Objective To observe a recombinant mutant of HBV core protein for dominant negative gene therapy against HBV encapsidation in vitro. Methods C gene and S gene of HBV were acquired through PCR and subcloned into pGEM T to construct pGEM T C and pGEM T S respectively. After digestion and ligation of these two plasmids, pGEM T CS was constructed. The cloned gene was inserted into pcDNA3.1 + to construct pcDNA3.1 + CS, which was identified by DNA sequencing. The recombinant plasmids were transformed into HepG2 cells, and screened with G418. The resistant HepG2 cell clones were chosen to test the expression of core surface protein by RT PCR, and the expressing HepG2 clones were cultured with 10% HBV DNA positive human serum for 72 hours. The intracellular HBV particles were extracted and the DNA was subjected to dot hybridization. Results The analysis showed that the HepG2 cells expressing mutant C protein had capabilities to resist HBV invasion in varied degrees. The mutant C protein had a dominant negative role in the encapsidation of HBV compared with the naive part of core protein. Conclusions The production of recombinant mutant core protein has a potential value for gene therapy against HBV infection.

Objective To summarize the imaging features of the thoracic canal stenosis caused by ossification of ligamentum flavum, and to evaluate its clinical results of a novel decompression of total laminotomy and autografting in-situ. Methods 31 patients underwent total laminotomy, autografting in-situ and decompression to treat the ossification of thoracic ligamentum flavum. In the study, there were 18 males and 13 females; the average age was 45.7 years (rang, 26 to 73). 3 patients were associated with cervical stenosis, 5 with lumbar stenosis, and 2 with both cervical and lumbar stenosis. 9 were associated with anterior compression by thoracic disc herniation or OPLL. The vertebrae affected were 1 level in 3 patients, 2 levels in 12, 3 levels in 11 and more than 4 levels in 5. 23 affected segments were located at the upper thoracic spine(T1-T4), 19 at the mid-thoracic spine(T5-T8) and 52 at lower-thoracic spine(T9-T12). All the patients underwent total laminotomy and autografting in-situ, 9 patients with thoracic disc herniation or OPLL received discectomy and internal fixation at one stage. The postoperative outcomes were evaluated according to Epstein criteria. Results The patients were followed up 6 to 63 months (mean, 15 months). The clinical outcomes were excellent in 14 cases, good in 7 cases, and fair in 3 cases, and the excellent and good rate was 87.5%. One case experienced neurological complication with Frankel C grade improved to Frankel D grade treated with neurotrophic drug and hyperbaric oxygen and to Frankel E grade at 5 months after operation. 2 presented with lower limb intravenous thrombosis, and CSF leakage happened in 1 case and cured with re-operation. The osteotomy of total laminotomy was bony healed at average of 5.5 months, and there was not new formed ossification anterior to re-implanted vertebral lamina and spinal cord compression. Conclusion MRI combined with CT scanning is a most useful imaging modality for the diagnosis of ossification of thoracic ligamentum flavum. The novel surgical procedure designed by the authors is safe and effective in treating ossification of thoracic ligamentum flavum.

Medial collateral ligament injury is the most common injury of the knee ligament. Most patients with conservative treatment can achieve preinjury activity level. Nonetheless,it's necessary to eval-uate the severity of the injury and judge the best time and details for surgery as serious injury of the medial collateral ligament will cause chronic knee stability and accelerate its degeneration. This paper summarizes the current status of diagnosis and treatment for medial collateral ligament injury of the knee,aiming to pro-vide guidance for dealing with this lesion.

Diffuse large B-cell lymphoma (DLBCL) is the most common type of non-Hodgkin lymphoma with highly heterogeneous clinical course. How to recognize patients with high risk is coming into the focus of the 57th American Society of Hematology (ASH) annual meeting. The International Prognostic Index (IPI) score as a classic prognostic system is challenged by variety of new prognosis systems. The tumor microenvironment plays an important role in progression of lymphoma, and its prognostic value draw much more attention. The value of PET-CT for prognosis of DLBCL has also been affirmed in this meeting.

BACKGROUND:Hydrolysis in vivo is the key mechanism of degradation in DL-polylactic acid (PDLLA). When it is combined with hydroxyapatite (HA), could the biodegradation and weight loss rate be improved? OBJECTIVE: To observe the changes in the interface and structure of HA/PDLLA composite after in vivo implantation into rabbit femoral defects.DESIGN: Randomized grouping and controlled observation.SETTING: Biomedical Materials and Engineering Research Center, Wuhan University of Technology.MATERIALS: Forty healthy adult Japan White Rabbits of 2.0-2.5 kg, either male or female were provided by the Animal Experimental Center of Hubei Province (No. SCXK. 2003-0005).METHODS: The experiment was conducted in Biomedical Materials and Engineering Research Center, Wuhan University of Technology from June 2005 to March 2006. ①The rabbits were randomly divided into two groups: HA/PDLLA group and PDLLA control group with 20 animals in each group. After anesthetized with ketamine and proazamine, the sample rods of HA/PDLLA and PDLLA were respectively implanted into the drilled bone cavities (φ5 mm × 8 mm) among condyles of femur sites of the rabbits, and the rod could be slightly higher than the surface of bone substance. The samples were covered by periosteum and skin, and then the skin and periosteum were repositioned. ②The complete implants and peripheral bone tissues were taken out respectively after 3, 6, 12 and 24 weeks implantation. The changes in the interface and structure of HA/PDLLA composite after in vivo implantation were observed by using scanning electron microscope (SEM, JSM-5610LV, Japan).MAIN OUTCOME MEASURES: Changes in the interface and structure of HA/PDLLA composite after in vivo implantation.RESULTS: Totally 40 rabbits were involved in the result analysis. After the materials were implanted, HA granules shed from the material surface, some fibroblasts grew into the tissue and a little new osteotylus was formed, indicating HA/PDLLA composite had capabilities of bone-formation and bone-connection. After 24 weeks implantation, the material was divided and wrapped by tissues, neogenetic bone tissue grew into the material, and the fracture healed well,indicating HA/PDLLA composite had good biocompatibility. As for biodegradable PDLLA polymer, hydrolysis in vivo is the most main mechanism of degradation; the degradation speed was decreased owing to being compounded with HA.CONCLUSION: HA/PDLLA composite has capabilities of bone-formation and bone-connection; the biocompatibility of the composite is improved accordingly on account of the decrease of the degradation speed. HA/PDLLA composite is suitable for clinical application as absorbable materials for internal fixation.

