Objective To evaluate the effects of aspirin-triggered lipoxin A4 (ATL) on lipopolysaccharide (LPS)-induced acute lung injury in mice.Methods Thirty male SPF BALB/C mice,aged 10-12 weeks,weighing 25-30 g,were randomly assigned into 3 groups (n =10 each):normal saline group (group NS),LPS group and ATL groups.ATL 0.1 ml was injected via the tail vein 1 h after intra-tracheal instillation of 3 mg/kg LPS in LPS group.In ATL group,ATL 0.2 mg/kg was injected via the tail vein 1 h after intra-tracheal instillation of 3 mg/kg LPS.At 24 h after instillation,the mice were sacrificed.Bronchoalveolar lavage fluid was collected for determination of the total cell count,proportion of the polymorphonuclear leukocytes,proportion of the mononuclear leukocytes,and concentrations of the total protein,TNF-αt,IL-6,monocyte chemoattractant protein-1 (MCP-1) and IL-10.Lungs were removed for determination of myeloperoxidase (MPO) activity,phosphorylation of p38 mitogenactivated protein kinase (p38 MAPK),c-Jun N-terminal kinase (JNK),and extracellular signal-regulated kinase 1/2 (ERK1/2) in lung tissues and for microscopic examination.The pathological changes of lungs were scored.Results Compared with NS group,the lung injury scores,total cell counts,proportion of polymorphonuclear leukocytes,and concentrations of TNF-α,IL-6 and MCP-1 were significantly increased,and the proportion of mononuclear leukocytes was decreased in LPS and ATL groups,and IL-10 concentrations were decreased,and the concentrations of the total protein,MPO activity,and phosphorylation of p38 MAPK,JNK and ERK1/2 were significantly increased in group LPS (P ＜ 0.05),and no significant change in the concentrations of the total protein,MPO activity,phosphorylation of p38 MAPK,JNK and ERK1/2 was found in group ATL (P ＞ 0.05).Compared with LPS group,the lung injury scores,total cell counts,proportion of polymorphonuclear leukocytes and the concentrations of the total protein,TNF-α,IL-6 and MCP-1 were significandy decreased,the proportion of mononuclear leukocytes and IL-10 concentration were increased,and MPO activity and phosphorylation of p38 MAPK and JNK were decreased (P ＜ 0.05),and no significant change in the phosphorylation of ERK1/2 was found in group ATL (P ＞ 0.05).Conclusion ATL can ameliorate LPS-induced acute lung injury by inhibiting activations of p38MAPK and JNK signal pathways in mice.

OBJECTIVETo evaluate the clinical features of patients with primary biliary cirrhosis (PBC) and positive expression of sp100 autoantibody in order to generate a clinical screening profile that may help to increase early diagnosis and timely initiation of therapy.

METHODSThe clinical data of 70 patients who were diagnosed with PBC by liver biopsy between January 2006 to December 2009 at the Second Affiliated Hospital of Kunming Medical University of Hepatobiliary and Pancreatic Medicine were retrospectively collected for analysis. The patients were divided according to expression of anti-sp100: positive patients, n = 12; negative patients, n = 58. The groups were comparatively analyzed for differences in clinical, biochemical, immunological, and histopathological parameters. Normally distributed data was compared by t-test, and non-normally data was compared by rank-sum test.

RESULTSThere was no significant difference in age among the sp100-positive and sp100-negative patients (51.6 +/- 9.5 vs. 50.0 +/- 14.7 years, P more than 0.05). The sp100-positive group had significantly more women (80.0% vs. 61.9%, X2 = 0.32, P more than 0.05) and more patients with atypical symptoms (18.2% vs. 13.8%) but the difference of the latter did not reach statistical significance. The sp100-positive group had significantly higher levels of alkaline phosphatase (ALP; 466 vs. 163 U/L, Z = 3.71), gamma-glutamyl-transpeptidase (GGT; 728 vs. 154 U/L, Z = 3.38), and immunoglobulin M (IgM; 4.25 +/- 2.86 vs. 2.81 +/- 2.15, t = 2.06, P less than 0.05). Forty of the total patients tested negative for antimitochondrial (AMA)-M2 antibodies, and eight of those were sp100-positive (20.0%) while 18 were antinuclear (ANA) antibody-positive (45.0%). There were significantly more AMA-M2-negative/ANA-positive patients than sp100-positive patients (P = 0.021). Anti-sp100 expression was not associated with the pathological stage of PBC (R1 = 5.500, P more than 0.05).

