Correlation of neutrophil to lymphocyte ratio to severity of coronary artery disease and in-hospital clinical outcomes in patients with acute coronary syndrome: A prospective observational study

Objective:To explore correlation of neutrophil-to-lymphocyte ratio(NLR)to severity of coronary artery disease(CAD)and in-hospital clinical outcomes in patients with acute coronary syndrome(ACS).Methods:In this prospective and observational study,we recruited 500 patients with ACS.For all the eligible patients,demographic details were collected,and laboratory parameters were evaluated.The CAD severity was evaluated in terms of the number of involved vessels.The NLR was calculated based on neutrophils and lymphocytes and the correlation of various risk factors and severity and outcome of CAD was performed.Results:77.2%of Patients was male,and 52%of the patients aged between 55-70 years.Based on the type of ACS,396 out of 500 patients had ST-elevation myocardial infarction.An ascending trend in the white blood cell levels and NLR value was noted as the severity of the ACS increased and the highest white blood cell levels and NLR was noted among classⅣpatients.The mean NLR value among the non-survivors were higher compared to the survivors(9.52±5.72 vs.4.76±2.36;P<0.01).Receiver operating curve showed that the cut-off NLR value was 5.76 with a sensitivity of 75.0%and a specificity of 77.3%.Conclusions:The NLR can be used as an independent prognostic marker in ACS.An elevated NLR value serves as a reliable predictor for short-term complications,notably in-hospital mortality.

Effects of different doses of dexmedetomidine on inflammatory factors and T lymphocyte subsets in elderly patients undergoing laparoscopic surgery

Objective:To investigate the effects of different doses of dexmedetomidine on inflammatory factors and T lymphocyte subsets in elderly patients undergoing laparoscopic surgery. Methods: from June 2016 to June 2017 in our hospital for laparoscopic surgery of 120 cases of elderly patients, were randomly divided into A group, B group, C group, D group of 30 cases, all patients underwent intravenous inhalational anesthesia, A, B, C in three groups were given dexmedetomidine before induction of anesthesia. Fixed loading dose of 0.5 μg/kg, after intubation respectively with dexmedetomidine 0.2 μg/(kg.h), 0.5 μg/(kg.h), 0.8 μg/(kg.h) after continuous infusion to surgery, D group at the same time with the same volume of physiological saline. Before induction of anesthesia (T0), 24 h after operation (T1), 72 h after operation (T2) of venous blood interleukin-1 were measured beta (IL-1 beta), interleukin -2 receptor (sIL-2R) and interleukin-6 (IL-6), tumor necrosis factor alpha (TNF-alpha) and T lymphocyte subsets (CD3+, CD4+, D8+).Results: the T1 of the 4 groups of sIL-2R, IL-6, IL-1 beta, TNF-alpha and T0 levels were significantly higher than, and A group, B group and C group than in D group, B group and C group than in A group, B group and C group had no significant difference;group IL-1, sIL-2R, IL-6, TNF-beta and alpha level of T1 decreased significantly, and A group, B group and C group than in D group, B group and C group than in A group, B group and C group had no significant difference. T1 of the 4 groups of CD3+, CD4+, D8+, CD4+/CD8+ and T0 decreased significantly, and A group, B group and C group than in D group, B group and C group than in A group, B group and C group had no significant difference;T2 D group CD3+, CD4+, D8+, CD4+/CD8+ lower than T1, and A group, B group, C group, CD3+, CD4+, D8+, CD4+/CD8+ higher than that of D group, B group, C group, CD3+ CD4+, D8+, CD4+/CD8+ higher than that of A group, B group, C group no significant difference.Conclusion: Dexmedetomidine can inhibit the inflammatory reaction and improve the immune function in elderly patients undergoing laparoscopic surgery, and the effect of middle and high dose is more significant.

