Clinicopathologic study of pimary paget disease in esophagus / 临床与实验病理学杂志

Purpose To investigate the clinicopathological and diagnostic characteristics of primary Paget disease (PD ) in esophagus. Methods The clinical presentation, histological observation and immunohistochemical staining were analyzed in four cases of primary PD involved esophagus and related literatures were reviewed. Results The patients were all male, aged from 61 to 74 years old. All the tumors were originated from the mucosa of the esophagus. Histologically, the Paget cells showed a single or small nesting and acinar distribution in the esophage-al mucosa. Adenocarcinoma in situ were seen in 2 cases and squamous cell carcinoma was seen in one of them. Immunohisto-chemically, the Paget cells were typically strongly positive for Ckpan, CKL, and CK7, while negative for CKH, CK5/6, CK14, p63. Conclusion Primary esophagus PD is rare. It can develop alone in esophagus or accompanied with adenocarcinoma in situ, invasive squamous cell carcinoma. The correct diagnosis need detailed pathological observation, immunohistochemical ev-idence and medical history.

Influence of donor mouse age on the establishment of murine acute graft versus host disease model after allogenic hematopoietic stem cell transplantation / 中国实验血液学杂志

This study was purposed to elucidate the influence of donor mouse age on the establishment of murine acute graft versus host disease (aGVHD) model after allogenic hematopoietic stem cell transplantation. The male mice with 2-week-old, 10-week-old and 18-week-old mice (BALB/cH-2Kb) were taken as donors. The 8-week-old mice (BALB/c, H-2Kd) were selected as recipients. Each group animals were irradiated with 7.5 Gy (60)Co for total body, the recipient mice were injected intravenously with 1 × 10⁷ bone marrow cells and 1 × 10⁷ spleenoctyes from various donors in 4-5 hours after irradiation. Mouse transplant characteristics and survival were observed every day. The white blood cell number in peripheral blood of each group were counted at day 5, 10, 15, 20, 25 and 30 after transplantation. Furthermore, the pathological damage in the liver, spleen, lung and intestines were evaluated by sectioning and in situ hematoxylin-eosin (HE) staining. The results showed that compared with the 2-week-old and 10-week-old donor groups, mice received bone marrow (BM) cells and splenocytes from 18-week-old mice showed higher incidence of aGVHD, lower clinical GVHD scores and suffered from diarrhea, ruffled hair, a hunched posture, and diminished body weight. In contrast, mice received BM cells and splenocytes from 2-week-old donor mice indicated attenuated GVHD symptoms and survived longer. The histo-pathological analysis in 18-week-old donor group demonstrated the most serious pathological damage in the liver, spleen, lung and intestines. It is concluded that the donor age has been confired to have an obvious influence on the establishment of murine aGVHD model. This study lay an important foundation for establishing animal models and may be helpful for further study.

Construction of mouse VCAM-1 expression vector and establishment of stably transfected MSC line C3H10T1/2 / 中国实验血液学杂志

This study was aimed to construct the mouse VCAM-1 expression vector, to establish the stably transfected MSC line and to investigate the effect of VCAM-1-modified mesenchymal stem cells (MSC) on the immunological characteristics of MSC. The cDNA of murine VCAM-1 gene was amplified by RT-PCR from the total RNA isolated from the mouse spleen; then the cDNA was inserted into the retrovirus vector PMSCVmigr-1; the recombinant plasmid was confirmed by restriction endonuclease experiments and sequencing, then designated as PMSCVmigr-1-mVCAM-1; the recombinant plasmid PMSCVmigr-1-mVCAM-1 was transfected into 293 cells by lipofecamin and the supernatant was collected to transfect MSC cell line (C3H10T1/2). Moreover, VCAM-1 expression on MSC was evaluated by FACS. Furthermore, the inhibitory effect of VCAM-1-MSC on lymphocytic transformation was tested by (3)H-TdR incorporation assay. The results indicated that the successful construction of recombinant retroviral expression plasmid of mouse VCAM-1 was confirmed by digesting and sequancing. After transfection of MSC with retroviral supernaptant, the high expression of VCAM-1 on MSC could be detected by flow cytometry. The MSC high expressing VCAM-1 could significantly inhibit the proliferation of Con A-inducing lymphocytes in dose-depentent marrer. It is concluded that recombinant retroviral encoding VCAM-1 (PMSCVmigr-1-mVCAM-1) has been successfully constructed and mouse VCAM-1 has been stably expressed in C3H10T1/2. MSC over-expressing VCAM-1 show more potent immunosuppressive effect on cellular immune reaction in vitro. Our data laid a foundation for the subsequent studying the effect of VCAM-1 transfecting into MSC on immune related disease study.

Effect of different irradiation doses on the establishment of murine cGVHD model after MHC matched spleen stem cell transplantation / 中国实验血液学杂志

This study was aimed to investigate the effect of different irradiation doses on the establishment of murine cGVHD model after MHC matched spleen stem cell transplantation. The male mouse BALB/c(H)-2d was totally irradiated with different radiation dose of (60)Co (TBI), then was infused with the same number of splenocytes from MHC matched DBA/2 male mice. After transplantation, the bodyweight, general appearance, hair changes, survival time and pathological damage were observed. The results indicated that compared to the control group (0 Gy) and the 7.0 Gy group, the mice irradiated with 7.5 Gy and 8.0 Gy showed cGVHD symptoms and obvious pathological damage. At the end of experiments (60 d after transplantation), all mice irradiated by 7.5 Gy survived while only 60% animals survived in the 8.0 Gy group. It is concluded that under infusion of 10(8) MHC matched splenocytes per mouse, 7.5 Gy irradiation is appropriate to efficiently establish cGVHD model. This study laid an important foundation for further studying the pathogenesis, biological characteristics, and intervention factors of cGVHD.

