Objective To investigate the relationship between the expressions of Twist and clinical pathological parameters in gastric carcinoma,to explore its significance in judging the prognosis of patients,and to analyze the expression of Twist proteins in the epithelial-mesenchymal transition (EMT) phenomenon.Methods Immunohistochemistry was used to detect the expressions of Twist and EMT relative proteins E-cad,N-cad in 120 cases of gastric carcinoma specimens and corresponding adjacent normal tissue.Analysis The relationship between Twist expression and clinical pathological parameters,and the Twist expression and E-cad,N-cad were analyzed.Results In gastric carcinoma,Twist,E-cad,N-cad positive rates were 59.1％(71/120),36.6 ％ (44/120) and 47.5 ％ (57/120),and in adjacent tissues,the positive rate were 17.5 ％ (21/120),93.3 ％ (112/120) and 11.6 ％ (14/120).There were significant differences in each index between tumor tissues and adjacent tissues (P ＜ 0.05).Patients with Twist positive expression showed higher incidence rates of lymph node metastasis (P ＜ 0.05) and distant metastasis (P ＜ 0.05) than those with Twist negative expression.The expression of Twist was significantly correlated in gastric carcinoma accompanied by low expression of E-cad and abnormal high N-cad expression (P ＜ 0.05).Conclusions Compared with the adjacent tissue,the expression of Twist,E-cad and N-cad have significant differences,and the expression of Twist was closely related to the biological characteristics in gastric carcinoma.The expression of Twist in gastric cancer have negative correlation with the EMT relative factor E-cad,while positive correlation with the N-cad.

ObjectiveTo analyze the distribution and density of Langerhans cells and dermal CD68 positive histiocytes in lesions of secondary keloid.MethodsTissue specimens were resected from the lesions of 30 patients with secondary keloid and normal skin of 14 human controls.Immunohistochemistry was performed to observe the expressions of CD68 and CD1a in these specimens.A micrometer was used to count the number of positively stained cells per unit area.The Student's t test was conducted for data analysis by using the SPSS software.ResultsThe density of CD1a+ Langerhans cells was (61 ± 49) cells/mm2 in the epidermis of secondary keloid lesions, (258 ± 61 ) cells/mm2 in the control epidermis,and(40 ± 65) cells/mm2 in the dermis of keloid lesions.CD68+ cells were absent in the epidermis of keloid lesions.Significant differences were observed in the density of CD1a+ Langerhans cells between the lesional and normal control epidermis(t =9.88,P ＜ 0.001 ) and in the percentage of CD68+ cells in nucleated cells between the superficial dermis of lesions and control skin(62％ ± 12％ vs.70％ ± 14％,t =2.66,P ＜ 0.05).The density of dermal CD68+ histiocytes was similar between the lesions and control skin ((287 ± 73) cells/mm2 vs.(290 ± 22) cell/mm2,t =0.02,P ＞ 0.05).Conclusions In keloid lesions,Langerhans cells decrease in the epidermis but increase in the dermis,CD68+ histiocytes are absent in the epidermis,and reduced in the dermis with a declined percentage in nucleated cells.

Objective To investigate the correlation of serum anti-M-type phospholipase A2 receptor (PLA2R) antibody with laboratory parameters of idiopathic membranous nephropathy (IMN) in adult patients with membranous nephropathy (MN),and to explore the role of anti-PLA2R antibody in the pathogenesis of IMN. Methods Forty-six adult patients with biopsy-proved glomerular diseases were involved in this study,including 20 cases with IMN,7 cases with IgA nephropathy (IgAN),6 cases with hepatitis B-associated membranous nephropathy (HBV-MN),6 cases with minimal change nephropathy (MCN),4 cases with focal segmental glomerulosclerosis (FSGS) and 3 cases with class Ⅴ lupus nephritis.Total RNA was extracted from human glomeruli and was reversely transcribed to the first-strand cDNA.The full-length human M-type PLA2R was amplified by PCR and the 605 bp product was subcloned into eukaryotic expression vector containing CMV promoter.The recombinant human M-type PLA2R plasmid vector was transiently transfected into human embryonic kidney (HEK) 239T cell line using the FuGene6 transfection reagent.Western blotting was used to detect serum anti-PLA2R antibodies.Correlations of antiPLA2R antibody level with laboratory parameters,including serum albumin,total cholesterol,Scr and 24-hour urine protein,of IMN patients were evaluated. Results Among 20 cases with IMN,15 cases showed positive anti-PLA2R antibodies (positive rate 75％).Of 7 cases with IgAN and 6 cases with HBV-MN,only 1 case showed positive anti-PLA2R antibody respectively (positive rate 14.29％ and 16.67％ respectively).Anti-PLA2R antibody was negative in other patients.The positive rate of anti-PLA2R antibody in IMN patients was significantly higher than that in patients with secondary MN and other types of glomerlonephritis (all P＜0.01).Furthermore,anti-PLA2R antibody level was positively correlated with 24-hour urine protein(r=0.803,P＜0.01) and negatively correlated with serum albumin in IMN patients (r=-0.816,P＜0.01). Conclusions The high positive ratio of anti-PLA2R antibody may indicate that it is the specific autoantibody in IMN.AntiPLA2R antibody is correlated with IMN disease severity,which indicates that it may be the pathogenic autoantibody in IMN.

