The producing condition and properties of chitinase secreted by Nomuraea rileyi strain CQ031021 were studied.The optimal conditions for the strain to produce chitinase are 6 days of 28℃ with the initial pH 6.0,and the liquid medium containing 2.0% glucose as its carbon source and 0.6% peptone plus 0.6% beef extract as nitrogen source after inoculating dosage 2mL suspension of conidia(1?107/mL).The optimal temperature and pH for enzyme activity is 50℃ and 6.0,respectively,while the activity can be enhanced by Tween-80 and inhibited by SDS.The enzyme activity is stable under 40℃ and in pH range of 5.5～6.5.

The endophytic microorganisms found widely in many kinds of plants mediate various effects to theirs hosts. In this study, seven different dominant endophytes (SDE01 to 07) isolated from a Hy-peraccumulator-Solanum nigrum L. were resistant to Cd2+, and the strain SDE06 survived even in the medium containing 80 mg/L of Cd2+. Bacteria strain SDE06 was identified as Bacillus sp.. The removal of Cd2+ of SDE06 in different conditions were studied. Under the optimal conditions, the incubating time was 36 h, the solution pH 6.0, the temperature was 37?C and the Cd2+ concentration of medium was 20 mg/L, the highest removal rate was up to 80.2% at this condition.

The rhizome of Panax japonicus var. major have been used as the natural medicinal agent by Chinese traditional doctors for more than thousand years. Most of the therapeutic effects of P. japonicus var. major had been reported due to the presence of tetracyclic or pentacyclic triterpene saponins. In this study, Illumina pair-end RNA-sequencing and de novo splicing were done in order to understand the pathway of triterpenoid saponins in this species. The valid reads data of 15. 6 Gb were obtained. The 62 240 unigenes were finally obtained by de novo splicing. After annotation, we discovered 19 unigenes involved in ginsenoside backbone biosynthesis. Additionally, 69 unigenes and 18 unigenes were predicted to have potential function of cytochrome P450 and UDP-glycosyltransferase based on the annotation results, which may encode enzymes responsible for ginsenoside backbone modification. This study provides global expressed datas for P. japonicus var. major, which will contribute significantly to further genome-wide research and analysis for this species.
Gene Expression Profiling ; Panax ; genetics ; Saponins ; biosynthesis ; Sequence Analysis, RNA

OBJECTIVETo explore the changes in the expressions of glial fibrillary acidic protein (GFAP) and growth- associated protein-43 (GAP-43) in retinal ganglial cells after neural transplantation.

METHODSThirty-nine rats were randomized into normal control group, nerve amputation group and nerve amputation with peripheral nerve transplantation group. Immunohistochemistry was used to detect the changes in the expressions of GFAP and GAP-43 at different time points after the operations, and real-time PCR was employed to detect the mRNA expressions of 13 genes in the retinal ganglial cells of the rats.

RESULTSImmunohistochemistry showed obviously increased GFAP expressions in the retina following the nerve amputation. GFAP expression was down-regulated while GAP-43 expression upregulated in the retinal ganglial cells after peripheral nerve transplantation. Real-time PCR results showed that 5 days after the operations, retinal GFAP and GAP-43 expressions increased significantly in the nerve amputation group and peripheral nerve transplantation groups as compared with those in the control group, but GAP-43 expression decreased significantly in the former two groups afterwards.

CONCLUSIONThe regenerated retina may adjust the production of GFAP. The retinal ganglial cells express GAP-43 during retinal regeneration. Up-regulation of the expression of GAP-43 provides the evidence for nerve regeneration following the nerve transplantation.

Animals ; Axons ; Female ; GAP-43 Protein ; genetics ; metabolism ; Glial Fibrillary Acidic Protein ; genetics ; metabolism ; Nerve Regeneration ; genetics ; Optic Nerve ; transplantation ; Optic Nerve Injuries ; metabolism ; Random Allocation ; Rats ; Retinal Ganglion Cells ; metabolism

OBJECTIVETo explore the effect of kallikrein-binding protein (KPB) in protecting retinal ganglion cells (RGCs) and promoting axonal regeneration following optical nerve injury in rats.

METHODSCrush injury of the optic nerve at 0.5-1.0 mm from the eyeball was induced in rats, which received subsequent KBP injection into the vitreous cavity (experimental group) and PBS injection (control group). At 7, 14 and 21 days after the injury, the rats were sacrificed and frozen sections of the eyeball were prepared to observe the structure and thickness of the retina and count the number of survival RGCs with HE staining. The optic nerves were collected for Western blotting to assess the effect of KBP on the RGCs and axonal regeneration.

