Objective To concentrate coagulation Factor Ⅺ(FⅪ)from human plasma.Methods FⅪwas isolated and purified from human plasma via two-step chromatography,including CM-Sepharose fast flow ion exchange and Heparin CL-6B affinity chromatography.Results The recovery rate of FⅪ was(21.02?5.04)%,the specific clotting activity of purified FⅪ was(17.59?1.96) U/mg,and the purification factor was(1162.29?129.64) fold(n=7).Conclusion The two-step chromatography is effective in concentrating FⅨ.

Background Retinal Müller cells are important gliocytcs and the source of retinal stem cells.Researching the biological behavior of Müller cells is of important significance to the study on retinal physiopathological process and stem cell therapy of retinal diseases.To establish a stable culture method of Müller cells is a solid basis of relative basic research.Objective This study was to establish a simple and stable method of isolation and culture of human retinal Müller cells and provide sufficient and high-quality Müller cell source.Methods Human retinal Müller cells were isolated from healthy human donor eyes.The mixture solution of hyaluronidase (100 U) and 0.25％ trypsin were used to digest chopped retinal tissue.The DMEM/F12 medium with 20％ fetal bovine serum (FBS) was added to stop the digestion process.RPMI1640 medium with 20％ FBS was used to culture the cell for 72 hours and then replaced the half medium.The cells were passaged by the RPMI1640 medium with 20％ FBS.The morphology of the cells were examied under the optical microscope,and the expressions of glial fibrillary acidic protein (GFAP),a marker of gliocytes,and glutamine synthetase (GS),a special marker of retinal Müller cells,were detected by immunochemistry and immunofluorescence technology.Results Human retinal Müller cells were successfully isolated by enzyme mixture solution of hyaluronidase (100 U) and O.25％ trypsin.The cells were adherent to walls 24 hours after primary culture and completely merged 9-10 days after culture.The cells showed oval in shape with abundant cytoplasm,and a part of cells presented with cone-shaped bulge bilaterally and ectasia in the posterior containing large nuclei.After cells passage,the cells were enlarged and grew toward polygonal shape.The positive expression of GFAP was observed in more than 95％ cells and strongly positive expression of GS was observed in more than 90％ cells by immunohistochemstry and immunofluorescent staining.Conclusions Human retinal Müller cells can be successfully isolated by hyaluronidase combined with trypsin digestion.Abundent and pure human retinal Müller cells can be obtained by successively using RPMI1640 medium with 20％ FBS and 10％ FBS.

Objective To study the potential role of PKC inhibitor Rottlerin in the treatment of asthma. Methods Balb/c mice were sensitized and challenged with ovalbumin(OVA) protein to construct murine experimental model of asthma. Then the mice were treated by means of injection of Rottlerin or phosphate buffered saline (PBS) into the abdominal cavity. Changes of the total serum IgE, pulmonary eosinophils, cytokines and pulmonary inflammation were investigated. Results Injection of Rottlerin at the dose 0.3 mg/kg into the murine abdominal cavity could inhibit the infiltration of pulmonary eosinophils and pulmonary inflammation and significantly decrease the production of total serum IgE and Th 2 cytokines as well. Conclusion Rottlerin can inhibit murine experimental asthma.

Objective To explore clinical effect of repairing soft tissue defect in forearm, hand and foot with free super-thin anterolateral thigh perforator flaps. Methods At first the site of perforator vessels were determined by Doppler, then the flaps were designed and harvested with the site as center; the fascia lata and subcutaneous fat were removed by sandhill-likely only the 4.0 cm × 3.0 cm - 3.0 cm×2.5 cm disc-like fascia lata and dermis layer were reserved. 15 traumatic soft tissue defects including forearm, hand and foot were repaired with the ree super-thin antemlateral thigh perforator flaps. Results No vascular crisis happened and all skin grafts survived in donor sites. 2.0 cm×1.2 cm of the distal of flap was necrosis in 1 case and it was healed by dress changing. 15 cases were followed up 3 months-2 years and the average is 6 months. The contour and texture of all flaps were good and two point discrimination (2-PD) was about 8-10 mm of. Conclusions The contour and texture of free super-thin anterolateral thigh perforator flap are good, the feeling of recipient site recovered well, it's less injury for donor site and there is no reshaping for flap. It is a fineness donor site for repairing soft tissue defects in hand and foot.