This study is to investigate the anti-allergic effect of anthocyanidin and to explore its possible mechanism. The experiments of passive cutaneous anaphylaxis reaction (PCA) and colorimetry were used to determine the effect of anthocyanidin on degranulation of mast cells in vivo. For in vitro study, various concentrations of anthocyanidin (100, 50 and 25 micromol x L(-1)) were added to the culture medium of mast cells cultured with 100 microg x L(-1) of dinitrophenyl (DNP) specific IgE overnight. The azelastine (100 micromol x L(-1)) was selected as the positive control. The antigen (DNP-human serum albumin, DNP-HAS)-induced release of degranulation was measured by enzymatic assay, histamine was determined by EIA, and interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) were measured by Western blotting, separately. In addition, the effects of anthocyanidin on phosphorylation of NF-kappaB, p38MAPK and Akt were observed by Western blotting. The results showed that treatments with anthocyanidin (100 and 50 mg x kg(-1)) were followed by a decrease in PCA of rats. Anthocyanidin (100 and 50 micromol x L(-1)) obviously suppressed the degranulation from mast cells, whereas results from anthocyanidin (100 and 50 micromol x L(-1)) group indicated significant inhibitory effect on histamine, the calcium uptake, TNF-alpha, IL-6, phosphorylation of NF-kappaB, p38MAPK and Akt of mast cells induced by antigen. Anthocyanidin may suppress the anaphylactic reaction by inhibiting the action of mast cells. NF-kappaB, p38MAPK and Akt at least in part contribute to this event.

Objective To evaluate the academic status of 16 Chinese core orthopaedic journals from 2005 to 2007 in terms of general citation frequency, impact factors, allo-citation rate and specialty influence index. Methods On the basis of the normalized data published by Chinese S& T Journal Citation Reports (CJCR), the general citation frequency, impact factors, allo-citation rate and specialty influence index of the 16 Chinese core orthopaedic journals from 2005 to 2007 were ranked respectively and sta-tistically analyzed. Results In terms of the above 4 indicators, Chinese Journal of Orthopedics ranked the first, followed by Chinese Journal of Trauma and Chinese Journal of Reparative and Reconstructive Surgery. Considering its short history, our journal, Chinese Journal of Orthopedic Trauma, made a rapid progress in academic status. Conclusion Generally speaking, the 16 Chinese core orthopaedic journals enjoy a high academic quality and relatively significant impact.

Objective To investigate the magnetic resonance imaging (MRI) characteristics of cerebral lesions,laboratory data and prognosis of patients with neuropsychiatric systemic lupus erythematosus (NPSLE) and evaluate the correlation of MRI findings,autoantibodies and prognosis of NPSLE.Methods Thirty-one patients with NPSLE admitted during Jan 1998 to Oct 2011 were enrolled into this study,including 2 males and 29 females.MRI characteristics of cerebral lesions,laboratory data and clinical outcomes were retrospectively analyzed.x2-test and Spearman's correlation were used for statistical analysis.Results Cranial MRI scans on admission were abnormal in 27 (87％) of the 31 patients,among which 17 cases showed diffuse manifestations,9 cases showed focal lesions in brain and 1 case showed brain atrophy.The percentage of cases with focal lesions in the dead group (86％) was significantly higher than that of the survival group (13％) (P＜0.01),while the percentage of diffuse lesion cases was significantly lower in dead group (P＜0.01).Although the mean lesion numbers in the dead group were fewer compared with the survival group,lesion numbers in brainstem (18％) (P＜0.01) or basal ganglia (29％) (P＜0.05) were much more in the dead group.The positive rate of anti-ribosomal P protein antibodies shown in the dead group (43％) was markedly higher than that in the survival group (29％) (P＜0.05).The positive rates of ACL in cases with diffuse lesions and cases with focal lesion were higher compared with cases showing normal MR images (P＜0.01).Furthermore,the positive rate of anticardiolipin antibodies (ACL) in cases with focal lesions (56％) were higher than cases with diffuse lesion (35％) (P＜0.05).There was no correlation between the seven types of autoantibodies and lesion locations such as basal ganglia,subcortical white matter,anterior and posterior horn of lateral ventricle,semiovale center,cerebral cortex,brainstem and cerebellum.Conclusion MRI findings and autoantibodies are correlated in patients with NPSLE and both of them may be used as prognostic markers for NPSLE.Focal lesions,location of brainstem and basal ganglia on cranial MR images,positive anti-ribosomal P protein antibody may predict an unfavorable clinical outcome of NPSLE; however,the correlation between ACL and prognosis could not be determined.

