ABSTRACT:Objective To prepare polymeric nanomicelles capable of simultaneously loading doxorubicin (DOX) and Bcl‐2 small interfering RNA (Bcl‐2 siRNA ) , and to explore their in vitro cytotoxicity and cellular uptake in MCF‐7 human breast cancer cells .Methods Copolymer poly (ethylene glycol )‐g‐polyethylenimine‐g‐poly(γ‐benzyl‐L‐glutamate) was synthesized by the combination of reductive amination and carbodiimide methods , and its chemical structure was verified by 1 H NMR .Empty and drug‐loaded copolymeric nanomicelles were prepared by dialysis method and characterized by transmission electron microscope and dynamic light scattering .The ability of the nanomicelles to compress Bcl‐2 siRNA was measured by by agarose gel electrophoresis method . The release profiles of DOX and Bcl‐2 siRNA from the nanomicelles were explored by means of fluorescence spectrometry and dialysis method .The in vitro cytotoxicity and cellular uptake of DOX and Bcl‐2 siRNA co‐loaded nanomicelles in MCF‐7 human breast cancer cells were characterized by MTT assay and confocal laser scanning microscopy , respectively .Results The critical micelle concentration of the copolymer was about 4 mg/L ,and the sizes of self‐assembled empty and drug‐loaded nanomicelles were smaller than 200 nm .The drug‐loading efficiency and drug‐loading content of DOX in the nanomicelles were 88 .7% and 15 .1% ,respectively .The DOX‐loaded nanomicelles could efficiently compress Bcl‐2 siRNA when an N/P ratio was ≥64 .The zeta potential of DOX and Bcl‐2 siRNA co‐loaded nanomicelles was +30 mV .The release behavior of the cargoes from the nanomicells was pH‐sensitive , and the release of Bcl‐2 siRNA was more sensitive to acidic pH than that of DOX . The nanomicelles could simultaneously deliver DOX and Bcl‐2 siRNA into MCF‐7 cells , and the co‐delivered DOX and Bcl‐2 siRNA significantly increased the cytotoxicity of DOX (P<0 .05) .Conclusion The polymeric nanomicelles can co‐load DOX and Bcl‐2 siRNA and deliver them into MCF‐7 cells , and DOX in combination with Bcl‐2 siRNA can synergistically inhibit the growth of MCF‐7 cells and promote cell apoptosis ,suggesting that the nanomicells may be a promising carrier for the co‐delivery for chemotherapeutics and genes .

Objective To observe and compare the atomic force microscopy (AFM) images of Yersinia pestis EV76 and the changes in the morphology of the bacteria treated with normal serum and F1 antibody from rabbit,and to explore the immunoassay method to detect Yersinia pestis by AFM. Methods The Yersinia pestis were treated with normal serum and F1 antibody from rabbit and control buffer. All the prepared samples were observed and analyzed by AFM. The changes in the cell surface structures were probed and characterized through sectional analysis,especially the changes of Ra and Rq value. Results The normal morphology of Yersinia pestis was oval in shape with a relatively smooth surface, the size dimension of which was about 1.1-1.3 μm in length with a section profile of 0.8-1.0 μm in width and 0.04-0.06 μm in step height. The step height of the bacteria treated with the normal serum and F1 antibody was obviously enlarged. The shape of the bacteria treated with F1 antibody changed irregularly. Furthermore, the surface of the bacteria was more roughened. Conclusion The morphological characters of Yersinia pestis has been acquired through its AFM images. The morphology of Yersinia pestis treated with F1 antibody has changed greatly, and the index of roughness can be regarded as the distinguished index to detect Yersinia pestis by AFM.

Objective: Analyze the ecological adaptable regions for the resource reservation and the expanding cultiva-tion of Quisqualis indica L. Methods: The Traditional Chinese Medicine Geographic Information System (TCMGIS) was used to analyze the ecological adaptable region. Results: The 17 provinces (city, municipalities) were the ecolog-ical adaptable region for Q. indica L. The 100% ecology similarity regions are mainly in 16 provinces (city, munici-palities), including Guangxi, Guangdong, Hunan, Fujian etc., which area accounts for 1 012 895.98 km2. Among these, the area in Guangxi province is the largest (195 313.64 km2), the second is Guangdong (150 276.36 km2), the following order is Hunan(128 925.29 km2), Fujian (101 471.16 km2) and Y unnan (100 660.83 km2). The 95%~100%ecology similarity regions is located in 16 provinces (city, municipalities), including Y unnan, Sichuan, Guizhou, Hubei province etc., which area accounts for 362 366.25 km2. The largest area is in Y unnan province (92 399.04 km2), followed by Sichuan(76 122.32 km2), Guizhou (43 902.38 km2), Hubei (31 760.32 km2) and Jiangxi (29 770.09 km2). Conclusion: The results are consistent with the survey results and the previous reports. To promote the develop-ment of Q. indica L. cultivation, the planting scientifically and quality evalution should be further conducted on the basis of the results by system analysis.

