Batch fermentation of lycopene by recombinant E. c ol i ZYL-2 at various temperatures ranging between 23℃～37℃ were studied in 7L fermentor. At 33℃, cell specific growth rate in earlier stage of culture was higher, and the time obtained maximum cell dry weight was shorter at other tempe rature. while after 9 h, lycopene specific production rate was higher at 28℃. Based on these results, a two-stage temperature control strategy was developed in which 33℃ was used for fermentation for the first 9 h and the temperaturew as switched to 28℃ after 9h. Using this temperature-shift strategy, the maxim al lycopene content and productivity reached 605.25?g L-1 and 28.82?g L-1h-1. The lycopene fermentation level obtained by the strategy was higher than those in single temperature-control experiments.

Objective To explore the MRI appearances of heterotopic gray matter. Methods The appearances of MRI of heterotopic gray matter(n=12) confirmed were analyzed retrospectively. Results In the 12 cases,3 lesions were nodular,9 lesions were lamellar. MRI can clearly show the lesions. On T1-WIs and T2-WIs,these lesions appeared isointense to the normal gray matter. 2 lesions associated with arachnoid cyst,2 lesions with schizencephaly and 1 lesion with dysgenesis of the corpus callosum and lipoma of midline. During contrast on MRI(n=5) ,all lesions show unenhancement. Conclusion The MRI have characteristics in diagnosis of heterotopic gray matter and is the best method.

Objective To observe the clinical efficacy of acupoint injection of mouse nerve growth factor in treating lumbar intervertebral disc herniation (LIDH).Method Totally 120 LIDH patients were randomized into a treatment group, control group 1, and control group 2, 40 cases in each group. Control group 2 was intervened by dehydration therapy; the treatment group was by acupoint injection of mouse nerve growth factor in addition to the intervention given to control group 2; control group 1 was by muscular injection of Trivitamin B in addition to the intervention given to control group 2. The common peroneal nerve and posterior tibial nerve conduction velocities and Visual Analogue Scale (VAS) were observed before and after treatment, and the clinical efficacies were compared among the three groups.Result The recovery and markedly effective rate was 97.5% in the treatment group, versus 92.5% in control group 1 and 85.0% in control group 2, and the rate in the treatment group was significantly different from that in control group 1 and 2 (P<0.05). Respectively 14 d and a month after intervention, the VAS scores were significantly different from that before treatment in the three groups (P<0.05). Respectively 14 d and 1 month after the intervention, the VAS score in the treatment group was significantly different from that in control group 1 and 2 (P<0.05). The common peroneal nerve and posterior tibial nerve conduction velocities a month after the intervention were significantly different from that before the intervention in the three groups (P<0.05). A month after the intervention, the posterior tibial nerve and peroneal nerve conduction velocities in the treatment group were significantly different from that in control group 1 and 2 (P<0.05). Conclusion Acupoint injection of mouse nerve growth factor is an effective way in treating LIDH.

