Primary intraosseous hematopoietic pseudotumor: clinicopathological analysis and 9-year follow-up of 3 cases / 南方医科大学学报

We analyzed the clinicopathological data of 3 cases of primary intraosseous hematopoietic pseudotumor (IHPT), which had been previously misdiagnosed as malignancies or metastases both clinically and pathologically. Two of the patients received close follow-up for 132 and 100 months, and one patient was lost to follow-up, and the tumors were confirmed to be benign in all the 3 cases. IHPT is a rare benign intraosseous solid lesion consisting of tissues resembling normal hematopoietic tissue, and can be easily misdiagnosed as malignancy. Understanding the clinicopathological features and the outcomes of the disease can facilitate the clinical decisions on individualized diagnosis and therapeutic regimens.

Culture and identification of tumor stem cells from surgically resected colorectal cancer tissues / 南方医科大学学报

OBJECTIVE@#To obtain cancer stem cells (CSCs) from surgically resected colorectal cancer specimens and identify their stem cell characteristics.@*METHODS@#Colorectal cancer tissue specimen obtained from a patient undergoing radical resection of colorectal cancer were implanted in nude mice, and the xenograft was harvested 1 month later to obtain purified tumor cells by enzyme digestion and adherent culture. The CSCs were screened by limiting dilution method and serum-free culture to identify their phenotypes. Soft agar colony assay was used to assess the proliferative ability of the CSCs and human colorectal cancer cell line SW480. The tumorigenic ability of the isolated CSCs and SW480 cells was evaluated by observing their subcutaneous tumor formation in nude mice. Western blotting and immunofluorescence assay were used to detect the immunophenotype of the CSCs and SW480 cells.@*RESULTS@#The primary cultured CSCs from clinical specimens of colorectal cancer underwent differentiation in the presence of serum in the culture. Soft agar colony formation assay showed that the CSCs had a colony formation rate above 50%, significantly higher than the rate of colorectal cancer SW480 cells (4.41%; < 0.01). In nude mice, subcutaneous injection of 500 CSCs was sufficient to result in subcutaneous tumor formation, while the injection of 500 SW480 cells failed to form any subcutaneous tumors. The CSCs expressed CD133 and CD44 but not CK7, while SW480 cells expressed CK7 but not CD133 or CD44.@*CONCLUSIONS@#CSCs can be derived by primary culture of cancer cells obtained from surgically resected colorectal cancer tissue followed by serum-free culture, and the CSCs obtained have self-renewal and differentiation abilities.

murine colonic adenocarcinoma cell line CT26.WT / 实用医学杂志

Objective To explore the antigen presentation of CT26.WT via intra-peritoneal injection. Methods The intra-peritoneal injection model was made via injecting cell suspensions in mice. The spleen was isolated from BALB/c mice toco-culture with CT26.WT to detect tumor-killed ability. Phenotype identification methods and CCK8 massy were used to measure the ability of antigen presentation and stimulate T lymphocyte proliferation. IHC was used to detect the expression of B7H4 in normal and tumor tissues. Results Along with the extension of intra-peritoneal injection, the surviving number of cells was increased, contrary to the apoptosis. DC cells failed in maturation and impaired in stimulating T lymphocyte proliferation. B7H4 was higher in tumor tissues. Conclusions With the extension of intra-peritoneal injection, the mature DC cells were scared in number, resulting in the impairement of antigen-presentation. Moreover, the higher B7H4 expression in tumor tissues led to the lack of second signals which may stimulate T cells. Consequently, the ability of T cells in killing tumor cells was decreased so that they escape immunosurveillance.

Serum follicle-stimulating hormone in combination with serum inhibin B evaluates spermatogenesis of azoospermic men / 南方医科大学学报

<p><b>OBJECTIVE</b>This study is in an attempt to evaluate the diagnostic significance to predict the spermatogenesis of azoospermic men in examination of serum follicle-stimulating hormone (FSH) combination with serum inhibin B (INHB).</p><p><b>METHODS</b>Quantitative examination of serum FSH and INHB was performed in 95 case of azoospermic men. According to their classifications of testicular biopsy with histopathological examination, there were 20 patients of Sertoli cell only, 25 of hypospermatogenesis, 18 of spermatogenic maturation arrest (complete or incomplete), and 32 of normal spermatogenesis. The association of serum FSH and INHB levels with histopathological classifications were analyzed by using statistical software.</p><p><b>RESULTS</b>Serum FSH, INHB and INHB/FSH levels of Sertoli cell only differed with statistical significance from hypospermatogenesis, spermatogenic maturation arrest and normal spermatogenesis (P<0.05). FSH, in which there were no statistical significance among the latter three classifications (P>0.05). Serum FSH, INHB and INHB/FSH levels were no relationship with maturation arrest (P>0.05), but were negatively related to the other classifications (P<0.05). INHB level less than 28.55 pg/ml predicted Sertoli cell only in a sensitivity of 97% and a specificity of 85%.</p><p><b>CONCLUSION</b>Serum FSH and INHB levels is ineffective to distinguish the spermatogenic classifications from azoospermic men, but they are available to confirm the disease of Sertoli cell only. The other abnormalities of azoospermic men is also dependent on bioptic histopathology to confirm the subtypes.</p>