BACKGROUND/OBJECTIVES: D. candidum is a traditional Chinese food or medicine widely used in Asia. There has been little research into the anticancer effects of D. candidum, particularly the effects in colon cancer cells. The aim of this study was to investigate the anticancer effects of D. candidum in vitro and in vivo. MATERIALS/METHODS: The in vitro anti-cancer effects on HCT-116 colon cancer cells and in vivo anti-metastatic effects of DCME (Dendrobium canidum methanolic extract) were examined using the experimental methods of MTT assay, DAPI staining, flow cytometry analysis, RT-PCR, and Western blot analysis. RESULTS: At a concentration of 1.0 mg/mL, DCME inhibited the growth of HCT-116 cells by 84%, which was higher than at concentrations of 0.5 and 0.25 mg/mL. Chromatin condensation and formation of apoptotic bodies were observed in cancer cells cultured with DCME as well. In addition, DCME induced significant apoptosis in cancer cells by upregulation of Bax, caspase 9, and caspase 3, and downregulation of Bcl-2. Expression of genes commonly associated with inflammation, NF-kappaB, iNOS, and COX-2, was significantly downregulated by DCME. DCME also exerted an anti-metastasis effect on cancer cells as demonstrated by decreased expression of MMP genes and increased expression of TIMPs, which was confirmed by the inhibition of induced tumor metastasis in colon 26-M3.1 cells in BALB/c mice. CONCLUSIONS: Our results demonstrated that D. candidum had a potent in vitro anti-cancer effect, induced apoptosis, exhibited anti-inflammatory activities, and exerted in vivo anti-metastatic effects.
Animals ; Apoptosis ; Asia ; Asian Continental Ancestry Group ; Blotting, Western ; Caspase 3 ; Caspase 9 ; Chromatin ; Colon ; Colonic Neoplasms ; Down-Regulation ; Flow Cytometry ; HCT116 Cells ; Humans ; Inflammation ; Methanol ; Mice* ; Neoplasm Metastasis ; NF-kappa B ; Up-Regulation

Numerous studies have shown that the health of spiral ganglion neurons is highly important for hearing. As a trophic factor of spiral ganglion neurons, neurotrophin 3 (NT3) is a potential candidate for prevention of spiral ganglion neuron degeneration in human. In our experiments, efficient transduction and long term expression of foreign gene of cochlea cells has been found with adenovirus carried lacZ gene (Ad-lacZ). A model of guinea pig deafness was made by intense noise exposure, which destroyed the entire organ of Corti in the middle part of the cochlea. Seven days after noise exposure, the animals were anesthetized and 1 10(8) recombinant adenoviral particles were injected into the scala tympani through the round window membrane. Animals inoculated with neurotrophin 3 adenovirus(Ad-NT3) were designated as the experimental group, animals inoculated with Ad-lacZ vector served as the control group. Four weeks after the inoculation of the virus, NT3 immunoreactivity was observed in the Ad-NT3 inoculated group. HE histochemical staining results showed that in the Ad-lacZ injected group, the neuronal degeneration was severer and the density of spiral ganglion neurons was significantly lower than those in the Ad-NT3 injected group. Our results demonstrate that with adenovirus-mediated overexpression NT3 may be developed into a new treatment to prevent secondary spiral ganglion degeneration following the damage to Corti organ.
Adenoviridae ; genetics ; Animals ; Cochlea ; pathology ; Gene Transfer Techniques ; Genetic Therapy ; Guinea Pigs ; Hearing Loss, Noise-Induced ; pathology ; In Vitro Techniques ; Neurotrophin 3 ; genetics ; Recombination, Genetic

OBJECTIVETo study and establish the GC fingerprint of essential oils of 10 batches the flower of Lonicera japonica.

