Objective To investigate the results of graft repair of peripheral nerve defects using small intestinal submucosa(SIS) with crude Schwann cells. Methods Thirty-six male SD rats were randomly divided into three groups,with 12 in each group.Firstly,12-mm gaps of sciatic nerve were made in rats and then treated respectively with porcine SIS(group A),SIS compounded with crude Schwann cells(group B) and auto-nerve grafting(group C) in the three groups.The grafts were harvested 16 weeks postoperatively to evaluate the results of nerve regeneration through histological examination, triceps surae weight,computerized imaging analysis,Trueblue retrograde tracing and transmission electron microscopy. Results Nerve regeneration was confirmed to have extended through the gaps according to the results of histological examination,Trueblue retrograde tracing and transmission electron microscopy.Furthermore,in the respect of triceps surae weight,quantity of axis cylinder per area and percentage of neural tissue,the results of group B and group C were superior to those of group A with a significantly statistical difference(P0.05). Conclusion SIS compounded crude Schwann cells was able to achieve the outcome of auto-nerve grafting and was a promising replacement graft.

Objective To observe adhesion and growth of Sehwann cells(SCs) on small intestinal submueosa(SIS) and study the bioeompatibility of SIS with SCs.Methods The SCs of SD neonate rat were isolated and cultured in vitro,then were seeded on prepared SIS.At different times,the adhesion,growth and proliferation of SCs on SIS were observed by phase contrast microscope,histological examination,scanning e- lectron microscope and transmission electron microscope.Results By phase contrast microscope,SCs grew well on the edge of SIS after 3 and 5 days.SCs adhered tightly on the surface of SIS after 5 days through histo- logical examination.By scanning electron microscope,on the surface of SIS,SCs grew and adhered actively, prominence of cells body were obvious.They connected end-to-end with each other or arranged in clusters. Protein granules were secreted on cells surface.By transmission electron microscope,SCs grew in good condi- tion and adhered tightly on the surface of SIS.At the interface of SCs and SIS,prominence was seen to contact with SIS in the bottom of cell body.Conclusion SCs are able to adhere and grow well on the surface of SIS.As a scaffold,SIS has good biocompatibility with SCs.

Objective To study the characteristics of dynamic susceptibility-contrast(DSC)MR perfusion curves,color images and perfusion values in pre-operative brain metastasis.Methods Twenty- eight brain metastases underwent DSC MR perfusion imaging by using a first-pass T_2~* echo-planar sequence. The patients' data were transferred to on-line workstation.Time-signal intensity curves,color perfusion maps and rCBV,rMTT values in both tumor parenchyma and peri-tumor edema were analyzed,and independent t- test was used and P0.05).Conclusion Different originated brain metastases have nearly same characteristics in DSC MR perfusion imaging.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; Female ; Hepatitis B virus ; physiology ; Hepatitis B, Chronic ; virology ; Humans ; Male ; Middle Aged ; Virus Replication

