Using the alkaline phosphatase-anti-alkaline phosphatase (APAAP) technique, plasma cells from multiple myeloma (MM, 23 cases), plasma cell leukemia (PCL, 2 cases) and reactive plasmacytosis (RP, 13 cases) were immunophenotyped with a panel of monoclonal antibodies (McAb). The results showed that McAbCD38 was strongly positive in high percentage of MM and RP cases and the CD9 was the next. 9/23 MM expressed CD10. Our results might indirectly support that CD10 is a malignant marker of MM with poor prognosis, a concept proposed by Durie. The results were (1) all RP but 1 acute monocytic leukemia related to RP were CD10 negative. (2) In our series 2 cases of plasma cell leukemia (PCL) expressed CD10; (3) 4 MM cases survived more than 2 years were CD10 negative. A few MM cases also expressed other surface markers of pre-B and B lymphocyte, such as CD19, CD20, CD22, HLA-DR, cytoplasmic mu chain. CD20 was positive in 4/21 MM and negative in all RP cases. 7/22 MM expressed HLA-DR, and 1/13 RP did so, among them there was a significant difference. HLA-DR seems to be another malignant marker of plasma cells. 1 MM expressed CD8, and 1 PCL highly expressed CD4 indicating PCL might be heterogeneous. Lymphoid stem cells may be involved in MM and PLC. We conclude that multiple myeloma cells have different immunophenotypes and CD10, CD20 and HLA-DR may help to differentiate MM from RP.

41 patients (35 with leukemia, 4 lymphoma, 1 myeloma and 1 neuroblastoma) were treated with autologous hematopoietic stem cells transplantation. 8 patients were treated with autologous bone marrow transplant without purging (ABMT) 15 with autologous bone marrow transplant and purging (pABMT) and 18 with autologous blood hematopoietic stem cells transplant (ABHSCT). The mean hematopoietic reconstitution time was 60, 53, and 29.5 days respectively and the mean GM-CFU recovery time was 37, 44, and 30.5 days respectively in these three groups. The infection complication rates were 37.5%, 26.6% and 16.6% and the relapse rates were 37.5%, 30% and 18.7% respectively in the three groups. The results indicated that the ABHSCT had an earlier hematopoietic reconstitution time, a lesser infection complication rate and a lower relapse rate. Comparing with conventional chemotherapy, all the three transplant groups had an obviously longer disease-free survival time and a better curative effect.

Transpial migration of implanted 5-HT neurons from the subarachnoid space into the spinal cord was studied in adult Wistar rats. Embryonic raphe tissue or cell suspension containing 5-HT cells was used as grafts. The implanted 5-HT cells were monitored by 5-HT immunohistochemical method. The results are as follows: (1) 10 d after cutting the spinal cord at lower thoracic level, 5-HT fibers disappeared in the transected spinal cord. (2) Raphe tissue was implanted into the subarachnoid space of the thoracic lumbar segment after the spinal cord was cut. One month later, 5-HT positive cells could be found in the transected spinal cord with fibers extending into both the grey and the white matters. (3) If the raphe cell suspension instead was implanted, a number of 5-HT positive cells appeared in the grey matter near the implanted region and the distribution of these cells in the grey matter was quite consistent with the implanted range of the cell suspension in the subarachnoid space. The 5-HT positive cells which had entered into the spinal cord sent out fibers and reestablished a new fiber network in the grey matter. (4) After implantation, the number of the 5-HT positive fibers in the transected grey matter became more and more sparsely distributed with increasing distance from the cell bodies and the 5-HT positive fibers reappeared in the white matter were much less than that in the grey matter. Present results show that the implanted 5-HT neurons are able to migrate transpially from the subarachnoid space into the spinal cord.

The synergetic radiosensitizing effect of isomers S-8932 and S-8933 on HeLa-S3 cell-line was investigated in vitro using cell colony forming assay. The results suggested that the combined use of the two isomers showed a lower cytotoxicity than their single use. The ID50 was 55.3 mM. Their synergetic radiosensitizing effect on HeLa-S3 cells under hypoxic condition was higher than either one used alone. The SER of the combined one at 4mM was 1.98 (D0 ratio) and 1.79 (D9 ratio) indicating a sensitizing effect at both low and high radiation doses. The type of the synergetic radiosensitization of the two compounds indicates that they are radiosensitizers of "alpha-beta" type according to the analysis of their molecular models. If the two isomers are used in combination in fractionation radiotherapy, less number of fraction and larger dose per fraction should be selected in the scheme of treatment.

