RNA interference (RNAi) has been proved as a novel approach for gene therapy. However, RNAi mono-therapy only aims at single gene, it therefore may ultimately fail to cure cancers caused by polygene variation. To overcome the deficiency of RNAi mono-therapy, "combinatorial RNA interference" (coRNAi) was put forward as a new strategy. By co-expressing the inducers of RNAi triggering single or multiple targets directly and other RNA- or protein-based silencers, coRNAi keeps target genes silent, prevents carcinogenic progression and induces apoptosis of tumor cells. This paper mainly reviews the major strategies of coRNAi and their applications in cancer gene therapy.
Animals ; Apoptosis ; Genetic Therapy ; methods ; Humans ; MicroRNAs ; genetics ; Neoplasms ; genetics ; pathology ; therapy ; Oncogenes ; RNA Interference ; RNA, Small Interfering ; genetics ; RNA, Small Nuclear ; genetics

Objective To retrospectively analyze 45 cases of oral and maxillofacial malignant tumors and to assess the efficacy and feasibility of oral and maxillofacial malignant tumors implanted with 125I radioactive seeds under guidance of ultrasound.Methods A total of 47 focuses in these 45 patients were determined with the size of these tumors by imaging study,the section was planed by ultrasound,the number and distribution of radioactive seeds were determined with the help of the particle treatment planning system,and were percutaneously implanted particles under guidance of ultrasound.The number and the distribution of particles were assessed by CT.Efficacy endpoints were reexamined and evaluated regularly by ultrasonic and CT according to the standards of WHO.Results The total percentage of efficacy was 70.2% (including complete remission,partial remission).The treatment effect of metastatic carcinoma of lymph node is superior to the parotid tumor.There was no serious complication during the period of implanting and 2 patients with oral ulcers were found after operation.Conclusion The oral and maxillofacial malignant tumor treated implanted with 125I radioactive seeds under guidance of ultrasound is very effective and safe,which is deserved to popularize.The ultrasound is the first choice as a guided method for oral and maxillofacial malignant tumors.

Objective:To investigate the adhesion,proliferation and differentiation of the stem cells from human exfoliated deciduous teeth(SHEDs)on 3 different types of hydroxyapatite(HA)composite scaffold materials.Methods:Pulp cells from human exfoliated de-ciduous teeth were harvested from impacted deciduous teeth by enzyme digestion,expanded and cultured.Cells were verified by immuno-histochemical methods and in vitro differentiation test.Then the cells were cultured on HA/beta tricalcium phosphate (HA/TCP),HA/collagen (HA/COL)and HA/poly-ethylene propylene lactide (HA/PLGA)scaffold respectively.Adhesion rate was examined at hour 4,6,8 and 10 of culture,proliferation was observed by MTT assay on day 1,4,7 and 10 of culture,respectively.The osteogenic dif-ferentiation was studied by alkaline phosphatase(ALP)test,Von Kossa staining and calcium content measur.Results:The attachment of SHEDs was significantly lower on the HA/COL than on the other 2 scaffolds(P <0.05).The ALP activity,mineralization and calci-um content were the highest on HA/PLGA,and the last on HA/COL(P <0.05).Conclusion:HA/PLGA scaffold is more effective in the promotion of the proliferation,attachment and differentiation of SHEDs than HA/TCP and HA/COL scaffolds.

OBJECTIVETo explore the mechanism of occupational medicamentosa-like dermatitis (OMDT) induced by trichloroethylene (TCE) and some immunity indexes in workers occupationally exposed to TCE.

METHODSThe blood samples from 8 cases with medicamentosa-like dermatitis in 1st, 2nd, 3rd, 4th and 5th weeks after admitting to hospital were examined for liver function, immunoglobulin and some complement indexes. Thirty nine workers occupationally exposed to TCE were investigated for urinary TCE and some immuno-complement indexes. The TCE concentrations of air in workplaces were monitored.

