Animals ; Hypoxia ; metabolism ; physiopathology ; Insulin-Like Growth Factor I ; genetics ; metabolism ; Male ; Muscle, Skeletal ; metabolism ; Myostatin ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley

OBJECTIVEThe study was to verify the feasibility of an improved method using reverse flow island flap nourished by the sural nerve nutrition vessel to repair severe frostbite of feet.

METHODSAt the proximal end of the principal flap, an auxiliary triangular skin flap of 6.5 - 7 cm in length was designed in order to cover the pedicle of the principal flap. This operation was performed on 13 patients (21 feet) with frostbite.

RESULTSAll the flaps survived well. Postoperative follow-up for 5 - 18 months demonstrated satisfactory results in all the cases. No ulcer happened.

CONCLUSIONSThe method is helpful to prevent constriction of the pedicle and ensure blood supply of the flap. It is an ideal treatment for severe frostbite of feet.

Adult ; Female ; Foot Injuries ; surgery ; Frostbite ; surgery ; Humans ; Male ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; Surgical Flaps ; blood supply ; innervation ; Young Adult

Glucose was transported by the large number of hexose transporters in yeast cells. There were 18 hexose transporter genes had been identified in Saccharomyces cerevisiae. However,as an excellent expression system,there was no information of these genes had been reported in Pichia pastoris. Based on high homologous recombination efficiency in yeast,we chose G418 resistance for screening,200 bp were cloned from the up and down sequences of HXT1 ORF respectively,then ligated to the 5′ and 3′ end of G418 resis-tance gene for recombination. After electroporation of GS115 spheroplast and screened through different G418 concentration plates,finally we obtained one HXT1 gene deletion mutant named GS115?HXT1. The growth rate and glucose consumption of this mutant were both lower than the wide type.

The rice water weevil (RWW) Lissorhoptrus oryzophilus Kuschel (Coleoptera: Curculionidae) is an invasive insect pest of rice Oryza sativa L. in China. Little is known about the interactions of this weevil with indigenous herbivores. In the present study, adult feeding and population density of the weevil, injury level of striped stem borer Chilo suppressalis (Walker) (Lepidoptera: Pyralidae) and pink stem borer Sesamia inferens (Walker) (Lepidoptera: Noctuidae) to rice, as well as growth status of their host plants were surveyed in a rice field located in Southeastern Zhejiang, China, in 2004 with the objective to discover interspecific interactions on the rice. At tillering stage, both adult feeding of the weevil and injury of the stem borers tended to occur on larger tillers (bearing 5 leaves) compared with small tillers (bearing 2~4 leaves), but the insects showed no evident competition with each other. At booting stage, the stem borers caused more withering/dead hearts and the weevil reached a higher density on the plants which had more productive tillers and larger root system; the number of weevils per tiller correlated negatively with the percentage of withering/dead hearts of plants in a hill. These observations indicate that interspecific interactions exist between the rice water weevil and the rice stem borers with negative relations occurring at booting or earlier developmental stages of rice.
Animals ; Coleoptera ; growth & development ; Oryza ; growth & development ; parasitology ; Population Density ; Weevils ; growth & development

Objective To improve the level of early detection and treatment of pyonephrosis. Methods This study included 41 cases(17 men and 24 women;mean age,49 years)of pyonephrosis.A variety of examinations,including urinary analysis,blood analysis,kidney nuclear medicine scan,ultrasonog- raphy,intravenous urography(IVU),and CT were used for the early diagnosis of pyonephrosis.Pereutaneous nephrostomy(PCN)drainage was done for the interim management of pyonephrosis,then phase 2 operation was performed in 28 cases.The double-J tube was placed in ureter by ureteroscope for drainage,and then phase 2 operation was done in 2 cases.Emergency operation was done in 10 cases.The remaining 1 case un- derwent ESWL after anti-infective therapy.Results Definite diagnosis of pyonephrosis before operation was made by invasive examinations in 31 cases(75.6%),and by percutaneous drainage in 4 cases;the other 6 cases were detected during operation.Only 6 cases(14.6%)underwent nephrectomy;the other 35 cases (85.4%)underwent kidney-sparing operation.Follow-up of 3 months to 9 years was available in 37 cases. No nephrectomy was needed in 33 cases with spared kidney.Serum creatinine was normal in the 4 cases un- dergoing nephrectomy.Conclusions The key to the treatment of pyonephrosis by kidney-sparing surgery is early diagnosis,timely drainage and relief of obstruction.Ultrasonography plays an important role in the early diagnosis of pyonephrosis,and CT has a high sensibility in the diagnosis.Pereutaneons nephrolithotomy (PCNL)secondary to drainage through pereutaneous nephrostomy was beneficial to the patients with kidney stones or upper ureter stones.

