Contributions of HO-1-Dependent MAPK to Regulating Intestinal Barrier Disruption

The mitogen-activated protein kinase (MAPK) pathway controls intestinal epithelial barrier permeability by regulating tight junctions (TJs) and epithelial cells damage. Heme oxygenase-1 (HO-1) and carbon monoxide (CO) protect the intestinal epithelial barrier function, but the molecular mechanism is not yet clarified. MAPK activation and barrier permeability were studied using monolayers of Caco-2 cells treated with tissue necrosis factor α (TNF-α) transfected with FUGW-HO-1 or pLKO.1-sh-HO-1 plasmid. Intestinal mucosal barrier permeability and MAPK activation were also investigated using carbon tetrachloride (CCl 4) administration with CoPP (a HO-1 inducer), ZnPP (a HO-1 inhibitor), CO releasing molecule 2 (CORM-2), or inactived-CORM-2-treated wild-type mice and mice with HO-1 deficiency in intestinal epithelial cells. TNF-α increased epithelial TJ disruption and cleaved caspase-3 expression, induced ERK, p38, and JNK phosphorylation. In addition, HO-1 blocked TNF-α-induced increase in epithelial TJs disruption, cleaved caspase-3 expression, as well as ERK, p38, and JNK phosphorylation in an HO-1-dependent manner. CoPP and CORM-2 directly ameliorated intestinal mucosal injury, attenuated TJ disruption and cleaved caspase-3 expression, and inhibited epithelial ERK, p38, and JNK phosphorylation after chronic CCl 4 injection. Conversely, ZnPP completely reversed these effects. Furthermore, mice with intestinal epithelial HO-1 deficient exhibited a robust increase in mucosal TJs disruption, cleaved caspase-3 expression, and MAPKs activation as compared to the control group mice. These data demonstrated that HO-1-dependent MAPK signaling inhibition preserves the intestinal mucosal barrier integrity by abrogating TJ dysregulation and epithelial cell damage. The differential targeting of gut HO-1-MAPK axis leads to improved intestinal disease therapy.

Expression and significance of PD-1 and PD-L1 in the specimens of epithelial ovarian cancer / 中华妇产科杂志

Objective:To examine the expression of programmed cell death 1 (PD-1) and its ligand (PD-L1) in epithelial ovarian cancer (EOC) tissues, and investigate the correlation among their expression, clinicopathological features and prognosis.Methods:The specimens of 180 patients with EOC treated in the First Affiliated Hospital of Dalian Medical University from October 2002 to December 2013 were confirmed by pathological examination. The pathological tissue specimens of subtypes ，included 120 cases of serous carcinoma, 30 cases of mucinous carcinoma, 20 cases of endometrioid carcinoma, and 20 cases of clear cell carcinoma. The normal paracancerous tissues of 50 cases randomly selected from the 180 patients as control group. Immunohistochemical SP method was used to detect the expressions of both PD-1 and PD-L1 in epithelial ovarian cancer tissues, and the relationships among their expressions,the clinicopathological parameters and prognosis were respectively analyzed.Results:（1） PD-1 was expressed in lymphocytes infiltrated in EOC tissues, and PD-L1 was expressed in the cell membranes of cancer tissues. In all EOC cases, 33 cases (18.3%, 33/180) of both PD-1 and PD-L1 were highly expressed, and only 1 (2.0%, 1/50) of control group showed high expression. There was statistically significant difference between two groups ( P<0.01). (2) Among the four subtypes tissue specimens of EOC, the high expression rate of PD-1 was 25.0% (30/120) for serous carcinoma, 3/15 for endometrioid carcinoma, 0 (0/30) for mucinous carcinoma, and 0 (0/15) for clear cell carcinoma. The high expression rate of PD-L1 was 23.3% (28/120) for serous carcinoma, 3.3% (1/30) for mucinous carcinoma, 2/15 for endometrioid carcinoma, and 2/15 for clear cell carcinoma. Both PD-1 and PD-L1 expressions in the four sub-types of tissue specimens were significantly different ( P<0.05). The high expression rate of both PD-1 and PD-L1 was 9.2% (8/87) in the early stage and 26.9% (25/93) in the late stage. There was a statistically significant difference between the two groups ( P<0.01). Similarly, the expression of both PD-1 and PD-L1 were significantly higher in the cases of high-grade EOC （type Ⅱ） than those of low-grade （type Ⅰ） and in the cases of EOC distributed bilaterally than that distributed unilaterally, and there were statistically significant differences ( P<0.05). (3) The Kaplan-Meier survival analysis showed that the survival time were respectively 35 and 36 months in the cases with high expressions of both PD-1 and PD-L1, and the survival time were the same as 61 months in the cases with low expression of both PD-1 and PD-L1, and the comparison was statistically significant ( P<0.05). Conclusions:The expression levels of PD-1 and PD-L1 in EOC tissues are higher than those in adjacent tissues, especially in serous carcinomas. The expression of both PD-1 and PD-L1 is higher in specimens of the patients with advanced stages. The results showed that the high expression of both PD-1 and PD-L1 is an indicator of poor prognosis of patients suffering from EOC.