RESULTSRGC counts and retinal thickness showed significant differences between the two groups. Western blotting also demonstrated a significant difference in the expression of the nerve regeneration marker protein GAP-43 between the two groups.

CONCLUSIONKBP offers protection on RGCs and promotes regeneration of the optic nerve axons after optic nerve injury in rats.

Animals ; Axons ; physiology ; Female ; GAP-43 Protein ; metabolism ; Nerve Regeneration ; drug effects ; physiology ; Neuroprotective Agents ; pharmacology ; Optic Nerve Injuries ; drug therapy ; Rats ; Rats, Sprague-Dawley ; Retinal Ganglion Cells ; drug effects ; physiology ; Serpins ; pharmacology

BACKGROUNDThe blood vessels of a transplanted organ are the interface between donor and recipient. The endothelium in the blood vessels is thought to be the major target for graft rejection. Endothelial cells of a transplanted organ can be of recipient origin after transplantation. In this study, we tested whether endothelial chimerism correlated with the graft rejection and cold ischemia.

METHODSWe studied the biopsy samples from 34 renal transplants of female recipients who received the kidney from a male donor for the presence of endothelial cells of recipient origin. We examined the tissue sections of renal biopsy samples by fluorescence in situ hybridization (FISH) for the presence of endothelial cells containing two X chromosomes using a biotinylated Y chromosome probe and digoxigenin labelled X chromosome probe, and then analyzed the relationship between the endothelial cell chimerism and the rejection and cold ischemia.

RESULTSEndothelial chimerism was common and irrespective of rejections (P > 0.05). The cold ischemic time of chimerism group was longer than no chimerism group ((14.83 +/- 4.03) hours vs (11.27 +/- 3.87) hours, P < 0.05).

CONCLUSIONSThere is no correlation between the percentage of recipient endothelial cells in vascular endothelial cells and the type of graft rejection. The endothelium damaged by ischemic injury might be repaired by the endothelial cells from the recipient.

Animals ; Biopsy ; Endothelial Cells ; pathology ; Female ; Graft Rejection ; Humans ; In Situ Hybridization, Fluorescence ; Kidney ; pathology ; Kidney Transplantation ; Male ; Mice ; Time Factors ; Transplantation Chimera ; Transplantation, Homologous

OBJECTIVETo breed estrogen receptor beta (ERbeta) gene knock-out female mice for studying postmenopausal osteoporotic fracture.

METHODSThree pairs of ERbeta gene knock-out mice were bred for 3 months, and 14 2-month-old female wild-type C57BL/6J mice with the same genetic background were paired at the ratio of 2:1 and mated with the male ERbeta gene knock-out homozygote mice. After further breeding to obtain sufficient number of mice, the genome DNA was extracted from the tail of the mice for genotyping by PCR. Ten 4-month-old female filial mice with ERbeta gene knock-out and 10 wild-type female mice were randomly selected and sacrificed, and the right proximal tibiae were removed and subjected to micro CT for measuring the parameters of trabecular bone histomorphometry.

RESULTSA total of 340 filial generation mice were reproduced in 2 months and genotypic identification revealed a proportion of ERbeta+ or + mice of 23.5%, ERbeta+ or - mice of 48.27 percent; and homozygous mutant (ERbeta- or -) mice of 28.3% (in which 54 were female). The MicroCT data revealed that the micro-architecture of the proximal tibiae was significantly different between ERbeta gene knock-out mice selected from the filial generation and wild type mice (P<0.05).

CONCLUSIONIt is feasible to breed ERbeta knock-out female mice by introducing female wild-type mice to pair and mate with ERbeta knock-out homozygote male mice. This approach allows breeding of sufficient number of female ERbeta knock-out mice as the animal models for studying the role of ERbeta.

