·AIM: To investigate the preparation of endostatin protein and its biologic activity on vascular endothelial cell.· METHODS: pBlast-hEndostatin and pBlast-Mcs were identified by digesting with Nhe Ⅰ and Sal Ⅰ, by PCR reaction, by sequencing, and by Alignments of PCR products with gene bank using NCBIBLAST software. The identified pBlast-hEndostatin as well as pBlast-Mcs were then purified with QIAGEN Endofree plasmid maxi kit.The purified plasmids transfected human fibroblasts. The expression of endostatin was detected by RT-PCR, Westem-Blot and immunohistochemistry. The endostatin prorein produced by transfected fibroblasts was purified by ultrafiltration and affinity chromatography. The inhibitory action of endostatin on human umbilical vein endothelium was measured by MTT assay.· RESULTS: pBlast-hEndostatin was found to contain human endostatin gene. Endostatin protein was produced by transfected fibroblasts. The inhibitory ratio of 2.5,5,10,20,40,80mg/L endostatin on human umbilical vein endothelium for 48h were 8.5%,13.1%,27.7%,38.1%,56.7%,63.8% respectively. IC50 value was 34.5mg/L.No inhibition action was found on fibroblasts.·CONCLUSIONS: Endostatin protein can be produced by the transfected fibroblasts. The produced endostatin has inhibitory action on human umbilical vein endothelium and has no inhibition action on fibroblasts.

OBJECTIVETo study the therapeutic effect of reverse island flap based on tibial plantar digital artery for soft tissue defect at the distal end of first toe.

METHODS18 cases with soft tissue defect at the distal end of first toe were treated with reverse island flap based on tibial plantar digital artery.

RESULTSThe patients were followed up for 6 to approximately 12 months (mean 9.5 months). All the flaps survived completely with good functional and cosmetic results.

CONCLUSIONSThe reversed island flap based on tibial plantar digital artery has a reliable blood supply and is adjacent to the recipient area. It is practical and suitable for soft tissue defect at the distal end of first toe.

Adult ; Female ; Hallux ; blood supply ; injuries ; Humans ; Male ; Middle Aged ; Skin Transplantation ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; blood supply ; Young Adult

Comparative analysis of variable region ORF14/15 genes of white spot syndrome virus (WSSV) genome in Guangxi Penaeus vannamei (P. vannamei) could provide useful information for the evaluation of genetic diversity and genetic evolutionary relationship among WSSV isolates from Guangxi, China and other places. Based on geographical and temporal considerations, 40 WSSV-positive P. vannamei samples were collected during the period between May 2010 and July 2013 from Beihai, Qinzhou, and Fangchenggang, which were the main P. vannamei production areas in Guangxi, and the variable region ORF14/15 genes of the WSSV genome from all infected samples were amplified by PCR and then subjected to cloning and sequence analysis. Pairwise and multiple alignment analysis was then conducted to evaluate the degree of genetic divergence between different strains. The variable region ORF14/15 genes from 25 of 40 WSSV positive samples were successfully cloned and sequenced; among the ORF14/15 genes of 25 WSSV-positive strains, 22 was 619 bp in length and 3 was 620 bp. All the 25 Guangxi strains carried a 5949-bp deletion in the ORF14/15 region relative to TH-96-II, which has the longest nucleotide sequence in this region; the deletion of Guangxi strains occurred in the middle region of ORF14/15 gene, with only 190 bp and 429 bp/ 430 bp at 5' and 3' ends, respectively, which were coincident with WSSV-IN-05-I in deletion length and position. Sixteen of 25 Guangxi strains had completely identical nucleotide sequences in the variable re gion, and the homology between other strains also exceeded 97.9%. There were single nucleotide substi tution, deletion, and insertion in the ORF14/15 region of Guangxi strains compared with other strains in GenBank. In the phylogenetic tree based on WSSV variable region ORF14/15, the Guangxi strains were closely related and formed a separate branch with Indian strain IN-05-I, but far from other strains in GenBank. The ORF14/15 gene of WSSV isolates in cultured P. vannamei in Guangxi has a large deletion in the middle of the variable region, and the Guangxi WSSV strains show no significant spatio-temporal differences; the Guangxi strains are closer in genetics to Indian strain IN-05-I than other strains in GenBank.
Animals ; China ; Cloning, Molecular ; Evolution, Molecular ; Genome, Viral ; genetics ; Genomics ; Penaeidae ; virology ; Phylogeny ; White spot syndrome virus 1 ; genetics

Adult ; China ; epidemiology ; Condylomata Acuminata ; epidemiology ; Female ; Humans ; Incidence ; Male ; Retrospective Studies ; Rural Population ; Urban Population ; Young Adult

BACKGROUNDRecent advances in real-time three-dimensional echocardiography (RT3DE) offer the potential to assess the left ventricular (LV) dyssynchrony simultaneously by analyzing the 17 segments time-volume curves. The purpose of this study was to test the feasibility and accuracy of RT3DE for quantitative evaluation of left ventricular systolic synchronicity.