Expression and significance of MAPK/ERK in the specimens and cells of epithelial ovarian cancer / 中华妇产科杂志

Objective To detect phosphorylated-extracellular signal-regulated kinase (p-ERK1/2) protein expression in epithelial ovarian cancer and cell lines,and to examine the effects of mitogen-activated protein kinase (MAPK) kinase (MEK) inhibitor AZD6244 on cell proliferation,apoptosis as well as cell cycle of ovarian cancer cells.To explore the function and significance of MAPK/extracellular signal-regulated kinase (ERK) signaling pathway in the development of ovarian cancer.Methods (1) A total of 104 cases of patients with ovarian cancer who accepted the treatment of gynecological surgery and being confirmed by pathological examination in First Affiliated Hospital,Dalian Medical University from January 2004 to December 2013 were selected.The expressions of p-ERK1/2 protein were detected by immunohistochemistry in ovarian cancer specimens,and the relationship between the expressions of p-ERK1/2 and the clinical features of patients was analyzed.(2) p-ERK1/2 and other related proteins were determined by western blot in various ovarian cancer cells,including SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3 treated with or without MEK inhibitor.The cellular proliferation,apoptosis and cell cycle of ovarian cancer cells after treatment with MEK inhibitor were analyzed by methyl thiazolyl tetrazolium (MTT) assay and flow cytometry,respectively.Results (1) The immunohistochemical method showed that p-ERK1/2 between low grade serous carcinoma and clear cell carcinoma were not significantly higher expressed (P＞0.05).However,a lower level of the p-ERK1/2 expression were observed among high grade serous carcinoma,mucinous carcinoma and endometrioid carcinoma (all P＜0.05).There was no significant correlation between the protein expression of p-ERK1/2 and patients' age,pathological stage of surgery,and preoperative serum CA125 level (P＞0.05).(2) Western blot showed that the protein p-ERK1/2 was widely expressed in various ovarian cancer cell lines such as SKOV3,OV2008,C13,A2780S,A2780CP,OVCAR4,OVCAR5,OVCAR8 and CAOV3.After treatment with AZD6244 (5,10 μmol/L),the level of p-ERK 1/2 in OVCAR5 and OVCAR8 decreased significantly in dose-dependent manner.Additionally,we found a reduction of the expression level of cyclin D 1,caspase-3 and appeared cleaved poly adenosine diphosphate ribose polymerase (PARP) in OVCAR5 and OVCAR8,compared with control groups.MTT assays showed that OVCAR5,OVCAR8 and A2780S were differently inhibited in the dose-dependent manner after being treated with different concentrations of AZD6244 (0,2.5,5,10,25,50 and 100 μmol/L,all P＜0.05).Further tested by flow cytometry,the results showed that AZD6244 (5,10 μmol/L) was able to induce the apoptosis of OVCAR5,OVCAR8 and A2780S,as well as G0/G1 phase arrest,both in a dose-dependent manner (P＜0.05).Conclusions As the main active and functional unit of MAPK/ERK signaling pathway,p-ERK1/2 protein is expressed in both the tissues and various ovarian cancer cell lines.AZD6244 could down-regulated the expression of p-ERK1/2 in ovarian cancer cells,accompanied by the decreased proliferation and increased cell apoptosis of ovarian cancer cells.In conclusion,MAPK/ERK signaling pathway might play a role in the development and progression of ovarian cancer,and may be provide a novel option for molecular targeted therapies of the disease.