RESULTSC3d-CIC and C3 of patients before admission were (92.86 ± 44.80) mg/L and 0.91 ± 0.19 mg/L, respectively. C3d-CIC and C3 of patients before discharge were (52.41 ± 17.75) mg/L and (1.14 ± 0.22) mg/L, respectively. There were significant differences between admission and discharge (P < 0.05). The average TCE concentration in 4 workplaces was (351.96 ± 36.72) mg/m(3), which was higher than the occupational exposure limits (OELs). The number of workers exposed to the TCE concentration-time weighted and TCA in urine over OELs were 28.21% and 56.41% of total subjects, respectively. The serum IgG and CIC levels of patients before discharge were (10.03 ± 1.21) mg/L and 103.50 ± 29.17 mU/L, which were significantly lower than those (17.21 ± 1.85) mg/L and (227.46 ± 111.67) mU/L of patients before admission (P < 0.01).

CONCLUSIONThe type II and III hypersensitivity may be associated with OMDT and the organ injure induced by TCE.

Adolescent ; Adult ; Complement System Proteins ; immunology ; Dermatitis, Occupational ; immunology ; Female ; Humans ; Male ; Occupational Exposure ; Trichloroethylene ; toxicity ; Young Adult

Objective To investigate the endurance or resistance of different bacteriophages to bubbling stress in different sampling solutions,to select the optimum sampling solution from three different ones and to select relatively stress-resistant bacteriophages from five different ones.Methods AGI-10(all glass impinger)was used as the representative for all the impingers that would bubble during operation to fulfill the bubbling experiment.Three different sampling solutions used,such as distilled water(DW),phosphatic buffer solution(PBS),and suspension medium(SM),were divided into two groups by adding olive oil(50 μl) or otherwise(0 μl).The impingers were operated 30 min at a flow rate of 7.0 L/min.The titers of bacteriophages and the volume of final sampling solutions were determined before the corrected survival probability was used to evaluate the stress resistance of several different bacteriophages.Results It was found that the survival probability of the same bacteriophage bubbling with different sampling solutions was different except for bacteriophage F2.The use of SM as the collection fluid was related to a high survival probability which remained unchanged between 50 μl and 0 μl olive oil.The corrected survival probability was 79%,77%,86%,50% and 71% for phage SM701,SM702,PhiX174,EcP1 and F2 respectively after 60 minutes of impingement at a flow rate of 7.0 L/min.Conclusion The endurance or resistance of different kinds of bacteriophages in the same sampling solution is different.SM might be an optimum sampling solution for phages.Bacteriophage SM701,SM702 and PhiX174 are more resistant to bubbling stress than EcP1 and F2.

Objective To investigate the improvement effects of sericin on the retinal oxidative stress and micro-inflammatory state in diabetic rats.Methods A diabetic rat model was established by using high-fat and high-sugar diet and intraperitoneal injection of streptozotocin.Then 24 diabetic rats were randomly divided into sericin treatment group and diabetic model group,with 12 rats for each group,and additional 12 normal rats with the same age were collected as a normal control group.Next,the rats in the sericin treatment group received sericin solution,while the other two groups was given the same amount of normal saline once a day for 35 days.After the agent intervention,the content of malondialdehyde (MDA) and glutathione (GSH) in the retina of all rats were detected by related kits.The expressions of nuclear factor erythroid 2-related factor 2 (Nrf2),heme oxygenase 1 (HO-1),nuclear factor-κB (NF-κB) and tumor necrosis factor-α (TNF-α) protein were detected by Western blot,and finally,the retinal morphology was examined by hematoxylin-eosin staining in the 3 groups.Results The content of MDA,NF-κB and TNF-α protein expression in the sericin treatment group was (4.145 ±0.282) mmol· gprot-1,0.232 ±0.027 and 0.761 ±0.058,respectively,which was significantly lower than that in the diabetic model group [(6.813 ± 0.446) mmol · gprot-1,0.334 ± 0.024 and 0.994 ± 0.084] (all P ＜ 0.05).The content of GSH,Nrf2 and HO-1 protein expression in rat retina in the sericin treatment group was (78.518 ± 4.317) mg · gprot 1,0.591 ± 0.054 and 0.954 ± 0.091,respectively,which was significantly higher than that in the diabetic model group [(59.890 ± 5.932) mg · gprot-1,0.351 ± 0.044 and 0.585 ± 0.054] (all P ＜ 0.05).The diabetic model group presented the disorder arrangement of the neurocyte in different levels in the retina,irregular and swollen inner limiting membrane,vacuoles in the ganglion cells,but in the sericin treatment group,the morphology of retinal layers was more regular and mildly disorderly arranged.The pathological damages in the retina were alleviated significantly.Conclusion Sericin can ameliorate oxidative stress and inflammation in diabetic retina,thereby delaying the development of diabetic retinopathy.

