Objective To investigate the phenotype,number and distribution of inflammatory cells in early and late stages of spontaneous regression of halo nevi,and to elucidate the immunological mechanisms for spontaneous regression of these nevi.Methods Halo nevi,their surrounding non-lesional skin,and normal control skin were examined by immunohistochemical staining with monoclonal antibodies to CD3,CD4,CD8,CD20,CD1a,CD56 and CD68.Staining results were observed and analyzed by the computer image analysis system,image-pro plus 6.0.Results The number of CD4+,CD8+,CD20+,CD1a+cells,along with the diameter of CD1a+and CD68+ cells was significantly increased in the lesions of early and late stage of spontaneous regression of halo nevi than in non-lesional skin and normal control skin(both P＜0.01).The ratio of CD8+/CD4+ cells in the lesions of late stage of spontaneous regression was also higher than that in the lesions of early stage (2.05∶1 VS 1.82∶1).A massive infiltrate of CD8+ cells was observed in the nests of nevus cells.ConclusionsCD4,CD8,CD20,CD1a,CD56 and CD68 positive cells are all involved in the spontaneous regression of halo nevi,and CD8+ cells may play a predominant role in this process.

Objective To observe colorectal tumor's growth in hyperglycemia mice and its vascular endothelial growth factor (VEGF)'s expression, insulin-like growth factor-1 (IGF-1)'s variation of blood through the experiment, then to ascertain whether type 2 diabetes mellitus (T2DM) danger factors to promote colorectal cancer happen and progress or not. Methods The mouse model of colorectal cancer combined T2DM was established. The volume of tumor was observed. After 5 weeks, all mice were executed and IGF-1 in the blood and the expression of VEGF in the tumor tissue was examined. Results The average tumor volume of colorectal tumor-diabetes group [(1628.5±882) mm3] were larger than that of colorectal tumor group [(1950.2±726) mm3] (P <0.05), and its expression IGF-1 of blood [(105.33±32.32) ng/ml] were higher than that of the control group [(69.83±25.57) ng/ml] and colorectal tumor group [(70.17±25.27) ng/ml] (P <0.05). The expression of VEGF [(70.0±11.5)%] in colorectal tumor-diabetes group were significantly higher than that of colorectal tumor group [(42.9±7.5)%] (P <0.05), too. Conclusion The model of T2DM and transplanted colorectal tumor can be duplicated successfully in ICR mice. Diabetes mellitus may be one reason of promoting colorectal cancer progress. Besides high blood glucose, its mechanism is the high level of IGF-1 which can inhibit apoptosis, promote cell differentiation and hyperplasia, and through inducing VEGF duplicating, heighten its expression, promote the tumor vessel growth, lead to tumor happen and metastasis.

The market and literature were studied to understand the existing situation of Magnoliae Officinalis Cortex goods, and the collected samples were analyzed, combined with the actual production, a new standard of Magnoliae Officinalis Cortex commercial specification and grade was drafted. Magnoliae Officinalis Cortex goods was divided into two categories according to the source in the old standard. Then each category was divided into four kinds of specifications according to the site. Each kind of specification was divided into several grades according to the length and weight. To judge the quality of Magnoliae Officinalis Cortex goods was mainly based on the appearance quality. In the new standard, the classification of commercial specification and grade is based on the thickness, magnolol and honokiol content. The goods of Magnoliae Officinalis Cortex can be divided into three specifications: Tongpu, Genpu and Doupu. Tongpu is divided into three grades, the remaining two are not graded.
Magnolia ; anatomy & histology ; chemistry

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007–2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R. edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R. heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Spotted fever caused by spotted fever group rickettsiae (SFGR) is found throughout China. During 2007—2008, 28 human SFGR isolates and 34 rat SFGR isolates including 15 isolates from Rattus fulvescens, 5 isolates from R.edwardsi, 7 isolates from Callosciurus erythraeus roberti and 7 isolates from Dremomys rufigenis) were obtained from L929 cell culture. Previous research indicated that the 62 strains of SFGR mentioned above shared not only the same serophenotype but also 100% of identity sequences of 16S rRNA, gltA, ompA, groEL and 17KD, which enabled us to apply multispacer typing (MST) to the 62 SFGR isolates in the study. Six primer pairs, which were used for typing of Rickettsia rickettsii and Rickettsia conorii, were chosen, and the results exhibited greater nucleotide polymorphisms among the 62 isolates tested. A total of 48 distinct genotypes were identified. The dominant genotype, represented by h3 isolates, accounted for 21.7% (13/60) of the isolates tested, and the remaining 47 genotypes were all unique. Phylogenetic analysis showed that all the 48 genotypes could be classified in the same clade, while the genetically related strain, R.heilongjiangensis, was close but not the same as the cluster. We concluded that the genetically diverse of spotted fever group rickettsiae strains are endemic in Chengmai County, Hainan Province, China.