Distrbution and risk factors analysis of infections caused by extended-spectrum β-lactamase-producing Escherichia coli / 国际检验医学杂志

Objective To analyze the specimen types,ward distribution and risk factors for infections caused by extended-spectrum β-lactamase(ESBLs)-producing-Escherichia coli(ECO) in recent two years,so as to provide bacteriological basis for both hospital infection control and clinical anti-infection treatment.Methods Non-repetitive 443 ECO strains isolated from the hospitalized patients in the Third People′s Hospital of Shenzhen were collcted,and the phoenix100 system was employed for bacterial identification and antimicrobial susceptibility tests.ESBLs-ECO was further confirmed by the double-disk synergy test,and the risk factors caused ESBLs-ECO were statistically analyzed.Results A total of 115 strains of ESBLs-ECO were identified among the 443 strains of ECO,which accounted for 26.0%.The ESBLs-ECO strains were mainly isolated from the sputum,urine,and blood specimens.Among the isolated ESBLs-ECO strains,20.9% were isolated from the department of Tuberculosis,13.9% from the department of pediatric,12.2% from the department of live disease,and 8.7% from the department of infection.The male sex,surgery and use of the third generation cephalosporins were independent risk factors of ESBLs-ECO infection.Conclusion The isolation rate of ESBLs-ECO in this hospital is high.It is necessary for the hospital to strengthen the control of nosocomial infections according to the risk factors.More attention should be payed on male patients,the standardization of surgical operation and disinfection,and the restriction of using the third generation cephalosporins,so as to reduce the incidene of ESBLs-ECO infections.

Effect of sitagliptin on insulin resistance and glucose variability in elderly patients with type 2 diabetes mellitus requiring high-dose insulin therapy / 重庆医学

Objective To study the effect of sitagliptin on insulin resistance and glucose variability in elderly patients with type 2 diabetes mellitus(T2DM) requiring high-dose insulin therapy.Methods A total of 100 elderly patients with T2DM failing to reach the standard application of large-dose insulin(＞60 U/d) treatment for three months or more [glycosylated hemoglobin (HbA1c) ＞8.0％] was randomly divided into sitagliptin group and pioglitazone group.Patients in sitagliptin group(50 cases) were treated with sitagliptin for oral use,100 mg each time,once a day,and patients in pioglitazone group(50 cases) were treated with pioglitazone for oral use,15 mg each time,once a day.The insulin dose was adjusted according to the blood glucose level in the two groups.Two groups were treated for 12 weeks.The indicators in both groups were compared,including fasting blood glucose (FBG),2 hours postprandial glucose (2hPG),glycosylated hemoglobin (HbA1c),24 hours glucose area under the curve (AUC),blood glucose coefficient of variation (CV),fasting C-peptide (FCP),2 hours postprandial C-peptide (2hPCP),fasting glucagon (FGG),2 hours postprandial glucagon(2 hFGG),cholesterol (TC),triglyceride (TG),systolic blood pressure (SBP),diastolic blood pressure(DBP),blood uric acid(BUA),daily insulin dosage(DID),body mass index(BMI),incidence of hypoglycemia and drug adverse reactions.Results After 12 weeks of treatment,the levels of FPG,2hPG,HbA1c,AUC,CV,FGG,2hFGG,TC,TG,SBP,DBP,DID and BMI in the sitagliptin group were significantly decreased than those before treatment(P＜0.05 or P＜0.01);The levels of FPG,2hPG,H bA1c,AUC,CV and BUA in the pioglitazone group were significantly decreased than those before treatment (P＜ 0.0 5 or P＜ 0.01);Compared with the pioglitazone group,the levels of 2 hPG,HbA1c,AUC,CV,FGG,2 hFGG,TC,TG,SBP,DBP,DID and BMI were significantly decreased in the sitagliptin group(all P＜0.05),and the levels of FCP and 2hPCP in the sitagliptin group were higher than those in the pioglitazone group(all P＜0.01).The incidence of hypoglycemia in the sitagliptin group was lower than that in the pioglitazone group(x2 =4.039,P =0.045).The incidence of adverse reactions in the sitagliptin group was lower than that in the pioglitazone grouP(x2 =3.979,P=0.043).Conclusion Sitagliptin combined with insulin is better than insulin combined with pioglitazone in elderly patients with T2DM requiring the application of high-dose insulin therapy,and the combining treatment could decrease insulin resistance,insulin dosage and the incidence of hypoglycemia.