Objective To investigate the CT findings of primary abdominal dedifferentiated liposarcoma(DDL),and to improve the diagnostic accuracy.Methods CT images of 23 cases with primary abdominal DDL confirmed by pathologically were analyzed retrospectively,and the CT findings were compared with pathological results.Results The masses in 20 cases out of 23 cases were located in retroperitoneal region,2 in abdominal cavity and 1 in the pelvic extraperitoneal space.The mean diameter of the masses was 26.5 cm.CT showed the fatty and non-fatty regions in the masses.In 2 1 cases,the non-fatty region was manifested as a single mass and was located at the edge or on the surface of fatty region.In 2 cases,the non-fatty region was manifested as multiple well-defined masses and was located within the fatty region.In 2 1 cases,the fatty region showed misty density with strip and reticular septa.In 2 other cases,the fatty region showed slightly low density,mingled with some fat tissue density,which showed no enhancement with mild enhanced internal septa.In 20 cases,the non-fatty region showed soft tissue density,with mild to moderate heterogeneous enhancement.In 3 other cases,the density of non-fatty region varied between liquid and soft tissue with patchy enhancement on delayed phase.In addition,stippled calcification was found in 4 cases.Conclusion The primary abdominal DDL has relatively specific CT findings,which different dedifferentiated components pro-duce different CT characterizes.Finding well-differentiated fat components around non-fatty mass may help to improve the accuracy of diagnosis and reduce the misdiagnosis.

ObjectiveTo compare X-ray,spiral CT and low does digital subtr,action angiongraphy technology of three dimensional reconstruction in 30 cases of the applications in the joint fractures,to study the application value of the digital subtraction angiography three- dimensional reconstruction in joint fracture.MethodsA retrospective analysis was made on 30 cases with joint fracture in Guangdong Provincial Guangzhou Development District Hospital,who were confirmed by the X-ray and spiral CT three-dimensional reconstruction and digital subtraction angiography three-dimensional reconstruction and the demonstrations three-dimensional reconstruction,spiral CT three-dimensional reconstruction and X-ray were compared.ResultsAll 30 cases fractures showed clear line,line of walk line,of the number of fracture,source and separation shift condition.Digital subtraction angiography could be clearly shows 30 cases in spiral CT could clearly show in 20 cases,10 cases not clearly show in,while X-ray clearly show in 10 cases,not show in 20 cases.Spiral CT showed clearly joint capsule and soft tissue.The SPSS 11.0 statistical soft package was used.Spearman correlation analysis,the single factor analysis of variance and the independent t test and the chi-square test were employed for data analysis,P ＜ 0.05 was considered statistical significance.Conclusions Digital subtraction angiography three-dimensional reconstruction can more clearly display fracture line,fracture pieces,walk line,source and separation shift condition compared with spiral CT and Xray essaminations,which is helpful for treatment options and prognosis estimation.