BACKGROUND: Previous researches indicate that ADp14ARF transfecting positive tumor cell line of p53 can inhibit the proliferation; in addition, the inhibitory effect is superior to transfection negative tumor cell line of p53. Whether simultaneous transfection of p14ARF and p53 genes can increase expression and accumulation of p53 and accelerate apoptosis of tumor cells needs further studies.OBJECTIVE: To construct double plasmid expression vector plRES-p14ARF-p53 by using gene engineering so as to observe the inhibitory effect on proliferation of osteogenic sarcoma cells.DESIGN: Randomized controlled observation.SETTING: Department of Orthopaedics, Union Hospital Affiliated to Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: The experiment was carried out in the Public Laboratory Platform, Immune Researching Room, Basic Medical College, Tongji Medical College, Huazhong University of Science and Technology from January 2005 to October 2006. Human osteogenic sarcoma MG-63 cells were provided by Cell Laboratory, Immune Researching Room, Tongji Medical College, Huazhong University of Science and Technology. plRES-p53 plasmid and plRES vector containing p53total-length gene order were provided by Wuhan Jingsai Biology Company.METHODS: Based on gene engineering, p14DNA (0.5 kb) was amplified from cultured L02 cells of normal human hepatic cells into plRES vector. Recombinant plasmid plRES-p14ARF-p53 was determined with polymerase chain reaction (PCR) and restriction enzyme and transfected into human osteogenic sarcoma MG-63 cells through mediation of liposome to screen positive clones. Otherwise, cells were divided into three groups, including blank control group (MG-63cells), blank vector control group (stably transfecting plRES-neo cells) and p14ARF-p53 group (stably transfecting plRES-p14ARF-p53 cells). ① DNA content and cycle of tumor cells were measured by using flow cytometry before and after transfection. ② Reverse transcription polymerase chain reaction (RT-PCR) and Western blot were used to detect quantitative and semi-quantitative expression of p53 and p14ARF protein in tumor cells after stable transfection. ③Thiazole blue chromatometry and growth curve were used to observe proliferation.MAIN OUTCOME MEASURES: ① DNA content and cycle of osteogenic sarcoma cells; ② expressions of p53 and p14ARF protein in tumor cells; ③ proliferation.RESULTS: Double plasmid expression vector plRES-p14ARF-p53 was constructed successfully. ① DNA content and cycle of osteogenic sarcoma cells: Flow cytometry demonstrated that tumor cells mainly stayed in G1 phase after transfection. ② Protein expression: RT-PCR and Western blot indicated that p14ARF and p53 gene independently expressed in target cell mRNA and protein, respectively. ③ Cell growth: At 24, 48, 72 and 96 hours after MG-63 transfection, inhibitory rates of tumor cells were 33.43%, 69.37%, 66.19% and 75.26%, respectively, which was significant difference as compared with blank vector control group (P ＜ 0.01).CONCLUSION: Wild p53 and p14ARF can synergistically inhibit the proliferation and accelerate the apoptosis of osteogenic sarcoma cells.

Objective To investigate the effectiveness and safety of combined heparin and acutobin in the treatment of acute inferior vena cava thrombus in rabbits. Methods The inferior vena cava thrombus model was established in 72 rabbits and they were randomly divided into three groups; heparin group(A) , group for combination of urokinase and heparin (B), group for combination of acutobin and heparin (C) ,each group including 24 rabbits. Drugs were administrated 3 days after thrombosis. Coagulation indexes were tested to assess their safety, and Doppler ultrasound was used to assess their effectiveness, on day 3, day 7, and day 10. Results The prolongation of prothrombin time ( PT) in group C was shorter than that in group B( P < 0. 05 ) , the fibrinogen ( FBG) value in group C was lower than that in group B (P < 0. 05 ) , the prolongation of PT in group B and group C was longer than that in group A (P < 0. 01), the FBG value of group B and C were higher than that in group A ( P < 0. 01 ), D-dimer ( D-D) value in group B and C gradually returned to normal range. There was no difference between the two groups (P > 0. 05). The thrombolytic effect in group B and C were better than that in group A, statistical difference was reached between groups B and A (P <0. 01), and the difference was statistically significant between groups C and A 10 days after administration (P < 0. 01). Thrombolytic effect was not different statistically between groups B and C (P > 0. 05). Conclusion Acutobin combined with heparin in the treatment of acute inferior vena cava thrombus in rabbits was effective and safe.

Animals ; Antioxidants ; physiology ; Dehydroepiandrosterone ; physiology ; Lipid Metabolism ; physiology ; Male ; Oxidative Stress ; drug effects ; Rats ; Rats, Sprague-Dawley

Objective To evaluate the changes in the expression of acetylated histone H3 (Ac-H3) and deacetylase silent information regulator 1 (SIRT1) in dorsal root ganglions in a rat model of negative phenotype neuropathic pain.Methods Forty-eight male Sprague-Dawley rats,weighing 250-300 g,were randomly divided into 2 groups (n =24 each):sham operation group (group S) and C-fiber dysfunction group (group CFD).The rats were anesthetized with 10％ chloral hydrate 3 ml/kg.C-fiber dysfunction was induced by exposing sciatic nerve to 8％ capsaicin for 30 min in group CFD.The thermal withdrawal latency (TWL) and mechanical withdrawal threshold (MWT) were measured before and on 1,3,7 and 14 days after CFD.Six rats were then sacrificed at each time and the lumbar segments (L5) of the dorsal root ganglions were removed for detection of SIRT1 mRNA expression (by RT-PCR) and Ac-H3 and SIRT1 protein expression (by Western blot).Results Compared with group S,TWL was significantly increased at 1,3,7 and 14 days after CFD,SIRT1 mRNA and protein expression was up-regulated and Ac-H3 expression was down-regulated at 3,7 and 14 days after CFD (P ＜ 0.05),while no significant change was found in MWT at each time point in group CFD (P ＞ 0.05).Conclusion The mechanism of negative phenotype neuropathic pain is related to up-regulation of deacetylase SIRT1 expression and decreased acetylation of histone H3 in rat dorsal root ganglions.