Difference of peripheral blood leucocyte percentages in malaria patients among Sysmex-XE5000,CellaVisionDM96 and manual microscope classification / 国际检验医学杂志

Objective To compare the difference of peripheral blood leucocyte percentages in malaria patients among Sysmex‐XE5000 ,CellaVisionDM96 and manual microscope classification ,and to verify the accuracy and reliability of the Sysmex‐XE5000 automatic blood corpuscle detection instrument for detecting the leukocyte classification in blood routine .Methods The leucocyte percentages data in 82 cases of malaria detected by using the Sysmex‐XE5000 in the Shenzhen Municipal Third People′s Hospital from January 2011 to December 2015 were retrospectively collected ;the peripheral blood smear in 82 cases of malaria obtained the percentages after the classification by the CellaVisionDM 96 ;then the peripheral blood smear was performed the leucocyte classifica‐tion by the manual microscopy for calculating the percentage .Results In the pairwise comparison of percentage obtained from the peripheral blood leucocyte classification by Sysmex‐XE5000 ,CellaVisionDM96 and manual microscopy ,only the monocytes percent‐age had statistical difference between CellaVisionDM 96 and manual microscopy (P0 .05) .Conclusion By comparing the peripheral blood leucocyte percentages data in malaria patients by Sysmex‐XE5000 ,CellaVisionDM96 and manual microscopic classification ,it is indicated that the leukocyte classification data by Sysmex‐XE5000 are accurate and reliable ,malaria parasite does not affect peripheral blood leukocyte classification ,but it is necessary to pay more attention to monocytes classification in CellaVision DM 96 classification .

Concomitance of P-gp/LRP Expression with EGFR Mutations in Exons 19 and 21 in Non-Small Cell Lung Cancers

PURPOSE: Traditional chemotherapy is the main adjuvant therapy for the treatment of non-small cell lung cancer (NSCLC). However, the emergence of multi-drug resistance (MDR) has greatly restricted the curative effect of chemotherapy. Therefore, it is necessary to find a method to treat MDR NSCLC clinically. It is worth investigating whether NSCLCs that are resistant to traditional chemotherapy can be effectively treated with tyrosine kinase inhibitors targeting epidermal growth factor receptor (EGFR). MATERIALS AND METHODS: The expression of P-glycoprotein (P-gp) and lung resistance-related protein (LRP) was detected by immunohistochemistry, and mutations in EGFR (exons 19 and 21) and Kirsten rat sarcoma viral oncogene homolog (KRAS) (exon 2) were detected by high-resolution melting analysis (HRMA) of surgical NSCLC specimens from 127 patients who did not undergo traditional chemotherapy or radiotherapy. A Pearson chi-square test was performed to analyze the correlations between the expression of P-gp and LRP and mutations in EGFR and KRAS. RESULTS: The expression frequencies of P-gp and LRP were significantly higher in adenocarcinomas from non-smoking patients; the expression frequency of LRP was significantly higher in cancer tissue from female patients. The frequency of EGFR mutations was significantly higher in well to moderately differentiated adenocarcinomas from non-smoking female patients. The frequency of EGFR mutations in the cancers that expressed P-gp, LRP, or both P-gp and LRP was significantly higher than that in cancers that did not express P-gp or LRP. CONCLUSION: NSCLCs expressing P-gp/LRP bear the EGFR mutation in exon 19 or 21 easily.

‘Response to injury’ teaching based on the concept of global minimum essential requirements / 中华医学教育探索杂志

Taking global minimum essential requirements(GMER)as standard,school of basic medical science in Wuhan University paid more attention to the cultivation of students' professional skills, innovation ability and communication ability. ‘Response to injury’integrated course was carried out. Based on the immunology,this course involves pathology and pathophysiology and explores the mecha-nism of the diseases' diagnosis and treatment. It took small classes,bilingual education,literature read-ing ,experiments and discussions in various forms . This reform aimed to improve the immunology , pathology and pathophysiology teaching and make our medical education meet the process of globalization.

Study on dynamic of chemical components of rhizoma alismatis and Evodia rutaecarpa in their compatibility / 中国中药杂志

This article studied the changes of chemical components in decoctions that made from the single medicinal herb of Rhizoma Alismatis or Evodia rutaecarpa, the co-decoctions made from different compatibilities of the two medicinal herbs, and that mixed decotions of two single medicinal herb decotions, by HPLC. The changes of chemical components of the codicotions were focused on. The relationships between the changes of chemical components of the codicotions and defferent compatibility ratios of medicinal herb were summarized.

