Objective · To investigate the histone methyltransferase capability of DOT1L-long form and its role in breast cancer metastasis.Methods · The existence of DOT1L-long form was confirmed by PCR, and the mRNA level of DOT1L was tested by real-time PCR. In HEK293T cells in which DOT1L canonical and DOT1L-long were overexpressed respectively, Western blotting was used to test the expression level of DOT1L and the histone methyltransferase capability. In the MCF10A cell line with inducible expression of DOT1L-long, real-time PCR was used to detect the mRNA level of epithelial-mesenchymal transition (EMT) marker, and transwell assay was used to detect the migration of breast cancer cells in which the expression level of DOT1L is low or high. Results · PCR demonstrated the existence of DOT1L-long form, and real-time PCR showed it widely exists in HCT116, T98G, MCF10A cells, etc. Western blotting showed the expression of DOT1L-long form and its H3K79 methyltransferase activity. In MCF10A cells in which overexpressed canonical DOT1L and DOT1L-long, mRNA levels of N-cadherin and fibronectine increased. Transwell showed canonical DOT1L and DOT1L-long both substantially increased the migration of breast cancer cells. Conclusion · The existence of DOT1L-long was confirmed and investigated, which is 202 amino acids longer than the canonical DOT1L, and is coded by a new exon, located between exon 27 and 28. Further, the DOT1L-long has H3K79 methyltransferase activity, and is able to promote breast cancer metastasis.

Objective To analysis and summarise the feature of conventional ultrasound(US) and contrast-enhanced ultrasound(CEUS) of anterior mediastinal tumor.Methods From April 2011 to March 2015,24 patients,diagnosed as anterior mediastinal tumor by chest CT and could be detected by conventional US,were enrolled in this study.Among them,there were 11 lymphomas,5 thymic carcinomas and 8thymomas.The US and CEUS,micro flow imaging(MFI) and time intensity curve(TIC) parameters were evaluated respectively.Results Compare with lymphoma and thymoma in US,there were significant difference between the tumor shape and internal echo respectively (P ＜0.05,P ＜0.05).Compare with lymphoma and thymoma in CEUS,there were significant difference between the display rate of microvascular and feature of enhancement respectively (P ＜ 0.05,P ＜0.05).The result of TIC in different tumors were analyzed respectively.The rise time of lymphoma was early than thymoma,but the half time of wash out of lymphoma was later than thymoma,there was significant difference between the two diseases respectively(P ＜0.05,P =0.01).Further more,the arrive time of invasive thymoma was later than noninvasive thymoma,there was significant difference between the two diseases (P ＜ 0.05).Conclusions CEUS could further assess the characteristic of microvascular perfusion in anterior mediastinal tumors,based on the evaluation of conventional US.It could have a potential clinical value and a development capacity for differentiation diagnosis.

Objective To evaluate the role of contrast-enhanced ultrasound (CEUS) on the the detection of hepatic metastases in comparison with conventional ultrasonography (US) and contrastenhanced computed tomography (CECT).MethodsNinety-seven patients with hepatic metastases underwent US,CEUS and CECT images.Their detection of hepatic metastases were compared.Results Hepatic metastases showed five enhancement patterns with CEUS,including bolus hyper-enhancement,peripheralrimenhancement,inhomogeneousenhancementwithnecrosis,hypo-enhancement,isoenhancement with liver parenchyma,all hepatic metastases showed dark defects in portal and delayed phase.The mean number of metastases at CEUS was greater than that of US (2.6±1.9 vs 1.6 ±1.2,P＜0.05).The detection of hepatic metastases was 53.4％ and 87.1％ respectively(P＜0.05).CEUS and CECT have no statistically significant difference in the detection of hepatic metastases (P＞0.05).ConclusionsCEUS can improve detection of hepatic metastases.

