To express and prepare polyclonal antibody of Human papillomavirus type 6b (HPV6b) E7 protein. a prokaryotic expression vector pGEX-4T-2/HPV6b E7 was constructed and GST-HPV6b E7 fusion protein was expressed as a soluble protein in E. coli. The expressed fusion protein was purified via Glutathione-Sepharose 4B column and thrombin cleavage in order to obtain HPV6b E7 protein. Polyclonal IgG antibody was prepared by immunizing New-Zealand rabbits with HPV6b E7 protein. Western-Blot and immunofluorescence analysis showed that the polyclonal IgG antibody could specifically recognize HPV6b E7 protein and its titer was identified. SDS-PAGE analysis demonstrated that large amounts of soluble GST-HPV6b E7 fusion protein was expressed in E. coli after 3.0-6.0 hours of IPTG induction. Polyclonal IgG antibody successfully prepared from immunized rabbits showed high titer and high specificity as confirmed by Western-Blot and immunofluorescence. The preparation of anti-HPV6b E7 polyclonal antibody will facilitate further research on the biological and immunological functions of HPV6b E7 protein.
Animals ; Antibodies ; immunology ; Escherichia coli ; genetics ; metabolism ; Female ; Gene Expression Regulation, Bacterial ; HEK293 Cells ; Humans ; Immunoglobulin G ; immunology ; metabolism ; Oncogene Proteins, Viral ; genetics ; immunology ; metabolism ; Rabbits ; Recombinant Fusion Proteins ; genetics ; immunology ; metabolism

Adolescent ; Female ; Humans ; Lymphoma, Large-Cell, Anaplastic ; pathology ; Neutrophils ; pathology ; Skin Neoplasms ; pathology

BACKGROUND: Magnetic nanoparticles have attracted tremendous attention for their diverse biomedical applications involving tumor hyperthermia, drug controlled release and magnetic resonance angiography. The interaction between biomaterials and cells is the major part in the in vitro study, and the evaluation of cellular uptake of magnetic nanoparticles is indispensable for tumor hyperthermia and drug delivery. OBJECTIVE: To review the progress of evaluation methodology of cellular uptake of magnetic nanoparticles. METHODS: The articles about magnetic nanoparticles and cellular uptake were retrieved from PubMed, SCI and Embase databases published during January 2010 to June 2017 by the first author using computer. The Mesh terms were "magnetic nanoparticles", and the key words were "cellular uptake", "internalization" or "endocytosis". The logical operator between them was "AND". RESULTS AND CONCLUSION: Qualitative quantitative assessment methods for cellular uptake of magnetic nanoparticles include Prussian blue staining, fluorescence microscopy, confocal microscopy,transmission electron microscopy. Quantitative assessment methods for cellular uptake of magnetic nanoparticles include inductively coupled plasma, magnetization measurements, colorimetry of Prussian blue staining. The development direction of the evaluation methodology is from qualitative to quantitative. As each method has its own merits and demerits, we should choose the right evaluation method based on the function and application of magnetic nanoparticles and then supply the right experimental basis for the biomedical application of magnetite nanoparticles.

BACKGROUNDThe etiology and pathology of functional constipation (FC) is unclear. Some researches suggest that psychological factors may be related to this common problem. This study aimed to investigate the relationship between attention processing and emotional status in FC patients compared with healthy controls.

METHODSWe investigated selective attention and emotional status in patients with FC using the counting Stroop task. Thirty-five FC patients (FCP group) and 24 healthy controls (HCs) underwent an event-related potential (ERP) study while performing the task. Response time, latency and amplitude of P300 were collected and compared between the two groups. The Hamilton Depression Rating Scale (HAMD-17), Hamilton Anxiety Scale (HAMA), Symptom Checklist-90 (SCL-90) and Eysenck Personality Questionnaire (EPQ) were administered prior to the counting Stroop task.

RESULTSScores for the FCP group differed significantly from those for the HCs on the HAMD-17, HAMA, SCL-90 (Global Severity Index, Positive Symptoms Total, dimensions of somatization, obsessive-compulsive, depression, anxiety, and psychosis), as well as extraversion and neurosis dimensions of the EPQ (P < 0.05). There was no significant difference in response time or accuracy rate between the two groups (P > 0.05). ERP results showed prolonged latency at F4, additionally found enhanced activation at F3, FC1 and T8, and decreased activation at sites FC6 and P7 in the patients with FC. Statistically, significant differences of P300 latency at site F4 (P < 0.05) and P300 amplitude at F3, F4, T8, P7, FC1 and FC6 (P < 0.05) between the patients with FC and the HC were revealed.