Chemical constituents of aerial parts of Gelsemium elegans / 中国中药杂志

<p><b>OBJECTIVE</b>To study the chemical constituents from the aerial parts of Gelsemium elegans.</p><p><b>METHOD</b>Compounds were isolated and purified by repeated column chromatography, as well as semiprep arative HPLC, and their structures were identified by physicochemical properties and spectroscopic methods, such as NMR and MS.</p><p><b>RESULT</b>Sixteen compounds were obtained and identified from G. elegans, including nine alkaloids: koumine (1), gelsenicine (2), 19-(Z)-akuammidine (3), gelsemoxonine(4), gelsemin (5), gelsevirine (6), humantenine (7), 11-methoxygelsemamide (8) and gelegamine D (9). Three megastigmane glycosides: (3R, 5S, 6S, 7E, 9R)-megastigman-7-ene-3, 5, 6, 9-tetrol-9-O-beta-D-glucopyranoside (10), (6R, 7E, 9R)-9-hydroxy-4, 7-megastigmadien-3-one-9-O-[alpha-L-arabinopyranosyl-(1 --> 6)-beta-D-glucopyranoside] (11) and (6S, 7E, 9R) -6, 9-dihydroxy-4, 7-megastigmadien-3-one-9-O-[alpha-L-arabinopyranosyl-(1 --> 6) -beta-D-glucopyranoside] (12). Two flavone C-glycosides: orientin (13) and isorientin (14); one iridoid glycoside, sweroside (15) and one fructoside, n-butyl-alpha-D-fructofuranoside (16).</p><p><b>CONCLUSION</b>Compounds 10-16 were isolated from the genus Gelsemium for the first time.</p>

Expression and clinical significance of hypoxia-inducible factor-1alpha and angiopoietin-2 in laryngeal squamous cell carcinoma / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To investigate the expressions and clinical significance of Hypoxia-Inducible Factor-1alpha (HIF-1alpha) and angiopoietin-2 (Ang-2) proteins in laryngeal squamous cell carcinoma.@*METHOD@#Immunohistochemical technique was used to detect the expressions of HIF-1alpha and Ang-2 protein in 52 laryngeal squamous cell carcinoma and 10 normal tissues.@*RESULT@#The positive rates of HIF-1alpha and Ang-2 proteins were significantly higher in laryngeal squamous cell carcinoma than those in normal tissues (P 0.05). The level of Ang-2 expression was also correlated with TNM stage, lymph node metastasis and histological differentiation of laryngeal squamous cell carcinoma. But not related to the age and sex. There was positive correlations between both expressions of HIF-1alpha and Ang-2 proteins (r = 0. 333, P < 0.01).@*CONCLUSION@#High expressions of HIF-1alpha and Ang-2 proteins in laryngeal squamous cell carcinoma may play an important role in tumor growth, invasion and metastasis.

CXC chemokine ligand 13 and CC chemokine ligand 19 cooperatively render resistance to apoptosis in B cell lineage acute and chronic lymphocytic leukemia CD23+CD5+ B cells.

CXCL13/CXCR5 and CCL19/CCR7 play a quite important role in normal physiological conditions, but the functions of both chemokine/receptor pairs in pathophysiological events are not well-investigated. We have investigated expression and functions of CXCL13/CXCR5 and CCL19/CCR7 in CD23+CD5+ and CD23+CD5- B cells from cord blood (CB) and patients with B cell lineage acute or chronic lymphocytic leukemia (B-ALL or B-CLL). CXCR5 and CCR7 are selectively expressed on B-ALL, B-CLL, and CB CD23+CD5+ B cells at high frequency, but not on CD23+CD5- B cells. Although no significant chemotactic responsiveness was observed, CXCL13 and CCL19 cooperatively induce significant resistance to TNF-alpha-mediated apoptosis in B-ALL and B-CLL CD23+CD5+ B cells, but not in the cells from CB. B-ALL and B-CLL CD23+CD5+ B cells express elevated levels of paternally expressed gene 10 (PEG10). CXCL13 and CCL19 together significantly up-regulate PEG10 expression in the same cells. We have found that CXCL13 and CCL19 together by means of activation of CXCR5 and CCR7 up-regulate PEG10 expression and function, subsequently stabilize caspase-3 and caspase-8 in B-ALL and B-CLL CD23+CD5+ B cells, and further rescue the cells from TNF-alpha-mediated apoptosis. Therefore, we suggest that normal lymphocytes, especially naive B and T cells, use CXCL13/CXCR5 and CCL19/CCR7 for migration, homing, maturation, and cell homeostasis as well as secondary lymphoid tissues organogenesis. In addition, certain malignant cells take advantages of CXCL13/CXCR5 and CCL19/CCR7 for infiltration, resistance to apoptosis, and inappropriate proliferation.

