Objective To develop neural stem cells(NSCs) which can stably express exogenous brain-derived neurotrophic factor(BDNF) in vitro. Methods NSCs from the subependymal zone of embry-onic day 14.5(E14.5) rat brain were purified by limiting dilution assay and then infected with supernatant of recombinant retrovirus pLXSN-BDNF and retrovirus pLXSN. The original copy numbers of exogenous gene templates from three groups NSCs(pLXSN-BDNF viral infection group, pLXSN viral infection group, control group) were detected by fluorescent quantitative PCR(FQ-PCR). ELISA assay was used for determining the protein contents of BDNF of supernatant from three groups NSCs for six days continually after seeded in 24-well plates in the same cell density. Results NSCs were purified successfully by limiting dilution assay.The original copy numbers of exogenous BDNF gene templates from pLXSN-BDNF viral infection group by FQ-PCR were (19.57±0.65) × 10~3 copies/μl, higher than those of another two groups(P < 0.05). The protein contents of BDNF of supernatant from NSCs of pLXSN-BDNF viral infection group was highest among three groups and compared with another two groups had statistical significance (P <0.05) . Conclusion The purified NSCs can be transduced exogenous BDNF successfully with supematant of recombinant retrovir-us pLXSN-BDNF which provide experimental evidences and laying foundations for further research of retinal transplantation and quantization investigation of gene therapy for optic nerve injury.

AIM:To make a survey on people suffering trachoma in Gansu province, and to provide evidence for developing trachoma control and prevention therapy.
METHODS: We chose the zone on the basis of relative information. Provincial Office of Blindness Prevention carried out the survey in 3 counties including Tange Township of Wushan, Xiqu Township of Minqin and Hulinjia Township of Jishishan from October 14, 2013 to November 23, 2013. One hundred and fifty primary school students were selected, including 72 boys and 78 girls aging from 5a to 10a with the average age of 7. 5y. The targeted students received the fast trachoma assessment by the adoption of simplified trachoma classification system which was recommended by the World Health Organization.
RESULTS: No case of active trachoma, trachomatous trichiasis and corneal disease were examined among 150 students.
CONCLUSION: The rate of trachoma is low in Gansu province. But we still cannot get the conclusion that there is no epidemic of trachoma in Gansu. And we need to further expand the survey scope to correctly assess the trachoma case and to provide reliable evidence for trachoma prevention and control.

ObjectiveTo observe the effects of bone marrow stromal cells (BMSCs) on vessel endothelial cells proliferation and microvessel formation in vitro.MethodsBMSCs and brain vessel endothelial cells were separated from adult and divided into co-culture group of BMSCs and endothelial cells, medium group of BMSCs, comparison group. Endothelial cells proliferation and microvessel formation were observed. ResultsEndothelial cells were promoted to proliferate and formate the microvessel in medium group and co-culture group. And the effect was prominence in co-culture group.ConclusionBMSCs can promote the proliferation and microvessel formation of endothelial cells.

To develop a cell-penetrating chimeric apoptotic peptide AVPI-LMWP/DNA co-delivery system for cancer therapy, we prepared the AVPI-LMWP/pTRAIL self-assembled complexes containing a therapeutic combination of peptide drug AVPI and DNA drug TRAIL. The chimeric apoptotic peptide AVPI-LMWP was synthesized using the standard solid-phase synthesis. The cationic AVPI-LMWP could condense pTRAIL by electrostatic interaction. The physical-chemical properties of the AVPI-LMWP/pTRAIL complexes were characterized. The cellular uptake efficiency and the inhibitory activity of the AVPI-LMWP/pTRAIL complexes on tumor cell were also performed. The results showed that the AVPI-LMWP/pTRAIL complexes were successfully prepared by co-incubation. With the increase of mass ratio (AVPI-LMWP/DNA), the particle size was decreased and the zeta potential had few change. Agarose gel electrophoresis showed that AVPI-LMWP could fully bind and condense pTRAIL at a mass ratio above 15:1. Cellular uptake efficiency was improved along with the increased ratio of W(AVPI-LMWP)/WpTRAIL. The in vitro cytotoxicity experiments demonstrated that the AVPI-LMWP/pTRAIL (W:W = 20:1) complexes was significantly more effective than the pTRAIL, AVPI-LMWP alone or LMWP/pTRAIL complexes on inhibition of HeLa cell growth. Our studies indicated that the AVPI-LMWP/pTRAIL co-delivery system could deliver plasmid into HeLa cell and induce tumor cell apoptosis efficiently, which showed its potential in cancer therapy using combination of apoptoic peptide and gene drugs.
Antineoplastic Agents ; chemistry ; Cell-Penetrating Peptides ; chemistry ; DNA ; chemistry ; Drug Delivery Systems ; HeLa Cells ; Humans ; Neoplasms ; drug therapy ; Particle Size ; Plasmids

