急性原粒细胞白血病部分分化型(M_(2b))组化特点的探讨(英文)

目的 利用各种细胞组织化学染色的特点 ,综合分析来鉴别诊断M2b。方法 对M2b患者的骨髓片进行组化染色 ,观察其阳性率、积分以及阳性物表现出来的形状。结果 M2b患者的异常中幼粒细胞大致可以分为空泡型和核旁淡染区型两种 ,其中核旁淡染区型细胞行组化染色后 ,在POX、SB、CE、AE、NBE染色中有其特异的团块状反应 ;空泡型异常中幼粒细胞 ,在POX、SB、CE中表现为强阳性反应 ,并且POX、SB的阳性率、积分在所有白血病中仅次于M3 ,CE的阳性率一定大于 50 %。AE、NBE、PAS、ACP这几种组化染色的观察 ,对M2b有辅助诊断作用。结论 对于M2b的诊断 ,不仅有其特异的形态学特征、免疫学标记、细胞遗传学、分子生物学 ,作为基础的组化染色对鉴别诊断M2b仍具有相当重要的意义。

甜菜幼苗叶片光合性能、渗透调节及活性氧对高硼胁迫的响应

为探究甜菜幼苗对硼毒害的响应,采用水培试验的方法,研究了不同硼浓度(0.05、0.25、0.50、2.50、5.00 mmol·L-1 H3BO3)条件下,高硼胁迫对甜菜幼苗叶片光合性能、渗透物质调节及活性氧代谢的影响。结果表明:高硼胁迫影响甜菜幼苗生长发育进程,硼处理第15~20 d时对幼苗影响最大。随着硼浓度的增加,叶绿素a、叶绿素b含量呈下降趋势,与对照相比差异显著,净光合速率与叶绿素含量规律表现一致,叶绿素a/b在0.50 mmol·L^-1时达到最大值;果糖及淀粉含量在高硼胁迫下增加,在2.50 mmol·L^-1时与对照相比均差异显著,分别比对照增加了114.5%、78.2%。蔗糖含量显著下降,与对照相比降低了31.8%~54.1%;当硼浓度达到0.50 mmol·L^-1时,丙二醛(MDA)、脯氨酸(Pro)含量显著增加。染色观察发现,随着硼浓度的增加,叶片中超氧阴离子与过氧化氢积累也逐渐增加,均在5.00 mmol·L^-1时累积最严重。研究表明,硼毒害使甜菜叶片光合能力下降,阻碍光合产物的运输,细胞内活性氧大量积累,对细胞产生氧化胁迫,进而抑制甜菜植株生长。本试验条件下,硼浓度0.50 mmol·L^-1可以作为甜菜幼苗高硼胁迫的临界值,超过该浓度,植株生长受到显著抑制。

pHGF对CBMC表面标志表达的影响

目的 :为了再次证明pHGF的免疫功能。方法 :应用免疫组织化学法测定pHGF对脐血单个核细胞表达CD4、CD8、HLA DR分子的影响。结果 :证明pHGF均能促进上述分子在脐血单个核细胞表面的表达。与pHGF诱导前相比较 ,CD4、CD8、HLA DR分子的表达增加了 3～ 4倍。结论

Nuclear β-catenin expression as a prognostic factor in advanced colorectal carcinoma

AIM： To investigate the changing pattern of β-catenin expression and its prognostic value in advanced colorectal cancer （CRC）. METHODS： Archival tumor samples were analyzed for β-catenin using immunohistochemistry （IHC） in 95 patients with advanced CRC. RESULTS： Membranous β-catenin expression was found in the normal colorectal epithelium. Almost 100% of CRC cases showed membranous and cytoplasmic expression, and 55 （58%） cases showed nuclear expression. In univariate （Kaplan-Meier） survival analysis, only the nuclear index （NI） was a significant predictor of disease free survival （DFS） （P = 0.023; n = 35）, with a NI above the median associated with longer DFS （34.2 too） than those with a NI below the median （15.5 too） （P = 0.045, ANOVA）. The other indices were not significant predictors of DFS, and none of the three tested indices （for membranous, cytoplasmic, or nuclear expression） predicted diseasespecific survival （DSS）. However, when dichotomized as positive or negative nuclear expression, the former was a significant predictor of more favorable DFS （P = 0.041） and DSS （P = 0.046）. CONCLUSION： Nuclear β-catenin expression provides additional information in predicting patient outcome in advanced CRC.