Effect of Bifidobacterium triple live bacteria on inflammatory factors, oxidative stress and T lymphocyte subsets in patients with ulcerative colitis

Objective: To explore the effect of Bifidobacterium triple live bacteria on inflammatory factors, oxidative stress and T lymphocyte subsets in patients with ulcerative colitis (UC). Method: A total of 90 patients with UC treated in our hospital were selected and randomly divided into the observation group and the control group each with 45 cases. The control group was given Mesalazin Enteric-coated Tablets 1 g/times, 4 times/d, continuous treatment for 1 months:when the clinical symptoms were stable, switched to 500 mg/times, 3 times/d, continuous treatment for 3 months. The observation group on the basis of conventional treatment to give Bifidobacterium triple live bacteria scattered adjuvant therapy (Bifico capsule), 0.84 g/time, 2 times/d, continuous treatment for 3 months. In the two groups, the levels of inflammatory factors (IL-6, IL-10, CRP, TNF-α), oxidative stress markers (MDA, SOD) and T lymphocyte subsets (CD3+, CD4+, CD8+, CD4+/CD8+) were compared pre and post treatment. Results: The levels of IL-6, CRP, MDA, TNF-α, CD8+ in both two groups were significantly decreased compared with treatment before;the levels of IL-10, SOD, CD3+, CD4+ and CD4+/CD8+were significantly increased compared with treatment before. After treatment, the levels of serum IL-6, CRP, TNF-α, MDA and CD8+ in observation group with the data (72.17±15.18) pg/mL, (21.52±10.21) mg/mL, (15.98±4.12) pg/mL respectively were decreased more significantly than those data in control group which were (86.55±17.26) pg/mL, (43.02±12.27) mg/L, (22.35±3.67) pg/mL, MDA level (5.89±0.56) nmol/mL in observation group was decreased more significantly than the level in the control group (6.75±0.68) nmol/mL;CD8+level (17.24±3.06)% in observation group was decreased more significantly than the level (19.01±2.62)% in the control group. After treatment, IL-10 level (70.21±6.03) pg/mL in observation group was increased more significantly than the level in the control group (56.48+ 8.67) pg/mL;SOD level (1.84±0.06) U/mL in observation group was increased more significantly than the level in the control group (1.32±0.05) U/mL;the levels of CD3+, CD4+, CD4+/CD8+ in the observation group (57.84±6.07)%;(36.78±4.32)%;(1.92±0.29) were increased more significantly than the level in the control group (54.93±6.87)%;(35.42±5.27)%;(1.89±0.12). Conclusion: Bifidobacterium triple live bacteria scattered adjuvant treatment of ulcerative colitis helps regulate oxidative stress and inflammatory cytokines inhibiting inflammatory reaction and improving enhance function, and can effectively improve the clinical symptoms of patients with UC.

Effects of Ribavirin combined with Clostridium butyricum powder on T lymphocyte subsets and inflammatory factors in child patients with rotavirus enteritis

Objective: To observe clinical efficacy of Ribavirin combined with Clostridium butyricum powder on child patients with rotavirus enteritis and analyze serum IL-6, IL-8, TNF-αconcentration and change of T lymphocyte subsets. Methods: A total of 90 patients with rotavirus enteritis were randomly divided into control group (n=45) and observation group (n=45). The control group was given conventional therapy combined with Ribavirin. The observation group was treated with Clostridium butyricum powder on the basis of control group. The changes of T lymphocyte subsets and inflammatory factors were measured before and after treatment in all subjects. Results: There was no significant difference in serum T lymphocyte subsets between the control group and the observation group before treatment. After treatment, the levels of serum CD3+, CD4+ and CD4+/CD8+ in both groups were higher than those before treatment, while CD8+ level was lower than that before treatment. The levels of serum CD3+, CD4+ and CD4+/CD8+ in the observation group were higher than those in the control group after treatment, while the CD8+ level was lower than that in control group in the corresponding period. There was no significant difference in the concentration of inflammatory factors between the control group and the observation group before treatment. After treatment, the levels of IL-6, IL-8 and TNF-α in the two groups were lower than those before treatment. The observation group was significantly lower than control group in corresponding period after treatment and difference was statistically significant. Conclusion: On the basis of conventional therapy and Ribavirin treatment, combined with Clostridium butyricum Powder for children with rotavirus enteritis can reduce concentration of inflammatory factors in peripheral blood and decrease expression of CD8+ T lymphocyte subsets level and improve the level of CD3+, CD4+ level. It is a potent effective drug for treatment of children with rotavirus enteritis.

