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Objective To explore the effects of long-term low-protein intake on nonspecific inflammatory responses in the liver and T lymphocytes in the spleen in middle-aged and older mice and underlying mechanisms.Methods Fourteen month-old female KM(Kunming)mice were randomly divided into the control group,the low-protein group,the high-protein group and the high-protein + rapamycin group.Additionally,Two month-old female KM mice served as the adult group and were fed regular chow.Animals in the high-protein + rapamycin group received injections of rapamycin intraperitoneally once every two days.Animals were sacrificed at the end of 3 months.Liver histology slices were prepared for the examination of pathological changes and detection of the expression of CD68 and mTOR(the mechanistic target of rapamycin).Spleen lymphocyte suspensions were prepared to count the percentages of CD4+ T and CD8+ T cells.Results Compared with the control group,the low-protein group and the high-protein-+-rapamycin group in histology slides showed regularly arranged hepatocytes,no obvious sinus hepaticus expansion and only mild time-related changes.Moreover,compared with the control group,the low-protein and high-protein + rapamycin groups were associated with increased percentages of CD4+T and CD8 + T cells in the spleen(all P＜ 0.05)and decreased expression of CD68 in the liver(P＜0.05),though the levels of mTOR were similar among the groups.Conclusions Long-term low-protein intake has beneficial effects in mitigating the reduction of T lymphocytes in the spleen and slowing age-related structural and nonspecific inflammatory changes in the liver,possibly through downregulation of the expression of the mTOR protein.
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Integrin alphavbeta3 plays a major role in various signaling pathways, cell apoptosis, and tumor angiogenesis. To examine the functions and roles of alphavbeta3 integrin, a stable CHO-677 cell line expressing the murine alphavbeta3 heterodimer (designated as "CHO-677-malphavbeta3" cells) was established using a highly efficient lentiviral-mediated gene transfer technique. Integrin subunits alphav and beta3 were detected at the gene and protein levels by polymerase chain reaction (PCR) and indirect immunofluorescent assay (IFA), respectively, in the CHO-677-malphavbeta3 cell line at the 20th passage, implying that these genes were successfully introduced into the CHO-677 cells and expressed stably. A plaque-forming assay, 50% tissue culture infective dose (TCID50), real-time quantitative reverse transcription-PCR, and IFA were used to detect the replication levels of Foot-and-mouth disease virus (FMDV) in the CHO-677-malphavbeta3 cell line. After infection with FMDV/O/ZK/93, the cell line showed a significant increase in viral RNA and protein compared with CHO-677 cells. These findings suggest that we successfully established a stable alphavbeta3-receptor-expressing cell line with increased susceptibility to FMDV. This cell line will be very useful for further investigation of alphavbeta3 integrin, and as a cell model for FMDV research.
Subject(s)
Animals , Mice , Animals, Suckling , CHO Cells , Cloning, Molecular , Cricetulus , DNA, Complementary/genetics , Disease Susceptibility/virology , Foot-and-Mouth Disease/genetics , Foot-and-Mouth Disease Virus/physiology , Integrin alphaVbeta3/geneticsABSTRACT
ObjectiveTo investigate the role of CD4+ CD25+ regulatory T cells(Treg) in peripheral blood and intestinal endotoxin (ET) in liver injury of rat severe acute pancreatitis (SAP).MethodsSixty male Wistar rats were randomly allocated into sham operation group(SO group) and severe acute pancreatitis group(SAP group).The rats in SAP group recevied the injection of Sodium Taurocholate(NaTc) into their far-end of bile-pancreatic duct and were sacrificed in 3-,6-and 12-hour,serum amylase(AMY) and alanine aminotransferase(ALT) determined by full-automatic biochemistry instrument,limulus reagent method is used to determine ET content in plasma,the proportion of Treg among peripheral lymphocytes was determined by Flow Cytometry(FCM),HE stain is used to observe pathological changes in liver and pancreas,the protein of Foxp3 activity was evaluated by immunohistochemistry staining,and the relationships between these indicators were analyzed using Pearson correlation analysis test.ResultsHistopathologic examination and the level of ALT revealed the occurrence of pancreatic inflammation and pathological changes of liver in SAP group.The percentage of Treg in SAP groups significantly increased at 3 h(2.26％ ±0.32％),6 h(2.36 ％ ±0.48％)and 12 h(2.80％ ±0.35％) comparing to the SO groups(P＜0.01) ; plasma ET levels compared with the SO group was statistically significant (P＜ 0,01 ),and 12 h (0.85 ± 0.11) compared to,3 h (0.74±0.11) and 6 h (0.78-±-0.07) was no significant difference (P＞0.05).The expression of Foxp3 protein on the livers were upregulated markedly.Pearson correlation analysis teat showed that quantities of Treg were positively proportional to the levels of ET(r=0.89,P＜0.01) after liver injury of SAP.ConclusionsSAP may lead to liver injury and the high plasma levels of ET and Treg in peripheral blood may play an important role in liver injury of SAP.
