To observe the expressions of matrix metalloproteinase-2 (MMP-2) and tissue inhibitor of metalloproteinase-1 (TIMP-1) in rats with pulmonary fibrosis (PF) induced by bleomycin, and to explore the mechanisms of Shenlong Decoction in preventing and treating PF.

Objective To investigate the optimal window width and window level for measuring the airway dimensions with spiral CT scan in Japanese white big-ear rabbits so as to lay the foundation for airway stenting in animal experiments. Methods Multi-slice spiral CT scanning of cervico-thoracic region was performed in 30 healthy adult Japanese white big-ear rabbits, the anteroposterior and transversal diameter of the thoracic trachea, the anteroposterior diameter of the right and left bronchus were measured with lung window, mediastinum window and special fat window separately. The revealing rate of the tracheal wall and the measuring results in different windows and levels were recorded and compared with the anatomical data. The differences of the relevant data were statistically analyzed. Results With lung window, the tracheal wall was well demonstrated, but the relevant data were smaller than that with mediastinum window. With mediastinum window, the data were bigger and the tracheal wall border appeared blurred. The results obtained with fat window were close to the actual anatomical data. Conclusion For accurately measuring the anteroposterior and transversal diameter of the thoracic trachea in Japanese white big-ear rabbits with multi-slice spiral CT scan, fat window should be adopted, which is helpful for the preparation of tracheal and bronchial stents.

Objective To establish a method of inducing dendritic cells(DC)from rat bone marrow cells in vitro,and identify the phenotype and function characteristics.Methods The rat bone malToW cells were collected and cultured in vitro under the condition of recombinant rat GM-CSF(rrGM-CSF)and recombinant rat IL-4(rrIL-4).After 2 weeks,the morphological character of DCs was observed under light microscope and scanning electron microscope.Expression of MHC-Ⅱ,CD80 and CD86 were detected by flow cytometry.The ability to stimulate allogenic T cells of the cultured DCs was detected by mixed lymphocyte reaction.Results DCs showed typical morphology with elongated dendritic processes under inversion microscope and scanning electron microscope.DCs at day 6 revealed immature phenotype,including MHC-Ⅱ(29.03 ±4.39)%,CD80(21.98±7.08)%and CD86(25.94±6.80)%.DCs at day 12 showed higher expression of MHC-Ⅱ(74.05±5.97)%,CD80(79.85±6.53)%and CD86(81.00±7.47)%,and stimulatory capacity of allogenic T cells,compared with that in DCs at day 6.Conclusion Matured DCs could be generated from rat bone marrow cells and attendance with rrGM-CSF and rrIL-4,which present the feasibility for further research on its application to allograft immunorejection.

Objective To investigate the applicability of clinical grade and a non-contact measurement method in evaluation of underneath eye wrinkles and to compare two methods.Methods A lot of 46 healthy Chinese women were recruited for this study.Underneath eye wrinkles severity was evaluated using clinical grade and a non-contact measurement method.The correlations were calculated for clinical grade and non-contact measurement parameters and age.The non-contact measurement parameters were classified by factor analysis.The correlations between age,clinical grade and factors were analyzed.Results The correlation coefficient between clinical grade in comparison to subject's age was 0.818.The parameters getting from non contact measurement were obviously correlated with age and clinical grade except SEr and SEsc;the correlation coefficients between parameters and age were-0.601 to 0.605;the correlation cofficients between parameters and clinical grade were-0.630 to 0.570.The non-contact measurement parameters could be classified into two factors;one represented wrinkle depth and roughness;the other represented wrinkle width and counts.These two factors were also obviously correlated with age and clinical grade.Conclusions Clinical grade and non contact measurement methods are both applicable in evaluation of underneath eye wrinkles.The parameters getting from two methods are obviously correlated with each other.