Observation on the Effect of Spontaneous Ventilation Single-Port Thoracoscopy and Conventional Thoracoscopy on T Lymphocyte Subsets and Inflammatory Factors in Patients with Non-Small Cell Lung Cancer

Objective:To explore the effects of two types of surgical approaches,spontaneous ventilation single-port thoracoscopy and conventional thoracoscopy,on T lymphocyte subsets and inflammatory factors in patients with non-small cell lung cancer.Methods:Fifty-eight patients with non-small cell lung cancer who underwent surgery in the Affiliated Hospital of Hebei University from January 2020 to December 2021 were selected as the study subjects.All of them underwent standard lobectomy and lymph node dissection.Among them,32 patients who underwent spontaneous ventilation single-port thoracoscopy were included in the study group,while 26 patients who underwent conventional thoracoscopy were included in the control group.The perioperative related indices,lung function,immune factor levels,and inflammatory factor levels were compared between the two groups.Results:In the study group,the perioperative incision length,intraoperative blood loss,operation time,and number of lymph node dissections were 4.28±2.01,77.89±12.02,87.21±16.11,and 20.69±4.45,respectively.In the control group,the perioperative incision length,intraoperative blood loss,operation time,and number of lymph node dissections were 6.32±2.45,84.98±16.98,95.88±14.89,and 21.45±4.36,respectively.There was no statistical difference between the study group and the control group in the number of lymph node dissections and intraoperative blood loss;the lung function in the study group was significantly better than that in the control group;the levels of T cell subsets CD4+,CD8+,CD4+/CD8+,CD16+CD56+of the study group were 46.36±5.87,30.98±4.12,1.19±0.23,and 17.41±6.25,respectively,while those in the control group were 35.78±4.12,34.14±3.87,1.04±0.24,and 12.45±5.56,respectively;the levels of immunoglobulin IgG,IgM,and IgA of the study group were 10.45±2.14,1.21±0.24,and 1.26±0.25,respectively,while those of the control group were 8.78±1.78,1.06±0.12,and 1.06±0.26,respectively,with statistical differences;the levels of CRP,IL-6,IL-8,and TNF-αof the control group were 21.87±4.26,98.01±9.58,111.03±9.96,and 123.05±9.77,respectively,while those of the study group were 19.47±3.89,89.12±8.96,104.32±9.12,and 112.98±9.16,respectively,in which the comparison was statistically significant.Conclusion:In the surgical treatment of non-small cell lung cancer,spontaneous ventilation single-port thoracoscopic surgery has several advantages,which include less trauma and bleeding.It reduces the impact of surgical trauma on the immune function of the body,improves the postoperative lung function and inflammatory stress of the body,as well as accelerates the recovery of patients.It is an alternative to open lung lobectomy for the treatment of lung cancer.

In vitro and in vivo analyses of a genetically-restricted antigen specific factor from mixed cell cultures of macrophage, T and B lymphocytes

An immunostimulatory factor was identified to be secreted by antigen-pulsed macrophages.This factor was able to induce the generation of antigen specific T helper lymphocytes in vitro as well as in vivo.Further in vitro experiments testing for the genetic restriction of this factor indicated that it is a geneticallyrestricted antigen specific factor (ASF).The Cunningham plaque assay was used to quantify the generation of T helper lymphocytes by measuring the number of plaque forming cells after sequential incubations of antigen-qulsed macrophages with T lymphocytes,and then spleen cells,and finally the TNP-coated sheep red blood cells.

Effect of cytotoxic T-lymphocyte antigen-4,TNF-alpha polymorphisms on osteosarcoma: evidences from a meta-analysis

Objective： Previous studies have investigated the role of cytotoxic T-lymphocyte antigen-4 （CTLA-4） and tumor necrosis factor-alpha （TNF-a） in carcinogenesis of osteosarcoma, but their results were inconsistent. We aimed to clarify the associations between CTLA-4, TNF-a polymorphism and osteosarcoma risk by using meta-analysis. Methods： We searched relevant studies without language restriction in PubMed, EMbase, Cochrane Library, Google Scholar databases, Chinese National Knowledge Infrastructure （CNKI） and conference literature in humans published prior to March 2013. The strengths of the associations between genetic variants and osteosarcoma risk were estimated by odds ratio （OR） with 95% confidence interval （95% CI）. Results： A total of seven studies with 1,198 osteosarcoma patients and 1,493 controls were selected. Four studies were eligible for CTLA-4 （1,003 osteosarcoma and 1,162 controls）, and three studies for TNF-a （195 osteosarcoma and 331 controls）. Pooled results showed that rs231775 polymorphism of CTLA-4 was associated with osteosarcoma risk （GG vs. AA： OR=1.63, 95% CI=1.24-2.13; GG ＋ GA vs. AA： OR=1.56, 95% CI=1.21-2.01; AA ＋ GA vs. GG： OR=0.83, 95% CI=0.71-0.97; G vs. A： OR=1.21, 95% CI=1.08-1.36）. No significant heterogeneity was observed across the studies. No significant associations were found between rs5742909 polymorphism of CTLA-4 or rs1800629 polymorphism of TNF-a and osteosarcoma risk. Conclusions： These results suggest that the rs231775 polymorphism of CTLA-4 may play an important role in carcinogenesis of osteosarcoma.

