Objective To investigate epigenetic silencing of the proapoptotic gene MAPK10 by methylation in B-cell lymphoma. Methods We examined MAPK10 expression and methylation in seven cell lines derived from B-cell lymphoma by semi-quantitative RT-PCR and methylation-specific PCR (MSP), respectively. Methylation status was further examined in 24 diffuse large B-cell lymphoma (DLBCL), 15 follicular lymphoma (FL) and 12 reactive hyperplasia lymph nodes (LRH) both by MSP and bisulfite genomic sequencing (BGS). Results MAPK10 is silenced or downregulated in all seven B-cell lymphoma cell lines mostly due to promoter methylation. MAPK10 methylation was frequently detected in DLBCL (17 of 24, 71%) and FL (15 of 15, 100 %), but not in 12 LRH tissues. Conclusion MAPK10 is frequently inactivated by tumor-specific methylation, and thus, is a potential biomarker.

Objective In Caucasian populations, the tumor cells of Epstein Barr virus (EBV)-positive classical Hodgkin lymphoma (cHL) patients more frequently express HLA class Ⅰ and HLA class Ⅱ molecules compared to EBV-negative cHL patients. HLA expression (in relation to EBV) in Asian cHL patients has not been previously investigated. Methods 145 cHL patients with formalin-fixed, paraffin embedded tissue blocks available from Beijing, China were randomly selected. Hematoxylin & Eosin-stained slides were used to reclassify the histological subtypes according to the WHO classification. EBV status was determined by visualization of EBER in tumor cells using in situ hybridization. Membranous expression of HLA molecules was detected by immunohistochemistry using antibodies HC-10 (class Ⅰ heavy chain) and antiβ2-microglobulin for HLA class Ⅰ, and CR3/43 for HLA class Ⅱ. Results EBV (+) tumor cells were observed in 40 % (58/145) of the cHL patients. As expected, the percentage of EBV(+) cases was much higher in the mixed cellularity subtype (71%) than that in the nodular sclerosis subtype (16 %) (P ＜0.001). The expression of HLA class Ⅰ was observed in 79 % of the EBV (+) cHL cases and in 30 % of the EBV (-) cases (P ＜0.001). For HLA class Ⅱ, 52 % of EBV(+) cHL cases were positive, compared to 43 % in EBV(-) cases (P =0.277). Conclusion The results in China population were similar to that in the Caucasian population for HLA class Ⅰ, but not for HLA class Ⅱ.

There are some major changes win the revised 2016 WHO Classification of Lymphoid Neoplasms.Based on the clinicopathological changes and genetic/molecular findings in the past years,the new classification clarified the diagnosis and clinical management of some very early stages of lymphoproliferative disorders,refined the diagnostic criteria for some lymphoid neoplasms,and further emphasized the significance of genetic/molecular studies in the diagnosis and clinical treatment of lymphomas.A small number of new provisional entities were added to the 2016 edition.

Objective To explore the clinical significance of changes of monocyte chemoattractant protein-1(MCP-1) in patients with pregnancy induced hypertension(PIH).Methods The MCP-1 levels of urine and blood were measured by enzyme linked immunosorbent assay(ELISA) in 96 patients with PIH and 49 normal pregnant women.Results The MCP-1 levels in blood and urine in patients with PIH were significantly higher than those in normal pregnant women,and were associated with the severity of the disease.The urine MCP-1 level was positively correlated with the levels of ?_2-microglobulin(?_2-MG),N-acety1-?-D-glucosaminidase(NAG),interleukin-6(IL-6) and retinal-binding protein(RBP),and was negatively correlated with creatinine clearance rate(Ccr).Conclusion The detection of MCP-1 is useful for examining the severity of PIH and the degree of renal injury.

Objective:To investigate the clinicopathological and immunohistochemical characteristics of soft tissue mesenchymal chondrosarcoma. Methods:The clinical material,pathological and immunohistochemical characteristics(reactiontoLCA,CD3,CD20,CD45RO,CD79a,CD99,NSE,S-100,Syn,CgA,CK7,CK19,EMA,Coll-Ⅱ,Sarcomeric-Actin,Desmin,Ki-67,P53) of 2 cases of soft tissue mesenchymal chondrosarcoma in Jishuitan Hospital between 1995 and 2005 were reviewed and followed up. Results:The two patients were both females. The tumors were located in the low extremity muscles. The main roentgenographical appearance was stippled calcification in tumor and calcification at the edge of the tumor. The histological characteristic features showed undifferentiated small cells together with islands of chondrosarcoma;there was hemangiopericytoma-like arrangement of small cells.The tumor cells were positive for CD99,NSE,Syn,CgA;The cells in chondroid matrix were positive for S-100;chondroid matrix was positive for Coll-Ⅱ. All tumor cells were negative for LCA,CD3,CD20,CD45RO,CD79a,Sarcomeric-actin,Desmin and CK7,CK19, and EMA. The patient with followed radiotherapy was alive. and the other without radiotherapy dead. Conclusion: Mesenchymal chondrosarcoma of soft tissue has the characteristics of primary mesenchyme which differentiates to congenital cartilage. The pathological characteristics of bimophic pattern and roentgenographical appearance of tumor are useful for diagnosis.The prognosis is poor.