CONCLUSIONSP100-positive PBC may show a bias towards the female sex, and may be characterized by enhanced serum levels of ALP, GGT, and IgM. Further clinical differences may manifest as the disease progresses, and changes in autoantibodies' expression and liver function markers should be carefully monitored in follow-up.

Adult ; Aged ; Antibodies, Antinuclear ; blood ; Antigens, Nuclear ; immunology ; Autoantibodies ; blood ; Autoantigens ; immunology ; Female ; Humans ; Liver ; pathology ; Liver Cirrhosis, Biliary ; immunology ; pathology ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

Child ; Humans ; Treatment Outcome

OBJECTIVE: To construct the recombinant adenoviral vector carrying the rat interleukin-10 (rIL-10) gene, and to investigate whether it is stably expressed in bone marrow mesenchymal stem cells. METHODS: The rIL-10 gene was amplified by PCR from template rIL-10 cDNA, and the recovered 656 bp rIL-10 DNA fragment was cloned into pcDNA3.1 to construct pcDNA3.1-IL-10. Then HEK293 cells were transfected with pcDNA3.1-IL-10 and adenoviral vector for homologous recombination, and sequencing and PCR were used to evaluate whether recombination was successful. HEK293 cells were lysed by repeated freeze-thaw cycles, and bone marrow mesenchymal stem cells were infected with the virus solution containing the rIL-10 gene. Western blot was used to measure the expression of rIL-10 in bone marrow mesenchymal stem cells. RESULTS: Sequencing and PCR verified that the rIL-10 adenoviral vector was successfully constructed, with a virus titer of 4×10 PFU/mL. The expression of IL-10 was detected after bone marrow mesenchymal stem cells were infected by the virus solution containing the rIL-10 gene. CONCLUSIONS The constructed rIL-10 recombinant adenovirus can mediate the stable expression of rIL-10 gene in bone marrow mesenchymal stem cells, which provides a basis for gene transplantation therapy of inflammatory bowel disease.
Adenoviridae ; Animals ; Bone Marrow Cells ; Genetic Vectors ; HEK293 Cells ; Humans ; Interleukin-10 ; Mesenchymal Stem Cells ; Rats ; Transfection

OBJECTIVETo observe the clinical effect of dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises for treatment of female stress urinary incontinence.

METHODSSeventy one cases were randomly divided into two groups. Thirty six cases in the observation group were treated with acupuncture on Zhongji (CV 3), Zigong (EX-CA 1), Chize (LU 5) etc. and tortoise-shell moxibustion on Shenque (CV 8) combined with pelvic floor muscle exercises; while thirty five cases in the control group were treated with only pelvic floor muscle exercises. The scores of the International Consultation Committee on Incontinence Questionnaire Short Form (ICI-Q-SF) and the Medical Outcomes Survey Short Form-36 (SF-36) were evaluated before and after treatment, and the scores of SF-36 were also compared with 35 cases in normal group.

RESULTSThe total effective rate of 91.7% in the observation group was higher than that of 77.1% in the control group (P < 0.05). The dimensions of SF-36 of stress urinary incontinence patients were remarkably lower than those of normal group (all P < 0.05). The scores of ICI-Q-SF were decreased while the scores of SF-36 were increased obviously after treatment in both the observation group and the control group, there were pronounced improvements on physiological function, pain, physical activity, social function and affection function in the observation group (all P < 0.05).

CONCLUSIONThe quality of life for female stress urinary incontinence patients may be poor, however the dog-day acupuncture and tortoise-shell moxibustion combined with pelvic floor muscle exercises can improve the symptoms of urinary incontinence and increase the quality of life of patients.