Effect of continuous blood purification on T lymphocyte subsets and inflammatory response in patients with severe sepsis

Objective:To investigate the effects of continuous blood purification (CBP) on T lymphocyte subsets and inflammatory response in patients with severe sepsis, and to provide scientific evidence for the treatment of severe sepsis patients. Methods:96 cases of severe sepsis patients from June 2013 to June 2016 in Yulin First People's Hospital were as the research subjects, randomly divided into observation group and control group, 48 cases in each group. The control group received routine treatment of sepsis, while the observation group adopted CBP therapy on the basis of the control group, fasting venous blood was collected 7 d before and after treatment respectively, and the change of IL-6, TNF-α, CRP levels, CD3+, CD4+, CD8+lymphocyte percentage and ratio of CD4+/CD8+in two groups were compared before and after treatment in two groups. Results:Before treatment, there were no significant differences in IL-6, TNF-α, and CRP levels between the two groups (P>0.05). After treatment, the IL-6 and TNF-αlevels of observation group and the control group were significantly decreased, CRP level in the observation group decreased significantly, and the IL-6, TNF-αand CRP levels were significantly lower in the observation group than in the control group (P<0.05). Before treatment, there was no statistical difference in the percentage of T cell subsets between the two groups (P<0.05). After treatment, the two groups of patients with CD3+, CD4+, CD8+and CD4+/CD8+were significantly elevated (P<0.05). After treatment, the CD3+, CD4+and CD4+/CD8+were significantly higher in the observation group than in the control group (P<0.05). Conclusions:CBP therapy can effectively reduce the level of inflammatory factors in patients with severe sepsis, reduce the inflammatory response, and can effectively improve the immune function of patients, and the effect is significant.

Liver tumor infiltrating lymphocytes: Comparison of hepatocellular and cholangiolar carcinoma

AIM: To investigate the role of tumor inf iltrating lym-phocytes (TIL) in primary hepatocellular and cholangio-lar carcinomas of the liver.METHODS: Immunohistochemical analysis was per-formed including antibodies to CD3, CD4, CD8, CD20, CD56 and TIA-1 in formalin-f ixed and paraff in-embed-ded tissue of 35 liver resection specimens of hepatocel-lular or cholangiocellular carcinomas. Semiquantitative evaluation was performed with emphasis on the area of the tumor itself and of the tumor/liver interface.RESULTS: All hepatocellular carcinomas showed in-filtration of lymphocytes predominantly around the tumor in the tumor/liver interface consisting mainly of CD3+ CD4+ T lymphocytes [164.3/10 high power f ields (HPF)] and in the tumor itself of CD8+ cells (54.9/10 HPF). Cholangiocarcinomas contained a heterogeneous amount of TIL, composed mainly of CD3+ T cells with a predominance of CD8+ cells in the tumor tissue (52.6/10 HPF) and of CD4+ cells in the interface region (223.1/10 HPF). CD56+ cells of the innate immune system were scarce. There was no significant difference between hepatocellular or cholangiolar carcinoma. No correlation with the clinicopathological data was seen. CONCLUSION: Liver TIL consists of intratumoral CD8+ T cells and peritumoral CD4+ T cells indepen-dent of histogenetic origin. Different functions of lym-phocytes in these regions seem possible.

