Objective: To express the fusion protein of hOct4 and cell penetrating peptides using prokaryotic expression systems, and to optimize its expression methods and observe the membrane penetrating ability of the fusion proteins. Methods: The pET-based prokaryotic expression system was constructed by genetic engineering, and the fusion plasmid was transferred into E. coli BL21(DE3) and Rosetta2(DE3). The protein was purified by Ni affinity chromatography and identified by Western blotting analysis. The penetrating ability of the Rhodamine-labelled fusion protein was investigated using BJ cells. Results: We successfully constructed pET21a(+)-hOct4-11R-His and pET21a(+)-EGFP-11R-His vectors. Fusion proteins hOct4-11-His and EGFP-11R-His were generated by transfering the plasmids into E. coli. The fusion protein was verified by Western blotting analysis and was detected in BJ cells. Conclusion: We have successfully generated EGFP-11R-His and hOct4-11R-His fusion proteins, and the proteins can effectively enter the BJ cells and locate around the nuclei.

Objective: To provide theoretical basis and experimental evidence for diagnosis and therapy of dental pulp indirect injuries by gunshot wounds. Methods: The levels of IL 8 in different stages were studied using immunohistochemical method to evaluate its functions in inflammation process of dental pulps. Results: IL 8 expression was detected by immunohistochemical assay in postwound conditions. The results showed that positive staining of IL 8 was observed predominantly on infiltrating inflammatory cells, vascular endothelial cells and some fibroblasts, which was consistent with the recruitment and activation of inflammatory cells, and with the severity duration of pulpal tissue injury and response. Conclusion: These results indicate that IL 8 is involved in regulating physiologic responses of dental pulp injuries in maxillofacial trauma by a projectile. [

Near infrared chemical imaging (NIR-CI) is an emerging technology for rapidly analyzing the critical quality attribute of Chinese materia medica (CMM). It integrates NIR spectroscopy with chemical imaging. In this paper, it provided a systematic introduction to NIR-CI, such as the core part of instrument, the reliability, transformation, analysis and application of high-dimensional data acquisition. In addition, current studies of NIR-CI application in pharmaceutical field were analyzed. Finally, future opportunities and challenges of NIR -CI applications in the quality control of CMM preparation were prospected.

Objective To obtain the short peptides mimicking antigenic epitopes of Trichinella spiralis ( T\^s\^ ), and explore their cross protective immunity against Schistosoma japonicum ( S\^j. ) in mice. Methods IgG antibodies were purified from sera of mice infected with T\^s\^ . The purified IgG was used to immunoscreen a phage random peptide library of 7 amino\|acid residues displayed as a fusion to protein of filamentous phage. Positive clones were obtained by affinity selection, the reactivity of each clone binding to specific IgG was detected by ELISA. Kunming mice were immunized subcutaneously three times with mixed phage clones. The mice were sacrificed 45 days after challenge. The worms and the liver eggs were counted. Results After three rounds of panning, the relevant phages had been enriched approximately 150 times in production as compared to those from the first round. Of 24 phage clones randomly selected from the third round biopanning, 21 clones were shown to actually bind to the specific IgG. As compared with the control group, the worm and the liver egg reduction rates in vaccination group were 42\^8% and 66\^3% ( P

This study was aimed to optimize bio-solid bidirectional fermentation conditions of Trametes robiniophia Murr. for rhubarb. Sulphuric acid-phenol colorimetry, magnesium-methanol colorimetry, and HPLC were used in the
content determination of polysaccharide, total anthraquinone, and 4 free anthraquinones. The drying rate and con-sumption rate were combined as indicators for the optimization of technical parameters such as medicinal dosage, temperature and amount of water. The results showed that when using 500 mL conical flask, the best fermentation conditions were medicinal dosage of 10 g, the temperature of 34℃, adding water of 120%. It was concluded that bidirectional fermentation of rhubarb increased the content of free anthraquinones. Among them, the content of chrysophanol with anti-oxidation effect increased significantly. The decreasing of combined anthraquinone can relieve the severe laxative effect of rhubarb.

Objective To explore the predictive value of microvessel density(MVD)and blood vessel invasion(BVI)in hepatic metastasis from early-stage rectal cancer.Methods MVD and BVI in the tumor tissue from 380 patients with stage I and II rectal cancer was determined by immunohistochemical S-P method with anti-CDIOS antibody and anti-CD34 antibody,respectively.Multinomial logistic regression was performed to analyze the predictive value of MVD and BVI in hepatic metastasis from early-stage rectal cancer.Results CD105 was expressed in newborn blood vessels,not in normal blood veseels.in the rectal cancer tissue.MVD was correlated with histological type and infiltration depth(P＜0.05).Besides histological type and infiltration depth,BVI was also correlated with histological grade.Multivariate analysis revealed that histological type,tumor infiltration depth,BVI,adjuvant therapy,and MDV were independent predictors of hepatic metastasis from rectal cancer.The risk of hepatic metastasis in patients with postive expression of either MVD or BVI or both were significant higher than that in patients with low expression of MVD and those without BVI expression[hazard ratio(95%CI),4.210(2.182-11.214)].Conclusion BVI and MVD are independent predictors of hepatic metastasis from stage I and II rectal cancer.Combined detection of MVD and BVI may help to predict the clinical outcome of patients with early-stage rectal cancer.