Four patients with solid tumors were treated with high dose chemotherapy and HLA-mismatched multi-cord blood transplantation. Of them, 3 reached complete remission, 1 got fair response. The side effects were tolerable. Little or no graft-versus-host reaction (GVHR) was observed. Donor cell engraftment was documented in 2 patients by cytogenetic analysis and determination of hemoglobin F of the peripheral blood. Recurrent diseases were confirmed clinically on day 210 (4 year-old girl with liposarcoma) and day 90 (11 year-old boy with non-Hodgkin's lymphoma). These results demonstrated that HLA-mismatched multicord blood transfusion could rescue myelo-suppression of patients treated with myeloablative chemotherapy. Therefore, cord blood may serve as a rich source of hematopoietic stem cells.

The attachment and growth property of a human carcinoma cell line (Anip), which has high pulmonary metastatic potential, were investigated in vitro on the lung- or liver-derived biomatrix in culture medium supplemented with serum (SSM) or free of serum (SFM). Either in SFM or SSM, the carcinoma cells showed a higher rate of adhesion to the lung biomatrix than that to the liver biomatrix, In SFM, cells plated on plastic plates ceased proliferating and died; whereas those on the biomatrices, particularly the cells on the lung biomatrix survived and proliferated. Moreover, in SSM, carcinoma cells on the lung biomatrix had the highest clonal growth efficiency. These results indicate that biomatrices may provide conditions favourable for cell growth, and may compensate in part the deficiency of nutrition and growth factors when serum is absent in the culture medium. The different influences exerted on the growth and attachment rate of tumor cells by biomatrices derived from different organs also help to clarify the mechanism of organ specificity of tumor metastasis.

The purpose of this study is to develop a new way for leukemia patients to tolerate an ablative chemoradiotherapy without BMT. Previous studies had demonstrated that only a few leukemia cells remained in the body during remission phase following chemotherapy and did not migrate to distant organs via the circulation until they had given rise to 3-4 x 10(5) new cells. The period required for such growth was about 20 days. However, hematopoietic stem cells migrate earlier than do leukemia cells. Therefore, alternate half body irradiation (HBI) within this period would kill the residual leukemia cells, but hematopoietic stem cells would migrate from shielded marrow to irradiated marrow and reconstruct hematopoiesis. BN rats were injected i.v. with 10 BNML subline LT12nl #15 cells, which had been transfected with the E. coli Lac-Z and Neo genes by retrovirus-mediated gene transfer. Cytochemical X-gal staining was used to identify the leukemia cells in agar culture. At day 9 after inoculation of leukemia cells, the rats were treated with cyclophosphamide (Cy 120 mg/kg), and 3 days later irradiated with 10 Gy (2 Gy/min, 6 MV, X-rays) on the upper half body, with the lower body shielded by a lead brick. They were then irradiated with the same dosage in the lower body with the upper half body shielded a week later. The preliminary results were as follow: At day 9 after inoculation, the cellularity of bone marrow (humerus, sternum, femur and tibia) was in the normal range, and the number of L-CFU ranged from 3.6 x 10(-5) to 9.0 x 10(-5) in agar culture.(ABSTRACT TRUNCATED AT 250 WORDS)

Three chimeric mice were produced by injecting the embryonic stem cell-CCE cells into the cavity of 3.5 day host blastocysts from Kunming and C57BL/6J mice. By analyses of glucose phosphate isomerase (GPI-1), the results show a contribution from CCE cells in heart tissue of female Kunming chimera. Breeding experiments demonstrated that all chimeric mice were fertile but none of 3 chimeras produced germ line transmission.