Upregulated CD133 expression in tumorigenesis of colon cancer cells

AIM: To analyze the upregulated CD133 expression in tumorigenesis of primary colon cancer cells. METHODS: Upregulated CD133 expression in tumorigenesis of colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) was analyzed by flow cytometry. Human colon cancer tissue samples were stained with anti-human CD133. SW620 cells were sorted according to the CD133 expression level measured by fluorescence-activated cell sorting. Spheroids of colorectal cancer cells were cultured with the hanging drop. Expression of CD133 and Lgr5 in spheroids of colorectal cancer cells and monolayer culture was detected by RT-qPCR. Spheroids of colorectal cancer cells were analyzed using anti-human CD133 with immunohistochemical staining. RESULTS: CD133 antigen was expressed in colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) as well as in primary and metastatic human colon cancer tissues. However, the CD133 was differently expressed in these cell lines and tissues. The expression levels of CD133 and Lgr5 were significantly higher in spheroids of parental, CD133hi and CD133-cells than in their monolayer culture at the mRNA level (P < 0.05). Immunohistochemical staining of spheroids of CD133-cells showed that CD133 was highly expressed in colorectal cancer cell lines. CONCLUSION: Upregulated CD133 expression plays a role in tumorigenesis colorectal cancer cells, which may promote the expression of other critical genes that can drive tumorigenesis.

Extramedullary plasmocytoma associated with a massive deposit of amyloid in the duodenum

We report a rare case of extramedullary plasmocytoma associated with a massive deposit of amyloid in the duodenum. A 72-year-old Japanese man was admitted to our hospital presenting with a 3-mo history of epigastric pain, vomiting and weight loss. On computed tomography （CT） a wall thickening of the fourth part of the duodenum was observed. Multiple biopsies obtained from the lesion showed infiltration of plasma cells and lymphocytes, but they were not conclusive. The patient underwent resection of the lesion and, on histopathological examination, the lesion consisted of a dense and diffuse infiltrate of plasma cells and a few admixed lymphocytes with reactive follicles extending to the muscular propria. An extensive deposition of amyloid was also observed. Immunohistochemical stains revealed that a few plasmacytoid cells showed λ light chain staining, though most were κ： light chain positive. These cells also were positive for CD138 and CD56 but negative for CD20 and CD79. The findings were consistent with extramedullary plasmocytoma associated with a massive deposit of amyloid in duodenum. A subsequent workup for multiple myeloma was completely negative. The patient showed no signs of local recurrence or dissemination of the disease after 12 mo follow-up. Because of the association of plasmocytoma and amyloidosis, the patient must be followed up because of the possible systemic involvement of the neoplasm and amyloidosis in future.

Increased expression of chondroitin sulphate proteoglycans in rat hepatocellular carcinoma tissues

