基于视窗聚类的人体外周血白细胞快速无重复自动扫描方法

为了实现人体外周血白细胞的快速无重复自动扫描,提出了一种新的聚类方法——视窗聚类法。由于人体外周血白细胞在血液涂片中分布稀疏且不均匀,传统的利用高倍镜直接逐行扫描的人工方法效率比较低下。用高倍物镜有目的地对有白细胞的区域进行拍摄可以提高白细胞的扫描效率。用低倍物镜预扫描血片,由于低倍物镜视场大,在低倍图中,用图像处理的方法可获得白细胞核心坐标。视窗聚类方法首先用最大最小距离聚类方法确定白细胞核心原始聚类中心,再按照高倍镜视窗大小对聚类中心进行调整,在确保各聚类视窗内无重复白细胞存在的同时,尽可能合并原始聚类中心,减少总的聚类中心个数。以新的视窗聚类中心组成高倍物镜扫描路径扫描血片,可保证高倍图像最大程度地包含尽可能多的白细胞,进而减少高倍图像的拍摄次数,实现了白细胞的快速无重复扫描。实验表明,用该方法获取110个无重复的白细胞图像仅需要拍摄高倍图像72~85幅,整个过程大概需要1.5~2.0min。同时,视窗聚类方法可以在无人监管的情况下完全避免白细胞图像的重复拍摄,实现了人体外周血白细胞的自动扫描。

Lymphangioma of the small bowel mesentery: A case report and review of the literature

Lymphangioma is a rare benign condition characterized by proliferation of lymphatic spaces. It is usually found in the head and neck of affected children. Lymphangioma of the small-bowel mesentery is rare, having been reported for less than 1% of all lymphangiomas. Importantly, it can cause fatal complications such as volvulus or involvement of the main branch of the mesenteric arteries, requiring emergency surgery. Moreover, the gross and histopathologic findings may resemble benign multicystic mesothelioma and lymphangiomyoma. Immunohistochemical study for factor Ⅷ-related antigen, D2-40, calretinin and human melanoma black-45 (HMB-45) are essential for diagnosis. Factor Ⅷ-related antigen and D2-40 are positive in lymphangioma but negative in benign multicystic mesothelioma. HMB-45 shows positive study in the smooth-muscle cells around the lymphatic spaces of the lymphangiomyoma. We report a case of smallbowel volvulus induced by mesenteric lymphangioma in a 2-year-and-9-mo-old boy who presented with rapid abdominal distension and vomiting. The abdominal computed tomography scan showed a multiseptated mass at the right lower quadrant with a whirllike small-bowel dilatation, suggestive of a mesenteric cyst with midgut volvulus. The intraoperative findings revealed a huge, lobulated, yellowish pink, cystic mass measuring 20 cm × 20 cm × 10 cm, that was originated from the small bowel mesentery with small-bowel volvulus and small-bowel dilatation. Cut surface of the mass revealed multicystic spaces containing a milky white fluid. The patient underwent tumor removal with small-bowel resection and end-to-end anastomosis. Microscopic examination revealed that the cystic walls were lined with flat endothelial cells and comprised of smooth muscle in the walls. The flat endothelial cells were positive for factor Ⅷ-related antigen and D2-40 but negative for calretinin. HMB-45 showed negative study in the smooth-muscle cells around the lymphatic spaces. Thus, the diagnosis was lymphangioma of the small bowel mesentery with associated small bowel volvulus.

Diagnostic role of 18F-fluorodeoxyglucose positron emission tomography for follicular lymphoma with gastrointestinal involvement

