Objective To investigate the effects of recombinant human augmenter of liver regeneration (rhALR) on renal inflammation in acute kidney injury (AKI) induced by renal ischemia reperfusion (IR). Methods SD rats were randomly divided into sham-operated group,IR group,rhALR1 group (100 μg/kg) and rhALR2 group (200 μg/kg).Both renal pedicles of rats were identified and occluded with microvascular clamps for 60 min to induce acute kidney injury (AKI).Blood urea nitrogen and serum creatinine levels were evaluated using a Hitachi 747 automatic analyzer. For histological examination, sections were stained with HE. The activity of myeloperoxidase (MPO) was detected by spectrophotometer.Expression of TNF-α,ICAM-1,MCP-1 was determined by Western blotting. Results Blood urea nitrogen,serum creatinine levels and the injury of kidney were improved significantly in rhALR group as compared with IR group (all P＜ 0.05).They were improved more significantly in rhALR2 group as compared to in rhALR1 group (all P＜0.05).The protein levels of TNF-α,ICAM-1,MCP-1 and the activity of MPO in kidneys from the sham-operated rats were low,and increased significantly after renal ischemia reperfusion injury (all P＜0.05).After treated with rhALR,the expression of TNF-α,ICAM-1,MCP-1 and the activity of MPO were decreased significantly in kidneys as compared to those in IR group (all P＜0.05),which decreased more significantly in rhALR2 group than those in rhALR1 group (all P＜ 0.05). Conclusions nhALR can protect kidneys from ischemia reperfusion injury in rats.The mechanism may be associated with the inhibition of renal inflammatory cells infiltration and down-regulated expressions of YNF-α,ICAM-1 and MCP-1 in the kidney.

Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells. The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells, and the relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells. By using magnetic active cell sorting (MACS), we isolated a population of CD44(+)/CD133(+) prostate cancer cells that display stem cell characteristics from PC3 cell line. Immunohistochemistry revealed positive expressions of CD44, CD133 and α(2)β(1)-integin in the isolated cells. CCK-8 analysis showed that isolated cells had a strong proliferative ability. The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing, indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line. Western blotting exhibited that the isolated cells had higher experession of Nanog, an embryonic stem marker, as compared with PC3 cells. Our study showed that Nanog might be helpful in sustaining the self-renewal and the undifferentiation of prostate cancer stem cells, and may serve as a marker for prostate cancer stem cells for isolation and identification.

Objective To establish a doxycycline-controlled immortalized pre-cartilaginons stem cells (IPCSCs) strains, clone parathroid hormone-related peptide[PTHrP(1-36)] gene and construct re- sponsive plasmid, pTRE-PTHrP (1-36). Methods Plasmid pTet-on was transfected into IPCSCs by using LipoinfectaminTM 2000 and then the stable clones were obtained by G418 screening. The doxycyc- line was added into the medium of monoclonal cells that were transiently transfected with plasmid pTRE- 2Hyg-Lue. The total RNA was extracted from PCSCs and the PTHrP(1-36) gene obtained by RT-PCR method. Then, the PTHrP (1-36) gene was subcloned to plasmids of Tet-responsive element with the se- lection marker of hygromycin pTRE-2Hyg to construct recombinant eukaryotic expression plasmid pTRE- PTHrP(1-36). After transferred into E. coli-DH5α, the clone was amplified, the recombinant plasm0ids were purified and identified by double-enzyme digestion. Results The doxycycline induced IPCSCs line was obtained, with 50 times higher than the non-induced cell line. Double enzyme digestion analysis and sequencing showed that the target gene was cloned into recombinant plasmid. Conclusions The induced IPCSCs line can be used to highly express alien genes. The responsive plasmid containing PTHrP (1-36) gene may be premising for rigorous control of PTHrp (1-36) gene expression.

OBJECTIVETo explore the effects of lead on the thyroid function of occupationally exposed workers.

METHOD157 workers occupationally exposed to lead in a smelting factory were investigated. The concentration of lead in air at workshop was measured by flame atomic absorption spectrophotometry (FAAS) and the levels of blood lead (PbB) by atomic absorption spectrophotometry (AAS), zinc protoporphyrin (ZPP) by ZnPP meter, as well as the indexes of thyroid function, thyroid-stimulating hormone (TSH), triiodothyronine (T(3)), thyroxin (T(4)), free T(3) (FT(3)), and free T(4) (FT(4)) in serum by radioimmunoassay.