METHODSTwenty-four patients with dilated cardiomyopathy (DCM) and twenty-five healthy volunteers were enrolled in this study. Full volume RT3DE was performed by using Philips IE33 with X3-1 probe. The global and 17-segmental time-volume curves were obtained by the on-line Qlab software (version 4.2). The time to minimal systolic volume in each segment (T(msv)) was taken to derive the following indexes of systolic asynchrony: T(msv) 16-SD, T(msv) 16-Dif, T(msv) 12-SD, T(msv) 12-Dif, T(msv) 6-SD and T(msv) 6-Dif, which meant the standard deviation or the maximal difference of T(msv) among the 16, 12 and 6 segments of the left ventricle respectively. The software also provided with each of the above parameters as a percentage of the cardiac cycle.

RESULTST(msv) 16-SD, T(msv) 12-SD and T(msv) 6-SD were all significantly larger in the DCM group than those of the control group [T(msv) 16-SD: (52.9 +/- 40.6) ms vs (8.8 +/- 6.2) ms; T(msv) 12-SD: (29.5 +/- 30.8) ms vs (6.9 +/- 4.0) ms; T(msv) 6-SD: (28.9 +/- 34.6) ms vs (7.0 +/- 4.7) ms, all P < or = 0.001]. T(msv) 16-Dif, T(msv) 12-Dif and T(msv) 6-Dif were also significantly larger in the DCM group. There were close negative relations between the LVEF determined by RT3DE and each of the indexes of systolic asynchrony, among which the indexes of T(msv)-16-SD% and T(msv)-16-Dif% correlated most closely (r = -0.703 and r = -0.701, respectively). The DCM patients had significantly larger EDV and ESV, with significantly reduced LVEF compared with the healthy subjects.

CONCLUSIONRT3DE provides a simple, useful and unique approach to assess the systolic synchronicity of all the left ventricular segments simultaneously.

Adult ; Aged ; Cardiomyopathy, Dilated ; diagnostic imaging ; physiopathology ; Echocardiography, Three-Dimensional ; Female ; Humans ; Male ; Middle Aged ; Stroke Volume ; Systole ; Ventricular Function, Left

Chronic myeloid leukemia (CML) is a malignant clonal disease derived from hematopoietic stem cells. CML stem cells were thought to be the root which could lead disease development and ultimately rapid change. However, a stable animal model for studying the characteristics of CML stem cells is currently lacking. This study was aimed to establish a transplanted human CML nude-mice model to further explore the biological behavior of CML stem cells in vivo, and to enrich CML stem cells in nude mice by series transplantation. The 4 - 6 weeks old BALB/c nude mice pretreated by splenectomy (S), cytoxan intraperitoneal injection (C) and sublethal irradiation (I) were transplanted intravenously with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase. Alternatively, 4 - 6 weeks old BALB/c nude mice pretreated by lethal irradiation were transplanted intravenously with 5 × 10(6) homologous bone marrow cells of BALB/c nude mice together with (5 - 7) × 10(7) of bone marrow mononuclear cells from CML patients in chronic phase simultaneously. The leukemic cells engrafted and infiltrated in organs and bone marrow of the mice were tracked by reverse transcription-polymerase chain reaction (RT-PCR), plastic-embedded biopsy and flow cytometry. The results of these two methods were compared. The results showed that human CML cells engrafted and infiltrating into the bone marrow of two nude mice pretreated with SCI could be detected. In spite of the low successful rate, results suggested the feasibility of this method by using BALB/c nude mice as a human CML animal model. In contrast, in nude mice pretreated by the lethal dose irradiation, CML cells in the bone marrow could not be found. It is concluded that human bone marrow CML cells can results in leukemia in nude mice pretreated by SCI. Thus this study provides a new strategy for establishment of CML animal models which deserves further elaboration.
Animals ; Disease Models, Animal ; Humans ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Male ; Mice ; Mice, Nude ; Mice, SCID ; Neoplasm Transplantation ; Neoplastic Stem Cells ; Transplantation, Heterologous

OBJECTIVETo observe the effect of the inducible nitric oxide synthase (iNOS) gene on androgen-independent prostate cancer DU145 cells in vitro.

METHODSThe iNOS gene was transfected into androgen-independent prostate cancer DU145 cells. The positive cells were selected as the transfected group for amplification, and an empty vector (EV) group and a control group were also set. The mRNA transcription of iNOS was analyzed by RT-PCR. The morphological changes of the cells were observed, the effect of iNOS transfection on the cell growth determined using the MTB method, and the apoptosis of DU145 cells detected by flow cytometry, followed by analysis of the effect of NOS inhibitors on the transfected cells.