Objective To construct a multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector and identify it preliminarily. Methods According to recombinant pcDNA3-p30-ROP2 restriction sites,HBV HBsAg gene sequences of primers were designed and synthesized to amplify target fragment,and then cloned into pcDNA3-HbsAg-p30-ROP2 expression vector. Af-ter sequencing,it was identified finally by restriction enzyme digestion and other molecular biology techniques. Results HBV HBsAg gene segment was amplified by PCR and the multi-gene recombinant pcDNA3-HBsAg-p30-ROP2 expression vector was constructed and identified to be correct as theoretical values. The PCR and restriction enzyme digestion results showed that HBsAg and p30-ROP2 gene in recombinant plasmid were confirmed by DNA sequencing. Conclusion The multi-gene recombinant pcD-NA3-HBsAg-p30-ROP2 expression vector is successfully constructed.

OBJECTIVETo assess the efficiency and safety of radiofrequency ablation (RFA) of recurrent hepatocellular carcinoma (RHCC) after hepatectomy and to investigate efficacy of RFA for patients with early and late phase recurrence, separately, setting 1 year as the cut-off between the early and late phases.

METHODSA total of 42 patients with 77 RHCC and a history of hepatic resection for hepatocellular carcinoma (HCC) underwent ultrasound-guided percutaneous radiofrequency ablation in our department and entered this study (RHCC group). The average diameter of RHCC was (3.8 +/- 1.4) cm (range, 1.5-6.6 cm). 21 of the 42 RHCC patients had Child-Pugh class A cirrhosis 19, class B and two, class C cirrhosis. The average interval between initial surgery and the diagnosis of recurrence was 22.8 months (range, 1-96 month). 42 RHCC patients were divided into two groups as early recurrence group including 20 patients with 40 RHCC, and late recurrence group including the other 22 patients with 37 RHCC according to the recurrence interval. During the same period 148 patients with 217 primary HCC were also treated by RF ablation and regarded as primary HCC group. The average diameter of primary HCC was (4.0 +/- 1.4) cm (range, 1.2-7.0 cm). Regular follow-up with enhanced CT was performed to evaluate the treatment results. Ablation was considered a success if no contrast enhancement was detected in the treated area on 1 month CT scans.

RESULTSThe ablation success rate, local recurrence rate, new tumor incidence and mean survival in RHCC group were 90.5%, 14.3%, 38.1% and (28.0 +/- 3.5) months, respectively, which were similar to the corresponding results of 87.2%, 16.2%, 37.8% and (39.0 +/- 2.1) month in primary HCC group. However, when further comparison was performed between early recurrence group, late recurrence group and primary HCC group, there were some significant differences. The incidence of new tumors in early recurrence group was significantly higher than that in late recurrent group (60.0% vs. 18.2%, P = 0.005); early recurrence group survived shorter than primary HCC group [(15.4 +/- 2.3) vs. (39.0 +/- 2.1) months, P < 0.005]. The survival time was similar between late recurrence group and primary HCC group. One case was found haemorrhage after RFA and recovered with conservative treatment. No major complications occurred in the remaining 41 patients.

CONCLUSIONSRF ablation is generally effective and safe in treating RHCC. And it's more effective in late recurrence than in early recurrence.

Adult ; Aged ; Carcinoma, Hepatocellular ; diagnostic imaging ; surgery ; Catheter Ablation ; Female ; Humans ; Liver Neoplasms ; diagnostic imaging ; surgery ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Retrospective Studies ; Survival Rate ; Time Factors ; Treatment Outcome ; Ultrasonography, Interventional

BACKGROUNDNanobone putty is an injectable and bioresorbable bone substitute. The neutral-pH putty resembles hard bone tissue, does not contain polymers or plasticizers, and is self-setting and nearly isothermic, properties which are helpful for the adhesion, proliferation, and function of bone cells. The aim of this study was to investigate the osteogenic potential of human bone morphogenetic protein 2 (hBMP2) gene activated nanobone putty in inducing ectopic bone formation, and the effects of the hBMP2 gene activated nanobone putty on repairing bone defects.

