein （CAE）， to analyze the role of MCU in the development of AP， and to provide a theoretical basis for clinical treatment. MethodsIn the in vivo experiment， wild-type male C57BL6/J mice， aged 4 weeks， were randomly divided into control group and AP group， with 6 mice in each group. The mice in the AP group were given intraperitoneal injection of CAE to establish a model of AP， and those in the control group were given intraperitoneal injection of an equal volume of normal saline. Serum and pancreatic tissue samples were collected after 24 hours of modeling. HE staining was used to observe pancreatic histopathological changes； Western Blot was used to measure the expression levels of MCU， glutathione peroxidase 4 （GPX4）， and acyl-CoA synthetase long chain family member 4 （ASCL4）； kits were used to measure the serum level of amylase. In the in vitro experiment， the human pancreatic ductal epithelial cell line HPDE6-C7 was co-cultured with CAE for 24 hours to establish an in vitro AP model， and the cells were divided into control group， CAE group， RR （an MCU activity inhibitor） group， CAE+RR group， Fer-1 （an ferroptosis inhibitor） group， CAE+Fer-1 group， Erastin （an ferroptosis inducer） group， and CAE+Erastin group. CCK-8 assay was used to observe the influence of different agents on cell viability； Western Blot was used to measure the expression levels of MCU， GPX4， and ASCL4； immunofluorescence assay was used to measure reactive oxygen species （ROS）， actin cytoskeleton， and monolayer permeability； kits were used to measure the concentrations of malondialdehyde （MDA）， glutathione （GSH）， Fe²⁺， and total iron. A one-way analysis of variance was used for comparison of continuous data between multiple groups， and the least significant difference t-test was used for comparison between two groups. ResultsIn the in vivo experiment， compared with the control group， the AP group had significant increases in pancreatic histopathological score， the serum level of amylase， and the expression levels of MCU and ASCL4， as well as a significant reduction in the expression of GPX4 （all P<0.05）. In the in vitro experiment， compared with the control group， the CAE group had significant increases in the expression levels of MCU and ASCL4， a significant reduction in the expression of GPX4， and significant increases in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability， as well as a significant reduction in the concentration of GSH （all P<0.05）， with the presence of actin cytoskeleton disruption. Compared with the CAE group， the CAE+RR group had a significant increase in the expression level of GPX4， a significant reduction in the expression level of ASCL4， and significant reductions in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant increase in the concentration of GSH （all P<0.05）， with alleviation of actin cytoskeleton disruption. Compared with the CAE group， the CAE+Fer-1 group had significant reductions in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant increase in the concentration of GSH （all P<0.05）， with alleviation of actin cytoskeleton disruption. Compared with the CAE group， the CAE+Erastin group had significant increases in the concentrations of Fe²⁺， total iron， and MDA， the green fluorescence intensity of ROS， and monolayer permeability and a significant reduction in the concentration of GSH （all P<0.05）， with aggravation of actin cytoskeleton disruption. ConclusionDuring the onset of AP， MCU mediates oxidative stress-induced ferroptosis and leads to the disruption of the pancreatic ductal epithelial barrier， which may be one of the possible pathogeneses of AP.