AIM:To investigate the expression of chondroitin sulphate proteoglycans(CSPGs)in rat liver tissues of hepatocellular carcinoma(HCC).METHODS:Thirty male Sprague Dawley rats were randomly divided into two groups:control group(n=10) and HCC model group(n=20).Rats in the HCC model groups were intragastrically administrated with 0.2%(w/v)N-diethylnitrosamine(DEN)every 5 d for 16 wk,whereas 0.9%(w/v)normal saline was administered to rats in the control group.After 16 wk from the initiation of experiment,all rats were killed and livers were collected and fixed in 4%(w/v)paraformaldehyde.All tissues were embedded in paraffin and sectioned.Histological staining(hematoxylin and eosin and Toluidine blue)was performed to demonstrate the onset of HCC and the content of sulphated glycosaminoglycan(sGAG).Immunohistochemical staining was performed to investigate the expression of chondroitin sulphate(CS)/dermatan sulphate(DS)-GAG,heparan sulphate(HS)-GAG,keratan sulphate(KS)-GAG in liver tissues.Furthermore,expression and distribution of CSPG family members,including aggrecan,versican,biglycan and decorin in liver tissues,were also immunohistochemically determined.RESULTS:After 16 wk administration of DEN,malignant nodules were observed on the surface of livers from the HCC model group,and their hepatic lobule structures appeared largely disrupted under microscope.Toluidine blue staining demonstrated that there was an significant increase in sGAG content in HCC tissues when compared with that in the normal liver tissues from the control group[0.37±0.05 integrated optical density per stained area(IOD/area)and 0.21± 0.01 IOD/area,P<0.05].Immunohistochemical studies demonstrated that this increased sGAG in HCC tissues was induced by an elevated expression of CS/DS(0.28±0.02 IOD/area and 0.18±0.02 IOD/area,P< 0.05)and HS(0.30±0.03 IOD/area and 0.17±0.02 IOD/area,P<0.01)but not KS GAGs in HCC tissues.Further studies thereby were performed to investigate the expression and distribution of several CSPG components in HCC tissues,including aggrecan,versican,biglycan and decorin.Interestingly,there was a distinct distribution pattern for these CSPG components between HCC tissues and the normal tissues.Positive staining of aggrecan,biglycan and decorin was localized in hepatic membrane and/or pericellular matrix in normal liver tissues;however,their expression was mainly observed in the cytoplasm,cell membranes in hepatoma cells and/or pericellular matrix within HCC tissues.Semi-quantitative analysis indicated that there was a higher level of expression of aggrecan(0.43± 0.01 and 0.35±0.03,P<0.05),biglycan(0.32±0.01 and 0.25±0.01,P<0.001)and decorin(0.29±0.01 and 0.26±0.01,P<0.05)in HCC tissues compared with that in the normal liver tissues.Very weak versican positive staining was observed in hepatocytes near central vein in normal liver tissues;however there was an intensive versican distribution in fibrosis septa between the hepatoma nodules.Semi-quantitative analysis indicated that the positive rate of versican in hepatoma tissues from the HCC model group was much higher than that in the control group(33.61%and 21.28%,P <0.05).There was no positive staining in lumican and keratocan,two major KSPGs,in either normal or HCC liver tissues.CONCLUSION:CSPGs play important roles in the onset and progression of HCC,and may provide potential therapeutic targets and clinical biomarkers for this prevalent tumor in humans.

Integration of human papillomavirus 18 DNA in esophageal carcinoma 109 cells

AIM:To detect human papillomavirus(HPV) DNA in esophageal carcinoma(EC) 109 cells and investigate the relationship between HPV and EC.METHODS:Genomic DNA and total RNA from EC109 cells were isolated.HPV DNA was detected by polymerase chain reaction(PCR) with the general primer sets of My09/11 and GP5 +/6 + for the HPV L1 gene and type-specific primer sets for HPV18 E6 and HPV18 E6-E7.Reverse transcription(RT) of mRNA isolated from EC109 cells was performed to produce a cDNA.And then a PCR-based protocol for the amplification of papillomavirus oncogene transcripts was used to analyze HPV18 DNA and integrated transcripts of HPV18 in the chromosomes of EC109 cells.The final nested PCR products were cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.RESULTS:HPV18 DNA was detected in EC109 cells by PCR using the general primer sets of My09/11 and GP5 +/6 + for HPV L1 and the type-specif ic primer sets for HPV18 E6 and E6-E7 to generate products of 450 bp,150 bp,335 bp and 944 bp,respectively.Approximately 600 bp of integrated HPV18-specific transcript was identified.The final nested PCR product of integrated HPV18 DNA was cloned into a pMD-18T vector and sequenced to analyze the chromosomal location of HPV integration.Sequence alignment showed that the HPV18 sequence from EC109 cells was identical to that of the encoded early protein E7-E1 of the standard HPV18 strain X05015,and another partial gene sequence was identical to a partial sequence of human chromosome 8.CONCLUSION:Integration of the HPV genome into the host cell chromosome suggests that persistent HPV infection is vital for malignant cell transformation and carcinogenesis.