Using Vibrio fluvialis Ⅱ as host bacteria, 19 bacteriophages have been isolated from the 76 samples which were collected from Haliotis discus hannai ～growing seawater in Dalian marine culture company Dalian, liaoning province from May in 1996 to August I 1997. Ultrastructure of 19 bacteriophages were observed with electron to Bradley the results showed that of these bacteriophages belonged to Bradley A type, they have hexagonal heads of bacteriophages were identified with VP1,VP2,VP4,VP8 as representatives respectively. The phages remain stable at pH6. 0～10. 0, moreover VP2,VP4 and VP8 are rather stable at basic pHs. Although the characterization of heat inactivation course of VP4 is different from others, four phages are sensitive to heat and inactivated at 80℃ in 5 minutes. One step growth experiment showed that the eclipse period of VP1,VP2,VP4,VP8 are sensitive to heat and the eclipse period of VP1, VP2, VP4, VP8 are 42, 30, 46, 28 minutes. In this experiment we have isolated at least four different types phages, it suggest that in fact there is a population of phages in the seawater environment. The result of this study provided a way to find the potential value of phages as an indicator of pathogenic microorganisms Vibrio fluvilis Ⅱ in marine environment.

Objective To observe the effect of δ-opioid receptor on proliferation and migration of human epidermal stem cells (hESCs) in vitro so as to offer treatment theory for skin injury.Methods hESCs from fresh foreskin tissues of normal young volunteers were isolated and cultured by enzyme digestion and differential adherence technique.Immunofluorescent staining was used to determine expression of integrin β1 and cytokeratin 19 (CK19) and flow cytometry was used for cell count.Second generation of cells were cultured for 5 consecutive days with keratinocyte serum-free medium (K-SFM) supplemented with 1 nmol/L (D-Ala2,K-Leu5)-enkephalin in Group A,with K-SFM supplemented with 1 nmol/L naltrindole and 1 nmol/L (D-Ala2,K-Leu5)-enkephalin in Group B,and with isolated K-SFM in Group C.Cellular division and proliferation were detected by MTT method.An in vitro 100 μm scratch-wound model was created on the confluent monolayer cells at 24 hours of incubation.Cells migrating from the wound margin were determined by inverted phase contrast microscope at 24,48,72,and 96 hours after wound formation,while wound closure rate was calculated at 72 hours.Results Primary cultured hESCs presented cobblestone-like shape after adherence growth,Immunofluorescence staining showed positive results for integrin β1 and CK19 and cell purity reached 95.6％.Moreover,MTT findings revealed proliferation of hESCs enhanced significantly in Group A,but lowered in Group B as compared to Group C (P ＜ 0.05).hESCs migrated from the wound margin in all groups at 24 hours.However,more migrated cells were seen in Group A than in Group C and less in Group B than in Group C.Rate of wound closure was (89.5 ±0.7)％ in Group A,(76.1 ±0.3)％ in Group B,and (81.1 ±0.6)％ in Group C at 72 hours,indicating significant differences among groups (P ＜ 0.05).Conclusion Activation of δ-opioid receptor promotes the proliferation and migration of hESCs in vitro and may be implicated in wound healing.

Objective To study the effect of propofol and midazolam on memory during target control infusion for sedation.Methods Forty healthy male volunteers were randomly devided into 4 groups,all subjects received target controlled infusion with propofol or midazolam for sedation.Group 1 and group 3 were sedated to OAA/S 1 or OAA/S 3 respectively by propofol;group 2 and group 4 was sedated to OAA/S 1 or OAA/S 3 by midazolam.All subjects learned a list of words before sedation and at the different sedation depths predicted.After recovery the explicit and implicit memory were tested using word stem completion method and process dissociation procedure.Results Compared with 0,there was no significant difference in explicit memory among all the 4 groups.While there was significant difference in implicit memory for group 3 and 4,and there was no significant difference in implicit memory for group1 and group2.Conclusion Sedative-hypnotic drugs not only deepen sedation,but also impair memory as serum concentration increases during target infusion.Drug-induced amnesia is independent of sedation.Propofol and midazolam have the same efficacy on memory at the same sedation scale.