Objective:To investigate the correlations between perfused lung volumes, visual scores (using perfusion SPECT/CT) and right-heart catheter (RHC) hemodynamic parameters in patients with chronic thromboembolic pulmonary hypertension (CTEPH).Methods:A total of 51 consecutive CTEPH patients (17 males, 34 females, age (59±12) years) in the First Affiliated Hospital of Guangzhou Medical University between March 2015 and July 2019 were retrospectively analyzed. All patients underwent lung perfusion SPECT/CT imaging and RHC examinations. Perfused lung volumes were determined using threshold-based (15%-85%) segmentation. Visual semiquantitative scoring in each lung segment was performed using Begic method. RHC hemodynamic parameters including pulmonary artery systolic pressure (PASP), pulmonary arterial diastolic pressure (PADP), mean pulmonary artery pressure (mPAP), pulmonary arteriolar wedge pressure (PAWP), pulmonary vessel resistance (PVR), cardiac output (CO), cardiac index (CI) were recorded. Spearman correlation analysis was used to evaluate the correlations between perfused lung volumes, visual scores and hemodynamic parameters.Results:There were significant correlations between perfused lung volumes (30%-70% threshold) and mPAP ( rs values: from -0.414 to -0.302, all P<0.05). Among them, perfused lung volumes under the threshold of 40% and 45% were moderately correlated with mPAP ( rs values: -0.414, -0.412, both P<0.05). Perfused lung volume (40% threshold) was moderately negatively correlated with PASP, PADP ( rs values: -0.402, -0.440, both P<0.05), and slightly negatively correlated with PVR ( rs=-0.352, P<0.05). Visual scores were slightly positively correlated with the PADP ( rs=0.311, P<0.05), while there was no correlation between visual scores and other RHC hemodynamic parameters ( rs values: from -0.201 to 0.275, all P>0.05). Conclusion:Perfused lung volumes based on threshold-based segmentation in lung perfusion SPECT/CT imaging can accurately reflect hemodynamic status and may provide useful information for severity assessment of CTEPH.

Objective:To explore the characteristics of self-referential network (SRN) functional connectivity in subjective cognitive decline (SCD) patients with normal and impaired metacognition.Methods:Seventy-one subjects were selected from Alzheimer's Disease Neuroimaging Initiative (ADNI) database, with 25 cognitively normal controls and 46 SCD patients. The metacognitive level of SCD patients was assessed by Everyday Cognition Scale (ECog), and then, they were divided into a metacognitively normal group ( n=25, metacognitive scores>-0.074) and a metacognitively impaired group ( n=21, metacognitive scores≤-0.074). Results of Geriatric Depression Scale (GDS), Montreal Cognitive Scale (MoCA), Mini‐Mental State Examination (MMSE), Rey Auditory Word Learning Test (RAVLT), Logical Memory Scale, expressions of pathological markers (cerebrospinal fluid β-amyloid protein [Aβ], total tau protein [t-tau] and phosphorylated tau protein [p-tau]), brain glucose metabolism, and functional magnetic resonance imaging (fMRI) were collected and compared among the 3 groups. Independent component analysis (ICA) was used to extract SRN and analyze the different brain regions among the 3 groups; Pearson correlation was used to analyze the correlations of SRN functional connectivity changes with cognitive scales and pathological markers. Results:No significant differences in demographic characteristics (age and gender), scores of GDS, MoCA and MMSE, or levels of Aβ, t-tau, p-tau and brain glucose metabolism were noted among the 3 groups ( P>0.05). The metacognitive scores in metacognitively impaired group were significantly lower than those in metacognitively normal group and cognitively normal controls ( P<0.05). Significant difference in the functional connectivity of bilateral anterior cingulate gyrus and bilateral orbitofrontal cortex was noted among the 3 groups (TFCE-FWE correction, P<0.01, voxel>100); compared with the cognitively normal controls, the metacognitively impaired group showed significantly decreased functional connectivity of bilateral orbitofrontal cortex, while the metacognitively normal group showed enhanced functional connectivity of bilateral orbitofrontal cortex (TFCE-FWE correction, P<0.01, voxel>100); compared with the metacognitively normal group, the metacognitively impaired group had statistically decreased functional connectivity of bilateral orbitofrontal cortex (TFCE-FWE correction, P<0.01, voxel>100). Further correlation analysis showed that difference value of functional connectivity of bilateral orbitofrontal cortex between metacognitively impaired group and cognitively normal controls was negatively correlated with RAVLT-immediate scores ( r=-0.445, P=0.043); difference value of functional connectivity of bilateral orbitofrontal cortex between metacognitively impaired group and metacognitively normal group was negatively correlated with RAVLT-immediate scores ( r=-0.463, P=0.034). Conclusion:SCD patients with different metacognitive levels have characteristic SRN functional connectivity changes; impaired metacognition may be an early feature of Alzheimer's disease.