Objective To study risk factors of cardiovascular disease (CVD) and status of bone mineral density (BMD) in women with hypoestrogenism.Methods From Jul 2011 to April 2013,a total of 256 women with hypoestrogenism in the First Affiliated Hospital of Shanxi Medical University were enrolled in this retrospective study,which were divided into four groups:133 women in ppausal group,25 women in premature ovarian failure (POF) group,67 women in menopausal transition group and 31 women in premature ovarian failure transition group.General statue,CVD risk factors and BMD were compared among four groups.General statue include menopausal period,menopausal symptoms (Kupperman Index),CVD risk factors include body mass index,blood pressure,waist circumference,waist-hip ratio,blood lipids and glucose,BMD include left hip,lumbar spine bone mineral density and T or Z value.Results (1) The median menopausal period were 3.4 years in postmenopausal group and 3.6 years in premature ovarian failure group,which did not show no statistical difference (P ＞ 0.05).Kupperman Index in four groups were 12 in postmenopausal group,9 in POF group,9 in menopausal transition group and 8 in premature ovarian failure transition group,which reached statistical difference (P ＜ 0.05).(2) The difference of body mass index (BMI),waist circumference,waist-hip ratio,diastolic blood pressure were no statistically significant among four groups(P ＞ 0.05) ; the systolic blood pressure in four groups were 120,110,110,110 mm Hg (1 mm Hg =0.133 kPa),their differences were statistically significance (P ＜ 0.05); the high-density lipoprotein (HDL-C) was 1.6 mmol/L in postmenopausal group,and 1.3 mmol/L in premature ovarian failure transition group,their differences were all statistically significance (P ＜ 0.05) ; the difference of the fasting plasma glucose (FPG) was not statistically different in 4 groups (P ＞0.05).(3) The abnormal rate of lower bone mass in lumbar spine were 57％ (46/81) postmenopausal group,8/15 in POF group,32％ (9/28) in menopausal transition group,12/19 in premature ovarian failure transition group,and osteoporosis was 9％ (7/81),3/15,1％ (3/28)and 0 respectively,their differences were statistically different (P ＜ 0.05) ; the abnormal rate of BMD of left hip and lumbar spine of 11/15 and 12/16 in POF group was higher than 65％ (53/81) in postmenopausal group.In the mean time,the abnormal rate of BMD of left hip and lumbar spine were,12/19 and 10/20 in premature ovarian failure transition group,which were significantly higher than 43％ (12/28) and 39％ (12/31) in the menopausal transition group.Conclusions The menopausal symptoms resulting from hypoestrogenism in natural postmenopausal women are mostly remarkable.The decrease of BMD in lumbar spine is more significant than that of left hip among postmenopausal women.Women with earlier menopause was prone to cause the changes of blood fat and abnormal of BMD,especially HDL-C decreased significantly compared with those natural postmenopause,it is more likely to cause CVD and osteoporosis.