METHODSTwenty four Kunming mice were randomly divided into two groups. The nanobone putty + hBMP2 plasmid was injected into the right thigh muscle pouches of the mice (experiment side). The nanobone putty + blank plasmid or nanobone putty was injected into the left thigh muscle pouches of the group 1 (control side 1) or group 2 (control side 2), respectively. The effects of ectopic bone formation were evaluated by radiography, histology, and molecular biology analysis at 2 and 4 weeks after operation. Bilateral 15 mm radial defects were made in forty-eight rabbits. These rabbits were randomly divided into three groups: Group A, nanobone putty + hBMP2 plasmid; Group B, putty + blank plasmid; Group C, nanobone putty only. Six rabbits with left radial defects served as blank controls. The effect of bone repairing was evaluated by radiography, histology, molecular biology, and biomechanical analysis at 4, 8, and 12 weeks after operation.

RESULTSThe tissue from the experimental side of the mice expressed hBMP2. Obvious cartilage and island-distributed immature bone formation in implants of the experiment side were observed at 2 weeks after operation, and massive mature bone observed at 4 weeks. No bone formation was observed in the control side of the mice. The ALP activity in the experiment side of the mice was higher than that in the control side. The tissue of Group A rabbits expressed hBMP2 protein and higher ALP level. The new bone formation rate and antibending strength of group A was significantly higher than those of group B and C. The defects in blank control were not healed.

CONCLUSIONSThe hBMP2 gene activated nanobone putty exhibited osteoinductive ability, and had a better bone defect repair capability than that of nanobone putty only.

Absorbable Implants ; Animals ; Bone Morphogenetic Protein 2 ; Bone Morphogenetic Proteins ; analysis ; genetics ; Female ; Male ; Mice ; Osteogenesis ; physiology ; Rabbits ; Transforming Growth Factor beta ; analysis ; genetics

Sudden death (SD) is a special kind of death owing to disease,which severely threatening the lives of community population.As the most common type of SD,sudden cardiac death (SCD) has always been a crucial content of identification and research in forensic pathology.This article reviews the research progress from the aspects of epidemiology,morphology,molecular pathology and virtual anatomy of SCD in forensic medicine,so as to provide a reference for the morphological identification,determination of cause of death,and integrated control of this kind of SD.

Objective To investigate the clinical pathologic features of a distinct variant of focal cortical dysplasia (FCD) characterized by neuronal loss of layer four.Methods Between 2005 and 2017,approximately 3 000 surgeries were performed for the treatment of intractable epilepsy at Xuanwu Hospital,Capital Medical University and Yuquan Hospital,Tsinghua University.Retrospective analysis of clinic-pathological data of patients with epilepsy surgery was made and histological manifestations of neuronal loss of cortical layer four were included in this study.Results In this cohort,25 patients (22 males and three females) were identified with early onset pharmaco-resistant epilepsy and regionally circumscribed neuronal loss of cortical layer four in surgical specimens from the occipital lobe.Histologically,except for neuronal loss in cortical layer four in all cases,glial scar lesions were found in some patients.Thus the histology of those cases can be subdivided into two groups:group A (13 cases):neuronal loss of cortical layer four without glial scar lesions;and group B (12 cases):neuronal loss of cortical layer four with glial scar lesions.Due to the prominent horizontal disorganization of cortical layering and lack of any other microscopically visible principle lesion,group A should be classified hitherto as FCD International League Against Epilepsy (ILAE) type Ⅰ b,however,group B with scar lesions and cortical dysplasia around the main leision,should be classified as FCD ILAE type Ⅲd.This retrospective analysis of clinical histories revealed a perinatal distress in 20 patients (80％),suggesting an acquired pathomechanism.Magnetic resonance imaging revealed abnormal signals in the occipital lobe in all patients,and signal changes suggestive of encephalomalacia were found in 18 patients.Surgical treatment achieved favorable seizure control (Engel class Ⅰ and Ⅱ) in 18 patients (75％ among 24 available follow up).Comparion of the two groups with age at epilepsy onset (group A:5.00±2.76,group B:5.01±3.78),the proportion of perinatal distress (group A:11/13,group B:9/12) and the follow-up results (favorable seizure control of the two groups was 9/13,9/11 respectively) showed that there was no statistically significant difference between the two groups.Conculsion Neuronal loss of cortical layer four in the occipital lobe should be classified as a distinct variant of FCD ILAE type Ⅲd.