Influence of Shenqing Recipe on Morphology and Quantity of Colonic Interstitial Cells of Cajal in Trinitrobenzene Sulfonic Acid Induced Rat Colitis

Objective To observe the influence of Shenqing Recipe (SQR), a kind of Traditional Chinese Medicine, on the morphology and quantity of colonic interstitial ceils of Cajal (ICC) in trinitrobenzene sulfonic acid (TNBS)-induced rat colitis, and to investigate the possible mechanism of SQR in regulating intestinal dynamics. Methods Sixty rats were randomly divided into normal control, model Ⅰ , model Ⅱ, mesalazine, and high-dose, and low-dose SQR groups with 10 rats in each group. TNBS (10 mg) dissolved in 50% ethanol was instilled into the lumen of the rat colon of the latter five groups to induce colitis. On the 4th day after administration of TNBS, each treatment group was administered one of the following formulations by enteroclysis gavage once a day for 7 days: 600 mg·kg^-1·d^-1 mesalazine, 2.4g^-1·d^-1 SQR, and 1.2g^-1·d^-1 SQR. Model Ⅱ rats received normal saline solution. After 7 days colonic samples were collected. While the colonic samples of model I group were collected on the 3rd day after TNBS administered. Ultrastructure of ICC in the damaged colonic tissues was observed with transmission electron microscope. Expression of c-kit protein in colonic tissue was determined by immunohistochemical staining and Western blot. Results The ultrastructure of colonic ICC in the rat model of TNBS-induced colitis showed a severe injury, and administration of SQR or mesalazine reduced the severity of injury. Similarly, the expression of c-kit protein of TNBS-induced colitis rat model was significantly decreased compared with the normal control group (P〈0.05). Treatment with SQR or mesalazine significantly increased the expression of c-kit protein compared with the administration of control formulations (P〈0.05), especially the high-dose SQR group. Conclusion SQR could alleviate and repair the injured ICC, and improve its quantity, which might be involved in regulating intestinal motility.

Granular cell tumor of the urinary bladder: A case report

We reported a case of a GCT of the urinary bladder and review the literature.A 23-year-old female presented with dysuria that had lasted for the previous 6 months.MRI revealed a 3×2.5 cm global mass in the anterior wall of urinary bladder.Cystoscopy showed a semispherical tumor approximately 3 cm in diameter that was covered with normal bladder mucosa and extended from the bladder neck to the anterior wall of the bladder.The patient underwent transurethral resection of the tumor.Histological examination and immunohistochemical staining showed a granular cell tumor(GCT).There were no features suggesting a malignant phenotype.On 6 months follow-up,the patient has remained free of bladder recurrence.We recommend careful pathologic assessment for establishing the appropriate diagnosis and either a conservative or aggressive surgical treatment for benign or localized malignant GCT of the urinary bladder,respectively.

ESTABLISHMENT OF A HUMAN T－LYMPHOMA CELL LINE（H－TL90） AND ANALYSIS OF ITS BIOLOGICAL CHARACTERISTICS

We established a human T-lymphoma cell line from the cancerous ascrtes of a male patient with prostate cancer which was named H-TL90. This cell line was characterized by its histological features, and by chromosomal and immunological analysis. Immunophenotypic analysis revealed that the cells expressed surface antigen CD3- CD4- CD7+ CD8-. Biological analysis revealed that the cell can promote lymphoryte proliferation. This suggested that the cell line has an autosecretion function. Cytogenetic analysis revealed that H-TL90 was a hyperdiploid with 47 chromosomes and had characteristic transloration between chromosome 3 and 11, and the deletion of the long arm of chromosome 6. These resuhs demonstrated the H-TL90 cell line can be a useful model for the study of human T-lymphoma.

Polytene chromosomes of an Indian Himalayan black fly Simulium (Nevermannia) praelargum (Diptera:Simuliidae)

High quality polytene chromosome maps （n=3） of a Himalayan Simuliid Simulium praelargum Datta, 1973 are presented and represent the first cytological description of a taxon found in the feuerborni group, subgenus Nevermannia. Polytene chromosomes one （I） and two （II） are metacentric, chromosome three （III） is submetacentric with the length of each chromosome occupying 37.25 %, 31.36 % and 31.34 % of the total complement length, respectively. Typical simuliid diagnostic intergeneric chromosomal markers are found within the polytene complement of this species. The nucleolar organizer （N.O.） is found at the base of the short arm of chromosome one （IS）, the Ring of Balbiani （R.B.）, double bubble （D.B.） and triad occur in the short arm of chromosome two （IIS）, the Parabalbiani Ring （EB.） and grey band （gb） occur in the long arm of chromosome two （IIL） and the Blister （BL） and Capsule （Ca） occur in the short arm of chromosome three （IIIS）.Terminal bands at the end of IIIS are heterochromatinized and present atypically with respect to other simuliid fauna. Populations studied so far are unique among the Simuliidae in that they exhibit chromosome structural monomorphism. These high resolution polytene chromosome maps will form the basis for future cytological characterization and phylogenetic comparisons amongst members of the feuerborni group