The GFP(green fluorescence protein)-streptavidin(SA) bi-functional fusion protein was generated and characterized in order to demonstrate novel platform for efficiently and durably modifying the cell surface with SA-tagged bi-functional proteins.The GFP-SA/pET24d construct was generated and expressed in BL21(DE3) host bacteria at the high level.The recombinant protein GFP-SA was purified through the Ni-NTA affinity chromatography,and then refolded.After biotinylation B16 tumor cells were modified with GFP-SA bi-functional fusion protein and then subjected to fluorescent microscopy and FACS analysis.The effect of surface modification on the viability and growth of B16.F10 tumor cells was evaluated by MTT staining.The GFP-SA recombinant fusion protein was expressed in BL21(DE3) at about 20 % of total bacterial proteins.The GFP-SA bi-functional fusion protein exhibited the bi-functionality,i.e.,SA-mediated high-affinity binding to biotinylated cell surfaces and GFP-emitted green fluorescence.The cell surface modification with GFP-SA bi-functional fusion protein did not affect the viability and growth of the modified B16.F10 tumor cells significantly.The GFP-SA bi-functional fusion protein was obtained and could be displayed efficiently on the surface of the biotinylated B16.F10 tumor cells through the specific and tight interaction between streptavidin and biotin,thus can be used as good trace protein and experimental control in the development of surface-modified tumor vaccine.

The aim of this study was to investigate the feasibility of the human cord blood adult stem cells (ASCs) to differentiate into hepatocytes in vitro induced by combined stimulation with hepatocyte growth factor (HGF), stem cell factor (SCF) and leukemia inhibitory factor (LIF). The adult stem cells were obtained through density gradient centrifugation and magnetic activated cell sorting (MACS). The adult stem cells were cultured in DMEM with HGF (10 ng/ml)+SCF (10 ng/ml)+LIF (10 ng/ml) in induced group I. In induced group II the enriched cells were cultured in DMEM with SCF (10 ng/ml)+LIF (10 ng/ml) and the undifferentiated cells acted as the control group without the factors. The morphology of cells was observed by the inverted phase contrast microscopy; the expression of albumin (Alb), human hepatocyte cytokeratin (CK18) and alpha-fetoprotein (AFP) were detected by immunofluorescence, immunohistochemistry and RT-PCR assay in the 21-day culture. Alb secreted by hepatocytes in the medium was determined by radioimmunoassay (RIA) at day 7, 14, 21, 23 and 25. The results showed that the shapes of ASCs changed and their sizes and number increased in the course of culture in group I. After being induced for three weeks, the cells turned round and resembled hepatocyte-like cells. The mRNA for Alb could be detected by RT-PCR in the differentiated adult stem cells in group I, and the mRNA for AFP was poorly detected by RT-PCR at day 21. Alb and CK18 were positive through immunofluorescence and immunohistochemistry at day 21, compared with group II and the control group. In group I, Alb in the medium significantly increased, compared with control group, and reached the highest level at day 21, then decreased at day 23. It is concluded that under some definite inducing conditions, human cord blood adult stem cells can differentiate into hepatocyte-like cells and HGF plays a critical role during the course.
Adult Stem Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Fetal Blood ; cytology ; Hepatocyte Growth Factor ; pharmacology ; Hepatocytes ; cytology ; Humans ; Leukemia Inhibitory Factor ; pharmacology

OBJECTIVETo investigate the clinical value of matrix assisted laser desorption ionization-time of flight-mass spectrometry (MALDI-TOF-MS) in detecting K-ras gene mutation.