Selectively frequent expression of CXCR5 enhances resistance to apoptosis in CD8(+)CD34(+) T cells from patients with T-cell-lineage acute lymphocytic leukemia.

We investigated CD4(+)CD34(+), CD8(+)CD34(+), CD4(+)CD34(-), and CD8(+)CD34(-) T cells from cord blood and from typical patients with T-cell-lineage acute lymphocytic leukemia and T-cell-lineage chronic lymphocytic leukemia in terms of expression and functions of CXCR5/CXCL13. We found that CXCR5 was selectively frequently expressed on T-cell-lineage acute (chronic) lymphocytic leukemia (T-ALL) CD8(+)CD34(+) T cells, but not on T-ALL CD4(+)CD34(+), CD4(+)CD34(-), and CD8(+)CD34(-) T cells. CXCR5 was rarely expressed on all types of CD34(+) and CD34(-) CB or T-CLL T cells. CXCL13/B cells attracting chemokine 1 induced significant resistance to TNF-alpha-mediated apoptosis in T-ALL CD8(+)CD34(+) T cells, instead of induction of chemotactic and adhesive responsiveness. A proliferation-inducing ligand expression in T-ALL CD8(+)CD34(+) T cells was upregulated by CXCL13/BCA-1 (B-cell attracting chemokine 1). The CXCR5/CXCL13 pair by means of activation of APRIL (A proliferation-inducing ligand) induced resistance to apoptosis in T-ALL CD8(+)CD34(+) T cells in livin-dependent manner. In this process, cell-cell contact in culture was necessary. Based on our findings, we suggested that there were differential functions of CXCR5/CXCL13 in distinct types of cells. Normal lymphocytes, especially naive B and T cells, utilized CXCR5/CXCL13 for migration, homing, maturation, and cell homeostasis, as well as secondary lymphoid tissue organogenesis. Meanwhile, certain malignant cells took advantages of CXCR5/CXCL13 for infiltration, resistance to apoptosis, and inappropriate proliferation.

CC chemokine ligand 25 enhances resistance to apoptosis in CD4+ T cells from patients with T-cell lineage acute and chronic lymphocytic leukemia by means of livin activation.

We investigated CD4 and CD8 double-positive thymocytes, CD4(+) T cells from typical patients with T-cell lineage acute lymphocytic leukemia (T-ALL) and T cell lineage chronic lymphocytic leukemia (T-CLL), and MOLT4 T cells in terms of CC chemokine ligand 25 (CCL25) functions of induction of resistance to tumor necrosis factor alpha (TNF-alpha)-mediated apoptosis. We found that CCL25 selectively enhanced resistance to TNF-alpha-mediated apoptosis in T-ALL and T-CLL CD4(+) T cells as well as in MOLT4 T cells, but CD4 and CD8 double-positive thymocytes did not. One member protein of the inhibitor of apoptosis protein (IAP) family, Livin, was selectively expressed in the malignant cells at higher levels, particularly in T-ALL CD4(+) T cells, in comparison with the expression in CD4 and CD8 double-positive thymocytes. After stimulation with CCL25 and apoptotic induction with TNF-alpha, the expression levels of Livin in these malignant cells were significantly increased. CCL25/thymus-expressed chemokine (TECK), by means of CC chemokine receptor 9 (CCR9) ligation, selectively activated Livin to enhance resistance to TNF-alpha-mediated apoptosis in c-jun-NH(2)-kinase 1 (JNK1) kinase-dependent manner. These findings suggested differential functions of CCR9/CCL25 in distinct types of cells. CD4 and CD8 double-positive thymocytes used CCR9/CCL25 for migration, homing, development, maturation, selection, cell homeostasis, whereas malignant cells, particularly T-ALL CD4(+) T cells, used CCR9/CCL25 for infiltration, resistance to apoptosis, and inappropriate proliferation.