Hereditary spastic paraplegia is a heterogeneous group of genetic neurodegenerative disorders of the nervous system. It is classified into four subtypes based on the mode of inheritance; and among them, most autosomal recessive hereditary spastic paraplegia cases are due to type SPG11 and SPG15 gene mutations. Autosomal recessive hereditary spastic paraplegia cases with SPG30 gene mutation have never been reported in China. Herein, we present our experience with a case of hereditary spastic paraplegia with SPG30 gene mutation in our hospital from North East China. In this patient we detected a missense mutation of c.499 C>T (p.Arg167Cys) in gene KIF1A, a causative gene of type SPG30.

OBJECTIVE:To investigate the improving effect of phenylethanoid glycosides (PhGCs) from Tibetan medicine Phlomis younghusbandii on rats with acute high-altitude cerebral edema. METHODS:60 Wistar rats were randomly divided into a normoxia control group (isometric sterile water for injection),a hypoxia model group (isometric sterile water for injection),a dexamethasone group(4 mg/kg),and three groups of PhGCs at high(400 mg/kg),middle(200 mg/kg)and low(50 mg/kg)dos-es,with 10 rats in each group. The rats were given drugs,ig,6 d before the establishment of models. On the 4th day of administra-tion,ig,the rats in all groups except the normoxia blank group were placed in a simulated 8 000 m altitude plateau environment for 72 h hypoxic exposure to establish the rat models of high-altitude cerebral edema. Following HE stain,the pathological changes in rats’brain tissues were observed under the light microscope. Dry-wet proportion method was used to determine the water con-tents in rats’brain. The content of MDA and the activities of SOD and GSH in rats’brain tissues were detected. Enzyme-linked im-munosorbent assay was adopted to determine the contents of IL-1β and TNF-α in rats’serum and brain tissues. RESULTS:Com-pared to the rats in the normoxia control group,those in the hypoxia model group showed obvious brain edema,and thickened lacu-nas around cells and vessels and inflammatory cell infiltration, higher water contents and MDA and weaker activities of SOD and GSH in brain,and higher contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). Compared to the rats in the hypoxia model group,those in the groups of PhGCs at high,middleand low dosages demonstrated less inflammatory cell infiltration and lower water contents in brain tissues,in which the groups of PhGCs at high and middle dosages demonstrated lower content of MDA and stronger activities of SOD and GSH in brain tissues, and lower contents of IL-1β and TNF-α in serum and brain tissues. There were statistically significances (P<0.01 or P<0.05). CONCLUSIONS:PhGCs can obviously alleviate the acute cerebral injury in rats which is caused by acute hypoxia and has im-provement effect to some degree on the rats with acute high-altitude cerebral edema.

Objective To observe the effects ofTrametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on the migration and invasion of human breast cancer MCF-7 cells and relevant factors; To discuss relevant mechanism of action.Methods Breast carcinoma cell line MCF-7 was used as research subject in this experiment. Control group, Isatidis Radix group,Trametes robiniophila Murr group, andTrametes robiniophila Murr and Isatidis Radix group were included in the experiment. The effects ofTrametes robiniophila Murr and Isatidis Radix bidirectional fermentation products on MCF-7 cell proliferation were measured by MTT method. Cell scratch assay, transwell assay and adhesion assay were used to measure the effects ofTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products on the migration, invasion and adhesion capability of MCF-7 cells, respectively. The effects ofTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products on the mRNA expression of MMP-9 and Vimentin were measured by RT-PCR.Results Compared with Isatidis Radix group andTrametes robiniophila Murrgroup, Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products could significantly inhibit the proliferation, migration, invasion and adhesion capability of MCF-7 cells (P<0.05). Similarly,Trametes robiniophila Murr and Isatidis Radix bidirectional fermentation products reduced the mRNA expression of MMP-9 and Vimentin (P<0.05).ConclusionTrametes robiniophila Murrand Isatidis Radix bidirectional fermentation products may down-regulate the expression of MMP-9 and Vimentin to inhibit the migration, invasion and adhesion capabilities of MCF-7 cells.

Objective To investigate whether IL-24 can inhibit the growth of pancreatic cancer in immune-competent mice,which could be mediated by tumor-targeting oncolytic adenovirus (ZD55-IL-24).Methods Crystal violet staining assay was used to evaluate the anti-proliferative influence of ZD55-IL-24 on Panc02.The tumor growth curve,the tumor formation curve,TUNEL assay,cytotoxic T lymphocyte (CTL) killing assay and serum cytokines detection were used to assess anti-tumor efficacy of ZD55-IL-24 for pancreatic cancer in immune-competent mice.Results ZD55-IL-24 significantly inhibited the growth of pancreatic cancer cells in vitro.Lower tumor formation and slower tumor growth were identified in ZD55-IL-24 group,compared with ZD55-EGFP group or PBS group.ZD55-IL-24 induced pancreatic cancer cells'apoptosis,and mediated anti-cancer immunity by increasing killing responses of CTL to cancer cells and up-regulating serum γ-IFN and IL-6 levels.Conclusion ZD55-IL-24 can induce anti-tumor immunity and suppress pancreatic cancer growth in immune-competent mice.