ObjectiveTo analyze the clinical characteristic of the nasal type extranodal NK/T cell lymphoma(ENKL)as well as the therapeutic effects for different treatment methods and element that would affect prognosis.MethodsReview and analyze the 27 patients with pathologically confirmed ENKL patients received radiotherapy (3 cases),chemotherapy (9 cases),combination of radiotherapy and chemotherapy (15 cases),3 of whom received auto-HSCT.Detailed B symptoms,lactate dehydrogenase (LDH),performance status(PS)scores,international prognostic index(IPI),Ann Arbor stage and therapeutic modality were included in univariate analysis.ResultsThe average overall survival time was 32 (2-42) months.The 1,2and 3 year overall survival (OS) rates were 79.5 ％,71.6 ％,53.7 ％,respectively.The 2-year overall survival of patients who was early stage (stage Ⅰ + Ⅱ ) was 83.3 ％,and those who was advanced stage(stage Ⅲ+Ⅳ) was 62.3 ％ (P =0.368).Among the patients in early stage,after radiotherapy or chemotherapy,4 cases achieved OR (overall response) which is complete remission (CR) or partial remission (PR),9/10 in combination of radiotherapy and chemotherapy achieved OR. While among those in their advantage stage,the OR in chemotherapy is 2 cases,2/5 in combination of radiotherapy and chemotherapy.In univariate analysis,age and short-term response were main factors of survival(P ＜0.05).ConclusionTo treat the nasal ENKL in the early stage,radiotherapy can achieve a better effect,while combination of radiotherapy and chemotherapy is an important method for patients in advantage stage.Age and short-term response may be.prognostic factors of nasal type ENKL.

Objective: To construct and identify the eukaryotic expression vector targeting the CD40 gene in rats.Methods: Two sequences corresponding to the rat CD40 gene were designed on Ambion's Web.The two complementary oligonucleotide strands of DNA fragments were synthesized by chemosynthesis.After annealing of the complementary strands,the DNA fragments were connected to the polyclone sites of the pSilencer 4.1-CMV neo vector,followed by transformation,amplification,plasmid extraction and identification of the recombinant plasmids by BamH Ⅰ and Hind Ⅲ digestion and DNA sequence analysis.Results: The connections between the DNA fragments encoding CD40-targeted siRNA and the pSilencer 4.1-CMV neo vector were correct,as confirmed by agarose gel electrophoresis and DNA sequence analysis.Conclusion: The two recombinant plasmids expressing siRNA of the rat CD40 gene were successfully constructed,which prepared the groundwork for future research on the role of the CD40 gene in homograft rejection.

Objective: To examine expression patterns of focal adhesion kinase (FAK) and its activated form, phosphorylated FAK (pFAK),in human osteosarcoma and to investigate the correlation of FAK expression with clinicopathological parameters and prognosis. Functional consequence of manipulating FAK protein levels was also investigated in human osteosarcoma cell lines. Methods: Immunohistochemical staining was used to detect FAK and pFAK levels in pathologically archived materials from 113 patients with primary osteosarcoma. Kaplan-Meier survival and Cox regression analyses were used to evaluate prognoses. The role of FAK in cytological behavior of MG63 and 143B human osteosarcoma cell lines was studied via the FAK protein knockdown with siRNA. Cell proliferation, migration, invasiveness, and apoptosis were assessed using cell counting kit-8, Transwell, and Annexin V/PI staining methods. Results: Both FAK and pFAK were overexpressed in osteosarcoma patients. Tumor cells exhibited cytoplasmicity and occasional membranous immunoreactivity for FAK. A total of 42 cases (37.17%) mainly showed expressed pFAK in cytoplasm of osteosarcoma cells. No overexpression staining of anti-FAK and anti-pFAK antibodies was observed in normal cancellous bone tissues or negative controls. Significant differences were observed in overall survival between FAK-/pFAK- and FAK+/pFAK- groups (P=0.016), FAK+/pFAK- and FAK+/pFAK+ groups (P=0.012), and FAK-/pFAK- and FAK+/pFAK+ groups (P<0.001). All groups showed similar metastasis-free survival. Cox proportional hazard analysis showed that FAK expression profile is an independent indicator of both overall andmetastasis-free survival. siRNA-based knockdown of FAK significantly reducedmigration and invasion of MG63 and 143B cells and affected proliferation and apoptosis in osteosarcoma cells. Conclusion: Osteosarcoma malignancies in vitro and in vivo were correlated with overexpression and phosphorylation of FAK. These findings suggest that FAK plays an important biological role in osteosarcoma carcinogenesis. This study provides a better understanding of diagnostic and prognostic relevance of FAK overexpression and phosphorylation in osteosarcoma patients. Therefore, FAK and pFAK can be used as independent predictors of overall and metastasis-free survival in osteosarcoma patients.