Correlation of STAT3, CEA in lung adenocarcinoma cell A549

Objective: The purpose of this study was to analyze the relationship between signal transducer and activator of transcription 3 (STAT3) and carcinoembryonic antigen (CEA) in lung adenocarcinoma cell A549, and to explore the value of STAT3 on early diagnosis of lung adenocarcinoma. Methods: The expression of CEA, STAT3 mRNA and it's protein in human lung adenocarcinoma cell A549 and normal human lung cells MRC-5 were tested by immunohistochemistry staining (PV) and quantitative real time fluorescent PCR. The correlation between STAT3 and CEA in human lung adenocarcinoma cell A549 was analyzed. Results: The protein and mRNA levels of STAT3, CEA in lung adenocarcinoma cell A549 were apparently higher than those in normal human lung cells MRC-5. The levels of STAT3 mRNA and it's protein were positively correlated with CEA in lung adenocarcinoma cell A549. Conclusion: STAT3 have the same value in diagnosis of lung adenocarcinoma.

Different morphologic features of rat cochlea progenitor spheres and their implications

Objective: To detect the different morphologic features, developmental regulation, potential of proliferation and differentiation of neonatal rat cochlea progenitor spheres. Methods: We isolated the cochlea sensory epithelium cells from neonatal rats and cultured them in nonadherent conditions to acquire different morphologic spheres. Then we observed the diameter and compositional change of cell colonies in distinct sphere types on day 3, 6, 9 and 12, and summarized the regularity of development and their conversion. We also detected the proliferative activity of distinct spheres by immunohistochemical staining of Abcg2, Nestin and BrdU. After induced spontaneous differentiation, the spheres were detected in the changes of the marker of hair cell, MyosinVIIA; by immunocytochemical staining, we revealed the potential of how different spheres were converted into hair cell-like cells. Results: The acquired three types of suspended spheres are solid, transitional, and hollow. There's morphologic significance among them and they can covert into the other type of spheres among them. The ability of self-renewing and proliferation in distinct spheres vary and all of them have the potential of spontaneously differentiation into hair cell-like cells. Conclusion: All the type of spheres not only has the potential of proliferation and differentiation, but also hasthe potential of spontaneous differentiation into hair cell-like cells. Distinct types of cell spheres neither originate from different progenitor cell subcolonies nor are different stages of the same cell spheres. Solid spheres are most practically useful.

急性未分化细胞白血病一例报告

急性未分化细胞白血病(acute undifferentiated leukemia,AUL)是指增殖的原始细胞在形态学、细胞化学和免疫表型上缺乏足够证据将其归类于髓系还是淋系起源的一类急性白血病[1]。该病白血病性原始细胞无任何形态学分化和成熟特征,无特异的系列抗原标记,发病率低,对该病的报道也较罕见,本研究报道1例急性未分化细胞白血病。

Expression of hunchback during oogenesis and embryogenesis in Locusta migratoria manilensis(Meyen)

hb(hunchback) is a contributing factor in anteroposterior axial patterning of insects.Although the hb function in Locusta migratoria manilensis has been investigated,its expression pattern remains unknown.Here,the mouse polyclonal antibody was produced against Hb fusion protein,and then its expression pattern during oogenesis and embryogenesis of L.migratoria manilensis was examined by immunohistochemical staining.Hb protein was detected in the oocyte nucleus which was positioned centrally within the developing oocyte.The oocyte nucleus gradually moved to the posterior end of the egg along with the oocyte maturing.In freshly laid eggs,Hb formed gradient at the posterior end of the egg,and then hb was expressed as a band in the middle of the blastodisc.As the blastodisc differentiated into the head and trunk,the expression region became wide,which would develop into spatial gnathal and thoracic segments.With abdominal segmentation,the expression domain in the gnathal and thoracic region became faint and eventually faded out,while the Hb expression domain appeared at the posterior growth zone in a discontinuous expression manner.The hb expression pattern of L.migratoria manilensis is greatly similar to that of other locusts,such as Schistocerca americana and another L.migratoria.Compared with other insects,hb expression is conserved in the gnathal and thoracic domains,while divergent in oogenesis and abdomen.