Effect of intercellular adhesion molecule-1 on the migration in vitro of murine mesenchymal stem cells and its related mechanism / 中国实验血液学杂志

This study was aimed to investigate the effect of intercellular adhesion molecule-1 (ICAM-1) on the migration in vitro of the murine mesenchymal stem cells (MSC) and its related mechanisms. The migration ability of murine MSC (C3H10T1/2), ICAM-1 transfected MSC (C3H10T1/2-MIGR1-ICAM-1) and empty vector-transfected MSC (C3H10T 1/2-MIGR1) were assayed in vitro by using the transwell system. Briefly, the cells were seeded on the membrane with 8 µm aperture and the fetal bovine serum was used as the chemotactic agent to induce MSC migration. The transmigrated cells were stained by crystal purple as well as DAPI for 8 h and 12 h respectively. The absolute cell numbers were counted and the migration rates of MSC were evaluated in each group. To explore the potential mechanisms which control the migration of MSC, the specific chemical inhibitors of MAPK pathway (SB203580, PD98059 and JNK inhibitor II) were added to the transwell system and the alteration of the MSC migration ability were evaluated at 12 h. The results showed that the migration ability at 8 h and 12 h of the ICAM-1-transfected MSC increased. Both absolute cell number and migration rate of MSC were significantly up-regulated by ICAM-1. Furthermore, the promoting effect of ICAM-1 on migration was partially suppressed by the inhibition of JNK/SAPK pathway. The transmigrated cell numbers and the migration rate decreased with the addition of JNK inhibitor II. However, the ICAM-1 promoting migration of MSC was not suppressed by the inhibitors for ERK/MAPK and p38/MAPK pathway did not work in the present study. It is concluded that ICAM-1 can induce mouse MSC migration in vitro, and the promoting effect is partially dependent on the activation of JNK/SAPK pathway.

Latest research progress on pathogenesis of chronic graft versus host disease and its related problems / 中国实验血液学杂志

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is an effective method for the treating of malignant diseases of hematopoietic system or non-malignant proliferative diseases, but the occurrence of graft-versus-host disease (GVHD) limits the success rate of hematopoietic stem cell transplantation. Moreover, chronic graft-versus-host disease (cGVHD) is the main factor affecting the long-term survival rate and life quality of recipient after hematopoietic stem cell transplantation. In this article, the latest research progress of the pathogenesis of cGVHD and related problems are reviewed from the thymus, cytokines, T lymphocyte subsets, B lymphocytes and its secreted antibody.

ICAM-1 regulates differentiation of MSC to adipocytes via activating MAPK pathway / 中国实验血液学杂志

This study was aimed to explore the molecular mechanism of the regulatory effects of ICAM-1 on the differentiation of mesenchymal stem cells (MSC) to adipocytes. The murine MSC cell line C3H10T 1/2 was treated with the supernatants contained plasmid MIGR1-ICAM-1 and MIGR1-ICAM-1/MSC (high expression of ICAM-1), the activation of the pathway was detected by Western blot. The ICAM-1 modified MSC and its control cells named MIGR1/MSC were cultured in adipocyte medium with or without the inhibitors of the ERK, P38, and JNK pathway. Oil-red-O staining was used to detect the lipid accumulation, and the expression of C/EBPα and PPARγ in differentiation of MSC to adipocyte were examined by real-time-PCR. The results showed that the overexpression of ICAM-1 stably activated the ERK, P38, and JNK pathway in MSC. Inhibiting of the activation of ERK pathways by chemical inhibitors up-regulated the mRNA expression level of C/EBPα and PPARγ in MIGR1-ICAM-1/MSC while inhibiting of P38 pathway resulted in lower mRNA expression of the transcription factors. Consistent with the mRNA expression, the lipid droplets were getting smaller and number of adipocytes increased when P38 pathway was inhibited, while bigger lipid droplet and increased quantity of adipocytes were identified in MIGR1-ICAM-1/MSC with the addition of ERK pathway inhibitor. It is concluded that ICAM-1 may suppress MSC differentiate into adipocyte via activating ERK pathway, while it can maintain the adipogenesis of MSC though P38 pathway.

Clinical and pathological analysis of 41 cases of acute leukemia combined with intracranial hemorrhage / 中国实验血液学杂志

This study was aimed to summarize the clinical and pathological features of patients with acute leukemia combined with intracranial hemorrhage. The clinical and pathological data of 41 adult patients diagnosed as acute leukemia in our hospital from 1953 to 1990 year were analyzed retrospectively. The results showed that there were 35 cases of AML, 6 cases of ALL; 9 cases in clinical hematologic remission, 32 cases in non-remission, 3 cases of AL with hypertension, 2 cases of AL with diabetes, 4 cases of AL with sepsis, 19 cases with WBC ≥ 100×10(9)/L; the pathologic examination showed 4 cases of AL accompanied with disseminated intravascular coagulation, 10 cases with prothrombin time INR ≥ 1.5, 26 cases with multifocal intracranial hemorrhage, 7 cases with single intracranial hemorrhage, 8 cases with diffused spotting intracranial hemorrhage; the examination also showed that 84 hemorrhage foci were found in 41 cases of AL, among them 46 foci located under cerebral cortex, 23 foci in cerebellum, 6 in basal ganglia, 5 foci in pons, 2 foci in thalamus, 2 foci in spinal cord. It is concluded that the intracranial hemorrhage is a major cause resulting in death of AL patients which should be think highly, and the diagnosis and treatment should be conducted through comprehensive analysis.