Cell cycle progression is tightly regulated in hematopoietic stem cells. The cycle state decides cells' fates, which includes self-renewal, proliferation and differentiation. Proper cell cycle regulation is a pivotal element for the maintenance of hematopoiesis homeostasis. HTm4 is a newly identified specific cell cycle regulator of the hematopoietic cell. Through interacting with KAP-CDK2 complex, it arrests cells in G(0)/G(1) phase. K562 is a human chronic myelogenous leukemia cell; it could be induced to megakaryoblast by phorbol 12-myristate 13-acetate (PMA). Such differentiation must be associated with cell cycle change. To further clarify HTm4's function in hematopoietic cell cycle regulation, K562 cells were treated with PMA. Cell cycle change was analysed using flow cytometric system. And during the induction process gene expression of HTm4 as well as CycleE and CDK2, which are responsible for G(1) to S transition, were analysed using semi-quantitative RT-PCR. The C-terminal domain of HTm4 protein has been shown to be important for HTm4's binding with KAP-CDK2 complex. To determine its impact on HTm4's function, HTm4 and C-terminal truncated HTm4 (HTm4-ct) were transfected into K562 cells using Tet-Off regulation expression system. Their influence on cell cycle was observed. The results showed that PMA induced both expansion and differentiation of K562 cells as measured by cell number count and NBT staining respectively. During PMA treatment, G(0)/G(1) cell proportion and HTm4 expression displayed coordinated change, which suggested that HTm4 might drive K562 cells out of cell cycle but was not involved in the quiescence maintenance. Additionally, transfection of HTm4 caused G(0)/G(1) arrest in K562 cells, while transfection of HTm4-ct did not. It is therefore suggested that the C-terminal domain is important for the function of HTm4 in cell cycle regulation.
Cell Cycle ; physiology ; Cell Cycle Proteins ; genetics ; physiology ; Cells, Cultured ; Gene Expression Regulation ; Hematopoiesis ; physiology ; Hematopoietic Stem Cells ; cytology ; physiology ; Humans ; K562 Cells ; Membrane Proteins ; genetics ; physiology ; Tetradecanoylphorbol Acetate ; pharmacology ; Transfection

Objective To investigate the effects of physical exercises on cardiac function and plasma N-terminal pro-brain natriuretic peptide (NT-proBNP) levels in hypertensive patients combined with diastolic cardiac dysfunction.MethodsA total of 66 essential hypertension patients who had abnormal left ventricular relaxation and normal systolic function were assigned to the intervention group ( n =33 ; doing physical exercises once a day,5 days a week) or control group (n =33 ).All the patients received standard treatment.At 6 months,body weight,blood pressure,heart rate,NT-proBNP,and echocardiography were measured.ResultsAt 6 months,body weight [ (68 ± 7 ) kg vs (72 ± 8 ) kg ],systolic blood pressure [ (135.4 ±5.1) mm Hg (1 mm Hg =0.133 kPa) vs (141.9 ±5.2) mm Hg ],diastolic blood pressure [ (81.1 ±4.0) mm Hg vs (84.7 ±4.6) mm Hg],New York Heart Association class (1.4 ±0.3 vs 1.8 ±0.4),NT-proBNP level [ (526 ± 126 ) ng/L vs (741 ± 189 ) ng/L] were significantly decreased in the intervention group when compared with the control group ( all P ＜ 0.05 ) although left ventricular ejection fraction (LVEF) (62.9 ±6.7 vs 59.0 ±5.6) and E/A ratio ( 1.1 ±0.3 vs 0.9 ±0.3) were significantly increased ( both P ＜ 0.05).ConclusionPhysical exercises could play a role in reduced blood pressure and body weight and improved cardiac function in hypertensive patients with diastolic cardiac dysfunction.

Objective To investigate the effect of microRNA-223-3p (miR-223-3p) on megakaryocytic differentiation and maturation, and explore the potential mechanism .Methods The endogenous expression of miR-223-3p during megakaryocyte ( MK) differentiation was detected by real-time PCR.Flow cytometry further indicated that alteration of miR-223-3p in human cell lines exerted effects on MK differentiation and maturation .By performing integrative bioinformatic analysis, the potential miR-223-3p target gene, MYH10,was identified.Real-time PCR, luciferase reporter assay and flow cytometry revealed that MYH10 was a direct target of miR-223-3p.Results Endogenous expression of miR-223-3p was in-creased with the differentiation and maturation of MK .The expression of megakaryocytic surface markers CD41 and CD61 and the ploidy were significantly increased in K562 and Meg-01 cells after transfection with miR-223-3p mimics.The expression of MYH10 decreased with the increase in miR-223-3p.Using a luciferase reporter assay ,we demonstrated that MYH10 was a direct target of MiR-223-3p.Furthermore, direct downregulation of MYH10 promoted MK polyploidization . Conclusion MiR-223-3p might regulate the polyploidization of MK by targeting MYH10.