OBJECTIVETo investigate the expression and significance of survivin, ki-67 and apoptosis index in patients with advanced gastric adenocarcinoma.

METHODSImmunohistochemical SP method for survivin expression as well as cell proliferative index (ki-67) and apoptosis index (TUNEL) was conducted on 120 gastric adenocarcinomas.

RESULTSThe survivin was detected in the cytoplasm of carcinoma cells in 59 (49.17%) of the 120 gastric adenocarcinomas, in 32 (64.00%) of the lymph node metastasis, and in 21 (17.50%) of the 120 basal layer in normal gastric mucosa, respectively. The mean proliferative index (ki-67) in primary tumors was 7.55%, which was significantly lower than the mean proliferative index of 8.34% observed in lymph node metastasis. The mean apoptosis index in primary tumors was 1.16%, which was significantly higher than the mean apoptosis index of 0.89% observed in lymph node metastasis. The frequency of survivin expression was significantly higher in lymph node metastasis than in primary gastric adenocarcinoma. Expression of survivin was significantly correlated with histological subtypes, the depth of invasion, or lymph node metastasis (P < 0.05). There was negative correlation between weighted survivin score and apoptosis index (P < 0.05), but no correlation with proliferative index.

CONCLUSIONThe high level expression of survivin might be a referenced indicator in evaluating differentiation of tumor and in predicting lymph nodes metastasis and estimating apoptosis index.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Apoptosis ; Female ; Gastric Mucosa ; chemistry ; pathology ; Humans ; Immunohistochemistry ; In Situ Nick-End Labeling ; Inhibitor of Apoptosis Proteins ; Ki-67 Antigen ; analysis ; Lymph Nodes ; chemistry ; pathology ; Male ; Microtubule-Associated Proteins ; analysis ; Middle Aged ; Neoplasm Proteins ; Stomach Neoplasms ; metabolism ; pathology

Objective::To establish differential metabolites between different varieties of Angelica sinensis, and provide reference for breeding, introduction, regional cultivation and ecological cultivation of new varieties of A. sinensis. Method::Comprehensive non-target metabonomics analysis was conducted for five new varieties of A. sinensis collected at the same time from the same origin: Mingui No. 1 (MG1), Mingui No. 2 (MG2), Mingui No. 4 (MG4), Mingui No. 5 (MG5), and Mingui No. 6 (MG6). The 50% methanol extract of each variety was taken, and then the differential metabolites among varieties were found by using ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS), software Progenesis QI, principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and non-targeted metabonomics analysis. Differential metabolites were identified based on precise molecular weight, secondary fragments, KEGG database, HMDE database and related literature information. Result::The results of UPLC-Q-TOF-MS and multivariate statistical analysis showed that there were significant differences in the metabolites of five Angelica varieties. As compared with MG1, the contents of chlorogenic acid, ferulic acid, tryptophan and ferulic aldehyde were significantly lower in MG2, MG4, MG5 and MG6, while the contents of ligustilide, coumarin, bovine keratin, palmitin, protocatechualdehyde and linolenic acid were significantly higher (P<0.05). The results of multivariate statistical analysis showed that the metabolites of MG2 and MG5 were similar with those of MG6, but were significantly different from those of MG4.In addition, 38 distinct metabolites were identified, involving 7 potential targeted metabolic pathways. Different varieties of A. sinensis could regulate the synthesis of their metabolites through phenylpropanoid biosynthesis and sesquiterpene-like compounds metabolism, lipid metabolism, amino acid metabolism, carbohydrate metabolism, nucleotide metabolism, carotenoids, linolenic acid, linoleic acid and some other metabolic pathways. Conclusion::UPLC-Q-TOF-MS and Progenesis QI metabonomics techniques were used to compare the chemical constituents of different varieties of A. sinensis from the overall level. The differences and their regularities were found, which could provide reference for quality control, variety sorting, identification, breeding and ecological planting of A. sinensis.