Bile Duct Neoplasms ; surgery ; Bile Ducts ; pathology ; surgery ; Bile Ducts, Intrahepatic ; pathology ; surgery ; Biliary Tract Diseases ; surgery ; Biliary Tract Surgical Procedures ; methods ; trends ; Cholecystectomy, Laparoscopic ; methods ; trends ; Cholelithiasis ; surgery ; Humans

Objective To search for a good method for generation of hematopoietic stem cells (HSC) by embryonic stem cells (ESC),and to investigate the potential of HSC derived from ESC to reconstruct hematopoiesis in vivo.Methods Using a three-step method to induce a mice ESC line, E14.1-into HSC.And identifying HSC by flow cytometry analysis cell markers with CD34 +/Sca-1+, then HSC (1×109L-1)from ESC differentiation were injected into severe combined immunodefieency (SCID) mice for observing teratoma formation.To validate function of HSC by colonngenic cells assay and to reconstitute the hematopoiesis in lethally irradiated mice.Results Combining to use more hematopoietic stimulating factor to availably promote the El4.1 cell into embryoid body (EBs) with abundant hematopoietic progenitors.EBs were induced after 14 day to fast differentiate for HSC with peak percentages of CD34+/Sca-1+ cells reached to (13.72±2.07)%.To harvested ceils from EBs by day 14 for second -step HSC differentiation, percent of CD34+/Sca-1+ cells rise to(24.62±2.50) % after day 16 induction.Cloning forming units (CFU) analysis showed that more Erythro -myeloid cloning generation was observed at this period.Cells obtained in the second step are subsequently plated on monolayer of mice bone marrow stromal cells, in the presence of TPO, FLt3 ligand and superoatant of mice fetal liver stromal cells, cultured for additional 15 days, followed fast expansion of CD34+/Sca-1+ to maximally (58.64±4.20 ) % with more CFU-E, CFU-GM and CFU-GEMM population.Wright-Giemsa stain showed that its had the character of hematopoietic progenitors.Cells from the third-step were injected into SCID mice, but no teratomas were recovered in 2 mices after 6 weeks.Positive selection of CD34+/Sca-1+ cells by magnetic sorting from the third - step differentiation were transplanted into 7 lethally irradiated female mice while predominant hematopoietic reconstitution were observed in 10 days after transplantation, with 71.4% successful engraftment rate.And 3 recipients showed that the cell population of the peripheral blood leukocytes,red cells, hemoglobin approached normal index at 40 days after transplant, hut followed relative'slow renew in platelet count.Survival rate of transplant group is 43.0%, compared to 100% mortality in control mice.Karyotyping assays confirmed female mice with XY.Conclusions The results showed that the three-step differentiation and the culture conditions described here good support differentiation of ESC into HSC.HSC derived from ESC were safe without teratomas formation in body, and its can reconstruct hematopoiesis.

Hepatectomy ; methods ; trends ; Humans ; Liver ; surgery

OBJECTIVE: To observe the chemical groups changing in rat kidney with regard to fatal hyperthermia by Fourier transform infrared microspectroscopy (FTIR-MSP) and to provide a new method to diagnose fatal hyperthermia. METHODS: Rats were sacrificed by hyperthermia, brainstem injury, massive hemorrhage and asphyxiation and divided into groups. The renal samples were dissected immediately after death. The data of infrared spectroscopy in glomerulus were measured by FTIR-MSP. RESULTS: The absorbances of 3290, 3070, 2850, 1540 and 1396 cm(-1) significantly increased (P < 0.05), and the ratios of Al650/A3290 and A1650/A1540 significantly decreased (P < 0.05) in group of hyperthermia. CONCLUSION FTIR-MSP can analyze the changes of chemical groups of kidney as an auxiliary diagnosis for discriminating hyperthermia with other causes of death.
Animals ; Fever/mortality* ; Fourier Analysis ; Kidney/metabolism* ; Microspectrophotometry ; Rats ; Spectroscopy, Fourier Transform Infrared/methods*

OBJECTIVETo evaluate and generalize the application of intermaxillary fixation screw in the jaw fracture.

METHODS41 cases of jaw fracture have been treated with intermaxillary fixation screw.

RESULTSBoth function and appearance have recuperated in 40 cases except 1 case has light malocclusion.

CONCLUSIONSApplication of intermaxillary fixation screw advanced the traditional therapy.

