R04, a Gram-positive bacterium, which can use biphenyl as the sole carbon source, was isolated from soil contaminated with oil in northern China. The bacterium has high biphenyl degradation efficiency and also can degrade polychlorinated biphenyl congeners, Aroclor1221 and Aroclor 1242. The bacterium was identified as Rhodococcus pyridinovorans by the method of 16S rDNA gene sequencing (accession No. AY072745).

The directed molecular evolution of enzyme in vitro can not only improve the efficiency of evolution, but also evolve enzyme according to the investigator's desire. This review summed up the feasible methods of this novel technique.

Objective To investigate the role of microsatellite instability(MSI) in the carcinogenesis and development of gastric cancer (GC). Methods MSI was examined in 36 gastric cancer specimens obtained endoscopically and during surgery and in 30 intestinal metaplasia specimens using PCR SSCP methods. Results MSI was detected in 15 of 36 GC and 9 of 30 intestinal metaplasia specimens at one or more loci. MSI was positive in all three cases of early GC. The incidence of MSI in well differenciated GC was significantly higher than that in poorly differentiated GC (66.7% vs 26.3%, P

Bone Marrow Cells ; Cell Line ; Cell Proliferation ; Humans ; Mesenchymal Stromal Cells ; cytology ; T-Lymphocytes ; cytology ; T-Lymphocytes, Regulatory ; cytology

Contraindications ; Humans ; Laryngeal Neoplasms ; classification ; pathology ; surgery ; Laryngectomy ; methods ; Neoplasm Staging

BACKGROUND:The treatment of infected nonunion is a difficult problem in orthopedic trauma, which needs to completely eliminate foci of osteomyelitis to control infection, repair soft tissue defect and close the wound, repairs bone defects to promote healing and protect limb function as much as possible. Autogenous micromorselized bone can be easily obtained and recombinant human bone morphogenetic protein-2 has variety of sources. Their mixed grafting mode is safe and reliable in repairing of bone defects, which are suitable for repair of infected nonunion. ＜br> OBJECTIVE:To study the operating method and clinical results of repairing infected nonunion using autogenous micromorselized bone grafting mixed with bone morphogenetic protein-2. ＜br> METHODS:Twenty-three cases underwent infected nonunion repair with autogenous micromorselized bone (l ium, long bone metaphyseal cancellous bone) mixed with bone morphogenetic protein-2, including 7 cases of femoral bone defect caused by osteomyelitis after debridement and external fixation and 16 cases of tibial bone infected nonunion. Eleven of 23 cases continued to use the external fixation because of scar contraction or poor conditions of soft tissue, and the other 12 cases switched to use internal fixation. X-tray was taken to evaluate the bone healing at the 1st, 3rd, 6th and 12th months after operation. The function of the affected extremity was assessed by Johner-Wruhs grade. ＜br> RESULTS AND CONCLUSION:Al of the cases were averagely fol owed up for 13.4 months. The bone defects and bone fracture healed in al cases without infection, and the wounds were al sealed. The excellent and good rate according to Johner-Wruhs grade was 95%. These findings indicate that the bone grafting mode of autogenous micromorselized bone mixed with bone morphogenetic protein-2 is characterized as variety of sources, high safety and reliable osteogenic ability in repair of bone defects.

Acute Disease ; Adult ; Humans ; Male ; Occupational Diseases ; diagnosis ; therapy ; Phosphines ; poisoning ; Poisoning ; diagnosis ; therapy

OBJECTIVETo explore the effect of drug-containing serum of Chinese herbal compounds [Xiongshao Capsule (XS, for activating blood) and Huanglian Capsule (HL, for dispelling toxin)] on tumor necrosis factor-alpha (TNF-alpha)-induced adherence between human umbilical vein endothelial cells (HUVECs) and polymorphonuclear neutrophils (PMN), inflammatory reaction and expression of related proteins in mitogen-activated protein kinase (MAPK) pathway.

METHODSThirty-two rats were randomly divided into four groups (8 in each group) using random digit table: the blank control group treated with distilled water, the test group I treated with Chinese herbal compound of XS (0.135 g/kg), the test group II treated with Chinese herbal compound of HL (0.135 g/kg), and the test group Ill treated with Chinese herbal compound of XS (0.135 g/kg) and HL (0.135 g/kg). All medication was given by gastrogavage once a day for a week. Rats' blood serum was harvested 1 h after the last administration to prepare drug-containing serum. HUVECs were exposed to TNF-alpha (100 ng/mL) to induce cell injury model and incubated with corresponding drug-containing serum (10%) for 24 h. Normal rats' serum was given to cells in the blank control group and the model group, while XC + HL containing serum was given to cells in the rest 3 groups. The adherence of HUVECs and PMN cells was detected by using rose bengal strain. Levels of E-selectin, intercellular adhesion molecule-1 (ICAM-1), and interleukin-1beta (IL-1P) in the supernatant of cultured HU-VECs were determined by ELISA. Protein expressions of mitogen-activated protein kinases p38 (p38MAPK) and extracellular signal-regulated kinase 1/2 (ERK 12) were determined by Western blot.