E2 gene of classical swine fever virus (CSFV) was cloned into secretory pPIC9K Pichia pastoris expression vector. After being linearized by digestion, the vector was transformed into Pichia pastoris by electroporation to integrate with the genome, the transformants with high copies were screened by G418 and were induced to express with methonal. The results of SDS-PAGE and Western blot demonstrated that the supernatant of the induced P. pastoris culture contained protein E2. The results of the study on the immunological activity indicated that the protein E2 expressed in P. pastoris can elicit animal bodies to produce antibodies against protein E2.
Animals ; Antibodies, Viral ; immunology ; Antigens, Viral ; genetics ; immunology ; Classical swine fever virus ; genetics ; immunology ; Cloning, Molecular ; Gene Expression ; Pichia ; Rabbits ; Swine ; Viral Envelope Proteins ; genetics ; immunology

As the new type cornea ulcer renovation material, the biological amnion is to be implanted into the human body for a long time, a subchronic toxicity study in rats is made to evaluate its possibility of subchronic toxicity. The study is based on the requirements of "Biological Evaluation of Medical Devices, Part 11: Tests for systemic toxicity and Part 6: Tests for local effects after implantation". After the implantation of examples to be tested, animals were observed daily for mortality and 92 days later the possible subchronic toxicity was evaluated. And a necropsy was conducted and the selected organs were excised, weighed, and processed histologically. Body weights, organ weights, organ/body weight ratios, hematology values and clinical chemistry values were analyzed statistically. Results show that daily clinical observation, body weights, necropsy findings, organ weights and organ/body weight ratios were within acceptable limits in test and control treatment groups. There were no obvious changes in histopathology, hematology values or clinical chemistry values in either male or female rats and no notable differences between the biological amnion and the control amnion. This study proves that, the cornea ulcer renovation material, the biological amnion does not induce subchronic toxicity.
Amnion ; transplantation ; Animals ; Biological Products ; toxicity ; Corneal Ulcer ; surgery ; Female ; Male ; Materials Testing ; Rats ; Rats, Wistar ; Toxicity Tests, Subchronic

OBJECTIVETo explore the mechanism of Flt3 receptor-interacting lectin (FRIL) maintains quiescence of hematopoietic stem cells (HSCs) in vitro.

METHODSCord blood CD34+ cells were cultured in suspension medium supplemented with or without FRIL and FL. Cells were collected at different time points and the expression of some cell cycle regulators, especially those involved in G0/G1 phase regulation were detected on mRNA and protein level.

RESULTSThe expressions of G0/G1 phase related cyclins or CDKs were undetectable in the newly isolated CD34+ cells, expressions of Cyclin D3, CDK6 and P27 were the lowest in FRIL cultured group after 3d's culture (FRIL group: 483 +/- 63, 553 +/- 39, 0.312 +/- 0.030; FL group: 2437 +/- 52, 3209 +/- 98, 0.787 +/- 0.024; BLANK: 914 +/- 105, 1497 +/- 55, 0.616 +/- 0.029, respectively), but the expression of P53 was the highest in FRIL group (FRIL group: 4.476 +/- 0.159; FL group: 0.581 +/- 0.099, BLANK: 2.167 +/- 0.114). The expression of positive regulators of cell cycle in FRIL group were the same as that of FL group and blank group or lower.

CONCLUSIONFRIL preserves HSCs effectively in vitro through the mechanisms of down-regulation of cyclin D3 and CDK6 and activation of P53. P27 is mostly involved in the differentiation of HSCs.

Antigens, CD34 ; Cell Cycle ; drug effects ; Cell Cycle Proteins ; genetics ; metabolism ; Cells, Cultured ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; drug effects ; metabolism ; Humans ; Mannose-Binding Lectins ; pharmacology ; Plant Lectins ; pharmacology ; RNA, Messenger ; genetics