AIM:To investigate the capacity for 18F-fluorodeoxyglucose(18F-FDG) positron emission tomography(PET) to evaluate patients with gastrointestinal lesions of follicular lymphoma.METHODS:This retrospective case series consisted of 41 patients with follicular lymphoma and gastrointestinal involvement who underwent 18F-FDG-PET and endoscopic evaluations at ten different institutions between November 1996 and October 2011.Data for endoscopic,radiological,and biological examinations performed were retrospectively reviewed from clinical records.A semi-quantitative analysis of 18F-FDG uptake was performed for each involved area by calculating the maximum standardized uptake value(SUVmax).Based on the positivity of 18F-FDG uptake in the gastrointestinal lesions analyzed,patients were subdivided into two groups.To identify potential predictive factors for 18F-FDG positivity,these two groups were compared with respect to gender,age at diagnosis of lymphoma,histopathological grade,pattern of follicular dendritic cells,mitotic rate,clinical stage,soluble interleukin-2 receptor levels detected by 18F-FDG-PET,lactate dehydrogenase(LDH) levels,hemoglobin levels,bone marrow involvement,detectability of gastrointestinal lesions by computed tomography(CT) scanning,and follicular lymphoma international prognostic index(FLIPI) risk.RESULTS:Involvement of follicular lymphoma in the stomach,duodenum,jejunum,ileum,cecum,colon,and rectum was identified in 1,34,6,3,2,3,and 6 patients,respectively.No patient had esophageal involvement.In total,19/41(46.3%) patients exhibited true-positive 18F-FDG uptake in the lesions present in their gastrointestinal tract.In contrast,false-negative 18F-FDG uptake was detected in 24 patients(58.5%),while false-positive 18F-FDG uptake was detected in 5 patients(12.2%).In the former case,2/19 patients had both 18F-FDG-positive lesions and 18F-FDGnegative lesions in the gastrointestinal tract.In patients with 18F-FDG avidity,the SUVmax value of the involved gastrointestinal tract ranged from 2.6 to 17.4(median:4.7).For the 18F-FDG-negative(n = 22) and-positive(n = 19) groups,there were no differences in the male to female ratios(10/12 vs 4/15,P = 0.186),patient age(63.6 ± 2.4 years vs 60.1 ± 2.6 years,P = 0.323),presence of histopathological grade 1 vs 2(20/2 and 17/2,P = 1.000),follicular dendritic cell pattern(duodenal/nodal:13/5 vs 10/3,P = 1.000),mitotic rate(low/partly high,14/1 vs 10/3,P = 0.311),clinical stage according to the Ann Arbor system(stages ⅠE and ⅡE/other,15/7 vs 15/4,P = 0.499),clinical stage according to the Lugano system(stages Ⅰ and Ⅱ-1/other,14/8 vs 14/5,P = 0.489),soluble interleukin-2 receptor levels(495 ± 78 vs 402 ± 83,P = 0.884),LDH levels(188 ± 7 vs 183 ± 8,P = 0.749),hemoglobin levels(13.5 ± 0.3 vs 12.8 ± 0.4,P = 0.197),bone marrow involvement(positive/negative,1/8 vs 1/10,P = 1.000),detectability by CT scanning(positive/negative,1/16 vs 4/13,P = 0.335),and FLIPI risk(low risk/other,16/6 vs 13/6,P = 0.763),respectively in each case.CONCLUSION:These findings indicate that it is not feasible to predict 18F-FDG-avidity.Therefore,18FFDG-PET scans represent a complementary modality for the detection of gastrointestinal involvements in follicular lymphoma patients,and surveillance of the entire gastrointestinal tract by endoscopic examinations is required.

Concomitant chemoradiation using gemcitabine in locally advanced hepatocellular carcinoma

Objective:Hepatocellular carcinoma (HCC) is among the most common and rapidly increasing cancers in Pakistan.There is currently no standard management for advanced HCC.The aim of the study was to assess response rate and toxicity of concomitant gemcitabine and external radiation therapy (ERT) in locally advanced HCC.Methods:Sixty patients were enrolled.Gemcitabine,70 mg/m2 was given weekly during ERT.ERT was delivered with 60Co beam up to 30.6 Gy,1.8 Gy/fraction.Tumor response was assessed by computed tomography (CT) at eight weeks.Complete and partial response (CR and PR),progressive and stable disease (PD and SD) were assessed.Hematological,gastrointestinal and hepatic toxicities were assessed weekly.Results:No CR was seen.PR,SD and PD were seen in 33%,40% and 27% of patients respectively.Grade 3/4 toxicity for total leukocyte count and platelets was observed in 7% and 27% patients.Grade 3/4 toxicity for liver failure,bilirubin,aspartate aminotransferase,alanine aminotransferase and alkaline phosphatase was seen in 35%,28%,38%,24% and 43% patients respectively.Conclusion:The study showed that concomitant gemcitabine and ERT is a feasible option with moderate toxicity in advanced HCC.

Cxcl16 Interact With SARS-CoV N Protein In and Out Cell

Our study investigated the host cell protein which can interact with SARS-CoV N protein, and explored the functional connections. The eukaryotic expression vectors pEGFP-N1/SARS-CoVN and pdsRed2-N1/ CXCL16 were constructed and used to co-transfect HEK293FT cells by the calcium phosphate method. The HIS-tagged fusion protein SARS-CoVN-GFP was then built and purified for the binding assay in vitro. The co-localization of SARS-CoVN and CXCL16 in the cytoplasm of HEK293FT cells was also shown using confocal laser scanning microscopy. It is suggested that their interaction might be through direct combination. Under a fluorescence microscope, it was observed that the purified fusion protein SARS-CoVN-GFP was attached to the cell membrane of CXCL16-transfected cells, indicating that SARS-CoVN and CXCL16 can be mutually combined.