Competition between TRAF2 and TRAF6 Regulates NF-κB Activation in Human B Lymphocytes

Objective To investigate the role of TNF receptor-associated factor 2 (TRAF-2) and TRAF6 in CD40-induced nuclear factor-κB (NF-κB) signaling pathway and whether CD40 signaling requires TRAF2. Methods Human B cell lines were transfected with plasmids expressing wild type TRAF2 or dominant negative TRAF2,TRAF2-shRNA,or TRAF6-shRNA. The activation of NF-κB was detected by Western blot,kinase assay,transfactor enzyme-linked immunosorbent assay (ELISA),and fluorescence resonance energy transfer (FRET). Analysis of the role of TRAF-2 and TRAF-6 in CD40-mediated NF-κB activity was examined following stimulation with recombinant CD154. Results TRAF2 induced activity of IκB-kinases (IKKα,IKKi/ε),phosphorylation of IκBα,as well as nuclear translocation and phosphorylation of p65/RelA. In contrast,TRAF6 strongly induced NF-κB activation and nuclear translocation of p65 as well as p50 and c-Rel. Engagement of CD154-induced nuclear translocation of p65 was inhibited by a TRAF6-shRNA,but conversely was enhanced by a TRAF2-shRNA. Examination of direct interactions between CD40 and TRAFs by FRET documented that both TRAF2 and TRAF6 directly interacted with CD40. However,the two TRAFs competed for CD40 binding. Conclusions These results indicate that TRAF2 can signal in human B cells,but it is not essential for CD40-mediated NF-κB activation. Moreover,TRAF2 can compete with TRAF6 for CD40 binding,and thereby limit the capacity of CD40 engagement to induce NF-κB activation.

Increased presence of effector lymphocytes during Helicobacter hepaticus-induced colitis

AIM:To identify and characterize drosophila mothers against decapentaplegic(SMAD)3-dependent changes in immune cell populations following infection with Helicobacter hepaticus(H.hepaticus).METHODS:SMAD3 -/-(n=19)and colitis-resistant SMAD3+/-(n=24)mice(8-10 wk of age)were in-fected with H.hepaticus and changes in immune cell populations[T lymphocytes,natural killer(NK)cells,T regulatory cells]were measured in the spleen and mesenteric lymph nodes(MsLNs)at 0 d,3 d,7 d and 28 d post-infection using flow cytometry.Genotypedependent changes in T lymphocytes and granzyme B+ cells were also assessed after 28 d in proximal colon tissue using immunohistochemistry.RESULTS:As previously observed,SMAD3 -/-,but not SMAD3+/-mice,developed colitis,peaking at 4 wk post-infection.No significant changes in T cell subsets were observed in the spleen or in the MsLNs between genotypes at any time point.However,CD4+and CD8+/ CD62Llo cells,an effector T lymphocyte population,as well as NK cells(NKp46/DX5+)were significantly higher in the MsLNs of SMAD3-/-mice at 7 d and 28 d post-infection.In the colon,a higher number of CD3+cells were present in SMAD3-/-compared to SMAD3+/-mice at baseline,which did not significantly change during infection.However,the number of granzyme B+cells,a marker of cytolytic lymphocytes,significantly increased in SMAD3-/-mice 28 d post-infection compared to both SMAD3+/-mice and to baseline values.This was consistent with more severe colitis development in these animals.CONCLUSION:Data suggest that defects in SMAD3 signaling increase susceptibility to H.hepaticus-induced colitis through aberrant activation and/or dysregulation of effector lymphocytes.

Characteristics of Lymphocyte Nuclear Factor-κB Signal Transduction Kinase Expression in Aging Process and Regulatory Effect of Epimedium Flavonoids

Objective：To study the characteristics of lymphocyte nuclear factor kappa B（NF-κB） signal transduction kinase-related molecular mRNA differential expressions at various month age segments in aging process and the intervening effect of Epimedium flavonoids（EF） on it.Methods：Sixty SD rats were divided into six groups,according to animals＇ age,i.e.,the 3 days（d） group,the 4 months（m） group,the 10 m group,the 18 m group,the 27 m group,and the 27 m＋EF group.RNA was extracted from separated splenic lymphocytes. Adopting NF-κB signal path functional genome oligonucleotide gene-chip（128 related genes）,the integral characteristics and differences of NF-κB signal transduction kinase-related mRNA expressions were determined, and the intervening effect of EF was examined.Results：The mean level of the NF-κB signal transduction kinase-related mRNA expressions in rats＇ splenic lymphocytes lowered with aging;the highest expression was presented at 3 d after birth,and then,it lowered gradually,with the lowest level at 18 m or 27 m.After EF intervention,the expression level was raised to the 10-18 m level in the aged rats.Conclusion：The changing rules of lymphocyte NF-κB-signal-transduction-kinase-related mRNA expressions in various stages of aging are helpful for selecting the well time for preventing and intervening aging,and will also give a hint to the molecular index for assessment of senility retarding researches.