CONCLUSIONSThe findings suggest that patients with FC are more susceptible to depression and anxiety, as well as somatization, obsessive-compulsive tendencies and other neurotic personality characteristics. Patients may employ psychological defense mechanisms to avoid the pain of depression and anxiety. ERP results imply there may be some brain dysfunction and attention deficits in patients with FC.

Adolescent ; Adult ; Aged ; Anxiety ; etiology ; physiopathology ; Constipation ; complications ; physiopathology ; Depression ; etiology ; physiopathology ; Female ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVEThis study compared the differences in clinical features between chronic aplastic anemia (CAA) and myelodysplastic syndrome (MDS) in children in order to provide a basis for the differential diagnosis of the two diseases.

METHODSA retrospective study of 23 cases of CAA and 9 cases of MDS from September 2007 to September 2010 was performed. The clinical data including routine blood test results, reticulocyte counts, serum lactate dehydrogenase level, serum ferritin level, cytological examination of bone marrow, bone marrow CD34+ cell counts, bone marrow chromosome and FISH test results were compared between the CAA and MDS groups.

RESULTSNeutrophils, reticulocytes, and serum ferritin and lactate dehydrogenase levels increased in the MDS group compared with those in the CAA group. There were significant differences in bone marrow blast cell counts and dyshematopoiesis phenomena of three lines blood cells between the CAA and MDS groups. The bone marrow CD34+ cell counts and the rate of chromosomal abnormalities detected in bone marrow cytogenetic analysis in the MDS group were significantly higher than those in the CAA group.

CONCLUSIONSThere are differences in the results of laboratory examinations and morphological and cytogenetic examinations of bone marrow between the children with CAA and MDS. The differences are useful to the differential diagnosis of the two diseases.

Anemia, Aplastic ; genetics ; pathology ; Bone Marrow Examination ; Child ; Child, Preschool ; Chromosome Aberrations ; Chronic Disease ; Female ; Humans ; Male ; Myelodysplastic Syndromes ; genetics ; pathology

OBJECTIVETo explore the feasibility a modified method for repairing hand soft tissue defects using abdominal pedicle flaps.

METHODSBetween January 1997 and October 2010, abdominal pedicle flaps were used to repair hand soft tissue defects in 108 cases. A strategy of segmented skin-flap ligations was adopted for early pedicle cut-off with early separation of the combined fingers, donor site closure with auxiliary flaps and application of thin flap and full-thickness skin graft.

RESULTSLigation of the pedicle site only was performed in 68 cases with an average time of pedicle cut-off of 10 days. Pedicle cut-off and separation of the combined fingers were simultaneously performed in 40 cases without flap necrosis. The healing rate of the donor sites reached 96.8%.

CONCLUSIONThe method is simple, safe and reliable and allows simultaneous pedicle cut-off and separation of the combined fingers to ensure good functional and appearance recovery of the hand with soft tissue defect.

Abdominal Wall ; Adolescent ; Adult ; Burns ; surgery ; Child ; Female ; Hand Injuries ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Soft Tissue Injuries ; surgery ; Surgical Flaps ; Young Adult

OBJECTIVETo discuss a method for reconstruction of deeply burned of extremities' functional regions with local skin flaps.

METHODSNinety-eight cases (with 126 place) of deeply burned wound of extremities were reconstructed with fasciocutaneous flaps.

RESULTSThe survival rate of 162 flaps was 93.2%. The movement of 112 joints (88.9%) was restored to normal, and the movement of 14 joints was mild limitation, none of the cases was severe obstacle.

CONCLUSIONSThe method is simple and with satisfactory results, which can decrease the rate of disability suffered from burn.

Adolescent ; Adult ; Burns ; surgery ; Child ; Extremities ; surgery ; Fascia ; Female ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Surgical Flaps ; blood supply

Objective To discuss the technique and technique-related issues of percutaneous kyphoplasty(PKP).Methods The study involved 69 vertebrae in 51 cases of painful osteoporotic vertebral compressive fractures.Under X-ray fluoroscopy monitoring,the fractured vertebral bodies were treated by kyphonplasty with inflatable balloon.The preoperative and postoperative vertebral height and Cobb angle in radiography were measured and analyzed.Results All patients tolerated the procedure well with dramatic pain relief within 72 hours after the procedure.No clinical complication was found.The loss heights of the anterior and mid portion of the vertebral body reduced from 15?4mm and 11?4mm preoperatively to 10? 4mm and 6?3mm postoperatively,respectively.Cobb angle corrected averagely from 22??6? preoperatively to 12??4?.There was significant difference between preoperative and postoperative measures (P

BACKGROUNDRecently, bilateral transpedicular kyphoplasty with a single balloon was performed for osteoporotic vertebral body compression fractures (OVCFs) to reduce the fees of the operation, but the time of operation and radiation exposure are longer. The aim of this study was to determine the safety and effectiveness of a single balloon cross-midline expansion via unipedicular approach in kyphoplasty for OVCFs.