Animals ; Breeding ; DNA ; analysis ; Estrogen Receptor beta ; genetics ; Female ; Gene Knockout Techniques ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Knockout

Objective To evaluate the clinical safety and efficacy of GuidezillaTM guide extension catheter during complex coronary percutaneous coronary intervention(PCI). Methods A total of 13 patients were included in the studywho presented with complex coronary lesions during PCI. Stents or balloons could not be delivered to these complex lesions with conventional approach and GuidezillaTM guide extension catheters were used. Results Among these 13 patients,there were 7 CTO lesions,7 calcified lesions, 6 tortuous lesions,8 diffuse lesions,8 distal lesions and 8 lesions with pre-existing stents. PCI success rate was 100% .In one patient,GuidezillaTM guide extension catheter was obstructed which were relieved after stent implantation. There was no major adverse cardiac event(MACE) during follow-up.Conclusions The GuidezillaTM guide extension catheter was safe and effective for PCI of complex coronary artery lesions.

OBJECTIVEThis study was aimed to investigate the association between serum against Epstein-Barr virus (EBV) antibodies levels and nasopharyngeal carcinoma (NPC) patients' prognosis.

METHODSBlood samples from 140 primary NPC patients without metastasis were collected before and after treatment. The titers of VCA/IgA and EA/IgA were detected by immunoenzyme assay, and the levels of NA1/IgA and Rta/IgG were detected by enzyme-linked immunosorbent assay (ELISA). All patients received consequent follow-up and long-term efficacy and survival assessment.

RESULTSPost-treatment serum levels of VCA/IgA, EA/IgA, NA1/IgA and Rta/IgG in NPC patients significantly decreased than those before treatment, while had significantly higher than those in control individuals (P < 0.05). Patients in remission had significantly lower pre-treatment serum levels of VCA/IgA and EA/IgA than patients with progression (P < 0.05). None of serum levels of VCA/IgA, EA/IgA, NA1/IgA and Rta/IgG was associated with the 3-year overall survival (P > 0.05). The progression-free survivals were significantly lower in patients with higher pre-treatment VCA/IgA (> or = 1 : 320) and EA/IgA (> or = 1:80) levels than in those with lower VCA/IgA ( < 1 : 320) and EA/IgA (< 1 : 80) levels, respectively (61.8% vs. 86.5% , 61.3% vs. 86.5%, P < 0.001). Cox regression model analysis demonstrated that pre-treatment serum VCA/IgA level was an independent risk factor for progression-free survival (HR = 3.80, P = 0.001).

CONCLUSIONAnti-EBV VCA/IgA and EA/IgA might provide information regarding the prognosis of NPC patients.

Adolescent ; Adult ; Aged ; Antibodies, Viral ; blood ; Antigens, Viral ; immunology ; Capsid Proteins ; immunology ; Carcinoma ; Child ; Female ; Herpesvirus 4, Human ; immunology ; Humans ; Immunoglobulin A ; blood ; Male ; Middle Aged ; Nasopharyngeal Neoplasms ; mortality ; virology ; Prognosis

Conditions of disuse such as bed rest, space flight, and immobilization result in decreased mechanical loading of bone, which is associated with reduced bone mineral density and increased fracture risk. Mechanisms involved in this process are not well understood except the suppression of osteoblast function. To investigate the effect of simulated weightlessness on mRNA level of extracellular matrix proteins, osteoblasts were rotated in horizontal plane as a model of simulated microgravity. Primordial osteoblasts of rats were grown for 2 d and then rotated for 24, 48 and 72 h, respectively. After isolating total RNA in cells, reverse transcription polymerase chain reaction (PT-PCR) was made to examine the mRNA level of osteopontin (OPN) and osteonectin (ON). Meanwhile, the levels of alkaline phosphatase (ALP) and osteocalcin (BGP) in the cultured medium were measured to evaluate the calcific function of cell. The expression of OPN and ON mRNA fell significantly after rotating for 24, 48 and 72 h, respectively. The contents of BGP descended significantly, meanwhile, the activity of ALP also showed a degressive tendency. Horizontal rotation decreased the expression of ON and OPN as well as diminished the secretion of BGP and ALP, which affected the calcific function of osteoblast. The results obtained suggest that depression of extracellular matrix proteins expression plays a key role in bone loss during weightlessness.
Animals ; Animals, Newborn ; Cells, Cultured ; Extracellular Matrix Proteins ; biosynthesis ; genetics ; Osteoblasts ; cytology ; metabolism ; Osteonectin ; biosynthesis ; genetics ; Osteopontin ; RNA, Messenger ; biosynthesis ; genetics ; Rats ; Rats, Wistar ; Rotation ; Sialoglycoproteins ; biosynthesis ; genetics ; Skull ; cytology ; Weightlessness Simulation