Combined evaluation of serum follicle-stimulating hormone, inhibin B, chromosome karyotyping and AZF microdeletion of Y-chromosome for predicting outcomes of testicular sperm aspiration in azoospermic patients / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the value of combined evaluation of serum follicle-stimulating hormone (FSH), inhibin B (INHB), chromosome karyotyping and AZF microdeletion of Y-chromosome (AZF-MD-Ych) in predicting the success of testicular sperm aspiration (TESA) in azoospermic patients.</p><p><b>METHODS</b>A total of 262 azoospermic patients were divided into two groups with normal (n=162) and abnormal (n=100) serum FSH levels. INHB levels, INHB/FSH ratio, chromosome karyotype patterns of the peripheral lymphocytes, and AZF-MD-Ych were compared between the two groups. Among the patients receiving TESA, the success rate of the procedure was compared between the two groups after excluding abnormalities in INHB, chromosome karyotype and AZF-MD-Ych.</p><p><b>RESULTS</b>Significant differences were found between the two groups in serum INHB level, INHB/FSH and chromosome karyotypes (P<0.05), but not in AZF-MD-Ych (P>0.05). After excluding the abnormalities in chromosome karyotypes, AZF-MD-Ych and INHB, sperms were obtained successfully by TESA from 61.82% (34/55) of patients with normal FSH but from none of those with abnormal FSH (P<0.01).</p><p><b>CONCLUSION</b>A combined evaluation of serum FSH, INHB, chromosome karyotypes and AZF-MD-Ych can effectively predict the success of TESA in azoospermic patients, and abnormalities in all the 4 indices suggest a very low success rate of sperm retrieval by TESA.</p>

A simple method for isolating and culturing single cancer stem cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop an effective method for isolating and culturing single cancer stem cells.</p><p><b>METHODS</b>The capillary glass tube was stretched on fire and connected to a sterile plastic tube to prepare the single cell separation apparatus. Single SW480 cell clone spheres in serum-free culture were marked with CD133 and CK7, and the single cancer stem cells were separated and cultivated in 96-well plates or microdrop covered by paraffin.</p><p><b>RESULTS</b>SW480 cell clone formation rate was about 1.04%, and the cell clone spheres highly expressed CD133 with low CK7 expression. The isolation of the single cancer stem cells showed a success rate of 98.99% using the separation device. The cell division profile was comparable between the cell cultures in microdrop and 96-well plates in the initial 2 cell divisions (P>0.05), whereas prolonged cell division occurred afterwards in the microdrop culture as compared to 96-well plate culture. The cell population expansion of the single cancer stem cells was similar between microdrop culture (11.5%, 22/192) and 96-well plate culture (9.2%, 17/184) (P>0.05).</p><p><b>CONCLUSIONS</b>Single SW480 cells can develop into cancer stem cell spheres. Microdrop culture is convenient and stable, and can be the primary choice for single cancer stem cell culture.</p>

Allogeneic hematopoietic stem cell transplantation for blastic plasmacytoid dendritic cell neoplasms / 白血病·淋巴瘤

Objective To investigate the effect of allogeneic hematopoietic stem cell transplantation (allo-HSCT) with intensified conditioning regimen followed by rapidly tapering immunosuppressants and sequential minimal residual disease (MRD)-guided donor lymphocyte infusion (DLI) post-transplantation on outcome of blastic plasmacytoid dendritic cell neoplasm (BPDCN).Methods Two cases of BPDCN from January 2009 to May 2011 in Nanfang hospital were diagnosed according to 2008 WHO classification of tumours of haematopoietic and lymphoid tissues.Case 1 initially presented with typical cutaneous involvement and was promptly diagnosed with CD+4CD+56LCA+TdT+CD+43 BPDCN by skin biopsy.Case 2 was recognized as acute lymphocyte leukemia and acute non-lymphocytic leukemia,which was diagnosed to BPDCN at recurrence through flow cytometry analysis.Total-body-irradiation plus cyclophosphamide based intensified conditioning regimen were followed by allo-HSCT from sibling donor.Graft-versus-host disease (GVHD) prophylaxis consisted of cyclosporine and methotrexate.Anti-thymocyteglobulin was included additionally for haploid donor allo-HSCT.Multi-color labeling flow cytometry was performed to monitor MRD.Rapidly tapering of prophylactic immunosuppressants and sequential MRD-guided donor lymphocyte infusion (DLI) were performed to control relapse of primary malignancy.Results Two cases of BPDCN received allo-HSCT from sibling donor after intensified conditioning regimen.Both patients achieved complete remission and complete donor engraftment.Case 1 survived refractory acyclovir-resistant Epstein-Barr virus viremia benefiting from preemptive treatment with rituximab and DLI-induced grade Ⅳ acute GVHD,but died of thrombotic microangiopathy mixed with diffuse alveolar hemorrhage and sepsis on +243 days.Case 2 relapsed just 2 months after allo-HSCT despite DLI and rapidly tapering of CsA,died of sepsis followed by diffuse intravascular coagulation on +101 days.Conclusion BPDCN is characterized with typical cutaneous and/or bone marrow involvement with CD4+CD+56CD+123CD+43 blastic plasmacytoid dendritic cell and highly aggressive clinical course.Allo-HSCT seems to be a promising treatment for early phase of aggressive BPDCN aided with MRD monitoring and DLI,but it deserves more intensive researches to promote outcome of advanced staged BPDCN.