Objective To investigate the effects of baicalein and acitretin on the apoptosis in a human cutaneous squamous cell carcinoma cell line, SCL-12. Methods Cultured SCL-12 cells were treated with different concentrations of baicalein (3.125, 6.25, 12.5 μmol/L) and acitretin (2.5, 5.0, 10.0 μ mol/L), alone or in combination, for 48 hours. Subsequently, cell proliferation was detected by MTT assay, and cell apoptosis by ELISA as well as annexin V-FITC and propidium iodide double staining. Real-time quantitative RT-PCR was used to detect the expression of Fas mRNA in SCL-12 cells. Results The cell proliferation of SCL-12 cells was inhibited by baicalein and acitretin alone or in combination. The combination of baicalein and acitretin at the three tested concentrations, except for that of baicalein at 3.125 μmol/L and acitretin at 2.5 μmol/L, more strongly inhibited the proliferation of SCL-12 cells compared with baicalein or acitretin alone, and the inhibitory effect was in a dose-dependent manner. The early apoptosis rate was 9.39% ± 1.52%, 20.86% ± 2.16%,36.85% ± 3.26% in SCL-12 cells treated with baicalein of 3.125 μmol/L, acitretin of 5.0 μmol/L alone and their combination, respectively, significantly higher than that in untreated cells (4.39% ± 0.64%, all P <0.05); the induction of apoptosis in SCL-12 cells by the combination of baicalein and acitretin was stronger than that by baicalein or acitretin alone (F = 138.44, P < 0.05). Baicalein and acitretin alone or in combination significantly increased the mRNA expression of Fas in SCL-12 cells, and the effect of their combination was stronger than that of baicalein or acitretin alone. Conclusions Baicalein and aeitretin could inhibit the growth of and induce the apoptosis in SCL-12 cells, and the effect is enhanced by the combination of baicalein and acitretin, which may be associated with the upregulation of Fas expression in SCL-12 cells.

BACKGROUND: Experimental autoimmune encephalomyelitis has become the most classical animal model for multiple sclerosis. However, the experimental autoimmune encephalomyelitis model of China presented one-way course of disease. By using proteolipid protein 139-151 and proteolipid protein 178-191, relapse remitting experimental autoimmune encephalomyelitis models may be induced in SJL/J mice which were susceptible to immune, which have similar clinical situation, course and histologicallterations to multiple sclerosis.OBJECTIVE: To establish the relapse remitting experimental autoimmune encephalomyelitis mouse model induced by proteolipid protein, which has similar clinical situation, course and histological alterations to multiple sclerosis.DESIGN: Completely randomized controlled study.SETTING: The centre of Neuro-information, and Neurological Institute,General Hospital of Chinese PLA.MATERIALS: The study was carried out at the Laboratory of Neuro-pathology, General Hospital of Chinese PLA, from February to June 2004.Sixty female SJL/J mice with 8-12 weeks old were selected and randomly divided into proteolipid protein 139-151 group and proteolipid protein-178-191 group with 30 in each.INTERVENTIONS: After injected with proteolipid protein-139-151 or proteolipid protein-178-191, the models of relapse remitting experimental autoimmune encephalomyelitis were induced, and the body weight and neurological signs of each female SJL/J mouse were viewed. The tissue morphological changes of models were observed with hematoxylin and eosin and uxol fast blue stain.MAIN OUTCOME MEASURES: The neurological symptoms and signs,features of relapse and remitting and the perivascular inflammatory cell infiltration, demyelinated lesion of the model of experimental autoimmune encephalomyelitis mouse induced by two proteolipid protein peptides.RESULTS: All 60 mice entered the final analysis. ① Significant neurological symptoms, signs and features of relapse and remitting was manifested in the model of experimental autoimmune encephalomyelitis mouse induced by two proteolipid protein peptides. Obvious phenomena of perivascular inflammatory cuffing, satellitism, predominant perivascular inflammatory cell infiltration and demyelinated lesion were found in spinal and cerebral tissue. ②Changes of body mass: Before immunity, the body mass of mice in two groups was( 17. 84 ± 2.59) g and (17. 88 ± 0.52) g respectively. Onset of relapse of the mice in proteolipid protein-178-191 group was earlier and faster, their body mass had no distinctive change after immunization and the mean body mass was(23.52 ± 2.37) g till the 60th day. Meanwhile, Onset of relapse of the mice in proteolipid protein-139-151 group was later and slower. After the immunity, the body mass of mice was little decrease, and the body mass was (16. 70 ±0.46) g on the 60th day. ③ Neural functional scores: The highest functional scores in the two groups were not different(3.86 ± 1.10vs 3.71 ±1.05, t=0.49, P=0.628).CONCLUSION: The two different antigenic peptides of proteolipid protein can all cause the autoimmune response of central nervous system. Both models have the same characters of relapse and remitting and the severity has no significant difference. But compared with proteolipid protein 139- 151 group,onset and recover of the experimental autoimmune encephalomyelitis of the mice in proteolipid protein 178-191 group were earlier, as well as weight variance was larger, which maybe due to the different structure of two peptides.

