The temperature and salinity ranges for growth of twelve strains of marine Bdellovibrio-and-like organisms (BALOs) which affiliated with four different phylogenetic clusters in the family Bacteriovoracaceae and the lytic ability of them to six kinds of common shrimp pathogen vibrios were studied. The morphological characteristics of four representative strains were examined with transmission electron microscope. The results showed that the strains within the same cluster or sub-cluster had identical growth temperature or salinity. The growth temperature ranges of clusters IV, VI, IX and X strains were 20?C～35?C, 15?C～35?C, 15?C～40?C, 10?C～40?C, respectively, and the optimum growth temperature was about 30?C or 35?C. The growth salinity ranges of clusters IV, VI, IX and X strains were 5‰～40‰, 2.5‰～30‰, 5‰～60‰ and 5‰～60‰, and the optimum salinities were about 10‰, 5‰, 20‰ and 20‰, respectively. Three of the six kinds of vibrios tested were lysed by all the BALOs, while the others could only be lysed by some of the BALOs. However, differential lysis to some of the vibrios was also observed among BALOs within the same cluster. The four strains of different clusters all exhibited typical BALOs morphology, hav-ing vibrioid cell with a single polar flagellum.

OBJECTIVETo investigate the effect of Hepatitis B Virus X Protein (HBx) on the expression of lipid metabolism-related genes and its role in pathogenesis of hepatocyte fatty degeneration.

METHODSHepatitis B Virus X gene eukaryon expression vector pIRES2-eGFP-HBx was transfected into HepG2 cells to establish HepG2/HBx cell model for HBx expression. HepG2 cells transfected with pIRES2-eGFP (HepG2/pIRES2 cell) and non-transfected were used as controls. At 24, 48 and 72 hours after transfection, the expression of green fluorescent protein (GFP) was observed by fluorescence microscope and the triglyceride(TG) content was detected. RT-PCR and Western blot were applied to detect the levels of sterol regulatory element binding protein-1 (SREBP-1), liver x receptor alpha (LXRalpha) mRNA and the levels of HBx, LXRalpha and fatty acid synthase (FAS) protein. At 24, 48 and 72 hours after transfection, the expression of GFP was found in HepG2/HBx and HepG2/pIRES2 cells, and increased gradually. The expression of HBx was detected only in HepG2/HBx cells, and was increased with time after transfection (F = 32.21, P less than 0.01). These suggested successful obtaining of HepG2-HBx cell model for HBx expression.

RESULTSAt 24h, 48h and 72h after transfection, the expression levels of LXRalpha mRNA (0.386+/-0.055, 0.505+/-0.071, 0.649+/-0.058 ) and SREBP-1 mRNA (0.395+/-0.055, 0.548+/-0.047, 0.795+/-0.058), as well as the levels of LXRalpha protein(0.178+/-0.036, 0.263+/-0.047, 0.347+/-0.058) and FAS protein(0.436+/-0.055, 0.608+/-0.053, 0.827+/-0.046) in HepG2-HBx group were dramatically higher than those in the controls at the same time points (all P less than 0.05/0.01), and were gradually increased with time (all P less than 0.05/0.01). A positive correlationship was observed between HBX protein level and the LXRalpha, SREbP-1 mRNA and LXRalpha, FAS protein levels. The difference of TG content between HepG2/HBx group and control groups was not statistically significant (P more than 0.05).

CONCLUSIONSHBx-LXRalpha-SREBP-1/FAS pathway suggested regulating transcription and expression of lipid metabolism-related genes, which might be one of the important molecular mechanism causing hepatocyte fatty degeneration.

Carcinoma, Hepatocellular ; metabolism ; Fatty Acid Synthase, Type I ; metabolism ; Hep G2 Cells ; Humans ; Lipid Metabolism ; genetics ; Liver Neoplasms ; metabolism ; Liver X Receptors ; Orphan Nuclear Receptors ; metabolism ; Sterol Regulatory Element Binding Protein 1 ; metabolism ; Trans-Activators ; genetics ; Transfection

This paper proposes a one-way micro valve with a simple structure and a simulation design for the engineering capsule. We have now got its design parameter selection method and its mechanic characteristic from experiments.
Capsules ; Computer Simulation ; Drug Delivery Systems ; methods ; Equipment Design ; Infusion Pumps, Implantable ; Technology, Pharmaceutical ; instrumentation ; methods

RNA interference (RNAi) is applied in the gene therapy of malignant tumors as an effective technique to degrade its homologous mRNA specifically. This technique has been developed quickly in broad prospects. Molecular imaging plays an important role both in gene imaging and in molecular signal transduction. It is used as a direct approach to study the onset and development of disease. The technique of RNAi can be used for preparing molecular probes, for making earlier and specific diagnosis, and for targeted gene therapy of malignant tumors.
Animals ; Genetic Therapy ; Humans ; Molecular Imaging ; methods ; trends ; Molecular Probe Techniques ; Neoplasms ; diagnosis ; genetics ; therapy ; RNA Interference

