OBJECTIVEEffects of ginsenoside Rb1, Rg1 and Re on neurotrophic factor signal transduction pathway using liposome-mediated transfection of eukaryotic cells approach.

METHODThe injury model was established by treating SH-SY5Y cells with 0.6 mmol x L(-1) of corticosterone (CORT) by 24 h. SH-SY5Y cell were pretreated with CORT for 30 min followed by co-treated with 120,60 and 20 micromol x L(-1) of Rb1, 120, 80 and 40 micromol x L(-1) of Rg1 and 120, 80 and 40 micromol x L(-1) of Re for 24 h. Cells viability was determined by Cell Counting Kit (CCK) assay. CREB expressing Luciferase reporter gene was constructed and transfected with plasmid containing hRaf, hcAMP, hAkt, hCaMK gene into human embryonic kidney (HEK293) cells using liposornal transfection reagent lipofection 2000. The expression of CREB before and after it addion of Rb1, Rg1 and Re was examined by Luc assay system and Western blotting.

RESULTCompared with normal control group, CORT significantly decreased the viability of SH-SY5Y cells to 67.21% (P < 0.01). CCK results show that Rb1 (60 micromol x L(-1)), Rg1 (80 micromol x L(-1)) and Re (80 micromol x L(-1)) on SH-SY5Y cells have significant protective effect (P < 0.01). Lucassay and Western blotting results show that the gene and protein levels of CREB increased significantly through the pathway of Raf and Akt with Rb1 and Rg1 (P < 0.01), Re can increase significantly the gene and protein levels of CREB through the pathway of Raf and CaMK II.

CONCLUSIONRb1, Rg1 and Re protects SH-SY5Y cells from CORT-induced damage and the neuroprotective mechanism may be associated with the Raf-CREB, Akt-CREB and CaMK II -CREB pathways.

Calcium-Calmodulin-Dependent Protein Kinase Type 2 ; genetics ; metabolism ; Cell Line ; Cell Survival ; drug effects ; Cyclic AMP Response Element-Binding Protein ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Genes, Reporter ; Ginsenosides ; pharmacology ; Humans ; Panax ; chemistry ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Signal Transduction ; drug effects ; raf Kinases ; genetics ; metabolism

Objective To study the effects of miR?21 on the proliferation,apoptosis and MMP2 expression of HeLa cells. Methods The miR?21 or the miRNA scramble control was transfected into Hela cells. The cell proliferation was detected by the Celltiter?GloTM assay 72 h after the transfection,and the apoptosis was evaluated by the Caspase3/7 Glo? Kit. The MMP2 RNA expression was quantified by quantitative real time PCR. Results The proliferation of HeLa cells transfected with miR?21 was significantly increased compared to that of the cells transfected with miRNA scramble control. The caspase 3/7 activity in HeLa cells transfected with miR?21 was downregulated compared to that of the cells transfect?ed with miRNA negative control. The MMP2 RNA expression in HeLa cells transfected with miR?21 was increased significantly compared to the cells transfected with miRNA negative control. Conclusion miR?21 can significantly promote the proliferation of HeLa cells. In addition,it ex?hibits the anti?apoptosis effect in HeLa cells. The transfection of miR?21 significantly increased the MMP2 RNA expression,which suggests that miR?21 may promote the tumor invasion and might be a therapeutic intervention target.

