Objective:To construct the eukaryotic expression vector of interleukin-18 gene and carry out analysis of pVAX1-IL-18 sequence . Methods:The recombinant eukaryotic expression vector pVAX1-IL-18 was constructed by inserting interleukin-18 gene into the eukaryotic expression vector pVAX1 with molecular cloning technique . It was confirmed by restrictive enzymes(BamHⅠ/ EcoRⅠ) digestion and analysis of DNA sequencing . Similar nucleotide sequences encoding IL-18 of pVAX1-IL-18 were looked up by Blast means in NCBI GenBank. Results:Restrictive enzymes digestion analysis and DNA sequencing results revealed that the interleukin-18 gene was cloned into the eukaryotic expression vector pVAX1 successfully .The sequence of encoding IL-18 of pVAX1-IL-18 was exactly the same as the reported sequence of encoding human IL-18 in GenBank . Conclusion: The recombinant eukaryotic expression vector pVAX1-IL-18 has been constructed successfully, which lay the foundation for pVAX1-IL-18 as a genetic adjuvant of DNA vaccine.

Increasing evidences have shown that intestinal microbiota may contribute to the etiology of colorectal cancer(CRC). Studies have shown some microbial species which can increase the risk of CRC. Intestinal bacteria can promote carcinogenesis by affecting DNA integrity,regulating immune reaction,inducing inflammation,increasing cell proliferation and altering stem cell dynamics. Modulation of intestinal microbiota may become an approach to prevent or treat CRC. This article reviewed the effect and mechanism of intestinal bacteria flora on CRC.

Objective To explore the possibility of human neutrophil peptide 1 (HNP1) gene engineering, we construct the eukaryotic expression vector carrying HNP1 gene. Methods With RNA extracted from human neutrophil cell as template, cDNA encoding mature HNP1 was amplified by RT-PCR, and then it was inserted into vector pMD18-T. After restriction endonuclease digestion and DNA sequencing confirmation the gene was subcloned into eukaryotic expression plasmid pcDNA3.1/V5-His-TOPO to construct a recombinant expression plasmid pcDNA3.1/V5-His-TOPO/HNP1, then the recombinant plasmid was transfected into COS-7 cells by lipofectamine, and the expressed product was identified by biotin-avidin enzyme-linked immunosorbent assay ( BA-ELISA). Results The sequence of HNP1 completely matched those published in GenBank, thus eukaryotic expression vector pcDNA3.1/V5-His-TOPO/HNP1 was constructed correctly. The ELISA results showed that the eukaryotic expression plasmid pcDNA3.1/V5-His-TOPO/HNP1 could temporarily express HNP1 in COS-7 cells. Conclusion The successful construction and expression of pcDNA3.1/V5-His-TOPO/HNP1 pave the way for the stable expression HNP1 in mammalian engineering cells.

Objective To observe the dynamic changes of high-sensitive C-reactive protein (hs-CRP) and visfatin in patients with acute traumatic injury of brain. Method A total of 120 patients with equal number in each gender ( n = 60) and with average age of (43.2 ± 6.2) years were admitted and treated by the neurosurgical department of ICU from August 2009 to June 2010. All patients were eligible to the diagnostic criteria of craniocerebreal injury. The clinical conditions of patients were assessed with Glasgow coma scale (C CS) at admission,and as per the scores of GCS, the patients were classified into mild degree (13- 15, n = 40), moderate degree (9 - 12, n = 40) and severe degree (3 - 8, n = 40). Another 60 subjects from those asking for health care by physical examination as control with equal number in each gender and their average age was (42.2±6.7) years.Blood samples were collected from fasted patients within 12 hours, 1d, 3d, 7d and 15 days after admission, and the levels of hs-CRP and visfatin in peripheral blood were detected. Results The levels of hs-CRP and visfatin were significantly higher in brain injury group than those in control group on the admission day (both P ＜ 0.01 ),and they both had positive relationships with severity of injury. The level of hs-CRP increased to peak on the first day of admission and visfatin increased to the peak on the 3rd day after admission. There was a correlation between levels of hs-CRP and visfatin ( r = 0.63, P ＜ 0.01 ). Conclusions hs-CRP and visfatin levels are related to the severity of acute traumatic injury of brain.