METHODThe essential oils were extracted by steam distillation from ten batches of the flowers, then separated by capillary gas chromatography.

RESULTThe similarity of 10 batches of the flower of L. japonica was high.

CONCLUSIONThe method was reliable, and can be used to evaluate the quality of the flower of L. japonica.

Chromatography, Gas ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Flowers ; chemistry ; Gas Chromatography-Mass Spectrometry ; Lonicera ; chemistry ; classification ; Oils, Volatile ; chemistry ; isolation & purification ; Phylogeny ; Reproducibility of Results

The aim of the study is through observing the morphology of the prepared influenza virus (H1N1) with transmission electron microscopy (TEM) and atomic force microscopy (AFM) to explore the application of AFM on the research of the external character of viruses and provide a new, simple and efficient technique for the study of the viral morphology. TEM image was obtained by negatively stained influenza virus with 1% Phosphotungstic Acid; AFM image applied the tapping mode to influenza virus without any further treatment in air at room temperature, and the morphology parameters, including length (diameter), Ra and Rq are calculated by sectional analysis. The shapes of influenza virus A are spherical, filamentous or other pleomorphous particles observed by both AFM and TEM. TEM image of influenza virus A is two-dimensional image, and viral surface has visible spikes, while AFM exhibits the three-dimensional image that can be described with several quantifiable indexes through sectional analysis. AFM phase images show viral surface clearly which is characterized by rugged feature and gear-like protuberance. As compared with TEM, AFM is a new research tool for viral morphology study with the advantages of simple sample preparing, visible interface and is intuitionistic for researchers. The surface characteristic parameters of viruses provided by AFM can be served as the main quantifiable indexes for viral morphological study.
Influenza A Virus, H1N1 Subtype ; ultrastructure ; Microscopy, Atomic Force ; Microscopy, Electron, Transmission

This paper studies the vein extraction technique based on the susceptibility weighted imaging (SWI) and introduced an improved self-adaptive threshold method based on the vessel enhancing diffusion. The approach employs the combination indicator of the local gray character, the global gray character and the tubular information of the vein. It first applies the vessel enhancing diffusion filter to enhance the continuity of the vein, increases the detection rate of tiny vein and suppresses the nucleus areas. And then it uses the improved self-adaptive threshold method to extract the vein. The results demonstrate that this approach can solve the problem above and extract the vein from the SWI image accurately.
Cerebral Veins ; anatomy & histology ; pathology ; Diffusion Magnetic Resonance Imaging ; methods ; Humans ; Image Enhancement ; methods ; Imaging, Three-Dimensional

OBJECTIVETo investigate the dynamic expression of Heat shock protein 70 (Hsp70) in the lungs and plasma of rats with pulmonary fibrosis induced by silicon dioxide (SiO2).

METHODSForty-eight Wistar rats were randomly divided into the control group exposed to normal solution and group exposed to SiO2 (50 mg/ml) with intratracheal injection. Each group was divided into four subgroups. The animals of SiO2 group and control group were sacrificed and lungs were collected on the 7th, 14th and 28th days after exposure, respectively. The left lung tissues were examined with the histopathologic HE staining. The expression and localization of Hsp70 protein in the lung tissues were examined with western blot assay and immunohistochemistry, respectively. The expression levels of Hsp70 protein in the plasma were measured by ELISA.

RESULTSThe expression of Hsp70 in lung tissues of SiO2 group increased on the 7th day and reached the peak value on the 14th day then decreased, but still was significantly higher than that of the control group, the expression of Hsp70 in plasma of SiO2 group still was significantly higher than that of the control group (P < 0.05). The maximum expression level of Hsp70 in plasma of SiO2 group on the 21st day after exposure was 0.216 ± 0.027 µg/ml.

CONCLUSIONThe expression levels of Hsp70 protein in the lung tissues and plasma of the group exposed to SiO2 significantly increased, which were associated with the process of pulmonary fibrosis. It was suggested that Hsp70 protein may play an important biological role in the pulmonary fibrosis induced by SiO2.

Animals ; HSP70 Heat-Shock Proteins ; blood ; metabolism ; Lung ; metabolism ; pathology ; Male ; Pulmonary Fibrosis ; chemically induced ; metabolism ; Rats ; Rats, Wistar ; Silicon Dioxide ; toxicity