OBJECTIVETo determine whether ischemia-induced bone marrow-derived EPCs mobilization is impaired in diabetic mice and the association with vascular endothelial growth factor (VEGF) release post ischemia.

METHODSC57Bl/6 mice were injected with 40 mg x kg(-1) x d(-1) streptozotocin for 5 days to induce diabetes and mice with fasting glucose > 10 mmol/L were included to DM group, control mice were injected with placebo. Two months later, hindlimb ischemia was induced by left femoral artery dissection and ligation. The ischemia was visualized by tetrazolium dye staining and pre-mortem angiography. The percentage of c-Kit(+)/Sca-1(+)/flk-1(+) early EPCs in peripheral blood mononuclear cells (PBMCs) was detected by flow cytometric analysis on days 0 (pre-ligation), 1, 3, 5, 7 and 14 days post-ligation. The plasma VEGF level was measured with a standardized ELISA-Kit.

RESULTSCirculating EPCs number was significantly lower in diabetic mice than that in control mice (0.60% +/- 0.03% vs. 0.95% +/- 0.09%, P < 0.001) and the plasma VEGF was undetectable in all animals before ligation Similar EPCs kinetics following induction of hindlimb ischemia were shown in both groups. However, EPCs mobilization was significantly impaired in diabetic mice compared with control mice within 3 days post ischemia (day 1: 1.16% +/- 0.20% vs. 1.80% +/- 0.32%, P < 0.05; day 3: 1.38% +/- 0.34% vs. 2.37% +/- 0.52%, P < 0.05). In parallel, plasma VEGF increase post ischemia was significantly less in diabetic mice than that in control mice (day 1: 73.1 pg/ml +/- 18.6 pg/ml vs. 128.5 pg/ml +/- 44.2 pg/ml, P < 0.05).

CONCLUSIONOur data suggest that ischemia-induced bone marrow-derived EPCs mobilization is impaired in diabetic mice, which may be related to the insufficient release of plasma VEGF post ischemia.

Animals ; Bone Marrow Cells ; cytology ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; Endothelial Cells ; cytology ; metabolism ; Endothelium, Vascular ; physiopathology ; Hematopoietic Stem Cell Mobilization ; Ischemia ; metabolism ; physiopathology ; Mice ; Mice, Inbred C57BL ; Stem Cells ; cytology ; metabolism ; Vascular Endothelial Growth Factor A ; metabolism

High mobility group A2 protein (HMGA2), an architectural factor, is highly expressed in various cancer types including lung cancers. It is a candidate target for cancer therapy. RNAi is an effective gene silencing method with low cost and less time-consuming. It is possible to exploit this technology in therapy. Here, 5 siRNAs targeting Hmga2 gene (HMGA2 siRNA1-5) were designed and synthesized. MTT assay, colony formation assay, transwell assay and flow cytometry were used to evaluate the effects of these siRNAs on lung cancer cell lines (NCI-H446 and A549). Results from cell proliferation, clone formation, migration and apoptosis showed that HMGA2 siRNA1, 3, 5 could affect these aspects for both lung cancer cell lines. Among the five siRNAs, HMGA2 siRNA5 showed the greatest inhibition effects. The inhibition effects of HMGA2 siRNA5 are sequence specific and are not due to the induction of interferon response. Taken together, siRNAs targeting Hmga2 gene are potential candidates for lung cancer gene therapy.
Apoptosis ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Colony-Forming Units Assay ; Gene Silencing ; Genetic Therapy ; HMGA2 Protein ; genetics ; metabolism ; Humans ; Interferons ; metabolism ; Lung Neoplasms ; genetics ; metabolism ; pathology ; Point Mutation ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo explore the way to induce mesenchymal stem cells (MSCs) to differentiate into dopaminergic neurons in vitro.