RESULTSThe workers with higher level of blood lead (> 2.88 micro mol/L) showed lower levels of T(3) [(1.54 +/- 0.39) nmol/L] and FT(3) [(5.50 +/- 1.26) pmol/L] than those with lower blood lead level [PbB: (1.92 approximately 2.88) micro mol/L group, T(3): (1.71 +/- 0.45) nmol/L, FT(3): (6.12 +/- 1.64) pmol/L, P < 0.05]. There was no obvious effect of length of service on thyroid hormone of exposed workers.

CONCLUSIONHigher level of blood lead may cause certain damage to thyroid function by inhibiting deiodination of T(4). No obvious relation between length of service and thyroid function was found.

Adult ; Female ; Humans ; Lead ; blood ; toxicity ; Male ; Middle Aged ; Occupational Exposure ; Thyroid Gland ; drug effects ; physiology

1,3-propanediol (1,3-PD) is an important material for chemical industry, therefore, there is much interest in the production of 1,3-PD. The gene dhaT encoding 1, 3-propanediol dehydrogenase ( PDOR) of Citrobacter freundii was amplified by PCR. Sequence analysis of the similarity at the nucleotide and amino acid level between the gene encoding C. freundii PDOR and that of C. freundii ( U09771 ) were 78% and 90% , respectively. The recombinant plasmid pSE-dhaT was constructed by inserting dhaT gene into expression vector pSE380 and then transformed E. coli JM109. The recombinant strain was induced by IPTG to express dhaT. Further more the recombinant enzyme was purifed from recombinant E. coli by Ni-nitrilotriacetate affinity chromatography followed by Sephacral S-300 gel filtration. A single obvious protein about 42kDa could be obtained by the sodium dodecyl sulfate-polyacrylamide gel electrophoresis of recombinant enzyme. The purified enzyme was used to determined enzyme property on the substrate of propionaldehyde and 1, 3-PD. The optimal temperature and optimal pH of the purified enzyme were 37℃, 8.0 for reduction and 25℃, 10. 5 for oxidation, respectively; and the kinetic property of PDOR about Km and V max were 10. 05mmol/L, 37. 27?mol/min/mg for propionaldehyde and 1. 28mmol/L, 25. 55?mol/min/mg for 1,3-PD, respectively; The deduced dhaT gene product (388 amino acids) showed a specific reduction activity of 49. 50U/mg and oxidation activity of 79. 92U/ mg. There also have a putative iron-binding motif ( G-XX-H-X-X-A-H-X-X-G-X-X-X-X-X-P-H-G) as a fingerprint pattern in the recombinant enzyme, the motif is fully conserved among these 1, 3-propanediol dehydrogenase. It is beneficial to the researches of high producing 1, 3-propanediol by gene engineering strain.

Recent studies suggested that the prostate cancer may arise from prostate cancer stem cells that share some same characteristics with normal stem cells.The purpose of this study was to detect the differences of Nanog expression between PC3 prostate cancer cell line and its tumor stem cells,and fhe relationship was preliminarily examined between Nanog and prostate cancer and its tumor stem cells.By using magnetic active cell sorting (MACS),we isolated a population of CD44+/CD133+ prostate cancer cells that display stem cell characteristics from PC3 cell line.lmmunohistochemistry revealed positive expressions of CD44,C D 133 and α2β1-integin in the isolated cells.CCK-8 analysis showed that isolated cells had a strong proliferafive ability.The formation of the cell spheres in serum-free medium and holoclones in serum-supplied medium showed that the cells were capable of self-renewing,indicating that the isolated cells were a population of cancer stem-like cells derived from PC3 cell line.Western blotting exhibited that the isolated cells had higher experession of Nanog,an embryonic stem marker,as compared with PC3 cells.Our study showed that Nanog might be helpful in sustaining the self-renewal and fhe undifferentiation of prostate cancer stem cells,and may serve as a marker for prostate cancer stem cells for isolation and identification.

The proteins encoded by the Herpesviridae β-gene play a critical role in the replication stage of the virus. In this paper, phylogenetic analyses provided evidence that someβ-gene products, such as UL2 and UL23 from HSV1, have their homologous genes in its family, and also exist in prokaryotic organisms, indicating that these viruses appear to have been assembled over evolutionary time by numerous independent events of horizontal gene transfer.