The insulin complement with gene therapy has been used as an experimental treatment for insulin dependent diabetes (IDDM). In the present study, we constructed naked plasmid DNA vector encoding recombinant human preproinsulin gene (pCMV-IN), and injected the plasmids (100 microg/mouse) intramuscularly combined with electroporation, to achieve the in vivo transfer of insulin gene in streptozotocin (STZ)-induced diabetic C57 mice. The expression of vector-derived insulin mRNA was detected with RT-PCR in transfected local skeletal muscles. The plasma insulin was elevated significantly in pCMV-IN injected diabetic C57 mice, which was complemented to the level similar to the intact normal control. The protein expression lasted for at least 35 days after the plasmid injection. Gene therapy with pCMV-IN plasmids considerably decreased the blood glucose level in STZ-induced diabetic mice from d 7 to d 35 by about 6 mmol/L. The gene therapy also reduced the mortality of severe diabetic mice significantly from 100% to 37% at the 6th week. Our results indicate that the direct intramuscular injection of naked plasmids encoding human preproinsulin gene achieves the effective expression of insulin. The restoration of insulin decreases blood glucose and increases the survival in severe diabetic mice. The gene therapy might be provided as a practical therapeutic approach to IDDM.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; blood ; therapy ; Diabetes Mellitus, Type 1 ; blood ; therapy ; Electroporation ; Gene Transfer Techniques ; Genetic Therapy ; methods ; Genetic Vectors ; Injections, Intramuscular ; Insulin ; Male ; Mice ; Mice, Inbred C57BL ; Plasmids ; genetics ; Proinsulin ; genetics ; therapeutic use ; Protein Precursors ; genetics ; therapeutic use

BACKGROUND: Patients with osteoporosis are prone to develop fractures, and moreover some patients are first diagnosed with osteoporosis because of a fragility fracture. Therefore, it is critical to understand the correlation between osteoporotic medications and fracture healing. OBJECTIVE: To summarize the effect of anti-osteoporosis medications on osteoporotic fracture healing in order to promote its clinical application. METHODS: A computer-based online search of PubMed, CNKI, VIP and WanFang databases between January 2012 and July 2016 was performed to retrieve the related articles with the keywords of "osteoporotic fracture, healing, bone nutrition supplements, anti-resorptive agents, anabolic agents, dual effect agents, new targeted agents" in English and Chinese, respectively. Literature concerning the effect of anti-osteoporosis medications on fracture healing was selected, and the articles published lately in authoritative journals were preferred. RESULTS AND CONCLUSION: Most of anti-osteoporotic medications have no harmful influence on fracture healing, including bone nutrition supplements (calcium and vitamin D), anti-resorptive agents (bisphosphonate, denosumab, estrogen and selective estrogen receptor modulators, statins and calcitonin), anabolic agents (parathyroid hormone), and dual effect agents (strontium ranelate). Calcium and vitamin D are the basic drugs; anti-resorptive agents exert overt anti-osteoporotic effect; and the new targeted agents like cathepsin K inhibitor and sclerostin monoclonal antibody provide more choices for the therapy of osteoporotic fracture. Partial anti-osteoporotic agents inhibit the viability of osteoclasts, so their early application may be against fracture healing. The optimal time of anti-osteoporotic medications and the effect on acute and non-acute osteoporotic fractures need to be further explored.

OBJECTIVETo explore the impact of different warm ischemia time on structure and function of the non-heart-beating donor lung and to find out the feasibility of non-heart-beating donor in rat lung transplantation.

METHODSSixty Sprague-Dawley rats were randomly divided into 3 groups: heart-beating donor (HBD) group, non-heart-beating donor (NHBD) with 30 minutes of warm ischemia time (WIT) group and NHBD with 60 minutes of WIT group. Each group has 10 pairs (the donors and the recipients). The donor lungs of group HBD were flushed with low potassium dextran (LPD) solution at 4 degrees C after asystolia while the lungs of group NHBD-30 and group NHBD-60 remained ventilated at the room temperature for 30 and 60 minutes after asystolia and then were flushed with LPD solution. All the donor lungs remained inflated when they were stored in LPD solution at 4 degrees C for 4 hours. The recipient rat underwent left thoracotomy, and then orthotopic left lung transplantation. Followed by a right thoracotomy, the right pulmonary hilum were ligated with one-hour reperfusion and ventilation.

RESULTSAll the recipients in group HBD and group NHBD-30 survived the observation period of one hour with excellent gas exchange, whereas 4 of recipients in group NHBD-60 survived for 10 minutes after the ligation of right pulmonary hilum and 3 for 20 minutes. The pulmonary compliance, ultrastructure, energy metabolite and other markers revealed no significant differences between group HBD and group NHBD-30 (P > 0.05). But the differences between group NHBD-60 and other two groups were significant (P < 0.05).