To produce monoclonal antibody (mAb) specifically against human thrombomodulin (hTM), an immune-tolerizing procedure was employed to generate monoclonal antibodies specific to hTM. Female BALB/c mice were first immunized with CHO cells following at 10 min, 24 h, 48 h by intraperitoneal injection of different doses of cyclophosphamide (CP) 2 times at an interval of 2 weeks, thereby tolerizing the mice to common epitopes shared between CHO and CHO-TM5 cells. Subsequently the selected mice with the lowest titer of serum polyclonal antibody by cellular enzyme-linked immunoabsorbent assay (CELISA) were immunized with CHO-TM5 cells, which have stable high level expression of hTM, to produce antibodies specific to hTM 3 times at an interval of 2 weeks. On the third day after the third immunization, mouse with the highest titer of serum polyclonal antibody was sacrificed and spleen cells were harvested to prepare hybridoma cells with SP2/0 cells at the ratio of 10 to 1. Hybridoma cells were then cultured at 96 well plates for screening with CELISA. To improve probability to obtain specific mAb, CELISA was applied twice. The first CELISA was done with polyethylene ELISA plate with a monolayer of CHO-TM5 cells. The positive clones from the first screen were then selected by reacting with similar screening ELISA plate but having CHO cells monolayer instead. Only clones that were positive for the first screening and negative for the second screening were kept, and called as CHO-TM5 +CHO- hybridoma cells. BALB/c mice were intraperitoneally injected with the selected hybridoma cells. Ascites were collected and monoclonal antibodies were purified using FPLC, and its Ig class, subclass, and titer were then determined respectively. The specificity of the yielded mAb was identified with CELISA, flow cytometry, ABC immunohistochemistry and immunoblotting. Detection of CELISA showed that 100 mg/kg dose of CP could tolerize the mouse to common epitopes shared between CHO and CHO-TM5 cells. And CELISA also discovered that all hybridomas positive for CHO-TM5 cells were negative for CHO cells. Five lines of positive hybridoma cells had been obtained altogether and 2F7 was selected randomly for next investigation. The Ig subclass of the mAb 2F7 was IgG1 and the titer of ascitic mAb was 1?10-6. Furthermore, the content of ascitic mAb was 19.56 g/L and chromosome numbers is 98. Flow cytometry, CELISA and Western blotting assays demonstrated that mAb 2F7 could specifically recognize hTM expressed on CHO-TM5 and human umbilical vascular endothelial cells (HUVEC). Meanwhile, the tissue specificity of mAb 2F7 was also identified by immunohistochemical ABC staining. On the other hand, Western blotting assays indicated that mAb 2F7 could recognize the antigen protein with 105 ku molecular mass under reduction condition. Moreover, the dissociation constant of mAb 2F7, 1.22? 10-9 mol/L, indicated the affinity higher than some others. The results suggest that the immunotolerizing protocol provides a convenient general method for producing antibodies specific to desired protein isoforms. mAb 2F7 can specifically recognize the natural hTM expressed mainly on vascular endothelial cells, which will potentially useful for investigating the functions and clinic values of hTM.

Objective To elucidate clinical pathological features of primary mediastinal B-cell lymphoma (PMBL) and its difference compared with diffuse large B-cell lymphoma,not otherwise specified (DLBCL,NOS).Methods The clinical histories and pathological datas of 24 PMBL cases and 31 cases of DLBCL,NOS as the control group were collected.Immunohistochemical staining and a follow-up study was conducted.Results The distribution of gender showed significant difference when the age of onset of PMBL patients was obviously younger with the medial age of 30 years old (P ＜ 0.001).All cases presented as a huge mass in mediastinal site with compression symptoms.PMBL was similar to DLBCL in the morphology of tumor cells but fibrosis of various degrees was common,more than 70.8 ％ (17/24) cases had the collagen bundles split.CD23 positive rate (40.0 ％,6/15) in PMBL was significantly higher than the control group (3.2 ％,1/31)(P =0.003).Conclusion PMBL frequently occurs in young female people,mostly happens in mediastinal site and adjacent area,but rarely has distant dissemination.PMBL has the characteristics of various degrees of collagen fiber hyperplasia,and CD23 positive could be used to differentiate PMBL from DLBCL,NOS.

Background and purpose:Solitary plasmacytoma of bone(SBP) is a rare disease,reports releveant to this disease are rarely seen. The purpose of this study was to investigate the relation between the clinical features and the prognosis of solitary plasmacytoma of bone. Methods:We reviewed the data of 12 patients diagnosed with solitary plasmacytoma of bone from 1998 to 2007 in Peking University third hospital,the clinical features,treatment and prognosis of the patients were analyzed retrospectively. Results:The age ranged from 37-71 years(mean 49.6 years) ,the male/female ratio was 3 to 1. Immunophenotype analysis showed that 11(91.6%) cases were positive for CD79a,10(83.3%) positive forVS38C,and all negative for CD20. With 12 to 87 months follow-up(average 40?22 months) ,three cases(33%) developed to multiple myeloma,two of them died from infection,the median survival time was 73 months,the 3 year and 5 year survival rate were 90 percent and 75 percent respectively. Conclusion:Middle and old male are more likely to develop SP. The prognosis is good,but some of them can progress to multiple myeloma.

AIM: To detect sequence and mutation of K-ras oncogene in tissue and stool DNA of patients with colorectal cancer in order to provide a method of noninvasive and simple colorectal cancer diagnosis. METHODS: DNA was separated and purified from colorectal cancer tissue or stool of patient with colorectal cancer, then the K-ras gene was amplified by PCR and PCR products were cloned, the K-ras gene was sequenced, and the mutation was identified. The expression of color/colorectal cancer antigen was inspected by immunohistochemical technique. Stool sample of patient with colorectal cancer was detected with enzyme-linked immunosorbent assay (ELISA). RESULTS: K-ras gene sequence of the stool was completely same as that of the tissue of the patient;K-ras mutation was detected in one case. There was relativity between the mutation of K-ras gene and the pathology type of colorectal cancer and the expression level of colorectal cancer antigen in stool sample. CONCLUSION: It is feasible that colorectal tumors can be detected by a noninvasive method based on the molecular pathogenesis of the disease. Detecting K-ras gene mutations of stool DNA can provide bases for the screening, early detection, and prognosis to patients with colorectal cancer.