In order to study the multidifferentiation of medullary carcinoma of the thyroid gland (MCT), 24 cases of MCT were examined for the presence of immunoreactive calcitonin (CT), thyroglobulin (Tg), chromogranin A (CgA), somatostatin (SS), serotonin (5-HT), S-100 protein (S-100), neuron-specific enolase (NSE), vasoactive intestinal polypeptide (VIP), adrenocorticotrophin (ACTH) and neurofilament protein (NF) by using immunohistochemical ABC methods. Results showed that CT-immunoreactive cells were present in all tumors. Tg was present in three tumors. 23 cases contained CgA-immunoreactive cells. 14 tumors contained 5-HT-immunoreactive cells, 10 cases were immunoreactive to NSE and SS. 4 tumors contained VIP-immunoreactive cells and only one cases was positive for S-100. The demonstration of immunoreactivity for multiple antigens in 24 cases suggests that the origin of medullary thyroid carcinoma may originate from neuroectoderm cells potentially capable of producing numerous hormone substances. In addition, as the neoplastic cells in 12% of the tumors containing hormone substances as well as thyroglobulin, it is suggested that follicular epithelial differentiation and mixed medullary thyroid carcinoma may be more common than previously suspected. Recent studies indicate that mixed carcinoma of the thyroid may be derived from common stem cells in posterior branchia capable of differentiating into both follicular and parafollicular tumor cells.

In this study, 16 eligible patients with intermediate and high-grade non-Hodgkin's lymphoma were treated with a new high-dose DHACT regimen supported by rhG-CSF and peripheral blood stem cell (PBSC) rescue. PBSC were mobilized by rhG-CSF or rhGM-CSF. Single leukapheresis was performed and the PBSC were then frozen in liquid nitrogen. CFU-GM clonogenic assay for mononuclear cells and resuscitated progenitor cells done to calculate how many progenitor cells were alive after freezing. The DHACT chemotherapy was composed of carboplatin 600 mg/m2 on d1, Ara-C 1500 mg/m2 on d2, VM-26 100 mg/m2 on d3, 4, and dexamethasone 40 mg/d, on d1-4. Autologous PBSC was reinfused after 24 to 48 hours of chemotherapy. Recombinant human G-CSF at 300 micrograms administered daily on 2 successive days when the absolute neutrophil count was greater than 1 x 10(9)/L. Other supportive care procedures were standard for the unit. The median amount of PBSC reinfused into a patient was 0.9 x 10(8)/kg. The recovery rate of CFU-GM was 78% after cryopresevation. Within 7 to 9 days after high-dose DHACT chemotherapy, the WBC count and the platlet count arrived nadir, and then rose gradually with rhG-CSF injection. The median time for WBC count from nadir to > or = 1 x 10(9)/L was 4 days, and that for platelet count from nadir to > or = 50 x 10(9)/L was 7 days. Nine patients achieved complete remission and 5 patients achieved partial remission. The median follow-up on survival was 9 months. High-dose DHACT regimen supported by rhG-CSF and PBSC rescue is a safe and effective treatment for patients with advanced intermediate and high-grade non-Hodgkin's lymphoma.

Transferring beta-globin gene and its enhancer into human hematopoietic cells is the basis for applying beta-thalassemia gene therapy in clinical practice. We isolated ecotropic virus producing clones and amphotropic virus producing clones by using a replication-defective retrovirus vector containing beta-globin gene and its enhancer to transfect ecotropic packaging cell line phi-2 and amphotropic packaging cell line PA317. Then by ping-ponging supernatant infection between ecotropic producer clone and amphotropic producer clone, we obtained high titer amphotropic PA317 producer clones with the highest titer up to 5.9 x 10(6) CFU/ml. Human mononuclear bone marrow cells were infected repeatly with this high titer virus vector under stimulation of hematopoietic growth factors IL-3, IL-6 and SCF, Southern blot hybridization analysis showed that beta-globin gene and its enhancer had been integrated into the genome of human hematopoietic cells.