RESULTSDU145 cells transfected with iNOS secreted significantly more nitric oxide ([272.50 +/- 15.82] micromol/L) than those of the EV and control groups ([122.00 +/- 18.93] micromol/L and [121.00 +/- 6.98] micromol/L) (P < 0.05). The rate of cell apoptosis was markedly enhanced in the transfected group as compared with the EV and control groups ([42.78 +/- 2.01]% vs [30.65 +/- 1.46]% and [28.96 +/- 1.50]%, P < 0.05). MTP test indicated a slower growth of the DU145 cells in the former than in the latter two (P < 0.05). NOS inhibitors enhanced their growth, but with no significance (P > 0.05).

CONCLUSIONDU145 cells transfected with iNOS could secrete high-concentration nitric oxide, induce cell apoptosis, and suppress cell proliferation, which may provide a potentially effective gene therapy for advanced androgen-independent prostate cancer.

Androgens ; pharmacology ; Apoptosis ; genetics ; Cell Cycle ; genetics ; Cell Line, Tumor ; Cell Proliferation ; Genetic Vectors ; Humans ; Male ; Nitric Oxide Synthase Type II ; genetics ; Prostatic Neoplasms ; genetics ; pathology ; Transfection

OBJECTIVETo study the causes of orchiectomy in different age groups.

METHODSWe retrospectively reviewed the clinical data about 291 cases of orchiectomy performed between March 1993 and October 2014 and analyzed the causes of surgery and their distribution in different age groups.

RESULTSThe main causes of orchiectomy were testicular torsion (45.8%), cryptorchidism (32.5%) and testicular tumor (16.9%) in the patients aged 0-25 years, testicular tumor (42.4%), cryptorchidism (25.9%) and tuberculosis (10.6%) in those aged 26-50 years. Prostate cancer was the leading cause in those aged 51-75 years (77.6%) or older (84.0%)), and testicular tumor was another cause in the 51-75 years old men (10.2%). Prostate cancer, testicular tumor, cryptorchidism, and testicular torsion were the first four causes of orchiectomy between 1993 and 2009. From 2010 to 2014, however, testicular tumor rose to the top while prostate cancer dropped to the fourth place.

CONCLUSIONThe causes of orchiectomy vary in different age groups. The proportion of castration for prostate cancer patients significantly reduced in the past five years, which might be attributed to the improvement of comprehensive health care service.

Adolescent ; Adult ; Age Factors ; Aged ; Causality ; Child ; Child, Preschool ; Cryptorchidism ; surgery ; Humans ; Infant ; Infant, Newborn ; Male ; Middle Aged ; Orchiectomy ; statistics & numerical data ; Prostatic Neoplasms ; surgery ; Retrospective Studies ; Spermatic Cord Torsion ; surgery ; Testicular Neoplasms ; surgery ; Tuberculosis, Male Genital ; surgery ; Young Adult

DNA, Viral ; analysis ; Hepatitis B Core Antigens ; analysis ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Immunohistochemistry ; Liver ; virology ; Liver Cirrhosis ; virology ; Oligonucleotide Array Sequence Analysis ; methods

OBJECTIVETo investigate the stability and three-dimensional movements of the atlantoaxial joint after artificial atlanto-odontoid joint (AAOJ) arthroplasty by comparing with a conventional method.

METHODSAfter anterior decompression, 24 human cadaveric spinal specimens of C0-C3 were randomly divided into two groups: Group A receiving artificial AAOJ arthroplasty; Group B experiencing anterior transarticular screw (ATAS) fixation. Two groups underwent flexibility test in intact and instrumented states. Rotational angle of the C0-C3 segments was measured to study the immediate stability and function of anterior decompression with AAOJ arthroplasty compared with the intact state and ATAS fixation.

RESULTSCompared with the intact state, anterior decompression with AAOJ arthroplasty resulted in a significant decrease in the range of motion (ROM) and neutral zone (NZ) during flexion, extension and lateral bending (P less than 0.05); however, with regard to axial rotation, there was no significant difference in ROM and NZ (P larger than 0.05). Compared with anterior decompression with ATAS fixation, anterior decompression with AAOJ arthroplasty during flexion, extension and lateral bending, significant difference was found in ROM and NZ (P larger than 0.05); however, as for axial rotation, there was a significant increase in ROM and NZ (P less than 0.05).

CONCLUSIONThe self-designed AAOJ has an excellent biomechanical performance, which can restore excellent instant stability and preserve the movement of the atlantoaxial joint.

Adult ; Arthroplasty ; methods ; Atlanto-Axial Joint ; physiopathology ; surgery ; Biomechanical Phenomena ; Bone Plates ; Bone Screws ; Cadaver ; Decompression, Surgical ; Female ; Humans ; Joint Instability ; physiopathology ; surgery ; Male ; Middle Aged ; Range of Motion, Articular ; physiology