Objective To study the changes of subgroups of peripheral blood T lymphocytes in the patients infected with the 2009 pandemic influenza A ( H1N1 ) virus of different severity type. Method A total of 66 patients infected by H1N1 evidenced by RT-PCR admitted from September 2009 to January 2010 were divided into three groups: mild type ( B group, n = 47 ), cured patients of severe and critical severe type ( C group, n = 14) and died patients ( D group, n =5), according to the severity and prognosis. A total of 20 healthy volunteers served as control group( A group). Peripheral blood lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count were detected by flow cytometry at the different time points. Fever duration and H1N1 virus negative time were compared. Statistical analysis were performed by using SAS version 9.13 software and the data were processed with ANOVA and SNK test. Results Lymphocyte count, CD3+,CD4+ and CD8+ T lymphocyte count declined in the early period in all the groups, and there were significant differences compared with A group (P＜0. 05), while rised with the clinical progression in group B and C,and those of C group were lower than B group ( P ＜ 0.05 ), but those of D group were always low. Fever duration and H1N1 virus negative time were (4.4 ± 1.6) days vs. (4.4 ± 1. 4) days, ( 12.9 ± 3. 1 ) days vs.( 10.2 ± 2.6) days and ( 15.2 ± 7.3 ) days vs. ( 13.3 ± 2.9 ) days respectively, and there were significant differences among the three groups ( P ＜ 0.05 ). Conclusions The cellular immune function was seriously damaged when patients were infected with H1N1. Further more, the changes of lymphocyte count, CD3+ , CD4+and CD8+ T lymphocyte count were tightly related with the degree of severity and prognosis. These findings can be used for clinical diagnosis and treatment.

Objective: To investigate whether lipopolysaccharide induced parkin expression and mitophagy in macrophages.Methods:The murine peritoneal primary macrophages were aseptically isolated from Kunming mice and cultured in complete medium.The mitochondrial membrane potential of macrophages was detected by flow cytometry,after the cells were stimulated with 200 ng/ml LPS and labeled mitochondria with JC-1.The parkin mRNA level of macrophages was detected by RT-PCR, protein levels of parkin and autophagic related protein LC3 Ⅱ and LC3 Ⅰ were determined by Western blot.The distribution and co-localization of parkin with LC3 and mitochondria in macrophages were respectively observed by laser scanning confocal microscope, before and after the cells were treated with LPS.Results: Flow cytometry results after JC-1 staining showed that mitochondrial membrane potential in macrophages was declined after stimulation with 200 ng/ml LPS, and continuously decreased with prolonged treatment time.The mRNA levels of parkin were increased slightly within 6 h after LPS stimulation,but parkin proteins were increased significantly within 6 h after LPS stimulation.The results of parkin distribution showed that parkin was evenly distributed in the cytoplasm at normal status, but became the obvious punctate distribution after LPS stimulation in macrophages.Western blot results showed LC3 Ⅱ/LC3 Ⅰ levels were increased after LPS stimulation, indicating the appearance of macrophage autophagy.Confocal microscopy showed that there were co-localization of parkin,LC3 and mitochondrial in macrophages after LPS stimulation.Conclusion:Parkin expression is increased significantly and mediated mitochondrial autophagy in macrophages after LPS stimulation, which is involved in the clearance of damaged mitochondria,thereby playing a role in regulating macrophage inflammatory response.

ObjectiveTo probe into the clinical efficiency of reconstruction to person who had composite tissue defect of the floor of mouth and the lower part of face with the free forearm flap and free iliac bone in the same term.MethodsBefore the surgery, the surgeon should carefully evaluate the scope and shape of defects in 11 cases who had the composite tissue defects of the floor of mouth and the lower part of face, to design the individual forearm flap, so that it matches with the defect region. During surgery, the first resumption of defects using titanium plates forming the basic shape of mandible and the occlusal relationship,then used to re-sawing to take modeling of the iliac bone,transplanted free iliac bone and fixed after the inside of the titanium plate, thereby restoring the continuity of mandible missing.ResultsFree forearm flap and free iliac bone all survived in 11 cases, the success rate of 100％. The patients were followed up for 6 to 12 months, although the shape of restoration areas were different levels of fat, but eating, swallowing and other oral function had been well improved, 7/11 could enter normal diet, 4/11 to enter liquid diet, At the same time, the patients in social activities can communicate in a language daily, sensory function of skin flap and facial appearance has been satisfactory recovery.ConclusionConformal free forearm flap and free iliac bone is an ideal way to reconstruct the composite tissue defects of the floor of mouth and the lower part of face in the same period, not only safe, practical prognostic effect, and also can significantly improve the patients quality of life, which is worthy of reference and use.