Effect of charge at an amino acid of basic fibroblast growth factor on its mitogenic activity

The amino acid at the 119th position of human basic fibroblast growth factor(hbFGF),lysine(K119),is a critical component for its mitogenic activity.However,little is known about the effects of the characteristics of this residue including charge on the mitogenic activity of hbFGF.Herein,this basic residue was replaced with neutral glutamine residue and acidic glutamic acid residue to construct mutants hbFGF^(K119Q) and hbFGF^(K119E),respectively.The mutants were produced by BL21(DE3)/pET3c expression system...

B lymphocyte stimulator： a new target for treating B cell malignancies

Blymphocyte stimulator （BLyS） is also known as B cell activating factor belonging to the tumor necrosis factor （TNF） family （BAFF）, or TNF and apoptosis ligand-related leukocyte-expressed ligand 1 （TALL-I）.^1-3 As a member of the TNF family, BLyS plays an important role in B cell development, and has been implicated in autoimmune diseases and B cell malignancies.

Effects of Shen-Fu Injection (参附注射液) on Apoptosis of Regulatory T Lymphocytes in Spleen during Post-Resuscitation Immune Dysfunction in A Porcine Model of Cardiac Arrest

Objective： To investigate whether Shen-Fu Injection（参附注射液, SFI） reduces post-resuscitation immune dysfunction in a porcine model of cardiac arrest by modulating apoptosis of regulatory T lymphocytes（Treg） in the spleen. Methods： After 8-min untreated ventricular fibrillation and 2-min basic life support, 24 pigs were divided into 3 groups with a random number table, i.e. SFI group, epinephrine（EP） group, and saline（SA） group（8 in each group）, which received central venous injection of SFI（1.0 m L/kg）, EP（0.02 mg/kg） and SA, respectively. The same procedure without CA initiation was achieved in the sham-operated（sham） group（n=6）. After successful return of spontaneous circulation（ROSC）, apoptosis rate of splenic Treg was detected by flow cytometry; and the m RNA expression of forkhead/winged helix transcription factor（Foxp3） of splenic Treg was detected by real time-polymerase chain reaction; and the levels of interleukin-4（IL-4） and interferon-γ（IFN-γ） in porcine splenic Treg were detected by using enzyme-linked immunosorbent assay（ELISA）. Results： Compared with the sham group, the apoptosis rate of Treg was significantly decreased, and the levels of Foxp3 m RNA expression, IFN-γ, IL-4 and IFN-γ/IL-4 were increased in the SA group（P〈0.05 or P〈0.01）. Compared with the EP and SA groups, SFI treatment increased the apoptosis rate of Treg and reduced the levels of Foxp3 m RNA expression, IFN-γ and IFN-γ/IL-4（P〈0.05）. Conclusions： SFI has significant effects in attenuating post-resuscitation immune dysfunction by modulating apoptosis of Treg in the spleen.

Rapid mitogen-activated protein kinase activation by transforming growth factor α in wounded rat intestinal epithelial cells

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Trefoil factors:Tumor progression markers and mitogens via EGFR/MAPK activation in cholangiocarcinoma

AIM:To investigate trefoil factor(TFF) gene copy number,mRNA and protein expression as potential biomarkers in cholangiocarcinoma(CCA).METHODS:TFF mRNA levels,gene copy number and protein expression were determined respectively by quantitative reverse transcription polymerase chain reaction(PCR),quantitative PCR and immunohistochemistry in bile duct epithelium biopsies collected from individuals with CCA,precancerous bile duct dysplasia and from disease-free controls.The functional impact of recombinant human(rh) TFF2 peptide treatment on proliferation and epidermal growth factor receptor(EGFR) /mitogenactivated protein kinase(MAPK) signaling was assessed in the CCA cell line,KMBC,by viable cell counting and immunoblotting,respectively.RESULTS:TFF1,TFF2 and TFF3 mRNA expression was significantly increased in CCA tissue compared to disease-free controls,and was unrelated to gene copy number.TFF1 immunoreactivity was strongly increased in both dysplasia and CCA,whereas TFF2 immunoreactivity was increased only in CCA compared to diseasefree controls.By contrast,TFF3 immunoreactivity was moderately decreased in dysplasia and further decreased in CCA.Kaplan-Meier analysis found no association of TFF mRNA,protein and copy number with age,gender,histological subtype,and patient survival time.Treatment of KMBC cells with rhTFF2 stimulated proliferation,triggered phosphorylation of EGFR and downstream extracellular signal related kinase(ERK),whereas co-incubation with the EGFR tyrosine kinase inhibitor,PD153035,blocked rhTFF2-dependent proliferation and EGFR/ERK responses.CONCLUSION:TFF mRNA/protein expression is indicative of CCA tumor progression,but not predictive for histological sub-type or survival time.TFF2 is mitogenic in CCA via EGFR/MAPK activation.