Objective To explore the damage of spiral ganglion neurons(SGNs),the protective effects of different dosages of healthy ear compound (HEC)from traditional Chinese medicine(TCM) against age-induced SGNs degeneration and its possible mechanism in spiral ganglion neurons(SGNs)of C57BL/6J mice.Methods Totally 36 C57BL/6J mice just after ablactation were randomly divided into four groups.Normal control group (n =6)drank tap water daily from ablactation to 2 months old.Ageing-related SGNs apoptosis control group(n=12)drank tap water daily from ablactation to 7 months old.High dose TCM group(n=12)at drank 3.65 g/kg/d of HEC from ablactation to 7 months old.Low dose TCM group(n=6)drank 0.91 g/kg/d of HEC from ablactation to 7 months old.The animal cochleae were immediately removed at the termination of the experiment.In each group,the cochleae of 6 animals were used for paraffin embedding,slicing and toluidine blue staining to observe neuronal morphological changes.The caspase-3 mRNA expression study was performed by real-time PCR technique in 6 cochleae of High dose TCM group and ageing-related SGNs apoptosis control group.Results The morphological structure of cochlear SGNs represented healthy and normal density in normal control group at 2 months old.In contrast,amount or density of SGNs in cochlear basilar part was significantly damaged and reduced in ageing-related SGNs apoptosis control group at 7 months old(P＜ 0.001).But the high dose TCM group at 7 months of age was similar to the normal control group at 2 months old in morphological structure,amount or density of SGNs(P＞0.05).The low dose TCM group was significantly different from other 3 groups in amount or density of SGNs (P＜0.001).However,SGNs in the middle part and apical part showed integrity in each group.In addition,the expression level of caspase-3 in the cochlea of high dose TCM group was also obviously different with age-related SGNs apoptosis control group(P＜0.01) Conclusions Ageing-related damage of SGNs in C57BL/6J mice begins from the base of cochlea and progresses towards the apex.The HEC of TCM could significantly protect SGNs against age-induced apoptosis in SGNs.The efficacy of the high dose TCM is better than that of the low dose TCM.Its SGNs protective mechanisms might be related to involving the caspase-mediated cell apoptotic pathway.

Objective To evaluate the detection rate of multi-slice spiral CT (MSCT)signs and the clinical value of multi-planar reconstruction (MPR)in T1a and T1b peripheral lung cancer patients.Methods Eighty-seven cases with peripheral lung cancer proved by pathology were collected.The cases were divided into T1a and T1b group based on the TNM classification.The MSCT and MPR images were compared between the two groups.Results (1)Detection rate of the deep sublobe sign,spinous process sign, short spiculated sign,pleural indentation sign,vascular convergence sign,multi-nodule accumulation sign,vacuole sign and air bron-chogram sign,were 1 1.4%,20.0%,31.4%,60.0%,25.7%,45.7%,42.9% in T1a group and 42.3%,36.5%,57.7%, 80.8%,5 1.9%,25.0%,21.2% in T1b group,respectively.The difference were all statistically significant (P < 0.05)between T1a and T1b group except that of the spiculated sign (P = 0.098).(2)The detection rate of the sublobe sign,spinous process sign, spiculated sign,pleural indentation sign and vascular convergence sign were higher on MPR images than on axial thin-slice images in both T1a and T1b group.Conclusion The detection rate of the tumor-lung interface’s signs are lower in T1a than in T1b,the detec-tion rate of internal structure signs of the tumor are higher in T1a than in T1b in peripheral lung cancer patients.MPR has important value in early diagnosis of peripheral lung cancer.

OBJECTIVETo investigate the effect of spearmint oil on emphysema-like changes and the expression of tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta(IL-1beta), matrix metalloproteinase-9 (MMP-9) and tissue inhibitor of metalloproteinase-1 (TIMP-9) in lipopolysaccharide (LPS) treated rats.

METHODEmphysematous changes model was induced by intratracheal instillation of LPS once a week for up to 8 weeks in rats. Rats were divided into control, dexamethasone (0.3 mg x kg(-1)), and spearmint oil (10, 30,100 mg x kg(-1)) groups. Each group was treated with saline, dexamethasone, and spearmint of oil respectively for 4 weeks. Then total and different white blood cell counts in bronchoalveolar lavage fluid(BALF) were carried out. The pathologic changes of lung tissue such as alveolar structure, airway inflammation, and goblet cell metaplasia were observed by HE and AB-PAS staining. Expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9 were measured.

