OBJECTIVETo study the expression of gelatinases, including matrix metalloproteinase-9 (MMP-9, gelatinase B) and matrix metalloproteinase-2 (MMP-2, gelatinase A), in CD(34)(+) cells and leukemic cell lines, and explore the significance of gelatinase in migrating and homing capacity of CD(34)(+) cells, as well as the role of gelatinase in leukemia pathogenesis.

METHODSCD(34)(+) cells were isolated from umbilical cord blood and normal bone marrow by Mini MACS system. By zymogram analysis, MMP-2 and MMP-9 were detected in the serum free condition medium of CD(34)(+) cells and cell lines.

RESULTSOne brilliant band with molecular weight of 92 x 10(3) was detected in condition medium of cord blood CD(34)(+) cells. No band was detected in condition medium of bone marrow CD(34)(+) cells. Brilliant bands with molecular weight of 92 x 10(3) and 72 x 10(3) were detected in the condition medium of U937, KG-1a and HL-60 cell lines, but not in that of HEL, Namalva, CEM, K562 and LCL-H cell lines. In the condition media of J6-1 and J6-2 cells only the 92 x 10(3) band was detected.

CONCLUSIONSCord blood CD(34)(+) cells produced MMP-9, but bone marrow CD(34)(+) cells did not, partly explains the fact that cord blood CD(34)(+) cells possessed higher migrating capacity in comparison with bone marrow CD(34)(+) cells. The expression of MMP-9 and MMP-2 in leukemic cell lines varied.

Antigens, CD34 ; analysis ; Culture Media, Conditioned ; metabolism ; Electrophoresis, Polyacrylamide Gel ; Fetal Blood ; cytology ; enzymology ; immunology ; HL-60 Cells ; Humans ; K562 Cells ; Leukemia ; enzymology ; pathology ; Matrix Metalloproteinase 2 ; biosynthesis ; Matrix Metalloproteinase 9 ; biosynthesis ; Tumor Cells, Cultured ; U937 Cells

BACKGROUNDThe CRF07_BC recombinant strain has been one of the most predominantly circulated HIV-1 strains in China, it is therefore necessary and urgent to develop a relevant animal model to evaluate candidate vaccines targeting HIV-1 CRF07_BC. A highly replication-competent simian/human immunodeficiency viruses (SHIV) construct containing the Chinese CRF07_BC HIV-1 env gene with the ability to infect Chinese rhesus monkeys would serve as an important tool in the development of HIV vaccines. The aim of this study was to examine whether SHIV XJDC6431 with the env fragment from a Chinese HIV-1 isolate virus could infect the human and monkey peripheral blood mononuclear cell (PBMC), establish infection in Chinese rhesus macaque.

METHODSA SHIV strain was constructed by replacing the rev/env genes of SHIV KB9 with the corresponding fragment derived from the HIV-1 CRF07_BC strain. The infectious activity of the SHIV clones was determined in vitro in PBMCs from both non-human primate animals and humans. Finally, one Chinese rhesus macaques (Macaca mulatta) was infected with one SHIV via intravenous infusion.

RESULTSOne SHIV clone designated as SHIV XJDC6431, was generated that could infect macaque and human PBMC. The virus produced from this clone also efficiently infected the CCR5-expressing GHOST cell lines, indicating that it uses CCR5 as its coreceptor. Finally, the virus was intravenously inoculated into one Chinese rhesus macaque. Eventually, the animal became infected as shown by the occurrence of viremia within 3 of infection. The viral load reached 105 copies of viral RNA per ml of plasma during the acute phase of infection and lasted for 10 weeks post infection.

CONCLUSIONSWe conclude that SHIV XJDC6431 is an R5-tropic chimeric virus, which can establish infection not only in vitro but also in vivo in the Chinese rhesus macaque. Although the animal inoculated with SHIV XJDC6431 became infected without developing a pathologic phenotype, the virus efficiently replicated with a persistent level of viral load in the plasma. This suggested that the SHIV could be used as a tool to test candidate AIDS vaccines targeting the Chinese HIV-1 CRF_07BC recombinant strain.

