Objective To prepare a specific anti-lactoferrin single chain variable fragment(ScFv).Methods Anti-lactoferrin clones were screened from a 'naive' phage antibody library against the immobilized lactoferrin antigen,then the clones were transformed to the E.coli HB2151 to give soluble expression of antibody fragments.The culture supernatant containing ScFv was purified by immobilized metal affinity chromatography,and then determined with SDS-PAGE and ELISA.Results The results demonstrated that ScFvs were specific;they did not react with transferrin,lysozyme and bovine serum albumin in ELISA.The SDS-PAGE showed that the ScFvs had high purity through affinity chromatography and the molecular weight of them was about 32 kD.Conclusions The successful generation of the ScFvs against lactoferrin provides a basis for further study and clinical applications for dry eye and other ocular diseases.

Objective To investigate the role of CT and MRI in diagnosis of the pineal region germinoma.Methods 11 cases of the pineal region germinoma proved by operation and pathology. CT and MRI features were analysed retrospectively. Results In 11 cases on CT most tumors were hyperattenuation or isoattenuation. Enhancement in most of the tumors were homogeneous. The characteristic MRI appearances of the pineal region germinoma were as follows: (1) On T 1WI, the lesions were isointense or slightly hypointense while isointense or slightly hyperintense on T 2WI. The pineal region germinoma showed no edema. (2) The lesions showed homogeneous or inhomogeneous Gd-DTPA enhancement in most of the tumors.Conclusion CT and MRI of multiplanar imaging and Gd-DTPA enhancement are helpful in diagnosis and differential diagnosis of pineal region germinoma. The tumors have sex, location and imaging characteristics.

[Objective]To establish animal model of avascular necrosis of the femoral head in rabbits induced by glucocorti-coid, to further investigate the pathogenesis of glucocorticoid-induced osteonecrosis.[Method]Twenty NewZealand white rabbitswere used and randomly divided into two groups.Group A was injected with horse serum and prednisone. Group B was used asthe control.The concentration of TNF-?was measured by ELISA in rabbits serum of group A and group B.Femoral head speci-mens were studied by histopathologic examination and the expression of VEGF by immunohistochemical examination.[Result]The concentration of TNF-?in rabbits serum of group A was higher than that of group B(P

[Objective]To explore a new method for the therapy of avaseular necrosis of the femoral head with vascular endothelial growth factor(VEGF).[Method]Thirty New Zealand white rabbits were chosen as experimental animals and avascularnecrosis of the femoral head model of 22 rabbits were established.Animals were egually divided into three groups:VEGF group,natural repair group,normal group.The medicament was injected to the femoral head by percutaneous injection in group A and normal saline in group B.The repairing process of bone necrosis of femoral head were studied.[Result]Compared to the untreated model group(group C),the femoral head of VEGF groups showed significant new vessel formation,number of empty bone lacunae decreased and hematopoietic tissue improving in marrow cavity.X-ray film and CT revealed evident that veGF could enhance repairing of the femoral head.[Conclusion]Pereutaneous injection of VEGF to the necro femoral head could significantly enhance bone tissue angiogenesis and ameliorated repair of osteonecrosis,thus providing a potential method for the treatment of the femoral head necrosis.

AIM: This study was designed to investigate the apoptotic effect of doxycycline in THP-1 cells.METHODS: After differentiated by PMA, THP-1 cells were treated with doxycycline at different concentrations ranging from zero to 80 mg/L. The morphological changes of THP-1 cells were observed under light microscope. MTT assay were used to examine the effects of doxycycline on proliferation of THP-1 cells. Apoptotic THP-1 cells were measured by Annexin-V flow cytometry analysis and TdT-mediated dUTP nick end labeling assay.RESULTS: Treated with a certain concentration of doxycycline, differentiated THP-1 cells contracted and turn round, a number of cells were dead. MTT assay and positive Annexin V-FITC on cell membrane and TUNEL assay showed that doxycycline induced apoptosis in THP-1 cells in a dose-dependent manner.CONCLUSION: Doxycycline induces apoptosis in THP-1 cells in a dose-dependent manner.

BACKGROUND:Increasing studies have found that, percutaneous vertebroplasty and bone cement are characterized by few traumas, few bleeding, simple operation, rapid recovery, rapid pain relief, and effective efficacy. However, tumor recurrence, symptom improvement, bone cement leakage and other complications make people controversial. OBJECTIVE: To explore and summarize clinical outcomes and safety of percutaneous vertebroplasty and bone cement for treating symptomatic vertebral hemangioma. METHODS:Fifty-two cases patients of symptomatic vertebral hemangioma (including 21 male and 31 female, aged 16-63 years) were treated with percutaneous vertebroplasty and bone cement. The lesions were found at T5-T12 in 36 cases and at L1-L3 RESULTS AND CONCLUSION: Al 52 patients were folowed up for at least 6 months after the operation. The Visual Analogue Scale and Oswestry Disability Index were significantly increased at 1 day, 3 weeks and 6 months post-surgery, when compared to before surgery (P< 0.05); the modified Macnab evaluation was excelent in 40 cases, good in 7 cases, fair in 3 cases, and poor in 2 cases, and the excelent and good rate was 90%. Only one patient had recurrence at 6 months after surgery and one patient had the leakage of bone cement. No other complications were found. Percutaneous vertebroplasty and bone cement could effectively cure vertebral hemangioma and relieve the pain, which is a safe treatment. in 16 cases. Clinical outcomes such as Visual Analogue Scale, Oswestry Disability Index and modified Macnab criteria, as wel as complication before and after surgery were evaluated.