To observe the distribution of multiple hormones in nonsecreting islet cell tumors of the pancreas and to study their histogenesis, 9 pancreas nonsecreting islet cell tumor cases were studies using 12 kinds of antisera. The results showed that 4 cases were positive for insulin, 6 for glucagon, 1 for gastrin, 6 for somatostatin, 1 for gastrin 5 for calcitonin, 7 for neurotensin, 4 for ACTH, 3 for TSH, 5 for FSH and 2 for LH. It is therefore confirmed that these tumors synthesize and secrete peptide hormones and glycoprotein hormones. We believe that these endocrine cells originate from primitive multipotential stem cells.

When ES-5 cells were transfected with an exogenous porcine TGF-beta 1 gene, one can obtain clones of genetically modified ES cells with over-expression of the transfected gene. We called the genetically modified ES-5 cells as ES-T cells. When ES-T cells were used to study their differentiation in vitro by all trans-retinoic acid (RA), it was soon noticed that embryoid bodies of ES-T cells can exclusively differentiate into endothelial cells and vessel-like structures, but not in their parent ES-5 cells. The above result is the first indication that the differentiation of tubular structures in embryoid bodies of ES-T cells may somehow be related to TGF-beta 1. To demonstrate further the role of TGF-beta 1 in the formation of vessel-like structures, the cultured ES-5 cells in the presence of added rhTGF-beta 1 were closely followed in the course of their differentiation. We have, thus, demonstrated the promoting effects of exogenous rhTGF-beta 1 in the formation of vessel-like structures, morphologically similar to those structures derived from ES-T6 cells, during the differentiation of ES-5 cells, both in monolayer culture, in three dimensional collagen gel and in embryoid bodies cultured on gelatin-coated tissue culture wells. Addition of suitable amount of anti-TGF-beta 1 monoclonal antibody IgG (TB21) to the culture medium of embryoid bodies of ES-T6 cells could effectively abolish the formation of vessel-like structures induced by retinoic acid. The percentage of the inhibition was very high, giving a figure comparable to that of atypical vessel-like structures formed in the control embryoid bodies from their parent ES-5 cells. The flat epithelial-like cells and round cells differentiated from embryoid bodies of ES-T6 cells were stained rather strongly for laminin and type IV collagen by immunofluorescent procedure. The above results indicate clearly that TGF-beta 1 is a crucial factor in organizing the differentiated derivatives (endothelial-like cells and their immediate progenitor cells) from ES-T6 cells to form vessel-like structures, and that the role of TGF-beta 1 in vasculogenesis might be performed, in part, through the modulation of the composition and organization of the extracellular matrix. In addition, the enhanced expression of bFGF mRNA in derivatives differentiated from both ES-5 cells treated with rhTGF-beta 1 and ES-T6 cells were detected by Northern blot analysis. Thus, aside from its effects on extracellular matrix, TGF-beta 1 might also modulate the bioactivity of bFGF in relation to the growth of vascular endothelial cells in the present system.

Immunoglobulin heavy chain (IgH) gene rearrangement serves as a marker of clonality in B lymphoproliferative malignancies. In order to study the IgH rearranged gene in acute nonlymphoblastic leukemia (ANLL) patients we combine polymerase chain reaction (PCR) with Southern blot to detect 41 ANLL patients and 7 of them (17.1%) were found to have IgH rearrangement by PCR amplification. All these 7 positive cases were confirmed by Southern blot. The sensitivity of this method was 10(-4)-10(-5) level. In 12 patients with complete remission, 3 (25.0%) were found to have IgH rearranged gene. All these 3 cases had clinical relapse within 6 months. Our results show that IgH rearrangement not only may occur in lymphoblastic leukemia of B lineage, but also can be found in ANLL. The mechanism may be that in some ANLL patients, the leukemic transforming event might involve stem cells capable of both B cell and myeloid differentiation or ANLL might differentiate along different lineage with predominant appearance of one or the other subclone in the course of the disease.