RESULTBoth spearmint and dexamethasone decreased the destruction of pulmonary alveolus. The total and different white blood cell counts in BALF including neutrophile and lymphocyte of spearmint oil 100 mg x kg(-1) and dexamethasone group were significantly reduced, and the goblet cell metaplasia was also inhibited. Dexamethasone had inhibitory effect on the expression of TNF-alpha, IL-1beta, TIMP-1 and MMP-9. Spearmint oil 30, 100 mg x kg(-1) significantly reduced TNF-alpha and IL-1beta respectively. Spearmint oil 10, 30 and 100 mg x kg(-1) had no effect on the expression of TIMP-1, but could decrease the expression of MMP-9 significantly in lung tissues.

CONCLUSIONSpearmint oil has protective effect on rats with emphysematous changes, since it improves alveolar destruction, pulmonary inflammation, and goblet cell metaplasia. The mechanism may include reducing TNF-alpha, IL-1beta content and inhibiting overexpression of matrix metalloproteinase-9 in lung tissues.

Animals ; Azo Compounds ; pharmacology ; Bronchoalveolar Lavage Fluid ; cytology ; Goblet Cells ; drug effects ; Interleukin-1beta ; drug effects ; metabolism ; Leukocytes ; drug effects ; metabolism ; Lipopolysaccharides ; Lymphocytes ; drug effects ; metabolism ; Matrix Metalloproteinase 9 ; drug effects ; metabolism ; Mentha spicata ; chemistry ; Metaplasia ; Monocytes ; drug effects ; metabolism ; Neutrophils ; drug effects ; metabolism ; Phytotherapy ; Plant Oils ; therapeutic use ; Pulmonary Emphysema ; chemically induced ; drug therapy ; enzymology ; pathology ; Rats ; Respiratory System ; drug effects ; pathology ; Tissue Inhibitor of Metalloproteinase-1 ; drug effects ; metabolism ; Tumor Necrosis Factor-alpha ; drug effects ; metabolism

OBJECTIVE To investigate the effects of 1,8-cineol on lung functions and mechanism in asthmatic guinea pigs. METHODS The guinea pig model was performed by intraperitoneal injection of the 0.5 ml Al(OH)_3 gel containing OVA 20 μg. The guinea pigs were constructed by immunization of intraperitoneal injection on the 0 day and the 7th day, and the experiment was performed on the 28th day. The effect of 1,8-cineol 10, 30 and 100 ml·kg~(-1) on the airway resistance(R_(aw)) and dynamic lung compliance (C_(dyn)) of asthmatic guinea pigs 1 h after challenge of OVA. The changes in leukocyte and different kinds of leukocyte in bronchoalveolar lavage fluid (BALF) after the challenge of OVA have been studied. The levels of eosinophil cationic protein (ECP), interleukin(IL)-4, IL-8 and tumor necrosis factor (TNF)-α in lungs of guinea pigs were determined using enzyme-linked immunosorbent assay (ELISA). The changes in R_(aw) and C_(dyn) of asthmatic guinea pigs were investigated 17 h after challenge of OVA and inhalated methacholine (MCh). The changes in leukocyte and different kinds of leukocyte in BALF after the challenge of OVA have been studied. The levels of ECP, IL-4, IL-8 and TNF-α in lungs of guinea pigs were determined using ELISA. RESULTS 1,8-Cineol inhibited increase in R_(aw) and decrease in C_(dyn) from 1 to 30 min after challenge of OVA in model group. The levels of ECP, IL-4 and TNF-α in asthmatic model group were higher than those in normal control group(P<0.05). The levels of ECP, IL-4 and TNF-α of 1,8-cineol 100 mg·kg~(-1) group were significantly lower than those in asthmatic model group (P<0.01). The level of IL-8 of asthmatic model group didn't have any significant difference from that of control group. 1,8-Cineol 100 mg·kg~(-1) could significantly decrease the numbers of leukocyte and the percent of eosinophils in BALF. Seventeen hours after challenge of OVA, R_(aw) and C_(dyn) of asthmatic model group were higher than these of control group (P<0.05, P<0.01); 1,8-cineol 100 mg·kg~(-1) significantly inhibited the increase in R_(aw), compared with model group (P<0.05); 1,8-cineol 10, 30 and 100 mg·kg~(-1) improved the decrease in C_(dyn) after MCh-induced in model group which were challenged by OVA after 17 h; 1,8-cineol 100 mg·kg~(-1) could significantly decrease the numbers of leukocyte and the percent of neutrophils, the levels of ECP, IL-8 and TNF-α compared with asthmatic group. The level of IL-4 in asthmatic model group didn't have any significant difference from that in normal control group. CONCLUSION In the course of early stage of asthma, 1,8-cineol inhibites the asthma by decreasing the number of eosinophils and down-regulating the activity of EPO. In the course of later stage of asthma, 1,8-cineol inhibits or improves the aggravation and lasting states of asthma which is directly coursed by neutrophils accumulating in the BALF that related to the increase in IL-8.