基于视场边界最大最小聚类分析的快速扫描方法

针对用高倍镜头按一定规律路径会扫描到很多无法提取人体外周血白细胞的图像，为此提出一种基于低倍显微图像扫描定位细胞坐标、高倍显微图像提取细胞细节特征的外周血白细胞图像扫描新方法，有效提高了自细胞扫描速度，降低了细胞扫描重复率；提出了一种基于视场边界条件的改进型最大最小距离聚类算法，彻底避免了细胞重复扫描。实验表明，新方法扫描112个白细胞由传统方法的8～14min缩短到1．5～2．0min，减少了拍摄次数，提高了拍摄效率，避免了重复拍摄。

Effect of osteoprotegerin in combination with interleukin-6 on inhibition of osteoclast differentiation

Objective： To observe the effect of recombinant interleukin-6 （IL-6） and osteoprotegerin （OPG） on inhibiting bone absorption induced by receptor activa- tor for nuclear factor- rB ligand （RANKL） in murine osteo- clast precursor cells （OCPs） model. Methods： RAW 264.7 cells were solely treated with 50 ng/ml RANKL for 1 day, and then they were divided into three groups： RANKL （control group）, RANKL＋IL-6 （IL-6 group） and RANKL＋IL-6＋OPG （combination group）. These cells were harvested and investigated by means of HE stain- ing under light microscope after consecutive 9 days. Furthermore, staining tartrate-resistant acid phosphatase （TRAP）-positive multinucleated cells were detected by in- verted phase contrast microscope. The absorption pits of bone slices were observed under scanning electron microscope. Results： The number of mature osteoclast cells in control group was more than that in IL-6 alone or IL-6 com- bined with OPG group （P〈0.05）. Interestingly, this experi- ment has also demonstrated that there was a large number of TRAP-positive multinucleated osteoclasts （more than 3 nuclei） and several bone absorption formation in the con- trol group, whereas the outcome was completely different in both IL-6 group and IL-6＋OPG group （P〈0.05）. Conclusion： IL-6 can suppress the differentiation of mature osteoclasts as directly adding it into the RAW 264.7 cells induced by 50 ng/ml RANKL, and further the effect of osteolysis is remarkably reduced. When treatment with IL- 6 combined with OPG, a more effective strategy for the treat- ment of osteoporosis is reached.

Therapeutic effect of osteogenically induced adipose derived stem cells on vascular deprivation-induced osteonecrosis of the femoral head in rabbits

Objective： To explore the therapeutic ef- fect of osteogenically induced adipose-derived stem cells （ADSCs） on vascular deprivation-induced osteonecrosis of the femoral head （ONFH） in rabbit model. Methods： Vascular deprivation-induced ONFH was established by intramuscular injection of methylpre- dnisolone, and vascular occlusion of the capital femoral epiphysis by electrocoagulation in adult New Zealand white rabbits. Eight weeks afLer the establishment of vascular dep- rivation-induced ONFH, animals were randomly divided into three equal groups. In Group A （control）, no therapy was given. In Group B, core decompression was performed by drilling a hole （1.2 mm in diameter） from the outer cortex 2.5 cm distal to the proximal end of the greater trochanter. In Group C, 1 x 107 osteogenically induced ADSCs were resus- pended in 0.5 ml PBS, and then injected directly into the femoral head. Femoral head specimens were obtained at postoperative 8 weeks. The bone formation and three-di- mensional microstructure of the femoral head was evaluated by micro-computed tomography scans （u-CT）. Immunohistochemical analysis was performed to detect the expression ofosteocalcin. Angiogenesis and repair of the femoral head were observed histologically. Results： In trabecular bone at the proximal femur region, the trabecular volume was higher in Group C （130.70 mm3± 4.33 mm3） than that in Groups A （101.07 mm3±7.76 mm3） and B （107.89 mm3±8.6g mm3, P〈0.01）. Bone volume was significantly increased in Group C （40.09 mm3±6.35 mm3） than in Groups A （29.65 mm3±4.61 mm3） and B （31.80 mm3± 4.01 mm3, P〈0.01）. The trabecular number was higher in Groups C （1.58±0.25） than other two groups （1.15±0.18, 1.16± 0.21, P〈0.01）. Bone mineral density showed statistically significant difference between Groups C and A or B （375.38± 23.06） mg HA/ccm, vs （313.73 ± 19.30） mg HA/ccm and （316.09± 16.45） mg HA/ccm, P〈0.01）. Histological examination indicated that there was more new bone formation in Group C than in other groups. Conclusion： Treatment with autologous osteogenically induced ADSCs transplantation results in an enhanced osteogenesis and microstructure of the vascular deprivation-induced osteonecrosis in rabbits.