METHODSThirty-six patients with painful OVCFs (61 vertebrae) were enrolled in this research. Unilateral transpedicular puncture was performed under the fluoroscopy monitoring of an oblique angle down the pedicle. A single balloon was introduced through unipedicular approach. The final balloon position was in the midline of the vertebral body with the balloon cross-midline expansion and bone cement filled. Clinical outcomes were determined by comparison of the preoperative and postoperative visual analogue scale (VAS) and Oswestry disability index (ODI). Radiographic assessment included restoration of vertebral height and correction of kyphosis. Follow-up was conducted for 6 - 12 months (mean 9.2 months).

RESULTSThirty-six consecutive patients with 61 vertebrae were successfully operated on with a mean operation time of 37.4 minutes per vertebra. All patients had dramatic pain relief and functional recovery within 96 hours after the procedure with no surgery or device-related complications. VAS score improved from 7.27 +/- 1.02 preoperatively to 2.71 +/- 0.75 postoperatively (P < 0.01). ODI score was decreased from (71.14 +/- 10.94)% preoperatively to (26.56 +/- 6.35)% postoperatively. The average loss of anterior body height was (14.33 +/- 2.76) mm before procedure and (10.03 +/- 1.83) mm after procedure (P < 0.01), while the average loss of middle body height was (10.15 +/- 2.70) mm before procedure and (5.89 +/- 1.83) mm after procedure (P < 0.01). The kyphotic deformity was corrected from (23.43 +/- 5.00) degree to (16.16 +/- 2.77) degree (P < 0.01). The pain relief and functional recovery were substantial and maintained to the last follow-up without any re-collapse or adjacent level fracture.

CONCLUSIONSA single-balloon cross-midline expansion via unipedicular approach in kyphoplasty for OVCFs is an effective and safe procedure with less cost, less operation time and less radiation exposure when compared with the conventional kyphoplasty technique.

Aged ; Aged, 80 and over ; Female ; Fractures, Compression ; surgery ; Humans ; Male ; Middle Aged ; Osteoporosis ; complications ; Spinal Fractures ; surgery ; Treatment Outcome ; Vertebroplasty ; methods

Objective To analyze the clinical features and prognosis of pediatric acute myeloid leukemia (AML) with EVI1 gene positive.Methods The nested RT-PCR was performed to detect the EVI1 expression in pediatric AML patients from Jan.2009 to Dec.2011.The patients with EVI1 were investigated on clinical features,curative effects and prognosis.The differences between EVI1 (+) and EVI1 (-) patients were also analyzed.Results The frequency of EVI1 (+) expression was 15.65％ (13/83 cases)in pediatric AML,with the highest incidence in high-risk patients.EVI1 (+) was obviously associated with some unfavorable molecular genetic changes such as complex karyotype,MLL rearrangement,and monosomy 7.There were significant differences for EVI1 (+) group and EVI1 (-) group in the complete remission rate (45.5％ vs 79.3％,x2 =5.497,P ＜ 0.05) and complete remission(CR) rate after the first chemotherapy (18.2％ vs 63.8％,x2 =7.828,P ＜0.01).Although significant difference in death rate was not observed,EVI1(+) group had significantly higher early-death rate (45.5％ vs 8.6％,P ＜0.01).The EVI1(+) group also had lower 4 years event-free survival (EFS) [(21.2 ± 13.8) ％ vs (50.2 ± 9.1) ％,x2 =4.493,P ＜ 0.05] and lower 4 years overall survival(OS) [(32.4 ± 7.1) ％ vs (60.3 ± 10.9) ％,x2 =4.602,P ＜ 0.05] compared with EVI1 (-) group.But binary Logistic analysis did not identify EVI1 (+) as an independent unfavorable prognostic factor.Conclusions The pediatric AML with positive EVI1 expression had lower CR rate,higher early death rate and lower EFS.Positive EVI1 expression is related with an adverse outcome,but is not an independent poor prognostic factor.