This study is to investigate the effect of baicalin (BL) against oxidative injury stress of SH-SY5Y cells induced by H2O2 and the possible mechanism. SH-SY5Y cells were pre-incubated with baicalin (25, 50, and 100 micromol x L(-1)) for 12 h prior to exposure to H2O2 (150 micromol x L(-1)) for 24 h. The viability of SH-SY5Y cells was measured by MTT assay. The contents of LDH and NO were determined. The percentage of apoptotic cells was assessed by flow cytometry (FCM). The content of Caspase-3 was tested by immunofluorescence histochemical method. BL at 50 and 100 micromol x L(-1) separately increased the cell viability and up-regulated SIRT1, reduced the contents of LDH, NO, Caspase-3 and the apoptotic percentage of SH-SY5Y cells. This study results suggest that baicalin could inhibit the H2O2-induced neuronal apoptosis. The further mechanism studies show that baicalin inhibit apoptosis via reducing Caspase-3 expression and up-regulating SIRT1.
Antioxidants ; isolation & purification ; pharmacology ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Cell Line, Tumor ; Cell Survival ; drug effects ; Flavonoids ; isolation & purification ; pharmacology ; Humans ; Hydrogen Peroxide ; toxicity ; L-Lactate Dehydrogenase ; metabolism ; Neuroblastoma ; metabolism ; pathology ; Nitric Oxide ; metabolism ; Oxidative Stress ; drug effects ; Plants, Medicinal ; chemistry ; Scutellaria ; chemistry ; Sirtuin 1 ; metabolism ; Up-Regulation

Atropa belladonna is a medicinal plant and main commercial source of tropane alkaloids (TAs) including scopolamine and hyoscyamine, which are anticholine drugs widely used clinically. Based on the high throughput transcriptome sequencing results, the digital expression patterns of UniGenes representing 9 structural genes (ODC, ADC, AIH, CPA, SPDS, PMT, CYP80F1, H6H, TRII) involved in TAs biosynthesis were constructed, and simultaneously expression analysis of 4 released genes in NCBI (PMT, CYP80F1, H6H, TRII) for verification was performed using qPCR, as well as the TAs contents detection in 8 different tissues. Digital expression patterns results suggested that the 4 genes including ODC, ADC, AIH and CPA involved in the upstream pathway of TAs, and the 2 branch pathway genes including SPDS and TRII were found to be expressed in all the detected tissues with high expression level in secondary root. While the 3 TAs-pathway-specific genes including PMT, CYP80F1, H6H were only expressed in secondary roots and primary roots, mainly in secondary roots. The qPCR detection results of PMT, CYP80F1 and H6H were consistent with the digital expression patterns, but their expression levels in primary root were too low to be detected. The highest content of hyoscyamine was found in tender stems (3.364 mg x g(-1)), followed by tender leaves (1.526 mg x g(-1)), roots (1.598 mg x g(-1)), young fruits (1.271 mg x g(-1)) and fruit sepals (1.413 mg x g(-1)). The highest content of scopolamine was detected in fruit sepals (1.003 mg x g(-1)), then followed by tender stems (0.600 mg x g(-1)) and tender leaves (0.601 mg x g(-1)). Both old stems and old leaves had the lowest content of hyoscyamine and scopolamine. The gene expression profile and TAs accumulation indicated that TAs in Atropa belladonna were mainly biosynthesized in secondary root, and then transported and deposited in tender aerial parts. Screening Atropa belladonna secondary root transcriptome database will facilitate unveiling the unknown enzymatic reactions and the mechanisms of transcriptional control.
Alkaloids ; biosynthesis ; genetics ; metabolism ; Atropa belladonna ; genetics ; metabolism ; Gene Expression Regulation, Plant ; genetics ; Hyoscyamine ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism ; Scopolamine Hydrobromide ; metabolism ; Tropanes ; metabolism

Animals ; Cell Differentiation ; Female ; Hematopoietic Stem Cells ; cytology ; Hepatocytes ; cytology ; Male ; Mice ; Mice, Inbred BALB C

OBJECTIVETo investigate the protective effect and mechanism of curcumin derivatives B06 on myocardium from type 2 diabetic rats.