AIM: To investigate the differences in the distribution of SRY-related HMG box 5 (SOX5) gene single nucleotide polymorphisms (SNPs) among stable chronic obstructive pulmonary disease (COPD) patients, COPD with pulmonary hypertension (PH) patients and healthy controls, and to explore the association of the SOX5 SNPs in COPD-related PH.METHODS: From April 2013 to April 2015, 250 patients with stable COPD were enrolled continuous-ly in Ningxia People’s Hospital according to COPD treatment guidelines (2013 edition).All the patients received echocar-diography, and were divided into COPD with PH group [pulmonary artery systolic pressure (PASP)≥50 mmHg, n =103] and COPD without PH group (PASP <50 mmHg, n =147).The healthy persons (matched for age, sex, race and smoking index, n =127) were selected as control group at the same period.Genotyping of SOX5 gene rs10842262 and rs11046966
loci was performed using MassARRAY genotyping system ( Sequenom).Genotype frequencies were calculated.RE-SULTS: Age, sex and smoking index showed no significantly difference between control group and COPD group, neither between COPD with PH group and COPD without PH group.Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci between control group and COPD group was of significant difference (P<0.05).Genotype frequencies of SOX5 gene rs10842262 and rs11046966 loci showed no significant difference between COPD with PH group and COPD without PH group.CONCLUSION: SOX5 gene rs10842262 and rs11046966 loci may play an important role in COPD, but not in COPD-related PH.

High temperature liquid water (HTLW) has drawn increasing attention as an environmentally benign medium for organic chemical reactions, especially acid-/base-catalyzed reactions. Non-catalyzed hydrolyses of gallotannin and tara tannin in HTLW for the simultaneous preparation of gallic acid (GA) and pyrogallol (PY) are under investigation in our laboratory. In this study, the hydrolysis kinetics of gallotannin and tara tannin were determined. The reaction is indicated to be a typical consecutive first-order one in which GA has formed as a main intermediate and PY as the final product. Selective decomposition of tannin in HTLW was proved to be possible by adjusting reaction temperature and time. The present results provide an important basic data and reference for the green preparation of GA and PY.
Catalysis ; Hydrolysis ; Kinetics ; Tannins ; chemistry ; Temperature

The microbial transformation of buflomedil by Cunninghamella blakesleana AS 3.153 was studied, as well as a microbial model which can be used to mimic metabolism of buflomedil in mammal was established. Experiments were conducted to screen the capabilities of four strains of Cunninghamella species to transform buflomedil, in which C. blakesleana AS 3.153 was selected for a preparative biotransformation. Furthermore, the microbial model was established based on the transformation condition optimization. The parent drug and its metabolites produced by C. blakesleana AS 3.153 were detected by liquid chromatography-mass spectrometry method and three metabolites were identified while two of them were new found metabolites. Two major metabolites, para-O-desmethyl buflomedil and 12-C-oxidated buflomedil, were isolated by semi-preparative HPLC. Based on the comparison between different species, the microbial transformation of buflomedil by C. blakesleana AS 3.153 is more similar to the metabolism of buflomedil in human and Beagle dog than that in rat.
Adult ; Animals ; Biotransformation ; Chromatography, High Pressure Liquid ; Cunninghamella ; metabolism ; Dogs ; Female ; Humans ; Male ; Molecular Structure ; Pyrrolidines ; chemistry ; pharmacokinetics ; Rats ; Rats, Wistar ; Spectrometry, Mass, Electrospray Ionization ; Young Adult