To study the damage to the hearing of miners under the shaft caused by noise and its recovery, an experiment with imitated noise was carried out in guinea pigs. The animals were divided into five groups with exposure to imitated underground noise of 117 dB (A) for one hour for one, two, four, eight and 16 days, respectively, and one control. Thresholds of cortex reaction and evoked electric reaction of brain stem were detected before and after exposure. And, ultrastructural changes in cochlear hair cells were observed by scanning and transmission electron microscopy. Results showed that exposed noise level and length since removing from exposure had significant effects on changes in hearing threshold. Damage to hearing was aggravated with prolonged exposure, to various extent, from changes in temporary threshold shift and permanent threshold shift to recovery from it after removing from exposure. Intracellular structural changes in internal and external hair cells and their injury deteriorated gradually with increase of exposure levels, at worst, to changes in epithelial plate, collapse of Corti's organ and vacuolation, edema and degeneration of cells. It suggests it is important to protect workers form exposure to noise.

Small-cell esophageal carcinoma (SCEC) is not uncommon. From 1979 through 1994, 42 cases of SCEC were diagnosed among 711 patients with cancer of the esophagus, with a relative frequency of 5.9%. Differential diagnosis is difficult according to clinical symptoms and signs. However, SCEC differs greatly from squamous-cell carcinoma of the esophagus in histopathology and biologic behavior but is similar to small-cell lung cancer (SCLC). Surgical treatment alone of SCEC patients has a mean survival period of 11.2 months while patients treated with radiotherapy alone has a longer survival (22.8 months). No case was treated only with chemotherapy in this group of patients. Those received no treatment could survive 6.2 months only. Therefore, SCEC is a very aggressive tumor with poor prognosis. Histochemical and immunohistochemical study revealed both epithelial and neuro-endocrine differentiation potentials suggesting that SCEC probably originates from mulfipotential primitive stem cells in the esophageal mucosa.

To study the effect of angelica polysaccharide (AP) on proliferation and differentiation of hematopoietic progenitor cells for clarifying the hematonic mechanism of angelica sinensis.

To investigate the most appropriate condition for collecting stem cells during peripheral blood stem cells transplantation (PBSCT), we used colony forming unit-granulocyte, macrophage (CFU-GM) as index to observe the change of PBSC after chemotherapy in 12 patients with malignant hematopoietic tumor. The results showed that it was more suitable to collect PBSC when the number of leucocytes was 2 x 10(9)/L, neutrophils 0.8 x 10(9)/L and platelets 110 x 10(9)/L after chemotherapy for 2 weeks. After chemotherapy, the number of leucocytes were suppressed evidently. On the contrary, a lot of CFU-GM appeared during the recovery phase of peripheral hemogram. At the nadir of peripheral hemogram, the number of leucocytes correlated negatively with the number of CFU-GM (P < 0.05). The longer the time of chemotherapy, the less the CFU-GM that in the same patients. After chemotherapy the CD34+ cells which appeared early might not form CFU-GM, but the CD34+ cells at the recovery phase of peripheral hemogram had the same trend of change with CFU-GM.

To determine if the sera of patients with acute promyelocytic leukemia (APL) and all-trans retinoic acid (ATRA) inhibit the formation of colony-forming-unit of megakaryocyte (CFU-Meg).

Proliferative potentiality of adult rat salivary gland cells was studied by means of bromodeoxyuridine (BrdU) labeling in different time and ABC immunohistochemistry. The labeling rate of intercalated cells in different time after BrdU injection manifest the character of stem cell which is concordant with the theory of semipluripotential bicellular reserve cell suggested by Eversole and Batsakis. This initial labeling rate of intercalated cell is very low and it rises very rapidly to 78 times within 5 days. On the other hand, it also goes down very fast. We proved that the acinar cells and striated cells have the capacity of proliferation which is not so high as intercalated cells for self renewal that can complete the theory of semipluripotential bicellular reserve cell and is beneficial to explain why there is a variety of salivary gland neoplasms.