Forty-five guinea pigs were randomly divided into 2 groups: 15 in the control group and 30 in the experimental group. All animals were injected immediately with 400 mg/kg kanamycin for 10 days. At the same time, the experimental animals were given orally 10 mg/kg thyroxin every other day 5 times. All animals were decapitated 10 days after administration of drugs and measurement of ABR. Then, the specimens of cochlea were prepared for both SEM and TEM. The result of SEM showed that the degeneration and loss of stereocilium in each turn was significantly milder in the experimental group than in the control. Under TEM, outer and inner hair cells showed changes caused by kanamycin. In the experimental group, the mitochondrias were basically normal. The secondary lysosomes gathered under cuticular plate and supranuclear area. In the control group, the mitochondria showed pyknosis and high density, and multivesicular bodies were increased. The cytoplasm was swelling. There were many vacuoles produced by accumulation of liquid in the cytoplasm. Based on the findings of ultrastructural changes of cochlear hair cells, the mechanism of thyroxine against ototoxicity of kanamycin was discussed.

Enhancement of tumor cell damage by ADPRT inhibitor, 3-aminobenzamide (3AB) was investigated. 3AB could inhibit DNA strand break repair in HeLa S3 cells after treatment with bleomycin. 3AB was able to lower the clonogenic ability of HeLa S3 cells after bleomycin treatment and hyperthermia. 3AB could also prevent potentially lethal damage repair (PLDR) in HeLa S3 cells treated with bleomycin. This work presents some experimental evidence and theoretical background for the possible use of 3AB as a sensitizer in the treatment of cancer.

The cytotoxicities of 2 derivatives of artemisinin B and 2 derivatives of artemisic acid (designated as Compound A, B, C, and D) were investigated, using trypan blue dye exclusion test and colony-forming units assay. At the concentration of 5 micrograms.ml-1, the inhibition rates of these 4 compounds against murine leukemia cell line P388 were > 85%. When tested against human hepatoma cell line SMMC-7721 at 25 micrograms.ml-1, the inhibition rates of Compound A, B, C, and D were found to be 92.3%, 96.9%, 84%, and 82.1%, respectively, and 27%, 8%, 37.8%, 1.7% against normal human embryonic lung cell line WI-38, respectively. These 4 compounds all showed an inhibition rate of 100% against human gastric cancer cell line SGC-7901 at 50 micrograms.ml-1.

BALB/C mice (H-2d) receiving allogeneic C57 mice (H-2b) spleen cells via the portal vein (PV), followed by administration of cyclophosphamide (CY), abrogated the capability of rejecting allogeneic EL-4G- tumor cells (C57 origin). BALB/C mice received this combined treatment and BALB/C mice untreated or treated with either PV presensitization or CY injection were all exposed to 7.5 Gy total body irradiation, which was confirmed to be a lethal dose for BALB/C mice, then given intravenously 2 x 10(7) fetal liver cells from C57 mice (FLC57). The results indicated that the survival times of grafted BALB/C mice combined treatment were longer than those of untreated or either PV or CY treated, (PV + CY + 7.5Gy + FLC 57: MST = 99.5 +/- 12.6 day), v(N + 7.5GY + FLC57: MST = 36.5 +/- 9.8 day; PV + 7.5Gy + FLC57: MST = 12.2 +/- 0.2 day; CY + 7.5Gy + FLC57: MST = 27.6 +/- 5.5 day). The difference was statistically significant. The spleen cells from the grafted mice were assessed by indirect immunofluorescence with anti-C57 serum. The spleen cells from BALB/C mice with survival time over 100 days after grafting of FLC57 in combined treatment group were about 70-80 percentage of C57 positive cells.

The erythroid colony formating ability and the therapeutic efficacy were investigated in patients with chronic renal failure (CRF) before and after treatment with baoyuantang. The results showed that the number of CFU-E, BFU-E of bone marrow was found lower in CRF patients than that in normal controls, and there was inhibitory action in the CRF serum on the number of CFU-E, BFU-E of normal human bone marrow. After treatment with baoyuantang, the amount of Hb was increased and creatinine decreased in CRF patients, and its effective rate reached 83.3%. The above-mentioned inhibitory action in CRF serum was also reduced post-treatment. The results indicated that baoyuantang could increase the ability for proliferation and differentiation of erythroid progenitor and adjust anemia in CRF patient.