METHODSMSCs were obtained from rat bone marrow, cultured and passaged. MSCs used in this experiment had multipotency, which was indirectly proved by being induced to differentiate into chondrocytes and adipocytes. MSCs were cultured in medium containing 0.5 mmol/L IBMX for 2 days. Then the medium was replaced with induction medium, which contained GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments. The surface markers of the differentiated neurons, such as NSE, nestin, MAP-2a, b and TH were detected by immunocytochemistry and Western blot after MSCs were cultured in induction medium for 7 days and 15 days.

RESULTSMSCs differentiated into neural progenitors and expressed nestin after MSCs were incubated with medium containing IBMX for 2 d. After the medium was replaced with induction medium containing many inducing agents, MSCs differentiated into neuron-like cells and dopaminergic neuron-like cells and expressed NSE, MAP-2a, b and TH. The percentage of NSE-positive cells, MAP-2a, b-positive cells and TH-positive cells was 30.032 +/- 2.489%, 41.580 +/- 5.101% and 34.958 +/- 5.534%, respectively after MSCs were induced in medium containing GDNF, IL-1beta, mesencephalic glial-cell-conditioned medium and flash-frozen mesencephalic membrane fragments for 15 days.

CONCLUSIONMSCs can differentiate into dopaminergic neuron-like cells and are a new cell source for the treatment of neurodegeneration diseases and have a great potential for wide application.

Adipocytes ; cytology ; Animals ; Blotting, Western ; Bone Marrow Cells ; Carboxylesterase ; analysis ; Cell Differentiation ; Cells, Cultured ; Chondrocytes ; cytology ; Culture Media, Conditioned ; Dopamine ; analysis ; Intermediate Filament Proteins ; analysis ; Mesencephalon ; cytology ; Mesenchymal Stromal Cells ; cytology ; Nerve Tissue Proteins ; analysis ; Nestin ; Neurons ; cytology ; metabolism ; Phosphoprotein Phosphatases ; analysis ; Rats ; Rats, Wistar

The liver injury induced by Polygonum multiflorum Thunb. (PM) was investigated based on idiosyncratic hepatotoxicity model co-treated with lipopolysaccharide (LPS) at a non-hepatotoxic dose. Sprague-Dawley (SD) rats were intragastrically administered with three doses (18.9, 37.8, 75.6 g crude drug per kg body weight) of 50% alcohol extracts of PM alone or co-treated with non-toxic dose of LPS (2.8 mg·kg(-1)) via tail vein injection. The plasma alanine aminotransferase (ALT) and aspartate aminotransferase (AST) activities were assayed and the isolated livers were evaluated for histopathological changes. The dose-toxicity relationships of single treatment of PM or co-treatment of LPS were investigated comparatively to elucidate the idiosyncratic hepatotoxicity of PM. The results showed that no significant alterations of plasma ALT and AST activities were observed in the groups of solo-administration of LPS (2.8 mg·kg(-1), i.v.) or different dosage (18.9, 37.8 and 75.6 g·kg(-1), i.g.) of PM, compared to normal control group (P > 0.05); while significant elevations were observed in the co-administration groups of PM and LPS. Treatment with LPS alone caused slight infiltration of inflammatory cells in portal area but no evident hepatocytes injury. Co-treatment with LPS and PM (75.6 g·kg(-1), i.g.) caused hepatocyte focal necrosis, loss of central vein intima and a large number of inflammatory cell infiltration in portal areas. When further reduce the dosage of PM, significant increases of plasma ALT and AST activities (P < 0.05) were still observed in co-administration groups of LPS and PM (1.08 or 2.16 g·kg(-1)), but not in LPS or PM solo-administration groups. Nevertheless, the co-treatment of low dosage of PM (0.54 g·kg(-1)) with LPS did not induce any alteration of plasma ALT and AST. In conclusion, intragastric administration with 75.6 g·kg(-1) of PM did not induce liver injury in normal rats model; while the 2 folds of clinical equivalent dose of PM (1.08 g·kg(-1)) could result in liver injury in the LPS-based idiosyncratic hepatotoxicity model, which could be used to evaluate the idiosyncratic hepatotoxicity of PM.
Alanine Transaminase ; blood ; Animals ; Aspartate Aminotransferases ; blood ; Chemical and Drug Induced Liver Injury ; pathology ; Hepatocytes ; pathology ; Lipopolysaccharides ; Polygonum ; toxicity ; Rats ; Rats, Sprague-Dawley