OBJECTIVETo analyze a case of supernumerary marker chromosome (SMC) with combined genetic techniques and explore its correlation with the clinical phenotype.

METHODSThe SMC was analyzed with G-banded karyotyping, multiplex ligation dependent probe amplification (MLPA), fluorescence in situ hybridization (FISH), and single nucleotide polymorphism array (SNP-array).

RESULTSG-banding analysis indicated that the patient has a karyotype of 47,XX,+mar. MLPA showed that there were duplications of proximal 15q. FISH assay using D15Z4 probes indicated that the SMC was a pseudodicentric chromosome derived from chromosome 15. And SNP-array revealed that there were two extra copies of 15q11-13 region spanning from locus 20 161 372 to 29 071 810.

CONCLUSIONThe duplication of Prader-Willi/Angelman syndrome critical region probably underlies the abnormal phenotype of the inv dup(15) case with a BP3:BP3 rearrangement.

Adult ; Chromosome Banding ; Chromosome Disorders ; genetics ; Chromosomes, Human, Pair 15 ; genetics ; Female ; Gene Rearrangement ; Humans ; In Situ Hybridization, Fluorescence ; Karyotyping

Objective: To assess the diagnostic rate, safety and clinical application of percutaneous renal masses biopsy for advanced renal cell carcinoma patients.Methods: In this retrospective study, we collected the data of renal masses from the patients who underwent renal masses biopsy under ultrasound from April 2001 to December 2014 in Peking University First Hospital.A total of 75 patients who were undiagnosed or diagnosed with advanced renal cell carcinoma by the imageological method were enrolled in this study.The patient and lesion characteristics such as tumor size, pathology of tumor, histologic subtype, pathological grade, biopsied location and biopsied cores were recorded and analyzed.Results: Among all the 75 patients, biopsy was diagnostic in 64 cases (85.3%) and non-diagnostic in 11 cases (14.7%).Of the 64 diagnostic biopsies, 60 were malignant, including 37 (61.7%) renal cell carcinoma (RCC), 13 (21.7%) urothelial carcinoma and 10 (16.7%) other malignant masses.Of all the RCC subjects, 24 suffered from clear cell RCC, 5 papillary RCC, 3 collecting duct carcinomas, 1 unclassified RCC and 4 unknown subtypes.The 11 non-diagnostic biopsied samplings included inflammatory, blood and extrarenal tissue and normal renal tissue.The proportion of collecting duct carcinoma in RCC was 10.8% and the proportion of squamous carcinoma in urothelial carcinoma was 23.1%, which were both higher than the previous research findings.For the male and female groups, non-diagnostic yields were 6.5% and 30.4%, respectively (P=0.022).Of all the 75 patients, 13 renal cell carcinoma patients underwent the surgical treatment and got the results of postoperative pathology.Comparing preoperative biopsy pathological diagnosis with postoperative pathological diagnosis, we found the diagnostic correct rates for benign and malignant lesions, pathological subtype and pathological grade were 100%, 81.8% and 60%, respectively.Mild macroscopic hematuria occurred in 1 case after RMB and there were no serious complications in all the cases.Conclusion: Percutaneous renal masses biopsy under ultrasound with a high diagnostic rate which can define the histologic subtype of renal cell carcinoma.With targeted therapy, more and more patients whose evaluation suggests local advanced disease or metastatic tumors adopt renal tumor biopsy to define the histologic subtype, which could avoid unnecessary surgical treatment.