Aberration of CCR7 CD8 memory T cells from patients with systemic lupus erythematosus: an inducer of T helper type 2 bias of CD4 T cells.

Chemokine receptors are important in the entry of leucocytes into the inflammatory sites of systemic lupus erythematosus (SLE). CCR7(+) and CCR7(-) memory T cells exert different functions in homing, cytokine production and cytotoxicity. To determine whether differential expression and functions of the CCR7 occur in SLE patients, we examined CCR3, CCR4, CCR5, CCR7 and CCR9 on CD4(+) and CD8(+) T cells from normal and SLE subjects. Flow cytometry, real-time quantitative reverse transcription polymerase chain reactions and Northern blotting were used to detect the expression of chemokine receptors and cytokines; a chemotaxis assay was used to detect their functions. CD4(+) T-cell stimulation with syngeneic CCR7(+) CD8(+) CD45RO(+) T cells and dendritic cells (including transwell chambers) was used to induce cytokine expression. We demonstrated that CCR7 was selectively, frequently and functionally expressed on CD8(+) (94.8%) but not on CD4(+) (16.1%) T cells from patients with active SLE, whereas this phenomenon was not seen in normal subjects and in those whose SLE was inactive. CCR7(+) CD8(+) CD45RO(+) memory T cells from patients with active SLE, themselves T helper type 2 (Th2) biased, were inducers of Th2 bias in CD4(+) T cells in a cell-cell contact manner in vitro, meanwhile, the cells from both normal subjects and those whose SLE was inactive drove CD4(+) T cells into a regulatory T-cell-derived cytokine pattern. Our findings might provide new clues to understanding the functions of CCR7(+) CD8(+) CD45RO(+)'central' memory T cells in autoimmune diseases (such as SLE). We suggest that in the case of active SLE, CCR7(+) central memory T cells were able to enter peripheral blood and inflammatory sites from secondary lymphoid organs, were continuously expressing CCR7, and interacted with dendritic cells and functioned as CCR7(-)'effector' memory T cells, which were described in normal humans.

Highly up-regulated CXCR3 expression on eosinophils in mice infected with Schistosoma japonicum.

CXCR3, predominately expressed on memory/activated T cells, is a receptor for both interferon-gamma inducible protein-10/CXC ligand 10 (CXCL10) and monokine induced by interferon-gamma/CXCL9. We reported here that CXCR3 was highly up-regulated on infiltrating eosinophils in Schistosoma japonicum egg-induced granuloma in the mouse liver. It was also highly and functionally up-regulated on peritoneal exudate eosinophils in mice infected with S. japonicum. The phenomena were demonstrated at protein and mRNA levels using immunohisto- and immunocytochemistry evaluation of biopsy, flow cytometry and real-time quantitative reverse transcriptase-polymerase chain reaction technique, and verified by Northern blotting and chemotaxis assay in vitro. We also found that CCR3 expression on the infiltrating and peritoneal exudate cells was significantly decreased, CXCR4 expression was unchanged during the 42-day period of infection. We screened mRNA expression levels of the all known chemokine receptors in purified peritoneal exudate eosinophils and liver granuloma dominated by eosinophils. CXCR3 was highly and functionally up-regulated on peritoneal exudate eosinophils in mice infected with S. japonicum, meanwhile CCR3 was significantly and functionally down-regulated in these cells. The findings could lead to a better understanding of the chemokine receptor expression pattern of eosinophils at inflamed tissue sites caused by parasites. These could be also crucial for establishing a therapeutic strategy for eosinophilic inflammation via intervention in chemokine actions.

Selectively increased expression and functions of chemokine receptor CCR9 on CD4+ T cells from patients with T-cell lineage acute lymphocytic leukemia.