Patients with advanced liver cancer have poor basic conditions and poor prognosis due to limited treatment options. Sorafenib is the first-line drug approved by FDA for the treatment of advanced liver cancer, and it has multi-kinase inhibitory activity and can improve the survival time of patients with liver cancer. Recent studies have shown that sorafenib is also an inducer of ferroptosis and its toxicity on hepatoma cells partly depends on the induction of ferroptosis of tumor cells. The enhancement of sorafenib-induced ferroptosis can improve the therapeutic effect of sorafenib on liver cancer. At the same time, ferroptosis also plays an important role in the drug-resistance mechanism of sorafenib, and some key proteins involved in the pathways of ferroptosis can also be used to indicate the efficacy of sorafenib and the prognosis of liver cancer. This article reviews the latest research advances in the role of ferroptos in the treatment of liver cancer with sorafenib.

OBJECTIVE: To investigate the inhibitory effect of compound Daqiqi decoction(CDQD) combined with cisplatin on subcutaneously transplanted ovarian cancer in nude mice and its related mechanisms. METHODS: The 60 subcutaneously transplanted model of nude mice was established with human ovarian cancer cell line SKOV3, and divided into 6 groups randomly, each group of 10 nude mice, which were model group that was treated with saline, CDQD low dose group with the CDQD dosage of 15.16 g•kg-1, CDQD medium dose group with the CDQD dosage of 30.33 g•kg-1, CDQD high dose group with the CDQD dosage of 60.66 g•kg-1, cisplatin group with the DDP dosage of 3 mg•kg-1 and combined group that was treated with the CDQD dosage of 30.33 g•kg-1 and the DDP dosage of 3 mg•kg-1. Cisplatin was administered once every 3 d, and the remaining drugs were administered once a day. Then,the tumor-bearing mouse model was given the corresponding drugs for 14 consecutive days, and the tumor volume was measured every 3 d. After the end of treatment, the tumor was removed and weighed. The morphology of the tumor cells were observed by HE staining. The apoptosis of tumor cells was detected by TUNEL method. The mRNA and protein expression of miR939 and STAT3 and VEGFA and EGFR in tumor tissues were detected by real-time fluorescence quantitative PCR and Western-blot. RESULTS: The tumor volume and the tumor weight of the treated groups were all decreased(P<0.01). Compared with the model group, the tumor volume and tumor weight of the combined group were less than those of the other groups(P<0.01), and the apoptosis rate of the combined group was significantly higher than other groups(P<0.01).The expression of miR939 and STAT3 and VEGFA were down-regulated and the expression of EGFR were up-regulated in the treatment groups. Compared with the model group, the expression of MiR-939, STAT3 and VEGFA were down-regulated and the expression of EGFR was increased in the treatment group. MiR-939, STAT3, VEGFA expression in the combined group was the lowest(P<0.05), and the EGFR expression was highest(P<0.05). CONCLUSION: Studies have shown that CDQD can inhibit ovarian cancer subcutaneously transplanted tumor in nude mice, and its mechanism may be related to inhibition of MiR-939/STAT3 pathway activation, down-regulation of VEGFA, and up-regulation of EGFR expression. The inhibitory effect of CDQD on ovarian cancer tissues has a concentration dependence. And the combination of CDQD and DDP can enhance the anti-tumor effect of DDP and reduce the side effects of DDP on tumor-bearing mice.

Objective To investigate the protective effect of high mobility group box 1 (HMGB1) gene silence on astrocyte injury caused by oxygen-glucose deprivation/reoxygenation (OGD/R) in vitro. Methods Astrocytes were divided into control group, OGD/R group and HMGB1 siRNA group. Astrocytes in OGD/R group were cultured with glucose free DMEM under 1% O2 condition, then they were treated with OGD for 2, 4 and 6h respectively, and then reoxygenated for 24h. In HMGB1 siRNA group, astrocytes that transfected with HMGB1 small interference RNA (siRNA) lentivirus-vector were treated with OGD 6h, and then reoxygenated for 24h. After 24h reoxygenation, the mRNA and protein expression of HMGB1 in the astrocytes were determined by RT-PCR and Western blotting. HMGB1 protein in culture supernatant of astrocytes was determined by ELISA. The cell injury and survival rate were assessed by LDH activity (LDH%) and MTT assay. Results Compared with the astrocytes without transfection of HMGB1 siRNA lentivirus-vector, the protein expression of HMGB1 was suppressed by siRNA. Compared with the control group, with prolongation of the OGD time, the mRNA and protein expression of HMGB1 increased gradually (P<0.05) after OGD/R, and it further increased with elapse of time. OGD resulted in significant injuries with time extention, and the LDH% increased (P<0.05) with marked lowering of survival rate (P<0.05). Compared with the OGD/R group, cell injury in HMGB1 siRNA group alleviated remarkably, and survival rate was elevated significantly (P<0.01). Conclusion The expression of HMGB1 in astrocytes can be inhibited by siRNA, and over-expression of HMGB1 might be an important factor in OGD/R-induced cell injury.