CONCLUSIONSThe adoption of non-heart-beating donor could be a safe and effective method to expand the lung donor pool. The NHBD lung with 30 minutes of WIT may be suitable for lung transplantation in rat.

Animals ; Heart ; physiopathology ; Ischemia ; physiopathology ; Lung ; physiopathology ; ultrastructure ; Lung Transplantation ; Male ; Microscopy, Electron ; Models, Animal ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Time Factors ; Tissue Donors

OBJECTIVETo observe the effects of combined treatment with sansanmycin and macrolides on Pseudomonas aeruginosa and formation of biofilm.

METHODSMicro-dilution method was used to determine the minimal inhibitory concentrations (MICs) of sansanmycin, gentamycin, carbenicillin, polymyxin B, roxithromycin, piperacillin, and tazobactam. PA1 and PA27853 biofilms were observed under optical microscope after staining and under SEM after treatment with sansanmycin at different dosages and combined treatment with sansanmycin and roxithromycin. Viable bacteria in PA1 and PA27853 biofilms were counted after treatment with sansanmycin at different dosages or combined treatment with sansanmycin and roxithromycin.

RESULTSThe MIC of sansanmycin was lower than that of gentamycin and polymyxin B, but was higher than that of carbenicillin. Roxithromycin enhanced the penetration of sansanmycin to PA1 and PA27853 strains through biofilms. PA1 and PA27853 biofilms were gradually cleared with the increased dosages of sansanmycin or with the combined sansanmycin and roxithromycin.

CONCLUSIONSub-MIC levels of roxithromycin and sansanmycin substantially inhibit the generation of biofilms and proliferation of bacteria. Therefore, combined antibiotics can be used in treatment of intractable bacterial infection.

Animals ; Anti-Bacterial Agents ; administration & dosage ; pharmacology ; Bacterial Adhesion ; drug effects ; Biofilms ; growth & development ; Cercopithecus aethiops ; Drug Therapy, Combination ; Macrolides ; administration & dosage ; pharmacology ; Microbial Sensitivity Tests ; Oligopeptides ; administration & dosage ; pharmacology ; Pseudomonas aeruginosa ; drug effects ; physiology ; ultrastructure ; Uridine ; administration & dosage ; analogs & derivatives ; pharmacology ; Vero Cells

Type B Niemann-Pick disease is an autosomal recessive sphingolipidosis due to mutations in the sphingomyelin phosphodiesterase 1 gene (SMPD1). Here we present molecular findings for two sibling patients. One mutation V36A due to c.107T>C in exon 1 is a single nucleotide polymorphism and the other N522S due to c.1565 A>G in exon 6 is a novel missense mutation. This non-fatal missense mutation leads to –20% residual lysosomal acid sphingomyelinase activity in vitro and only results in hepatosplenomegaly without neurologic involvement.
Female ; Humans ; Middle Aged ; Mutation, Missense ; Niemann-Pick Disease, Type B ; genetics ; Polymorphism, Single Nucleotide ; Siblings ; Sphingomyelin Phosphodiesterase ; genetics

Objective To observe the expression levels of matrix metalloproteinase-2 and 9 (MMP-2, MMP-9) in wound tissue and wound effusion in patients with chronic pressure ulcers treated with an alternating pres- sure relief mattress,and to correlate these observations with wound healing.Methods A total of 24 patients with chronic pressure ulcers were recruited and divided into two groups :one treated with an alternating pressure relief mat- tress and the other without,in addition to conventional treatment for pressure ulcers.The expression of MMP-2 and MMP-9 in the wound tissue and its exudate were amplified with a reverse transeriptase-polymerase chain reaction (RT-PCR) and analyzed using quantitative gelatin zymography.Wound healing was also observed and assessed using the pressure ulcer scale for healing (PUSH).Results The wounds were observed to heal well in both groups,as indicated by a significant decrease in the PUSH scores.Statistical analysis revealed a significant difference between the two groups with regard to the PUSH score on the 21st day after initiation of the study.With the healing of the pressure ulcers,the expression both MMP-2 and MMP-9 decreased.On the seventh and the 21st days,the expression of MMP-2 and MMP-9 in patients treated with an alternating pressure relief mattress was significantly less than in those without the mattress.Conclusion MMP-2 and MMP-9 can be biomarkers for the healing of pressure ulcers. An alternating pressure relief mattress is helpful for treating pressure ulcers.