Objective To compare the clinical effect and safety of percutaneous vertebroplasty between unipedicular and bipedicular approach for patients with osteoporosis vertebral compressive fractures. Methods A retrospective study was carried out on 147 patients with single osteoporotic vertebral compressive fractures from 2008 to 2014. Ninety-five cases were treated by PVP via a unipedicular approach (unipedicular group) and 52 cases were treated by the bipedicular approach (bipedicular group). The operation time, Cobb, frequency of X-ray exposure,complications like bone cement and postoperative VAS score were compared. Results No significant differences on the VAS score, cobb and complications were found (P > 0.05). However, there were significant differences on the operation time and X-ray exposure frequency (P<0.05). Conclusions No significant differences on clinical effects were found between the unipedicular and the bipedicular approach to percutaneous vertebroplasty for osteoporosis vertebral compressive fractures. However, the unipedicular approach can shorten the operation time and reduce X-ray exposure.

Objective Percoll density gradient centrifugation and Ficoll-Hypaque density gradient cen-trifugation, which are frequently-used methods for separation of tumor-associated macrophages (TAMs) from solid carcinoma were compared, in order to find an effective way to separate TAMs from colorectal carcinoma (CRC). Furthermore, we studied the best adherence time of separating macrophage among mononuclear cells. Methods specimens were collected from CRC patients , after digesting into single cells , TAMs were separated from the same specimen by 100% Ficoll, 35% percoll and 25% combined with 65% percoll respectively. After these pre-liminary separation, the collected cells were purified a second time by adherence separation. The purity of TAMs were detected by immunofluorescence. Results TAMs purity from Ficoll-Hypaque density gradient centrifugation was 80.18%, statistically higher than that from Percoll density gradient centrifugations (54.33% and 10.93% re-spectively). Conclusion Compared to Percoll density gradient centrifugation, Ficoll-Hypaque density gradient centrifugation is a more effective and simple way to isolate TAMs from colorectal carcinoma , suggesting it can be wildly used in clinical and basic medical research. 2-4 hours is the best adherence time for isolating macrophage.

AIM: To determine the optimal protocol and condition in which macaca irus mesenchymal stem cells (MSCs) are induced to differentiate into neuron-like cells by cryptotanshinone in vitro. METHODS: MSCs from macaca irus bone marrow were generated in vitro and induced with cryptotanshinone. The morphological changes of MSCs were evaluated by microscope. The positive percentages of neurofilament (NF), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) expression were measured by immunocytochemistry with ABC staining. RESULTS: The result showed that MSCs were positive for CD29, CD44, CD105, CD166, and negative for CD34, CD71, CD80 and CD86. After induced with cryptotanshinone, MSCs began to display neuronal morphologies, such as contracted multipolar cell body and formed extensive networks. The percentages of positive NSE, NF expression were 68.3%?3.5%, 70.3%?1.5%, respectively. CONCLUSION: Macaca irus MSCs could be induced to differentiate into neuron-like cells in vitro by cryptotanshinone and might be applied in cell transplantation and gene therapy in nervous system disorders.

OBJECTIVETo explore the expression of MALAT 1 in nasopharyngeal carcinoma (NPC) and its biological function.

METHODReal-time PCR was used to detect the expression of MALAT1 in 5-8F, C666-1, CNE-1, CNE-2, HONE-1, 6-10B and NP69 cell lines. CNE-1 cells engineered with MALAT1 RNA interference (RNAi) and RNA activation (RNAa) techniques were examined for changes in cell proliferation, invasion and metastasis using CCK8 assay, colony formation assay, Transwell in vitro invasion assay and wound-healing assay ability.

RESULTSMALAT1 was highly expressed in 5-8F cells with a high metastatic potential, and lowly expressed in normal nasopharyngeal epithelium cells. Overexpression of MALAT1 by RNAa suppressed the expression of E-cadherin, promoted the expression of vimentin and enhanced the proliferation, invasion, and metastasis of CNE-1 cells.

CONCLUSIONMALAT1 can enhance the proliferation, invasion, and metastasis of CNE-1 cells.

Cadherins ; metabolism ; Carcinoma ; Cell Line, Tumor ; Cell Proliferation ; Genes, Neoplasm ; Humans ; Nasopharyngeal Neoplasms ; genetics ; pathology ; RNA Interference ; RNA, Long Noncoding ; genetics ; Vimentin ; metabolism