Objective To prepare a mesoporous silica-coated polypyrrole nanoparticles loaded with honokiol (PPy@MSN-HK) and evaluate their in vitro release behavior. Methods In this study, PPy@MSN-HK was obtained in three steps: First, prepared polypyrrole nanoparticles; Second, coated mesoporous silica shell on its surface; Third, absorbed honokiol. The TEM, particle size, zeta potential, drug loading, infrared spectroscopy, in vitro photothermal properties, and in vitro release characteristics were chosen as indexes to investigate its potential as antitumor nanocarries. The release profiles were analyzed by simulating factor (f2), and the dissolution profiles were fitted by a variety of commonly used mathematical models. Results The results showed that the prepared nanoparticles had uniform particle size and uniform size distribution. The average particle size was (220.4 ± 4.2) nm, polydispersity coefficient was 0.042 ± 0.010, zeta potential was (-21.1 ± 0.8) mV, drug loading was (2.58 ± 0.53)%, and entrapment efficiency was (75.04 ± 0.95)%, respectively. The results of in vitro photothermal experiments showed that with the constant laser power density, the temperature change value of nanoparticles suspension increased with the increase of nanoparticles concentration. This showed that PPy@MSN have a good photothermal effect. In vitro release test revealed that the two release curves were not similar, and fitting best with Ritger-Peppas eqution and Logistic eqution respectively. Conclusion The water solution method could be used to prepare PPy@MSN, which may provide a promising drug delivery strategy for tumor treatment.

Objective To study the in situ intestinal absorption characteristics of cyclovirobuxine D (CB) hydroxypropyl-β-cyclodextrin inclusion complex (CBHD) in rats, and to explore the effect of cytochrome P450 inhibitor ketoconazole (KET) on CB and CBHD in situ intestinal absorption. Methods Twenty-four male rats were randomized into CB, CBHD, KET+CB and KET+CBHD groups, with 6 rats in each group. In situ intestinal absorption was adopted in a rat model. One-way intestinal perfusion model was employed to investigate the absorption of CB and CBHD in the intestinal segments of rats and the effects of KET on CB and CBHD absorption. The concentration of CB was determined by highperformance liquid chromatography with fluorescence detector (HPLC/FLD; Lichrospher C18 column [250 mm×4.6 mm, 5 μm]). The mobile phase was methanol-water with volume ratio being 85: 15. The excitation wavelength was set at 231 nm, and emission wavelength was set at 385 nm. The column temperature was 25 °C, and flow rate was 1.0 mL/min. The injection volume was 20 μL. Results The specificity of HPLC/FLD method was good and the standard curve equation was A=106.7 C+41.861 (R2=0.999 08) based on the linear regression of CB concentration (C) with CB peak area (A), indicating that the CB mass concentration was linear in the range of 0.5 to 20.0 μg/mL. The intra-day precision of the 1.0, 5.0 and 10.0 μg/mL samples was 2.25%, 2.44% and 3.04%, and the inter-day precision was 4.22%, 2.00% and 2.50%, respectively. The precision was good and the method was in accordance with the requirements of methodology. The recovery rates of the 1.0, 5.0 and 10.0 μg/mL samples were 99.08%, 98.24% and 97.25%, respectively, which were also in accordance with the requirements of methodology. The intestinal absorption rate constant (Ka) values of CBHD with KET were 4.18, 5.05, 1.91 and 2.85 times those of CB, and the effective permeability (Peff) values were 4.92, 5.98, 2.19 and 3.24 times those of CB in the duodenum, jejunum, ileum and colon, respectively (all P<0.05). Conclusion KET can improve the intestinal absorption of CB and CBHD in rats.

Meibomian gland dysfunction(MGD)is mainly characterized by terminal duct obstruction and/or qualitative/quantitative changes in the glandular secretion, resulting in the alteration of tear film and ocular surface diseases. It covers several meibomian gland diseases ranging from congenital to acquire. With the progress of clinical research, recent studies indicate there are many risk factors related to MGD, including ophthalmic diseases, systemic diseases, therapeutic approaches(topical/systemic medication, surgery)and even environmental factors. Thus, a proper understanding of the various risk factors acting on MGD will be helpful to the clinical diagnosis and treatment of MGD.