Objective To establish the athrmic inouse model of breast cancer in normal position and imitated metastatic breast cancer. Methods Breast cancer cells MDA-MB-231-luc carrying luciferase gene was injected into the athymie mice.The optieal imaging in vivo system was used to observe the establislament of the model. Reseults The breast tumor emerged after we planted the MDA-MB-231-lue cells in the mammary gland fatpads,the volume and photon of the tumor increased during the second weekto the fifth week.After injection by the tail vein,the tumors mainly located in the lungs While after infusion in the left alrtrium.the tumolrs metastate to all over the body. Conclusions We succeeded in the establishment of the athymic mice model of breast cancer.in situ and imitated metastatic breast cancer by iniection into the vena caudalis and the left alttrium.The optical imaging in vivo system could distinctly show the formation of the tumors.

AIM: To explore the therapeutic effects of flunarizine combined with aspirin in the treatment of migrainous cerebral infaction. METHODS: 38 cases of patients diagnosed as migrainous cerebral infaction were respectively given flunarizine 10 mg combined with aspirin 100 mg every night for a month. The observed indices included the dysfunction scores of nervous system, the total classes of living ability, and the accumulating rate of platelet and viscosity of plasma before and after the treatment. RESULTS: Before the treatment the dysfunction scores of nervous system and the living ability of the patients were 13.51 ? 4.78 and 3.45 ? 1.13 , and after the treatment the values were 4.34 ? 1.85 and 1.79 ? 0.72 respectively (P

We detected the seroprevalence of Toxoplasma gondii (T.gondii) in the wild birds in northeast China.The wild bird's blood was collected from the cutaneous ulnar vein and the serum was isolated and used for detection of anti T.gondii antibody by modified agglutination test (MAT).Results showed that totally 179 birds' serum samples were collected.Twenty serum samples (11.17％) were positive with T.gondii antibody,which belonged to 9 orders,17 families and 31 species.The seroprevalence against T.gondii was about 5.26％ (1/19) in Columbiformes,9.09％ (9/99) in Passeriformes,14.29％ (3/21) in Falconiformes,15.00％ (5/22) in Piciformes,16.67％ (1/6) in Coraciiformes,and 25.00％ (1/4) in Anseriformes.Based on their feeding behavior,the seroprevalence was 12.00％ (3/25) in carnivorous wild birds,10.60％ (15/141) in omnivorous wild birds,and 15.38％ (2/13) in the wild birds feeding on aquatic animals or plants.These wild birds also can be sorted as migratory and sedentary (non-migratory) according to their migration habits,and the serum positivity was 11.67％ (14/120),and 10.71％ (6/59) respectively.The seroprevalence against Toxoplasma gondii in wild birds in northeast China is about 11.17％,which indicates a common infection of Toxoplasma gondii in wild birds.

Objective The high energy ion bombardment technique is applied to enhancing the adhesion of the tetrahedral amorphous carbon (TAC) films deposited by the filtered cathode vacuum arc (FCVA). Methods The abrasion method, scratch method, heating and shaking method as well as boiling salt solution method is used to test the adhesion of the TAC films on various material substrates. Results The test results show that the adhesion is increased as the ion bombardment energy increases. However, if the bombardment energy were over the corresponding optimum value, the adhesion would be enhanced very slowly for the harder material substrates and drops quickly, for the softer ones. Conclusion The optimum values of the ion bombardment energy are larger for the harder materials than that for the softer ones.

Objective To explore the clinicopathological characteristics of non-muscle-invasive bladder cancer patients association with chronic kidney disease (CKD).Methods Between Jan 2009 and Dec 2010,536 patients (390 males and 146 females with mean age of 63 years) underwent surgical treatment at our institute for pathologically proven non-muscle-invasive bladder cancer.The clinical and pathological data of these patients were reviewed,and the relationships of these factors and CKD were analyzed.Presence of CKD was confirmed in patients with estimated glomerular filtration rate (eGFR)＜ 60ml/(min · 1.73 m2)calculated using the Modification of Diet in Renal Disease Study equation.Results Of the 536 consecutive cases,57 patients (10.6％) had CKD.Compared to the patients without CKD,there were more females and older patients in the patients with CKD (52.6％ vs 24.2％ and 69 years vs 62 years,both P ＜ 0.05).The patients with CKD proned to have multiple bladder tumor (71.9％ vs 50.9％,P ＜ 0.05) and synchronous upper urinary tract urothelial carcinoma (7.0％ vs 2.3％,P ＜0.05).The history of bladder cancer and upper urinary tract urothelial carcinoma were also predominated in these patients (43.9％ vs 29.0％ and 40.4％ vs 6.5％,both P ＜ 0.05).ConclusionConcurrent CKD in non-muscle-invasive bladder cancer patients is associated with greater risk of multiple tumors in urinary tract,particularly in female patients.