METHODSSixty-one paraffin-embeded specimens of colorectal cancer were selected. MALDI-TOF-MS and regular sequencing were used to test the mutation of codon 12 and 13 in K-ras exon 2.

RESULTSOnly 47 specimens could be detected successfully in regular sequencing, while all the specimens were tested successfully in MALDI-TOF-MS. Fourteen specimens had K-ras mutation in regular sequencing (30.0%), while 22 specimens had mutation in MALDI-TOF-MS (36.1%). Six specimens with mutation were found in MALDI-TOF-MS but were wild-type in regular sequencing. Same mutation types from 14 specimens were confirmed by both regular sequencing and MALDI-TOF-MS. MALDI-TOF-MS was able to detect the mutation in 2 specimens that was not identified in regular sequencing.

CONCLUSIONSMALDI-TOF-MS is a feasible approach of K-ras gene mutation testing in colorectal cancer, which is less demanding in terms of specimen quality and is more sensitive as compared to regular sequencing.

Colorectal Neoplasms ; genetics ; Genes, ras ; Humans ; Mutation ; Sensitivity and Specificity ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods

AIMIn order to investigate the effect of mesenchymal stem/progenitor cells on proliferation and differentiation towards megakaryocytes of CD34+ cells from human umbilical cord blood in vitro.

METHODSAfter mesenchymal stem/progenitor cells were advancely planted in DMEM medium and grown up to 80%, then the CD34+ cells were added to culture with mesenchymal stem/ progenitor cells or without mesenchymal stem/progenitor cells in DMEM for 14 days with TPO + IL-3 + SCF, TPO + IL-3 + SCF + IL-11 respectively. After cultured for 14 days, mononuclear cells (MNCs) were counted by automatic cell analyzer. The number of CD41+ cells and platelets were detected by flow cytometry. Platelets function were assessed through platelet aggregation test which was induced by thrombin.

RESULTSAs compared with the control group, the number of MNCs of co-culture system was not increased significantly (P > 0.05), but the number of CD4+ cells and platelets were increased significantly (P < 0.05). The platelets were aggregated by thrombin induced which could be seen in microscope or flow cytometry.

CONCLUSIONIt is concluded that mesenchymal stem/progenitor cells may be promoted to induce the cord blood CD34+ cells to differentiate towards megakaryocyte in the culture medium.

Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; Cell Differentiation ; physiology ; Cells, Cultured ; Fetal Blood ; cytology ; Humans ; Megakaryocytes ; cytology ; Mesenchymal Stromal Cells ; cytology ; physiology

OBJECTIVETo explore the mechanism of reactive oxygen species (ROS) in manganese chloride (MnCl(2))-induced apoptosis in PC12 cells.

METHODSThe model that MnCl(2) induced apoptosis in PC12 cells was established. The apoptotic effect of MnCl(2) on PC12 cells was analyzed with the MTT, the flow cytometry and the DNA fragmentation. The production of ROS and ATP in MnCl(2)-induced apoptosis of PC12 cells was examined. The influence of MnCl(2) on the expression of bcl-xl, bax and the activity of Caspase 3 was also analyzed.

RESULTSMnCl(2) triggered PC12 cells apoptosis in a dose-and time-dependent manner (P < 0.01). The rate of apoptosis was significantly increased (P < 0.01) when MnCl(2) of 2 mmol/L induced PC12 cells for 36 hours. The production of ROS was increased (P < 0.001) and the quantity of ATP was decreased (P < 0.01) in PC12 cells with the same inducement of MnCl(2). The expression of bcl-xl was inhibited and the bax was activated in this process (P < 0.01). Caspase 3 was also activated (P < 0.01).

CONCLUSIONMnCl(2) induces apoptosis of PC12 cells, which is related to the increase of ROS, the inhibition of the mitochondria and the activation of Caspase 3.

Adenosine Triphosphate ; biosynthesis ; Animals ; Apoptosis ; drug effects ; Caspase 3 ; metabolism ; Chlorides ; toxicity ; DNA Fragmentation ; drug effects ; Manganese Compounds ; PC12 Cells ; Rats ; Reactive Oxygen Species ; metabolism ; bcl-2-Associated X Protein ; biosynthesis ; bcl-X Protein ; biosynthesis