Animals ; Chimera ; Genes, env ; HIV-1 ; genetics ; physiology ; Humans ; Macaca mulatta ; Proviruses ; genetics ; Receptors, CCR5 ; physiology ; Simian Immunodeficiency Virus ; genetics ; physiology

OBJECTIVEChimeric human/bovine immunodeficiency virus (HBIV) cDNA was constructed by replacing HIV tat and LTR with bovine immunodeficiency virus (BIV) tat and LTR to study the activity of BIV tat and LTR in the chimerae.

METHODSThe target fragments of BIV tat, LTR and HIV gag, pol, env were respectively amplified by using PCR and sequentially inserted into pBluescript SK(+) vector. The chimeric clone was transfected into human MT4 cells. The transcript and gene expression of the HBIV chimeric virus were detected by using RT-PCR and a reverse transcriptase assay, respectively.

RESULTSBIV tat mRNA and HIV gag mRNA were detected. The reverse transcriptase activity of the chimeric virus was analyzed in the fluctuation curve.

CONCLUSIONSIn chimeric HBIV cDNA transfected MT?4 cells, BIV tat and HIV gag were transcripted. The reverse transcriptase of the chimeric virus had biological activity. These data suggest that in MT4 cells, BIV LTR had promoter activity and BIV tat had the function of transactivation in the chimeric virus. The study of the chimeric virus with infectivity is in progress.

AIDS Vaccines ; Animals ; Cattle ; Cloning, Molecular ; DNA, Complementary ; genetics ; DNA, Viral ; genetics ; Genes, gag ; genetics ; Genes, pol ; genetics ; Genes, tat ; genetics ; HIV-1 ; genetics ; Humans ; Immunodeficiency Virus, Bovine ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Transcription, Genetic ; Transcriptional Activation ; Transfection ; Virus Replication

Objective: To dynamically evaluate left ventricular perfusion, global and local functional changes during left ventricular aneurysm (LVA) formation and to explore the relationship between the size of LVA and LVEF, LVESV, LVEDV by gated99mTc-MIBI SPECT (GSPECT) and gated18F-FDG PET metabolic (GPET) imaging in experimental pigs. Methods: LVA model was established by occlusion of left circumlfex artery (LCX) and placing an Ameroid constrictor at the proximal end of left anterior descending artery (LAD) in a total of 16 Chinese mini-pigs. At the 1st, 4th and 8th weeks of surgery, the changes of total perfusion defect (TPD), LVA formation and LVEF, LVESV, LVEDV were dynamically evaluated by GSPECT and GPET; the relationships between the size of LVA and LVEF, LVESV, LVEDV were analyzed respectively.Results: There were 5 pigs died in surgery and 2 died at the 1st week of modeling. According to golden (pathological) standard, 9 animals successfully ifnished the dynamic imaging study. At the 1st week of (basic) modeling, 4 animals formed large LVA, 2 formed small LVA at the apex and 3 without LVA formation. At the 4th and 8th weeks of modeling, dynamic imaging presented that the animals with large LVA had gradually increased range and degree of perfusion defect, LVEDV, LVESV, while gradually decreased LVEF; the above indexes were relatively stable in animals with small or none LVA. In addition, the size of LVA was related to LVEF (r=-7.26), LVEDV (r=0.855) and LVESV (r=0.825), allP<0.05. Conclusion: In experimental pigs, at the beginning of LVA formation, large range and severe perfusion defect may cause large aneurysm, the LV functional damage and remodeling may gradually increase and the prognosis is poor; in contrast, the animals with small or none LVA have better prognosis and usually without ventricular remodeling; which implies that in acute phase of LVA formation, the size of aneurysm may predict the trend of global LV systolic function and remodeling at the early stage.

OBJECTIVETo evaluate the therapeutic effect of Compound Xuanju Capsule on type III prostatitis.

METHODSA total of 242 patients with type III prostatitis diagnosed by the NIH criteria were randomly divided into an experimental and a control group of equal number, the former treated with Compound Xuanju Capsule + Tamsulosin Hydrochloride, and the latter with Quinolinone antibiotics + Tamsulosin and Hydrochloride, both for 6 months. After treatment, we assessed the therapeutic effects based on the NIH-CPSI scores and the improvement of relevant complications.

