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

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

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

Objective To study the effect of different nursing methods in double plasma exchange treatment on multiple myeloma. Methods 30 patients with multiple myeloma were randomly assigned to the experimental group (EG) and the control group (CG), 15 patients in each group. In the CG, the canal was washed by the priming fluids according to the machine requirements; patients were given the LMWHC at the beginning of the treatment. During the treatment, the canal was washed discontinuously by normal saline (NS).An improved priming method was adopted in the EG: the canal was washed with NS included heparin, patients were given the LMWHC. After 20 - 30 mins, the treatment started. Squeeze the artery pot discontinuously during the treatment; compare the clotting situation and treatment parameters of two groups. Results Level Ⅱ - Ⅲ clotting incidence rate of EG was 23.3% and of CG was75.0%. The difference between two groups was statistically significant (X2 =37.95,P ＜0. 05); TMP of CG were higher than those of EG[(17.32 ±5. 22)mm Hg vs (4.73 ± 3.27) mm Hg, t = 9. 76, P ＜ 0. 05]; total NS amount used, replacement plasma, waste plasma and treatment time of CG were lower than EG (P ＜ 0. 05). Conclusions Compared with CG, the priming methods, anticoagulation solutions and washing methods of EG have better control on the occurrence of clotting and require less amount of washing NS to reduce the burden on kidney without influencing the effect of the treatment.

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

Objective To study the inhibitory effect of Tilianin on non -small cell lung cancer cell lines (A549) and the associated mechanisms.Methods The control group and 10,20,40,80 and 160 μmol· L-1 experimental group were treated with 0,10,20,40,80 and 160 μ mol· L-1Tilianin for 24 h,cell proliferation and toxicity test kit (CCK8) was used to observe the proliferation of A549 cells.The apoptosis of A549 cells in each group was detected by flow cytometry.In the hypoxia model group,A549 cells were cultured in hypoxic incubator for 4 h.In the hypoxia experimental group of 10,20 and 40 μmol · L-1Tilianin,A549 cells were pretreated with 10,20 and 40 μmol · L-1Tilianin for 4 h,and then cultured in hypoxic incubator for 4 h.Real-time fluorescent quantitative polymerase chain reaction (qRT-PCR) was used to detect the gene expressions of vascular endothelial growth factor(VEGF) and HIF-1α (HIF-1α).The protein expressions of VEGF-A,HIF-1α and p-Akt were detected by Western blot.Results Compared with control group,Tilianin significantly inhibited the proliferation of A549 in a dose -dependent manner.The inhibitory rates of proliferation of 10,20,40,80 and 160 μmol · L-1 experimental groups were (5.83 ±1.67)％,(6.77 ±0.87)％,(12.26 ±0.23)％,(22.97 ±0.50)％ and (46.24±1.44)％,the difference was statistically significant (P ＜0.05).The apoptosis rates of 10,20,40,80 and 160 μmol · L-1Tilianin were (6.80 ±0.62)％,(14.70 ±1.36)％,(24.76 ±4.37)％,(39.26±6.42)％ and (62.31 ±1.79)％,respectively,compared with the control group,the differences were statistically significant (P ＜ 0.05).In the hypoxia model group,the gene expression of HIF-1α was 4.30 ± 0.26,VEGF expression was 6.02 ± 0.53,compared with the control group,the differences was statistically significant (P ＜0.05).In 10,20,40 μmol · L-1Tilianin hypoxia experiment group,the gene expression of VEGF were 4.73 ±0.20,2.31 ±0.09 and 1.47 ±0.16,respectively.The expression of HIF-1α were 3.01 ± 0.11,1.81 ± 0.13 and 1.03 ± 0.16.Compared with hypoxia model group,the differences was statistically significant (P ＜ 0.05).The expression of p-AKT protein in hypoxic model group was (106.47 ± 2.08)％,the expression of HIF-1αwas (204.31 ± 8.35)％,the expression of VEGF-A was 212.30 ±4.80.The expression of p-AKT protein in 10,20,40 μmol · L-1 hypoxic experimental group were (87.51 ±2.72)％,(75.18 ± 1.67)％ and (32.40 ± 0.86)％,the protein expression levels of HIF-1α were (182.54 ± 6.42),(90.95 ± 2.76)％,(15.03 ± 4.21)％,the protein expression of VEGF-A were (156.38 ± 5.12) ％,(79.62 ± 2.84) ％ and (13.72 ± 4.64) ％,with significant difference (P ＜ 0.05).Conclusion Tilianin has the effect of inhibiting the proliferation and inducing apoptosis of A549 in vitro through AKT / HIF-1α signaling pathway.