In hematopoiesis; the human CD 34 protein is a strict developmental stage-specific antigen that marks hematopoietic stem/progenitor cells, suggesting that it plays an essential role in hematopoiesis. More recently, it has been demonstrated that hematopoietic cells expressing the CD 34 antigen constitute various heterogeneous cell populations in which each CD 34+ subset was associated with commitment to a particular lineage, and differed from other's in reconstituting hematopoiesis. In this report, we have assessed the different functional subpopulation of CD 34+ hematopoietic stem/progenitor enriched from human bone marrow using Isolex TM 50 system according to the strategy on immunomagnetic separation of positive selection. CD 34+ cell population of high purity (> 90% CD 34+) was analyzed by means of double staining procedure of flurorescein conjugated monoclonal antibodies on the two-color FACAcan or FACS 440. Eight cell subsets of at least of bone marrow CD 34 hematopoietic stem/progenitor cells have been defined by undertaking a comparative coexpressing of CD 71, CD45, CD 33 and HLA-DR antigens on CD 34 hematopoietic stem/progenitor cells. The frequencies of various subsets in two illustrative are as following: 1). CD 34+/CD 71- and CD 34+/CD 71+ (23.43%-56.6% versus 33.4%-66.6%): 2). CD 34+/CD45- and CD 34+/CD45+ (80.8%-82.5% versus 8.1%-11.2%): 3). CD 34+/CD 33- and CD 34+/CD 33+ (20.4%-80.6% versus 14.6%-64.8%): 4). CD 34+/DR- and CD 34+/DR+ (6.3%-11.0% verus 82.8%-85.5%). Immunological double color staining of IGSS-APAAP was also used to further analyse the CD 34+ cell subsets as above-mentioned, the results were very similar to those obtained by FACScan. Our data indicate that CD 34+ hematopoietic cell fraction is far from being a uniform cell population, and many works regarding biological properties and regulation mechanism of different subsets of CD 34+ hematopoietic stem/progenitor cells are being carried out.

In the present study, the potential of fibroblast-mediated IL-6 gene therapy for accelerating hematopoietic reconstitution after syngeneic bone marrow transplantation (BMT) in BALB/c mice subjected to total body irradiation at a dose of 7 Gy was investigated. We observed that not only recovery of platelets, WBCs, CFU-GM or CFU-MK in bone marrow in mice treated with IL-6 gene therapy in combination with transplantation of 10(7) syngeneic bone marrow cells was faster than that in mice treated with IL-6 gene therapy in combination with transplantation of 10(6) or 10(5) syngeneic bone marrow cells, but also the number of CFU-S or survival rate in mice treated with IL-6 gene therapy in combination with transplantation of 10(7) syngeneic bone marrow cells was significantly higher. Nevertheless, the number of platelets, WBCs, CFU-GM or CFU-MK in bone marrow, CFU-S and survival rate in mice treated with IL-6 gene therapy in combination with transplantation of syngeneic bone marrow cells were elevated more significantly than that in mice transplanted with syngeneic bone marrow cells alone. These data demonstrated that IL-6 gene therapy could markedly augment hematopoietic reconstitution after syngeneic bone marrow transplantation and the more bone marrow cells transplanted, the better was the effect.

In the present study, we investigated the effects on hematopoiesis of inactivated vaccines prepared from mouse erythroid leukemia cells (FBL-3) transfected with IL-6 gene. After treatment with IL-6 gene-transfected FBL-3 cells, the platelet count in mice started to increase at day 4, reached its maximum at day 10 and lasted for 25 days. The neutrophil count also increased significantly, but WBC count remained unchanged. The CFU-GM and CFU-MK were elevated to some extent in bone marrow and spleen. These results indicated that in addition to augmentation of antitumor immunity, IL-6 gene-transfected tumor vaccine could promote hematopoietic functions in bone marrow and spleen, and elevate platelet and neutrophil counts in peripheral blood. This approach to gene therapy may be applicable in leukemia treatment, especially for thrombocytopenia related to leukemia or chemotherapy.