Aim To study the effect of simvastatin on the production of reactive oxygen species ( ROS ) and the secretion of interleukin-1 beta ( IL-1β) in oxidized low density lipoprotein ( oxLDL )-induced macropha-ges. Methods After the murine macrophage J774A. 1 was treated with 0,50,100,200 mg·L-1 oxLDL, the contents of aggregated lipid in macrophages were ob-served and determined by oil red O staining. Then, the oxLDL-primed macrophages were treated with 0 . 5 ,1 . 0μmol·L-1 simvastatin, the production of ROS was de-termined by flow cytometry and the expressions of pro-caspase-1 , cleaved caspase-1 and mature IL-1βon pro-tein level were determined by Western blot. Results The oil red O staining results showed that oxLDL could induce obvious lipid aggregation in macrophages, and reached the saturation point with 100 mg·L-1 concen-tration. Flow cytometry results indicated that oxLDL could induce the production of ROS in macrophages, up to 167% ± 0. 47%, and ROS level decreased to 139% ± 0. 97% in a dose-dependent manner after treatment with simvastatin. Western blot indicated that simvastatin could inhibit the expression of cleaved caspase-1 and mature IL-1β in macrophages triggered by oxLDL;compared with oxLDL group, the expression of cleaved caspase-1 and mature IL-1β decreased in simvastatin treated group, and all results had statistical significance ( P<0. 05 ) . Conclusion In the lipid ag-gregation model of macrophages induced by oxLDL, simvastatin can inhibit the production of ROS, caspase-1 activation, and secretion of IL-1β in macrophages.

BACKGROUND: The important pathological changes of acute respiratory distress syndrome (ARDS) is disruption of the lung alveolar-capillary membrane barrier and resultant pulmonary edema associated with a proteinaceous alveolar exudate. Bone marrow mesenchymal stem cells (BMSCs) are able to carry on dividing and renewing themselves, and can eventually develop into many other types of cells. This provides a new treatment for treating injury of lungs.OBJECTIVE: To investigate the prevention of endotoxin-induced acute lung injury in rabbit by BMSCs.DESIGN, TIME AND SETTING: The randomized controlled animal study was performed at the Central Laboratory of Tangdu Hospital from October 2007 to January 2008.MATERIALS: A total of 20 rabbits were used in this study. Two rabbits were utilized to culture BMSCs. Eighteen rabbits were randomly assigned to three groups, saline control group, acute lung injury group and cell transplantation group (n = 6). Endotoxin was purchased from Sigma, USA.METHODS: Rabbit BMSCs were isolated and cultured by the Ficoll method. At the third passage, BMSCs were harvested for use.In the acute lung injury and call transplantation groups, endotoxin was infused into the trachea to establish models of acute lung injury/ARDS. Thirty minutes following model establishment, 2 mL BMSC suspension (1 x 105) was infused into the right jugular vein in the cell transplantation group. An equal volume of saline was injected into the saline control and acute lung injury groups.MAIN OUTCOME MEASURES: Number of neutrophilic granulocyte, wet to dry weight ratio of lung tissue, protein content and pathological changes in lung tissue in bronchoalveolar lavage fluid were measured.RESULTS: The increase in wet to dry weight ratio indicated the existence of pulmonary edema. The increase in neutrophilic granulocyte number suggested severe inflammatory reaction. The increased protein content showed the damage to lung alveolar-capillary membrane barrier. Following 48 hours of transplantation, neutrophilic granulocyte number and protein content in bronchoalveolar lavage fluid was significantly decreased (P < 0.01), and wet to dry weight ratio was significantly increased (P < 0.01) in the acute lung injury group compared with the saline control group. Compared with the acute lung injury group,neutrophilic granulocyte number and protein content was significantly increased (P < 0.01), and wet to dry weight ratio was significantly diminished (P < 0.01) in bronchoalveolar lavage fluid in the call transplantation group. Hematoxylin-eosin staining suggested that pulmonary alveoli was normal in the saline control group, presented typical acute lung injury in the acute lung injury group, and the pathological changes were mild in the cell transplantation group.CONCLUSION: BMSC transplantation can significantly reduce endotoxin-induced acute lung injury.