OBJECTIVETo detect the expression of FABP7 in the placenta of pregnant mice and in HTR-8/Svneo cells.

METHODSReal-time PCR and immunofluorescence were used to detect FABP7 mRNA and protein expressions in the uterine and placental tissue of pregnant mice at different days of gestation. FABP7 expression was also detected in cultured HTR-8/Svneo cells using immunofluorescence assay. The mice were treated with E, Por their combination for 6 and 24 h and Fabp7 mRNA level in the uterus was detected with real-time PCR.

RESULTSAt 7.5-10.5 days of gestation, the pregnant mice showed positive expressions of Fabp7 mRNA in the uterus and placenta, and FABP7 protein was detected in the decidualized cells and trophoblast giant cells. The expressions of FABP7 were detected at both the mRNA and protein levels in cultured HTR-8/Svneo cells. In mice treated with P4 alone or with E+Pfor 6 and 24 h, the expression level of Fabp7 mRNA was upregulated in the uterus. Fabp7 upregulation was observed in mice at 24 h following Etreatment but not at 6 h.

CONCLUSIONFABP7 is expressed in trophoblast giant cells and decidual cells in the placental tissue of mice and in cultured HTR-8/Svneo cells, suggesting the involvement of FABP7 in placental development and in maintenance of pregnancy. Eand Pcan regulate the expression of FABP7 in mouse uterus.

OBJECTIVETo investigate the effect of BIX01294 (BIX), a methyltransferase inhibitor, on the migration and decidualization of the stromal cells in mouse uterus.

METHODSMouse endometrial stromal cells were isolated and cultured from the uterus of pregnant mice on day 3.5 of gestation. The migration and decidualization of mouse endometrial stromal cells treated with BIX at different concentrations were observed with wound healing assay and real-time PCR.

RESULTSThe migration distance of mouse endometrial stromal cells increased as the BIX concentration increased within the range below 15 µmol/L. Compared with the control cells, the cells treated with BIX (15 µmol/L) showed significantly increased migration distances, but increasing BIX concentration to 20 µmol/L did not further increase the cell migration distance and began to cause cell death. Compared with the control cells, the BIX-treated stromal cells exhibited significantly down-regulated expression of Ehmt2 mRNA, and 15 µmol/L BIX caused inhibition of decidualization in the stromal cells.

CONCLUSIONWithin a defined concentration range, BIX promotes the migration and inhibits decidualization of mouse uterine stromal cells by inhibiting the expression of Ehmt2 mRNA.

OBJECTIVETo investigate the significance of three dimensional visualization and virtual surgery system in living related donor liver transplantation surgery.

METHODSTwo patients suffered biliary calculi were scanned by 64 slice helical computer tomography (CT) on livers and the data were imported into medical image proceeding system (MIPS) for sequence. Man-made segmentation and true-up on the image from the data were carried out. Three dimensional (3D) models of the liver and the intrahepatic vessels were reconstructed by VTK software respectively. The models were exported with format STL from it and then were imported into the FreeForm Modeling System for smoothing and modifying. At last, living related donor liver transplantation were simulated with the force-feedback equipment (PHANToM).

RESULTSIt had great verisimilar image for the reconstructed 3D liver models with artery, hepatic vein, portal vein and bile duct. By seeing through liver, it had high fidelity and strong 3D effect for the intrahepatic artery, hepatic vein, portal vein and bile duct, and their spatial disposition and course and co-relationship were shown clearly. In the virtual surgery system, the virtual scalpel could be manipulated on 3D liver model with PHANToM. The simulating effect was the same as the clinic operation for living related donor liver transplantation.

CONCLUSIONSThe visualized liver model reconstructed is 3D and verisimilar, and it is helpful to design reasonable scheme for liver transplantation. It can improve the surgical effect, decrease the surgical risk, reduce the complication, enhance the communication between doctor and patient through designing surgical plan and demonstrating visualized operation before surgery.

Adult ; Computer Simulation ; Female ; Humans ; Image Processing, Computer-Assisted ; Imaging, Three-Dimensional ; Liver ; diagnostic imaging ; Liver Transplantation ; Living Donors ; Models, Anatomic ; Tomography, Spiral Computed ; User-Computer Interface