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Objective To investigate the changes of Th1/Th2 cytokines and its relationship with lipopolysaccharide (LPS) in pretreatment of relieving nonalcoholic steatohepatitis (NASH).Methods The 24 male Wistar rats were randomly divided into 3 groups: normal control group, liver injury group and LPS pretreatment group. The rats were given normal diet in normal control group,high-sucrose and high-fat diet both in liver injury group and in LPS pretreatment group, and the rats in LPS pretreatment group were given hypodermic injection of LPS 0. 5 mg/kg every other day. The level of plasma endotoxin (ET), activity of alanine aminotransferase (ALT), content of tumor necrosis factor-α (TNF-α) and interleukin-10 (IL-10) were determined. At the end of week 9, the rats were executed, and the liver tissue slices were prepared to investigate hepatic pathologic change by hematoxylin and eosin (HE) staining.Results The level of plasma ET was significantly higher in liver injury group than in normal control group. The level of plasma ALT and infiltrating lymphocytes in liver tissue were significantly lower in LPS pretreatment group than in liver injury group. The level of plasma TNF-α was significantly lower in LPS pretreatment group compared with liver injury group.In contrast, the level of plasma IL-10 was higher (P＜0. 05). Histology with HE staining showed that hepatocyte steatosis was obviously relieved with smaller lipid droplet in LPS pretreatment group than in liver injury group. Conclusions LPS pretreatment can alleviate high-sucrose and high-fat induced NASH. The disequilibrium of Th1/Th2 cytokines may be an important part of mechanism.
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ObjectiveTo study the curative effect and safety of low dose of urokinase (UK) combined with low molecular heparin calcium.MethodsSixty-four cases of sudden cardiac arrest patients were divided into treatment group and control group by random digits table with 32 cases each.Two groups were given cardiopulmonary resuscitation according to the 2005 international guide for cardiopulmonary resuscitation and emergency cardiovascular care.Early in the recovery,the patients in treatment group were pumped in vein with low dose of UK(200 000 U) and injected subcutaneous with low molecular heparin calcium (4100 U ) in 30 minutes.The rate of return of spontaneous circulation (ROSC),survival rate longer than 24 hours and 30 days in two groups and patients dying of bleeding or bleeding conditions in treatment group were observed.ResultsThere were 11 cases (34.4％) of ROSC in control group,and compared with 20 cases (62.5％) in treatment group,there was significant difference (P＜0.05).There were 5 cases (15.6％) of survival longer than 24 hours in control group,and compared with 13 cases(40.6％) in treatment group,there was significant difference (P ＜ 0.05 ).There were 2 cases (6.2％) of survival 30 days in control group,and compared with 8 cases (25.0％) in treatment group,there was significant difference (P＜ 0.01 ).Nobody had subcutaneous bleeding or other organ hemorrhage in control group.But there was I patient who had subcutaneous limited ecchymosis in the injection site in treatment group.The difference of fibrinogen before and after treatment in treatment group was statistically significant(P ＜ 0.01 ),but there was no significant difference in prothrombin time and platelet count before and after treatment in treatment group (P ＞ 0.05).ConclusionsIt is safe and effective in cardiopulmonary resuscitation with low dose of UK combined with low molecular heparin calcium.
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Objective To improve the success rate of insertion of gastric lavage canal in patientswith tracheal intubation. Methods 70 patients with organophosphoms pesticide intoxication who neededgastric lavage after tracheal intubation were randomized into the test group and the control group, 35 easesin each. In test group, lavage canal was inserted through bite block orally and the bedside was raised till 70to 80 in angle. While in the control group, lavage canal was inserted directly through mouth with the posi-tion of hypsokinesis. The success rate of intubation was compared between the two groups. Results One-time success rate was 94.2% in the test group,which was statistically different from that of the control group(37.1%), P < 0.01. The complication of intubation in the test group was also lower than that of the controlgroup,which had statistical difference. Conclusions Through improvement of position to raise the bedsidetill 70 to 80 in angle, the insertion of gastric lavage canal through bite block orally was much better than thenormal one and it is worth applying.
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To investigate the security of semen biologically, 15 bull semen samples were collected (of which 5 exhibited clinical signs of Foot-and-mouth disease) and identified by RT-PCR and virus isolation. The results indicated that the semen of the infected bulls were contaminated by Foot-and-mouth disease virus (FMDV), but FMDV was not detected in semen samples from those bulls not showing clinical signs of Foot-and-mouth disease (FMD). This is the first report of the presence of FMDV in bull semen due to natural infection in China. The analysis of the partial sequence of the VP1 gene showed that the virus strain isolated from semen has 97.9% identity with the virus isolated from vesicular liquid of infected bulls showing typical signs of FMD and belonged to the same gene sub-group.
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Chronologicaol order of acupuncture,as one of the in uential factors of acupuncture therapeutic e ect,has been commonly ignored at present.This article is to explain the rationality and necessity of chronologicaol order of acupuncture through following aspects.One is based on TCM theory,such as the'branch'and the'root',gentleness and urgency,qi and blood,reinforcing de ciency and reducing excess.The other is ndings of modern clinical and experimental researches.
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AIM:To investigate the effect of lipopolysaccharides(LPS)preconditioning on CCl4-induced liver injury and the change of LPS signal transduction.METHODS:The male Wistar rats were divided randomly into liver-injury group,which were injected with CCl4 5 mL/kg first,three days later were injected 0.3 mL 40% CCl4 and 60% olive oil. Animals in LPS preconditioning group were injected with LPS 0.5 mg/kg before the day CCl4 was given. Rats received high fat diet were as liver injury group,and normal control group received normal diet. The lymphocytes infiltrated in the liver tissue were counted. The endotoxin and ALT level in rat plasma,TNF-? content and expressions of TLR4,p38,p-p38,I??,NF-?? in the rat livers were also determined.RESULTS:The lymphocytes in liver slice and ALT level of the plasma in LPS preconditioning group were lower significantly than those in the liver injury group,and the expressions of TLR4,p-p38,NF-?? in the liver were the same. In contrast,the expression of I?? was higher.CONCLUSION:LPS preconditioning relieves obviously CCl4-induced chronic liver injury. The mechanism may be associated with change of signal transduction of LPS,which results in decrease of pre-inflammatory cytokines.