Inhibition of p38 mitogen-activated protein kinase attenuates experimental autoimmune hepatitis: Involvement of nuclear factor kappa B

AIM: To investigate the role of p38 mitogen-activated protein kinase (p38MAPK) in murine experimental autoimmune hepatitis (EAH). METHODS: To induce EAH, the syngeneic S-100 antigen emulsified in complete Freud's adjuvant was injected intraperitoneally into adult male C57Bl/6 mice. Liver injury was assessed by serum ALT and liver histology. The expression and activity of p38 MAPK were measured by Western blot and kinase activity assays. In addition, DNA binding activities of nuclear factor kappa B (NF-κB) were analyzed by electrophoretic mobility shift assay. The effects of SB203580, a specifi c p38 MAPK inhibitor, on liver injuries and expression of proin? ammatory cytokines (interferon-γ, IL-12, IL-1β and TNF-α) were observed. RESULTS: The activity of p38 MAPK and NF-κB was increased and reached its peak 14 or 21 d after the fi rst syngeneic S-100 administration. Inhibition of p38 MAPK activation by SB203580 decreased the activation of NF-κB and the expression of proin? ammatory cytokines. Moreover, hepatic injuries were improved significantly after SB203580 administration. CONCLUSION: p38 MAPK and NF-κB play an important role in an animal model of autoimmune hepatitis (AIH) induced by autoantigens.

Noni(Morinda citrifolia L.) fruit juice delays immunosenescence in the lymphocytes in lymph nodes of old F344 rats

Objective： Aging is associated with the development of diseases because of immunosuppression and altered functioning of the neuroendocrine system. The medicinal properties of Morinda citrifolia L. have been widely exploited for the treatment of age-associated diseases. This study aims to investigate the in vitro and in vivo effects of noni（M. citrifolia） fruit juice（NFJ） on neuro-immunomodulation in the lymph node lymphocytes of F344 rats.Methods： Lymphocytes isolated from axillary and inguinal lymph nodes of young（3–4 months） and old（18–21 months） rats were treated in vitro with different concentrations（0.0001%, 0.01%, and 1%） of NFJ for a period of 24 h. In the in vivo study, old（16–17 months） male F344 rats were treated with 5 mL/kg body weight of 5%, 10% and 20% of NFJ, twice a day, by oral gavage, and lymph node lymphocytes were isolated after 60 d. Concanavalin A（Con A）-induced lymphocyte proliferation, interleukin-2（IL-2）and interferon-γ（IFN-γ） production and expression of intracellular markers, such as phosphoextracellular signal-regulated kinase（p-ERK1/2）, phospho-γ AMP response element-binding protein,phospho-protein kinase B（p-Akt）, phospho-tyrosine hydroxylase（p-TH）, phospho-nuclear factor of κ light polypeptide gene enhancer in B-cells inhibitor-α（p-IκB-α） and phospho-nuclear factor-κB（p-NF-κB p65 and p50） were examined in the lymphocytes of lymph nodes.Results： NFJ increased Con A-induced lymphocyte proliferation, IL-2 and IFN-γ production, and p-ERK1/2 expression both in vitro and in vivo. In in vivo NFJ-treated old rats, lymph node lymphocytes showed increased expression of p-TH and Akt, nitric oxide production and decreased expression of p-NF-κB p65 and p50.Conclusion： These results suggest that the immunostimulatory properties of NFJ are facilitated through intracellular signaling pathways involving ERK1/2, Akt and NF-κB.