Objective To investigate a more pedect method for a nice outward appearance of a reconstructed thumb.Methods A free one-stage plasty second toe transfer for thumb reconstruction by interchanging the whole skin-nail flap from the great toe with another one from the second toe.Results There were 12 cases in this group,following-up 6-9 months in 8 cases,7 cases was excellent and 1 cases was good.The reconstructed thumb got a nice looking and more normal function while no blight to the great toe occurred.Conclusion It is an effective new procedure in ameliorating outward appearance of the reconstructed thumb by transferring the free moulded second toe.

OBJECTIVE: To evaluate the effect of Xiangdan Injection on mRNA expression of the endothelial vaso-active factors of patients with coronary heart disease and blood stasis. METHODS: Fifty-six patients were randomly divided into two groups:twenty-eight patients were treated according to the therapeutic guide for coronary heart disease as the control group and 28 were given the same treatment plus Xiangdan Injection as the treated group. The expressions of ET-1 and eNOS mRNA were examined with RT-PCR before experiment and ten days later. RESULTS: The positive rate of eNOS mRNA of the treated group increased, while the positive rate of ET-1 mRNA of the treated group decreased after ten day's treatment, with significant differences as compared with that before the experiment. Xiangdan Injection up-regulated the eNOS mRNA expression and suppressed the ET-1 mRNA expression. Changes of expression were not observed in the control group. CONCLUSION: Xiangdan Injection improves the endothelial function of patients with coronary heart disease and blood stasis by regulating the expressions of ET-1 and eNOS mRNA.

OBJECTIVETo study the effects of sodium butyrate (NaBu) on cell cycle checkpoint and the apoptosis sensitivity in U937 cells.

METHODSTwo mutant U937 cell lines, U937-ASPI3K (ATM negative) and U937-pZeosv2(+) (ATM wild-type), were used as the cell model system. Immunoprecipitation and kinase assay were used to examine the p38 MAPK and ERK1 kinase activities. Western blot was used to analyze the phosphorylation of Bad protein.

RESULTSU937-pZeosv2(+) pretreated with NaBu exhibited enhanced apoptotic response in a NaBu dose dependent fashion upon (137)Cs irradiation, which could be abolished by olomoucine (OLM), a p38 MAPK specific inhibitor. On the other hand, Cyclin dependent kinase 2 (CDK2) specific inhibitor CDK2-I and p34cdc2/cyclinB inhibitor alsterpaullone (ALP) failed to block the effects of NaBu. Similar results were also observed in U937-ASPI3K. The effect of irradiation on p38 MAPK and ERK1 was strikingly potentiated by NaBu. Furthermore, inactivation of irradiated Bad protein via phosphorylation on serine 136 was also enhanced.

CONCLUSIONNaBu is able to enhance the apoptotic response in U937 cells, which is mediated by p38 MAPK activation but not ATM status.

Apoptosis ; Butyrates ; pharmacology ; Carrier Proteins ; metabolism ; Humans ; MAP Kinase Signaling System ; Mitogen-Activated Protein Kinase 3 ; Mitogen-Activated Protein Kinases ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; U937 Cells ; bcl-Associated Death Protein ; p38 Mitogen-Activated Protein Kinases