In a total of 38 typical T-cell lineage acute lymphocytic leukemia (T-ALL) and T-cell lineage chronic lymphocytic leukemia (T-CLL) cases investigated, we found that CC chemokine receptor CCR9 was selectively and frequently expressed on T-ALL CD4+ T cells, was moderately expressed on T-CLL CD4+ T cells, and was rarely expressed on normal CD4+ T cells. These findings were demonstrated at protein and mRNA levels using flow cytometry and real-time quantitative reverse transcription-PCR technique and were verified by digital confocal microscopy and Northern blotting. Thymus-expressed chemokine, a ligand for CCR9, selectively induced T-ALL CD4+ T-cell chemotaxis and adhesion. Interleukin (IL)-2 and IL-4, together, down-regulated the expression and functions of CCR9 in T-ALL CD4+ T cells including chemotaxis and adhesion. It was also demonstrated that IL-2 and IL-4, together, internalized CCR9 on T-ALL CD4+ T cells and subsequently inhibited functions of CCR9 in these cells. Thymus-expressed chemokine mRNA was highly expressed in CD4+ T cells, involving lymph node and skin in T-ALL patients, and was expressed at moderate levels in lymph node and skin tissues in T-CLL patients. Our findings may provide new clues to understanding various aspects of T-ALL CD4+ T cells, such as functional expression of CCR9-thymus-expressed chemokine receptor-ligand pairs as well as the effects of IL-2 and IL-4, which may be especially important in cytokine/chemokine environment for the pathophysiological events of T-ALL CD4+ T-cell trafficking.

Multicentricity and its associated factors in renal cell carcinoma / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the incidence and associated factors of multicentricity in renal cell carcinoma (RCC) in Chinese patients.</p><p><b>METHODS</b>One hundred and two kidney samples from radical nephrectomy due to RCC were step sectioned at 3 mm intervals and examined. All tissue abnormalities were removed, stained and examined for multicentricity. Then, on each slice of the sample, both the parenchymal margin of 15 mm beyond the pseudocapsule and tissue around the renal sinus were continuously sectioned and examined for completeness of the pseudocapsule and vascular and lymph node invasion. The relationship between muliticentricity and other pathological parameters was evaluated.</p><p><b>RESULTS</b>The incidence of multicentricity was 15.7% (16/102); it was significantly lower in primary tumors < or = 4.0 cm than in tumors > 4.0 cm (4.9%, 2/41 vs 23.0%, 14/61; chi(2) = 6.055, P = 0.014). The incidence was 9.8% (8/82) in tumors without vascular invasion and 40.0% (8/20) in those with it (P = 0.003, Fisher's exact test). The incidence of multicentricity was 1.9% (1/53) in tumors with a complete pseudocapsule and 30.6% (15/49) in those without it (chi(2) = 15.885, P = 0.000). The grade, stage, subtypes and lymph node invasion of the primary tumor were not significantly associated with multicentricity. Multiple logistic regression analysis showed that pseudocapsular incompleteness and vascular invasion were two significant predictors of RCC multicentricity (P = 0.005 and 0.023).</p><p><b>CONCLUSIONS</b>The incidence of multicentricity of RCC in this group of patients was in accordance with published studies. Multifocality was significantly associated with tumor size, pseudocapsule completeness and vascular invasion. NSS should be limited to tumors less than 4.0 cm when the contralateral kidney is normal and careful long-term follow-up is necessary in tumors with positive vascular invasion and incomplete pseudocapsule.</p>

A study of multicentricity in renal cell carcinoma / 中华泌尿外科杂志

Objective To study the incidence of multi ce ntricity in renal cell carcinoma (RCC). Methods 102 kidn eys from radical nephrectomy for RCC were step sectioned at 3 mm intervals and a ny abnormal tissue were removed, stained and examined. The tissue 2.0 cm beyond pseudocapsule and tissue of renal hilum were continueously or interruptedly sect ioned and examined for any tumor invasion of the pseudocapsule, micro-multifoca l carcinoma and vascular invasion beyond pseudocapsule. The relationship between the pathological findings of the primary tumors and multicentricity was evaluat ed. Results The total incidence of multifocal carcinoma in this group was 15.7%.It was 4.9% in primary tumors 4 cm or less and 23.0% in tumors larger than 4 cm (P= 0.014). Vascular invasion and interrupted pseud ocapsule were two significant predictors of multicentricity of RCC (P=0.017 and 0.006). Conclusions In the RCC patients with normal contralateral kidneys, nephron-sparing surgery should be performed only in tumo rs 4 cm or less. In patients nephron-sparing surgery is imperative, even tumors 7 cm or less might be in consideration, but intensive followup is necessary due to the increased incidence of multicentricity.In most cases, a partial nephrect omy instead of tumor enucleation should be performed.