METHODSThirty-five male SD rats were randomly divided into 5 groups, normal control group (NC group), high fat group (HF group), high fat treatment group (FT group), diabetes mellitus group (DM group) and diabetes treatment group (DT group) (n = 7). The late four groups were fed with high fat food, after four weeks of high fat feeding, the rats from DM group and DT group were injected with low dosage of streptozocin intraperitoneally to induce diabetes mellitus, FT group and DT group were gavaged with curcumin derivatives B06 at the dosage of 0.2 mg/kg x d. The blood glucose and lipid were detected biochemically, blood insulin was assayed by ELISA and the insulin resistance index was calculated, the morphology of myocardium was observed by light and transmission electron microscopy, the protein expression of AMP-activated protein kinase alpha (AMPKalpha) and phosphorylated AMP-activated protein kinase alpha (p-AMPKalpha) in myocardium were tested by Western blot.

RESULTSThe level of blood glucose, lipid, insulin and the insulin resistance index were increased in HF group and DM group, but they were decreased after the treatment with B06. The expression of AMPKalpha and p-AMPKalpha were decreased, but they became increased after the treatment of B06. There were increased collagen fibers in interstitium and expansion of mitochondria in cytoplasm of myocardium from DM group, but they were ameliorated in B06 treatment group.

CONCLUSIONIt is suggested that B06 may relieve the damage of myocardium from type 2 diabetic rats and the increased expression of AMPKalpha and p-AMPKalpha may be involved in it.

AMP-Activated Protein Kinases ; metabolism ; Animals ; Blood Glucose ; Curcumin ; pharmacology ; Diabetes Mellitus, Experimental ; physiopathology ; Heart ; drug effects ; Insulin Resistance ; Male ; Myocardium ; pathology ; Rats ; Rats, Sprague-Dawley ; Streptozocin

OBJECTIVETo explore the possibility of Schwann cells transplantation to promote the repair of injured brain stem reticular structure in rats.

METHODSSchwann cells originated from sciatic nerves of 1 to 2-day-old rats were expanded and labelled by BrdU in vitro, transplanted into rat brain stem reticular structure that was pre-injured by electric needle stimulus. Immunohistochemistry and myelin-staining were used to investigate the expression of BrdU, GAP-43 and new myelination respectively.

RESULTSBrdU positive cells could be identified for up to 8 months and their number increased by about 23%, which mainly migrated toward injured ipsilateral cortex. The GAP-43 expression reached its peak in 1 month after transplantation and was significantly higher than that in the control group. New myelination could be seen in destructed brain stem areas.

CONCLUSIONThe transplantation of Schwann cells can promote the restoration of injured brain stem reticular structure.

Animals ; Antimetabolites ; Brain Injuries ; therapy ; veterinary ; Brain Stem ; injuries ; Bromodeoxyuridine ; Cell Transplantation ; methods ; veterinary ; Electrophysiology ; Rats ; Rats, Wistar ; Schwann Cells ; transplantation

BACKGROUNDGlaucoma is one of the leading causes of blindness in the world. Primary open-angle glaucoma (POAG) and primary congenital glaucoma (PCG) are subtypes of glaucoma. Myocillin is the first gene identified to be involved in POAG. Recently, myocillin mutation has been found in PCG. In this context, we reported a special glaucoma pedigree, which was composed of both PCG and POAG patients, and analyzed the mutation of myocillin in this pedigree.

METHODSThe family was composed of the parents, a son and a daughter. All members of the family underwent the complete ophthalmologic examinations. All coding exons 1 - 3 and flanking introns of myocilin gene were screened for sequence alterations by polymerase chain reaction and direct DNA sequencing.

RESULTSThe son was the proband, who was diagnosed as PCG in both eyes. The father was diagnosed as POAG in the right eye, the left eye was still normal. Both the sister and the mother of the proband had normal intraocular pressure without glaucomatous optic disc changes. The mutations in intron 2 of myocilin gene were detected in the family. While the proband and the father were homozygous, the mother and the sister were heterozygous for the mutation.

CONCLUSIONSHomozygous mutation in intron 2 of myocilin gene is involved in both POAG and PCG. It is suggested that the pathogenesis might be overlapping in POAG and PCG.

Cytoskeletal Proteins ; genetics ; Eye Proteins ; genetics ; Female ; Glaucoma ; congenital ; genetics ; Glaucoma, Open-Angle ; genetics ; Glycoproteins ; genetics ; Humans ; Introns ; Male ; Mutation ; Pedigree