Acupuncture Therapy ; Adult ; Aged ; Exercise Therapy ; Female ; Humans ; Middle Aged ; Moxibustion ; Muscle Contraction ; Muscles ; physiopathology ; Treatment Outcome ; Urinary Incontinence, Stress ; physiopathology ; therapy

OBJECTIVESTo study the relationship between lymphangiogenesis and lymphatic metastasis in mice bearing hepatic carcinoma and analyze the mechanism of the lymphatic metastasis.

METHODSHepatic carcinoma cell lines of high and low potentialities of lymphatic metastasis were injected into the footpads of Balb/c mice. Their metastases to lymph nodes were examined. The tumor tissues of each group were stained with 5'-nucleotidase-ALP to observe the lymphoangiogenesis. The total RNA of high and low metastatic potential cell lines were extracted for metastasis gene DNA array. The vascular endothelial cell growth factor C (VEGF-C) and VEGF-D of each cell line were detected using semi-quantitative RT-PCR and were further quantatively analyzed using real time PCR.

RESULTSThe para-common iliac a. and renal hilar lymph nodes metastases of the high metastatic potential cells were significantly higher than in the controls (P>0.05). The quantity of lymphatic vessels in the high metastasis group was significantly larger than that of the control group (P<0.05). The expressions of CD44, E-cadherin, HER2/neu, H-Ras and VEGF-C in the high metastasis group were higher than those in the low metastasis group shown by the cDNA micro array experiment but the expressions of nm23A, nm23-E4, p16ink4a, CD61 were lower. The VEGF-C expression was higher and the VEGF-D was lower in the high metastasis group compared to those of the low metastasis group shown by semi-quantitative RT-PCR. The secretion of VEGF-D was significantly lower and the ratio of VEGF-C/VEGF-D was significantly higher in the high metastasis group than the low metastasis group (P<0.05).

CONCLUSIONSThe lymphatic metastasis of hepatic carcinoma is related to lymphoangiogenesis. The changes of VEGF-C and VEGF-D expressions might be a cause influencing the lymphoangiogenesis. VEGF-C/VEGF-D might be an effective parameter in affecting lymphatic metastases.

Animals ; Liver Neoplasms, Experimental ; pathology ; Lymph Nodes ; pathology ; Lymphangiogenesis ; Lymphatic Metastasis ; Male ; Mice ; Mice, Inbred BALB C ; Neoplasm Transplantation ; Vascular Endothelial Growth Factor C ; metabolism ; Vascular Endothelial Growth Factor D ; metabolism

Adolescent ; Child ; Child, Preschool ; Colonoscopy ; Female ; Gastrointestinal Agents ; pharmacology ; Humans ; Infant ; Lactulose ; adverse effects ; pharmacology ; Male

OBJECTIVETo evaluate the clinical effect of proton pump inhibitor-based triple therapy combined with Saccharomyces boulardii in the treatment of Helicobacter pylori (Hp) infection among children in terms of Hp eradication rate and incidence of adverse events.

METHODSA prospective randomised controlled study was conducted on 240 children with a confirmed diagnosis of Hp infection. These patients were randomized into triple therapy (n=120) and probiotics groups (n=120). The triple therapy group received amoxicillin [40 mg/(kg·d), Tid], clarithromycin [15 mg/(kg·d), Bid] and omeprazole [0.7-0.8 mg/(kg·d), Qd], while the probiotics group received Saccharomyces boulardii (250 mg, Bid) in addition to triple therapy. The course of treatment was 14 days in both groups. The adverse events in subjects were recorded by their parents during treatment. Hp eradiation was evaluated by (13)C breath test at 4 weeks after treatment, and the eradication rate and incidence of adverse events were compared between the two groups.

RESULTSThe Hp eradication rates were 75.8% (91/120) in the triple therapy group and 85% (102/120) in the probiotics group (P>0.05). Compared with the triple therapy group, the probiotics group had nonsignificantly lower incidence of nausea, vomiting, and abdominal pain (P>0.05) and significantly lower incidence of stomatitis, constipation and diarrhea (P<0.05).