BACKGROUND:There are less reports about the external use of recombinant human granulocyte-macrophage colony stimulating factor (rhGM-CSF) hydrogel to repair thick skin graft donor sites. By now, relevant self-control studies have not been retrieved. OBJECTIVE:To observe the effect of rhGM-CSF on the repair of thick skin graft donor sites. METHODS:Sixty patients with burns and scar hyperplasia undergoing autologous thick skin grafting were enroled, 47 males and 13 females, aged 18-65 years. The thigh was selected as donor sites. According to the depth of donor sites, the patients were divided into 0.4 mm and 0.55 mm groups, with 30 cases in each group. Wounds on the symmetric areas with equal area and same depth were selected or wounds with same depth were selected and divided equaly. The wounds were randomly assigned into treatment group and control group. The treatment group was treated with rhGM-CSF hydrogel externaly; the control group was only given vaseline dressing. At postoperative 3, 7, 10, 14 days, the fresh dressing was changed. Then, the wound appearance, healing time, healing rate and adverse effects were observed in the two groups. RESULTS AND CONCLUSION:At 14 days after operation, the wound surface was smoother and the pigmentation was relatively less in the treatment group compared with the control group; the degree of wound pain
was less in the treatment group than the control group during dressing change (P < 0.05). At 10 and 14 days after operation, the healing rate and healing time were better in the treatment group than the control group (P < 0.05). No general malaise or hypersensitivity cases were reported, and local issue hyperplasia was also not found. Al the above indicate that the external use of the rhGM-CSF hydrogel can evidently shorten the healing time and improve the healing condition when it is applied in the thick skin graft donor sites.

Interdigitating dendritic cell sarcoma (IDCS)is a rare malignant tumor of the dendritic cell, derived from the hematopoietic tissue.The major clinical manifestation of IDCS is superficial lymphadenopathy, and the enlarged lymph nodes may appear in some atypical ereas,such as the lung,kidney,bladder and the pleura,etc.With the development of the pathological diagnosis and the application of immunohistochemical staining and electron microscopes,the case detection rate is apparently improved.With the high degree of malignant,rapid progress and poor prognosis of the disease,currently,surgical therapy is still the main approach to the treatment of IDCS.

Objective To investigate the metastasis mechanism by observing morphological distributions of lymphic vessels in peripheral areas of the different development of uterine cervix cancer. Methods Cancer tissues from the center, peripheral and normal areas of uterine cervix cancer respectively were collected. The paraffin sections and semithin sections which were stained with HE were applied to those tissues for exploring the configurations and distributions of lymphic vessels of the cancer under a microscope. And the ultrathin sections were applied to those tissues for exploring under a electronic microscope. Results Under the microscope, the basement membrane had been destroyed by cancer cell, which continued to infiltrate interstitial tissue. Lymphic vessels were increased and dilated in peripheral areas of uterine cervix cancer than those in normal areas. Moreover, the walls of lymphic vessels were hazy and broken. Conclusion The increase and morphologic changes of lymphic vessels in peripheral areas of uterine cervix cancer will play an important role in lymphatic metastasis.

ObjectiveTo investigate the improvement of speech in patients with increased blood pressure(BP) and stable blood pressure during speech therapy. MethodsAfter monitoring blood pressure with dynamic blood-pressure meter during speech therapy, patients were divided into increased BP group and stable BP group. Patients received two-month speech therapy, then their score changes in ABC examination of pre-treatment and post-treatment were compared. ResultsIn oral expression, score changes in increased BP group were significantly different from those in stable BP group (P<0.05). In listening comprehension, score changes had no significant difference between two groups(P>0.05). Conclusions Patients in increased blood pressure group progressed obviously in oral expression.