RESULTSCompared with the blank control group, HUVECs were seriously injured; PMN adherence amount significantly increased; levels of E-selectin, ICAM-1, and IL-1beta increased; expression levels of p-p38MAPK and p-ERK 1/2 in the supernatant of HUVECs significantly increased in the model group (all P < 0.01). Compared with the model group, HUVECs-PMN adherence amount decreased (P < 0.05); levels of E-selectin, ICAM-1, and IL-1 beta in the supernatant of HUVECs decreased (P < 0.01, P < 0.05); expression levels of p-p38MAPK and p-ERK 1/2 of endothelial cells decreased in the test group I, II, and III (P < 0.01).

CONCLUSIONSDrug-containing serums of activating blood, activating blood and dispelling toxin could attenuate TNF-alpha induced injury of HUVECs, inhibit HUVECs-PMN adherence and the release of adhesion factors. Its mechanism might be involved with protein phosphorylation of p38MAPK and ERK 1/2 in the MAPK pathway.

Animals ; Drugs, Chinese Herbal ; therapeutic use ; E-Selectin ; Endothelial Cells ; physiology ; Human Umbilical Vein Endothelial Cells ; Humans ; Inflammation ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-1beta ; Mitogen-Activated Protein Kinase 3 ; Neutrophils ; Rats ; Serum ; Tumor Necrosis Factor-alpha ; metabolism ; p38 Mitogen-Activated Protein Kinases ; metabolism

Objective To explore the feasibility of anti-85B and ESAT-6 monoclonal antibodies targeted contrast agent of CT by the murine acute tuberculosis animal model.Methods Preparation the targeted contrast agent of computed tomography by iodine atoms coupled with anti-85B and ESAT-6 murine monoclonal antibodies after purified.Calculate the label rate and the quality of 127Ⅰ of the targeted contrast agent solution,and dilute the contrast agent solution to the required concentration (5μg I/ml) to spare.There were twenty mice of acute tuberculosis animal model,which were divided into four groups by completely randomized digital table and each group was five animals.According to the different antibody named as 85B group and ESAT-6 group of targeted contrast agent,common contrast agent and blank control separately.The common contrast agent group was injected with diluents of iohexol,which was diluted into the same concentration with the targeted contrast agent.The control group was injected with antibody diluents pH 7.4 Phosphate Buffered Saline (PBS).All the animals were scanned before and after injection the contrast agent in different time,such as immediate,6 hours,12 hours and 24 hours.Observe the display and changes of the murine tuberculosis lesions,and measurement the CT value,which was regarded as evaluating mark.Enhancement ratio was also calculated.Two sample mean differences with t test and the multiple sample mean differences with ANOVA.Results The volume of anti-85B contrast agent solution was 2.52 ml,and the quality of antibody and 127Ⅰ were range from 210 to 255 μg and 10.5 to 16.6 μg respectively.The volume of anti-ESAT-6 contrast agent solution was 2.93 ml,and the quality of antibody and 127Ⅰ were 147 μg and 20.58 μg respectively.The lesions of the control group showed no visible density changes before and after injection of PBS.The CT value of the lung lesions in the targeted contrast agent group were gradually increased with time,and the lesion showed visible enhancement after the contrast injection twelve hours(t12),and also remained visible enhancement after injection twenty-four hours(t24)[85B group t12 =(-125.04 ± 13.58) HU,t24 =(-117.37 ± 12.28) HU and ESAT-6 group t12 =(-122.14 ± 19.01) HU,t24 =(-114.23 ± 17.08) HU],which is significant difference compared to the common contrast agent [t (85B-24 h) =4.05,t (ESAT-6-24 h) =6.39,P ＜ 0.05].Conclusions The targeted property of anti-85B and ESAT-6 murine monoclonal antibody contrast agent of CT had been partly proved by the acute tuberculosis animal model.Also provided an experimental basis for further study of tuberculosis targeted contrast agent.

Objective To investigate the relationship between expression of active form of caspase-3 and cell cycle in Fas-mediated apoptosis of B lymphocytoma cell line MML-1. Methods MML-1 cells were incubated with agonistic anti-Fas antibody for different time,and cell apoptosis was induced.Cell apoptotic rates were analysed by flow cytometry,and sensitivity of MML-1 cells to apoptosis was determined.The expression of active form of caspase-3 was analysed by double staining with PI-Triton X and FITC-active caspase-3.Cyclin A,B_1 and E were selected as cell cycle markers for S,G_2/M and G_1 phase of MML-1 cells,and the expression of active form of caspase-3 was detected by flow cytometry. Results The cell apoptotic rate reached 56% after induction by Fas for 6 h.After induction by Fas for 4 h,the active form of caspase-3 was mainly expressed in cells of G_1 phase,while rarely in cells of S and G_2/M phase.Cells with negative cyclin A and B_1 and positive cyclin E expressed active form of caspase-3. Conclusion The expression of active form of caspase-3 in MML-1 cells mediated by Fas might be cell cycle dependent.Cells entering into late G_1 and early S phase first express active form of caspase-3,and their sensitivity to Fas-mediated apoptosis is the highest.