RESULTSAll the 242 patients completed the treatment. The total effectiveness rate was 77.69% (94/121) in the experimental group, 71.56% (78/109) in those with complications. In comparison, it was only 47.10% (57/121) in the control group, 31.78% (34/107) in those with complications. Both the NIH-CPSI scores and the improvement of complications were significantly higher in the experimental than in the control group (P < 0.05).

CONCLUSIONCompound Xuanju Capsule has a good therapeutic effect on type III prostatitis.

Adolescent ; Adult ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; Male ; Middle Aged ; Phytotherapy ; Prostatitis ; drug therapy ; Treatment Outcome ; Young Adult

Objective To assese the healing of stoma after magnetic anastomosis for the reconstruction of biliary-enteric continuity under severe inflammation.
Methods Acute bile duct injury was constructed as a bile peritonitis model in mongrel dogs (n=32). Magnetic anastomosis (group A, n=16) and traditional suture anastomosis (group B, n=16) were performed to reconstruct the biliary-enteric continuity in one stage. Half of the dogs in each group were euthanized on the 30th postoperative day, and the other half on the 90th postoperative day to harvest the stoma region. The healing conditions of the stoma after the 2 anastomotic approaches were observed with naked eyes, under light microscope and scanning electron microscope.
Results The stoma leakage rate (50%versus 0%on the 30th postoperative day, 37.5%versus 12.5%on the 90th postoperative day, both P<0.05) and stenosis degree (13.9%±0.3%versus 7.1%±0.3%on the 30th postoperative day, 17.2%±0.4%versus 9.4%±0.4%on the 90th postoperative day, both P<0.01) were significantly higher in group B than in group A. Compared with traditional manual anastomoses, the histological analysis under light and electron microscope showed a more continuous stoma with more regular epithelium proliferation and collagen arrangement, less inflammation in group A.
Conclusions Magnetic anastomosis stent ensures better healing of the stoma even under the circumstance of severe inflammation.

Objectives： This study was designed to observe the histological localization of SNC73 gene expression in colorectal cancer, and to investigate their expression in different phases from normal epithelium, transitional mucosa, adenoma to carcinoma, and to discuss the relationship between SNC73 gene expression and the clinic factors of colorectal cancer. Method： With in situ hybridization, the expression of the SNC73 was determined in 29 patients with colorectal cancer group specimens as the above. Results： SNC73 gene expression was observed in normal mucosal tissue and some lymph follicles of the mucosa, but seldom in cancer tissue. The positive rate of SNC73 expression was 20.69％ in colorectal cancer; was 62.07％ in normal mucosa; was 31.03％ in adjacent tissue; and was 20.00％ in adenoma. Statistical difference was found between expressions in normal mucosa and those in colorectal carcinoma from the same patient (P<0.01). Conclusions： The positive rate of SNC73 expression in colorectal cancer is lower than that in corresponding normal mucosa. The biological function of SNC73 may be different from that of common immunoglobulins.

Female ; Hernia, Ventral ; surgery ; Humans ; Laparoscopy ; methods ; Middle Aged

Internal carotid arterial system is the main source of blood supply in brain. In forensic identi-fication practice, blunt injures of internal carotid arterial system usually cause complications such as throm-bus, aneurysm and arteriovenous fistula, etc. The deaths following delayed cerebral infarction or intracra-nial haemorrhage are not rare. At present, literature of deaths caused by blunt injuries of internal carotid arterial system mainly consist of case reports in China. This paper reviews related literature and case re-ports at home and abroad, and summarizes forensic medical features and identification method of the deaths caused by such injuries. The results show that blunt injures of internal carotid arterial system are related to the direct or indirect force on head and neck, which can result in exceed physiological range traction of head and neck, incision following basicranial fracture, etc. Such injuries are common in the cases as mechanical asphyxia, cervical manipulation, traffic accident and fall, etc. The artery should be examined carefully, and the relationship among injury, disease and death should be analysed correctly when no cause of infarction and hemorrhage was found in routine examination of such cases in forensic pratice. Because of the difficulty for exposing the artery completely in autopsy, angiography can be used to infer the location when necessary for improving the scientificity and reliability of the appraisal conclusion.