Objective To analyze the reproducibility of keratometry and astigmatism measured by the VERION Digital Guidance System and the comparability of VERION with iTrace,Lenstar LS900 and manual keratometer.Methods The keratometry of 62 cataract patients were measured using four different devices.The steep keratometry (Ks),flat keratometry (Kf),astigmatic magnitude,astigmatic axis,cylinder at 0-degree meridian (vector component,J0) and cylinder at 45-degree meridian (vector component,J45) from each machine were recorded and analyzed.The three repeated measurements and the results of VERION system with other three devices were compared to analyze the reproducibility and comparability of VERION system.Results Reproducibility:Intraclass correlation coefficients and Cronbach's alpha values were higher than 0.9 for Ks,Kf,astigmatic magnitude,astigmatic axis,J0 and J45 measured by the VERION system (all P ＜ 0.001).Comparability:The results of Ks and magnitude of astigmatism of VERION were larger than the iTrace (all P ＜ 0.05) in the paired-samples t test.There was no statistical difference for the rest of parameters (all P ＞ 0.05).The Bland-Altman graphs revealed the 95％ limits of agreement (LOA) of J0,J45 and the astigmatic axis between VERION and iTrace were (-0.31-0.35) D,(-0.25-0.31) D and-13.5 °-12.3 °,respectively;There was no statistical differences for all parameters except for J45 in the paired-samples t test between the VERION and Lenstar LS900 (all P ＞ 0.05).The Bland-Altman graphs revealed the 95％ LOA of J0,J45 and the astigmatic axis were (-0.25-0.31)D,(-0.27-0.36) D and-13.5°-11.0°,respectively;There were statistical differences for the results of Kf and magnitude of astigmatism between the VERION and manual keratometer (all P ＜ 0.05).The Bland-Altman graphs revealed the 95％ LOA of J0,J45 and the astigmatic axis between VERION and manual keratometer were (-0.38-0.35) D,(-0.41-0.42) D,-12.6°-16.4°,respectively.Conclusion The VERION system is a reliable system for the measurement of keratometry and astigmatism.The keratometry and astigmatic magnitude of the VERION system have a good agreement with the iTrace,Lenstar LS900 and manual keratometer.However,the astigmatic axis measurements are significantly different among the four devices.

BACKGROUND: Myelination following axonal regeneration is a key factor affecting the recovery of spinal cord injury. Oligodendrocyte survival directly affects the myelination following axonal regeneration. OBJECTIVE: To investigate the feasibility of differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into oligodendrocytes induced by neurotrophic factors. DESIGN, TIME AND SETTING: The cell molecular biology in vitro study was performed at the Laboratory of Department of Orthopaedics, Tongji Hospital from September 2006 to June 2007. MATERIALS: A total of 5 Sprague Dawley rats aged 2-4 weeks, of both gender were selected. Bilateral femur and tibia bone marrow was obtained to harvest BMSCs. METHODS: At passage 4, BMSCs were incubated in serum-free medium, supplemented with N2, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor for 48 hours, and incubated in medium containing 500 ng/mL insulin-like growth factor I and N2 for 3 days. MAIN OUTCOME MEASURES: Morphological changes were observed using an phase contrast microscope. Semiquantitative RT-PCR was utilized to detect specific marker mRNA expression of oligodendrocytes. Using neuron marker anti-microtubule-associated protein, astrocyte marker anti-glial fibrillary acidic protein, oligodendrocyte marker anti-galactocerebroside, anti-myelin basic protein antibody, immunocytochemical staining was performed to detect the positive rate of the differentiation of BMSCs into oligodendrocytes. RESULTS: Morphological changes in BMSCs during the differentiation into oligodendrocytes: After the induction, a majority of BMSCs presented the morphological characteristics of oligodendrocytes. Cytoplasm retraction towards nucleus, cell process extension towards outwards, and strong refraction were found. With the prolongation of time, several cell processes connected and formed a typical net-shape structure. Specific marker mRNA expression of oligodendrocytes: Following induction, specific strap of myelin basic protein mRNA and galactocerebroside mRNA could be detected. Positive rate of oligodendrocytes: During induction, the positive rates of galactocarebroside, myelin basic protein and microtubule-associated protein were 65%, 45% and 10%, respectively. CONCLUSION: The combination of epidermal growth factor, basic fibroblast growth factor and insulin-like growth factor can effectively promote the directional differentiation of BMSCs into oligodendrocytes.