Adolescent ; Adult ; Bone Screws ; standards ; Female ; Humans ; Jaw Fractures ; surgery ; Male ; Maxillary Fractures ; surgery ; Middle Aged ; Orthopedic Fixation Devices ; standards ; Reconstructive Surgical Procedures ; instrumentation ; methods ; Treatment Outcome

Objective To study the expressions of erythmpoietin(EPO)and its receptors(EPOR)in the injured brain tissue ofthe rats with traumatic brain injury(TBI).Methods A total of78 SD rats were randomly divided into three groups including control group(six rats),sham group(36rats) and fluid percussion injury group(36 rats).The rats were sacrificed at 6,24 hours,3,5,7 and 14days after TBI in the sham group and the fluid percussion injury group(six rats at each time point).Then,the injured brain tissues were removed for observation of the mRNA and protein expressions of EPO and EPOR by meaDiB of real-time PCR and Western blot. Results The expression of EPO was increased at 24 hours and reached the peak at day 3 after TBI.The hish expression level of EPO could maintain for two days or so.began to decrease at day 7 and recovered to normal at day 14 after Till.While the expression of EPOR reached the peak at 24 hours after TBI and maintained hish level at day14. Conclusions The expressions of EPO and EPOR show increase within 24 hours after TBI.In fact,the expressions of both factors are not in consistency,with more transient expression of EPO.

Objective To investigate the effect and significance of microglia/macrophage activation prior to cerebral ischemic preconditioning (CIP) in regulating toll-like receptors 2 (TLR2)/nuclear factor-kappa B (NF-κB) inflammatory signaling pathway in early stage after ischemic brain injury in rats.Methods Thirty healthy male SD rats were selected and divided into normal control group,sham operation group,ischemia group,intervention group and treatment group according to random number table,with six rats per group.A rat model of focal permanent cerebral infarct was established by occlusion of middle cerebral artery (MCAO).CIP was performed by local ischemia-reperfusion.Minocycline was used to inhibit microglia/macrophage activation after CIP.Features of microglia/macrophage activation after CIP were detected by immunofluorescence; mRNA expressions of predominant factors (NF-κB inhibitor α,IκB-α;tumor necrosis factor α,TNF-α) of TLR2/NF-κB inflammatory signaling pathway in parietal cerebral cortex by in situ hybridization method; death rate by Kaplan-meier survival curves; neurological deficits by a 5-point neurological scale; brain infarct size by triphenyl tetrazolium chloride (TTC) staining.Results Microglia/macrophage started activation at one hour after cerebral ischemic injury in preconditioning group and presented a significant increase at 12 hours.Speed and range of activation were higher in preconditioning group than in ischemic group.IκB-α mRNA in preconditioning group started expression at one hour.TNF-α mRNA in preconditioning group remained a low expression in 12 hours and had a significantly lower peak value as compared with that in ischemic group (P ＜ 0.05).CIP increased rat survival rate significantly,improved nerve function and reduced infarction size when compared with the ischemia group (P ＜ 0.05).Minocycline inhibited nerve protection by CIP significantly (P ＜0.05).Conclusion CIP induces rapid activation of microglia/macrophage in early period of rat cerebral ischemic injury and provides brain protection probably via inhibition of TLR2/NF-κB activity and inflammatory overreaction to cerebral ischemia.

The biochemical mechanism of degrading keratins by S.fradiae var S-221 was primarily studied.The compounds (Na_ 2 SO_ 4 , Na_ 2 SO_ 3 and sulfdryl acohol), which respecitively enhance specific activity of keratinase, activate keratinase intensively and mainly act on the disulfide bonds reductase in the keratinase, Na_ 2 SO_ 3 activates intensively both disulfide bonds reductase and polypeptide hydrolytase at 0.01 mol/L, whereas Na_ 2 S_ 2 O_ 3 , which acts on the disulfide bonds reductase, inhibits keratinase.On the condition that substrate, keratins exists, S.fradiae var S-221 is induced to produce exo-keratinase, which is a multiproteinase, containing disulfide bonds reductase, which is a key enzyme degrading keratins, then, with polypeptidic, hydrolytase, graduately hydrolyzates denatured keratins into polypeptides, oligopeptides and free amino acids, so that keratins have been decomposed completely.Sulfur in the keratins was transferred into sulfhydryl compounds, H_ 2 S and sulfates in the course of keratinolysine.

PCR-SSCP(single-strand conformation polymorphism) was used for studying the community changes of pronucleus microorganisms in various fermenting stages of Luzhou-flavor Daqu. The results showed:(1) The pronucleus microorganism's community was similar and also had the polymorphism in each simple of various fermentation stage;(2) Different microflora had complex ecology effects of coordination and the restriction;(3) The diversity indexes of different stage of Daqu microorganisms were around 1.69～2.01,and the composition of them was stable relatively;(4) The similarity indexes were 0.67～1.00,and much higher in the approaching stages.

Biometry ; Clinical Protocols ; Cryoultramicrotomy ; methods ; Diagnostic Techniques and Procedures ; Frozen Sections ; utilization ; Humans ; Orthopedics ; methods