The expression of epidermal growth factor receptor (EGFR) in cochlear hear cells was studied in normal and guinea pigs exposed to blasts. The effects of epidermal growth factor (EGF) and dexamethasone (DXM) treatment for blast hearing loss and the underlying mechanisms were also explored. Scattered expression of EGFR was seen in IHCs and OHCs in normal guinea pigs. Segmentally distributing positive reaction was also located in stereocillia of hair cells. Distribution of EGFR reaction was seen in the cytoplasm of IHC 24 hours after exposure to blasts, and in the stereocillia of IHC and the cuticular plate of OHC 72 hours postexposure. At one week EGFR reaction in hair cells increased obviously and part of OHC stereocillia also showed positive reaction. EGFR reaction reduced at two weeks, though positive reaction could still be found in the stereocillia of hair cells at one month. Combination of EGF and DXM administrations promoted hearing recovery significantly. Our results suggest that healing of injured hair cells may be to EGF.

Colony forming unit of granulocytes and macrophages from peripheral blood (PB CFU-GM) represents stem and/or progenitor cells from human blood. In this paper, the effects of histamine H2 receptor agonist 4-methylhistamine (4-MH) and its antagonist ranitidine (Ranit) on the growth of PB CFU-GM cultured in methylcellulose system were studied and their differential effects on normal PB CFU-GM and leukemic HL-60 cells were compared with the effect of the antineoplastic agent cytosine arabinoside (Ara-C). It was found that the histamine H2 receptor agonist 4-MH stimulated the growth of PB CFU-GM when 4-MH was added at the concentrations from 10(-9) mol.L-1 to 10(-6) mol.L-1 among which the dose 10(-8) mol.L-1 exerted most potent stimulating effect (the PB CFU-GM colony numbers was 137.68% +/- 8.20% vs the control, P < 0.01). In contrast, the antagonist Ranit showed inhibitory effect on the growth of PB CFU-GM when at the concentrations 10(-9)-10(-5) mol.L-1 cultured for 14 d in the same methylcellulose system. The inhibition rate was 23.73% +/- 1.16% (10(-9) mol.L-1) and 41.42% +/- 6.75% (10(-6) mol.L-1), respectively. Although both Ranit and Ara-C could inhibit the growth of PB CFU-GM in vitro, Ranit exerted much greater inhibition on HL-60 leukemic cells than on normal PB CFU-GM at the dose of 10(-6) mol.L-1 (100% inhibition for HL-60 and < 50% inhibition for PB CFU-GM). However, the inhibition rate of Ara-C for both HL-60 and PB CFU-GM was 100% at the intensive chemotherapeutic dose of 10(-5) mol.L-1. It would appear that the histamine H2 receptor agonist 4-MH possesses stimulating effect on the growth of PB CFU-GM similar to its effect on CFU-GM from bone marrow as documented before. It is suggested that the histamine H2 receptor antagonist Ranit has, to some extent, potential in the treatment of myeloid leukemia, especially when combined with antineoplastic agent Ara-C at suboptimal doses.

DDB is a hepatoprotectant and has been widely used for the treatment of chronic viral hepatitis in China. The drug markedly improved the abnormal liver function particularly in lowering the elevated serum transaminases in patients. It is known that there is a close correlation between primary hepatocarcinoma and chronic viral hepatitis. The aim of the present study is to evaluate the effect of DDB on hepatocarcinoma cell line. The results showed that the growth and clonogenicity of Bel-7402 human hepatocarcinoma cell line cultured with DDB were markedly inhibited. The nucleoles of the cells treated with DDB disappeared or their numbers and nucleus/cytoplasm ratio decreased under electron microscopic observation. DDB at the concentration of 10(-4) mol.L-1 significantly increased the contents of cAMP and calmodulin (CaM) in Bel-7402 hepatocarcinoma cells. DDB was also found to inhibit topoisomerase II activity of Bel-7402 hepatocarcinoma cells. These results suggest that the mechanism of inhibition of DDB on several phenotypes of Bel-7402 cell line may be related to its effect on cAMP and CaM content as well as topoisomerase II activity.