Metastatic cancer stem cells and anti-metastasis strategies / 中国肿瘤生物治疗杂志

Cancer stem cells (CSC) are capable of self-renewal and can proliferate into a heterogeneous bulk with cancer progeny population, which is the main reason for recurrence and metastasis of cancer. Metastatic cancer stem cells (MCSC) have the properties of CSC and the ability of metastasis. Metastasis happens at both the late and early stages of tumorigenesis. MCSC are different from CSC in origin, epithelial-mesenehyrnal transition (EMT), mesenehymal-epithelial transition (MET), and microenvironment of target organs (niche), etc. Therefore, MCSC is the foundation of cancer metastasis. Anti-metastasis strategies include killing CSC, blocking EMT and MET of CSC, inhibiting MCSC adhesion to microvessels, and destroying MCSC dependent-niche. This review introduces the possible sources, biological features of MCSC, the possible breakthrough in MCSC research, and MCSC-targeted anti-metastasis strategy, hoping to provide reference for researches about tumor metastasis mechanisms and anti-metastasis strategies.

Expression of caspase-3 and TRAIL receptors in CD4~+ and CD8~+ T cells of systemic lupus erythematosus patients / 中华风湿病学杂志

Objective To explore the expression of caspase-3 and tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) receptors in the CD4+ and CD8+ T cells of systemic lupus erythematosus (SLE)patients. Methods The CD4+ and CD8+T cells from peripheral blood of 20 SLE patients and 10 healthy volunteers were separated using magnetic cell sorting system (MACS), reverse transcription-polymerase chain reaction (RT-PCR) was used to test the expression of caspase-3 and TRAIL receptors in CD4+ and CD8+T cells of SLE patients and healthy volunteers. Results The CD8+T cells from SLE patients had significantly higher caspase-3 (P=0.003) and TRAIL-R2 (P=0.024) expression than those from healthy volunteers. Conclusion The TRAIL-R2 signal pathway may be a possible important pathway to the apoptosis of T cells in SLE patients.

Detection and significance of CD4 and CD8 subsets of T lymphocytes in the peripheral blood of systemic lupus erythematosus patients by magnetic activated cell sorting / 第三军医大学学报

Objective To investigate the abnormality of CD4 and CD8 subsets of T lymphocytes in the peripheral blood of systemic lupus erythematosus patients. Methods The CD4 + and CD8 + T lymphocytes from 20 SLE patients and 10 healthy controls were separated using magnetic activated cell sorting (MACS). Results The numbers of CD4 + T cells in the peripheral blood and CD4 +/CD8 + ratio of SLE patients were significantly lower than those in healthy volunteers(CD4,P=0.001;CD4/CD8 ratio,P=0.046). Conclusion There exist abnormalities in the CD4 and CD8 subsets of T lymphocytes in the peripheral blood of SLE,especially the decreased numbers of CD4 + T cells.

Long serial analysis of gene expression in the yeast phase of Candida albicans / 解放军医学杂志

Objective To construct a library of long serial analysis of gene expression (LongSAGE) and investigate the characteristics of gene expression in the yeast phase of C. albicans. Methods A LongSAGE library was constructed with long serial analysis of gene expression (LongSAGE). Results A total of 17 773 tags, representing specific 7 333 transcripts, were extracted from 1 000 sequence files in the yeast phase of C. albicans. Tags matching single gene accounted for 16.65%, tags matching multiple genes accounted for 6.59%, tags for hypothetical protein accounted for 16.27%, tags for genome and cosmid accounted for 1.64%, and novel tags accounted for 58.86%. Eleven tags in 33 tags whose expression frequencies exceeded 35 may be new expression sequences. Conclusion LongSAGE is not only a powerful method in investigating the gene expression profiling, allowing the analysis of the expression of thousands of transcripts in a quantitative manner without prior sequence information, but also in finding new genes.

Expression of SLAM gene in CD4~+ and CD8~+ T cells from SLE patients / 第三军医大学学报

Objective To explore the expression of SLAM gene in the CD4+ and CD8+ T cells from SLE patients.Method The CD4+ and CD8+ T cells from peripheral blood of 20 SLE patients and 10 healthy blood donors were separated using magnetic cell sorting system(MACS).We detected the expression of SLAM mRNA in CD4+ and CD8+ T cells by RT-PCR.Results The CD4+ and CD8+ T cells from SLE patients showed significantly higher SLAM expression than those from healthy blood donors.Conclusion The SLAM signal pathway may play an important role in contributing to the abnormal activation of T cells in SLE patients.