Objective To investigates the efficacy and tolerability of second line treatment with S-1 plus thalidomide in patients with advanced pancreatic cancer.Methods Sixty patients hospitalized in Department of Oncology of Cangzhou Central Hospital from July 2010 to October 2012 were included in this study.All the patients were diagnosed as having pancreatic carcinoma.The patients were randomly divided into two groups,one group was treated with S-1 alone,and the other group was treated with S-1 plus thalidomide.Then the efficacy and toxicity of two groups was evaluated.Results The disease control rates were 36.7％ and 46.7％ in the S-1 alone group and the S-1 plus thalidomide group,and the difference between the two groups was not statistically significant (P =0.31).The progression free survival (PFS) was 2.9 months and 3.3 months,and the difference between the two groups was statistically significant (P ＜ 0.05),the Karnofsky score,pain,sleep and weight improvement rate was 63.3％,46.7％,66.7％ and 53.3％ in combination group,which were significantly better than those in control group (30.0％,13.3％,30.0％ and 20.0％),and the difference between the two groups was statistically significant (P ＜ 0.05).The major adverse events were nausea,vomiting,fatigue and drowsiness,mainly of grades Ⅰ ～ Ⅱ.Conclusions S-1 plus thalidomide as second line treatment of pancreatic cancer can prolong the PSF of patients with advanced pancreatic cancer with excellent safety,and patients' quality of life is also improved.

Objective To investigate the effect of three Scutellaria baicalensis extracts (baicalin,baicalein and wogonin) on the apoptosis of a human cutaneous squamous cell carcinoma cell line,SCL-1.Methods Methyl thiazolyl tetrazolium (MTT) assay was performed to detect the proliferation of cultured SCL-1 cells treated with different concentrations (12.5,25,50,75 and 100 μmol/L) of baicalin,baicalein and wogonin for various durations (12,24 and 48 hours).Some SCL-1 cells were treated with baicalin,baicalein and wogonin of 12.5 μ mol/L respectively for 48 hours followed by the detection of cell apoptosis by double staining with annexin V-fluorescein isothiocyanate/propidium iodide in combination with enzyme-linked immunosorbent assay (ELISA),as well as estimation of cell cycle by flow cytometry.The 50％ inhibitory concentration was determined by line regression model and inner insert method,and statistical analysis was carried out by Student's t test,one-way analysis of variance (ANOVA),and Student-Newman-Keuls-q test.Results Baicalin,baicalein and wogonin all inhibited the proliferation of SCL-1 cells in a dose-and time-dependent manner (all P ＜ 0.01).Under the same conditions (treatment concentration and duration),baicalein showed the strongest inhibitory effect on the proliferation of SCL-1 cells,followed by wogonin and baicalin (all P ＜ 0.01).All the Scutellaria baicalensis extracts induced the apoptosis of SCL-1 cells and arrested them in G1-phase.The percentage of cells in G1 phase was 56.37％ ± 2.41％,74.23％ ± 2.02％ and 64.15％ ± 1.87％,and early apoptosis rate was 8.09％ ± 1.02％,24.13％ ± 0.76％ and 14.45％ ± 1.57％,in SCL-1 cells treated with baicalin,baicalein and wogonin of 12.5 μmol/L for 48 hours,respectively,compared to 45.04％ ± 1.93％ and 4.12％ ± 0.29％ in the untreated control cells respectively (F =83.29,186.37,respectively,both P ＜ 0.01).Similarly,there was a downward trend from baicalein to wogonin and baicalin in the effect on cell apoptosis and cell cycle arrest of SCL-1 cells.Conclusions Scutellaria baicalensis can inhibit the growth and induce the apoptosis of SCL-1 cells,which may provide new ideas for the treatment of cutaneous squamous cell carcinoma with traditional Chinese drugs.