This study was aimed to detect the gene expression profile changes between human acute leukemia cell line HL-60 and VCR-resistance HL-60, and to investigate the underlying mechanisms of MDR by using genechip technology. In experiments, mRNA were harvested using TrizoL reagent from these two cell lines, through RT-PCR, the biotinylated nucleotide were incorporated into the cRNA during the in vitro transcription reaction. The high quality RNA was hybridized to the gene expression array--human genome U133A developed by Affymetrix. It was scanned by G2500A GeneArray Scanner and the acquired image was analysed by a series of softwares. The results showed that 5,507 genes were differentially expressed between human acute leukemia cell line HL-60 and VCR-resistant HL-60. Compared with HL-60, 3,100 genes were up-regulated and 2,407 genes were down-regulated in VCR-resistant cell line. These genes were involved in different cell activities such as growth regulation and signal transduction. Among the genes with remarkable differential expression between the two cell lines, 435 were up-regulated and 605 were down-regulated. It is concluded that many different kinds of genes are involved in the mechanism of MDR and there is an intricate molecular network that controls the sensitivity of leukemia cells to the chemotherapeutic agents. Genechip is an efficient tool for parallel gene expression analysis.
Drug Resistance, Neoplasm ; genetics ; Gene Expression Profiling ; Gene Expression Regulation, Neoplastic ; Genome, Human ; HL-60 Cells ; Humans ; Oligonucleotide Array Sequence Analysis ; Vincristine ; pharmacology

OBJECTIVETo evaluate the predictive value of portal vein flow rate preoperative for portal vein thrombosis (PVT) after periesophagastric devascularization in hepatitis B cirrhosis-related portal hypertension.

METHODSFrom January 2007 to July 2008, 45 patients with portal hypertension caused by hepatitis B cirrhosis were performed splenectomy with peri-esophagogastric devascularization in the same medical group in West China Hospital of Sichuan University. The portal vein flow rate and the diameter of portal vein were measured with doppler sonography respectively before and after the operation. At the same time, the level of PT and PLT were detected. The weight of spleens were measured after operation.

RESULTSThirteen cases suffered from PVT postoperatively. Portal vein flow rate was significantly lower in patients with PVT postoperation than that in patients without PVT (P < 0.01). In patients with PVT (n = 13) postoperation, the preoperative portal vein flow rate was (19.5 +/- 5.3) cm/s. Among the 13 cases, there were 12 cases whose flow rate were lower than 25 cm/s, and 1 case whose flow rate was 32. 3 cm/s; In patients without PVT (n = 32), the preoperative portal vein flow rate was (9.6 +/- 8.0) cm/s. In patients with lower rate (n = 17), the incidence rate of PVT was 70.6%; in patients with higher rate (n = 28), the incidence rate of PVT was 3.6%. The incidence rate of PVT in patients with lower rate was significantly lower than patients with higher rate (P < 0.01). The diameter of portal vein in patients with PVT was significantly wider than patients without PVT. The diameter of portal vein was negative correlative with the portal vein flow rate. The value 25 cm/s was of diagnostic efficiency, the sensitivity was 92.3%, and specificity was 70.6%.

CONCLUSIONSThe portal vein flow rate preoperative can be used as an early predictor of portal vein thrombosis after periesophagastric devascularization in hepatitis B cirrhosis-related portal hypertension to give a guide to clinical work.

Adult ; Aged ; Blood Flow Velocity ; Female ; Humans ; Hypertension, Portal ; etiology ; physiopathology ; surgery ; Liver Cirrhosis ; complications ; Male ; Middle Aged ; Portal Vein ; diagnostic imaging ; physiopathology ; Postoperative Complications ; diagnosis ; etiology ; Preoperative Care ; Risk Factors ; Splenectomy ; Ultrasonography ; Venous Thrombosis ; diagnosis ; etiology