CONCLUSIONSTriple therapy combined with Saccharomyces boulardii cannot significantly increase Hp eradication rate, but can significantly reduce the incidence of stomatitis, constipation, and diarrhea during treatment.

Amoxicillin ; administration & dosage ; Child ; Child, Preschool ; Clarithromycin ; administration & dosage ; Combined Modality Therapy ; Drug Therapy, Combination ; Female ; Helicobacter Infections ; therapy ; Helicobacter pylori ; Humans ; Male ; Omeprazole ; administration & dosage ; Probiotics ; administration & dosage ; Prospective Studies ; Saccharomyces

OBJECTIVETo establish the restriction fragment differential display-polymerase chain reaction (RFDD-PCR) as an efficient technique for constructing and studying the gene expression profile of human tissues.

METHODSThe tissues of mamma adenocarcinoma (T), cancerometastasis lymph node (L) and normal mammary (N) from one mammary infiltrating ductal carcinoma case were collected, and the gene expression profile of each kind of tissue was constructed using RFDD-PCR technique at equal pace according to the operating manual of Qbio-gene Company. Then all fragments of the three gene expression profiles were separated and displayed by electrophoresis. With the use of gene database at the website http://www.Qbio-gene.com/display, the authors identified the names of the probable fragments by bioinformatics analysis. Through comparison of the three profiles, the numbers and types of most differentially expressed gene fragments were displayed.

RESULTSThe expression profiles of the three kinds of tissue have been constructed covering 1716 fragments of mammary adenocarcinoma, 1769 of cancerometastasis lymph nodes and 1922 of normal mammary tissue. Among these 5407 fragments, 39.39% were exactly the same. While 33.9% sequences of T and L showed differences in abundance or presence, 40.9% of T and N and 39.6% fragments of L and N were observed differentially expressed. These differentially expressed gene fragments were found to relate with metastasis, differentiation, inflammation and so on.

CONCLUSIONRFDD-PCR is an efficient technique for research in human diseases genomics as a mass screening for complete gene expression profile with high-flux. Through comparison among three or more profiles, the screening for candidate genes of a certain disease can be accomplished, and there is probably a chance to identify novel gene or expressed sequence tag.

Adenocarcinoma ; genetics ; Breast Neoplasms ; genetics ; Carcinoma, Ductal, Breast ; genetics ; Computational Biology ; Electrophoresis ; methods ; Female ; Gene Expression Profiling ; methods ; Humans ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; methods

OBJECTIVETo analyze the relationship between the prognosis of patients with oropharyngeal squamous cell carcinoma (OSCC) and human papillomavirus (HPV) infection in OSCC.

METHODSSixty-six patients with oropharyngeal carcinoma who met the enrollment criteria during the period from January 1999 to December 2009 were retrospectively reviewed. The presence or absence of HPV oncogenic types in OSCC specimen was determined by multiplex polymerase chain reaction (PCR). Overall survival (OS) and disease specific survival (DSS) for HPV-positive and HPV-negative patients were estimated by Kaplan-Meier analysis. Cox regression model was used for multivariate analysis.

RESULTSHPV-DNA was detected in 11(16.7%) of all specimens. Among them, 7 were positive for HPV-16, 1 for HPV-16/11, 1 for HPV-35, 1 for HPV-58/52, and 1 for HPV-33/52/54. With the follow-up of 3-78 months (a median of 24.5 months), patients with HPV-positive tumors had significantly better overall survival (χ2=5.792, P=0.016) and disease specific survival (χ2=4.721, P=0.030), the 3-year OS and DSS were 90.0% vs 52.4% and 90.0% vs 56.4%, respectively. Multivariate analysis by Cox regression model showed that HPV infection and nodal status were both independent prognostic factors for patients with OSCC (P<0.05).

CONCLUSIONSPatients with HPV-positive OSCC have significantly better prognosis than patients with HPV-negative tumors. HPV infection is an independent prognostic factor.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; diagnosis ; virology ; Female ; Human papillomavirus 16 ; Humans ; Male ; Middle Aged ; Oropharyngeal Neoplasms ; diagnosis ; virology ; Papillomavirus Infections ; pathology ; Prognosis ; Retrospective Studies