One hundred and twenty-eight cases of gastric carcinoma were examined with immunohistochemical technic for carcinoembryonic antigen (CEA), human chorionic gonadotropin (HCG), serotonin, gastrin and lysozyme. CEA were observed in 105 cases. Twenty-four cases were positive for HCG, 53 cases for serotonin, 31 cases for gastrin, 89 cases for lysozyme. Sixty-nine cases exhibited more than two hormones or one hormone and lysozyme simultaneously in different cells of the same tumor. Ultrastructurally, sometimes three types of secretory granules were noticed. The electron dense granules in the lysozyme-containing tumor cells were similar to those of Paneth's cells in intestinal metaplasia. The positive rates of the above three hormones, lysozyme and multi-marker expression in diffuse type carcinoma were higher than those in intestinal type, and 42/44 cases of the diffuse type carcinoma were histologically undifferentiated carcinomas or signet-ring cell carcinomas. Lymph node metastasis occurred more frequently in those carcinomas with hormone or lysozyme positivity. These findings suggest that these neoplastic endocrine cells and Paneth's cells have originated from multipotential differentiation of neoplastic stem cells in the stomach, reflecting the state of the gene activity in the tumor cells.

We have studied the function of dried and prepared Rehmannia glutinosa f. hueichingensis in treating hemorrhagic anemic mice and in fostering bone marrow hematopoietic cells CFU-S, CFU-E. The results show that Rehmannia glutinosa f. hueichingensis can promote the recovery of blood deficient animals, RBC. Hb expedite the multiplication and differentiation of CFU-S, CFU-E, and thus proves markedly helpful to the generation of blood.

The killing of human leukemia cells by cloned human LAK cell was investigated with 51Cr release, semisolid agar colony-formation and MTT assay. Human LAK cells were generated and cloned from normal peripheral blood mononuclear cells stimulated with IL-2. Cloned LAK cells showed significant cytotoxicity against human erythroleukemia cell line K 562, promyelocytic leukemia cell line HL 60 and T-lymphoblastic leukemia cell line Jurkat in standard 4-hr 51Cr release assay and the lytic percentage were 67.1%, 58.4% and 53.1% with E/T ratio of 40:1. Moreover, cloned LAK cells could also inhibit the 6-day spontaneous colony-formation of K 562 and HL 6 C cells in semisolid cultures when the LAK cells were preincubated with target cells for 4 hours in liquid medium at 37 degrees C. The degree of colony inhibition was 91% for K 562 and 96% for HL 60, which was quantitatively greater than that determined by 51Cr release at the same E/T ratio of 40:1. In addition, with MTT assay, cytotoxicity of supernatant of cloned LAK cells 3 days in culture against K 562 and HL 60 cells was observed. Our data indicated that cloned human LAK cells could kill both human leukemia cells and their stem cells and the mechanism may involve direct lysis and/or inhibition mediated by LAK cells and indirect inhibition of some soluble factors released from LAK cells.

Ginseng was said to be benefit for anemia in TCM. Proliferation effects of total saponins of panax ginseng (TSPG) on hematopoietic progenitor cell in normal individuals and 29 patients with aplastic anemia (AA) were observed by bone marrow culture of BFU-E, CFU-E, CFU-GM in vitro compared with methyltestosterone (MT). The results showed that TSPG might prompt proliferation of normal progenitor cells at the concentration of 20 micrograms/ml. The number of BFU-E, CFU-E and CFU-GM had increased by 37.8 +/- 2.9%, 31.4 +/- 2.9% and 33.3 +/- 4.0% over the controls respectively; furthermore TSPG was still useful to BFU-E, CFU-E growth without Epo in vitro, although the colony numbers were very lower. Otherwise MT was useless to CFU-GM. 14 of the 29 patients with AA who responded to MT showed sensitivity to TSPG in marrow culture (the rising rate of colony formation exceeded 30%), but immune-mediated AA (patient's PBMNC suppressed normal hematopoiesis) and stem cell-decreased AA (few of colony was formed) showed almost no expression for TSPG activity because of immunological suppression system and absence of progenitors.

Aplastic anemia can be classified distinctively three types as progenitor depletive; immunosuppressive and androgenic sensitive. Using bone marrow culture in vitro which had been accomplished in our laboratory, 53 patients with aplastic anemia were also classified according to TCM term in three groups as Yin deficiency, Yang deficiency and both Yin and Yang deficiency, and the correlation was observed between TCM classification and lab character of these patients. The results showed that the number of CFU-GM, CFU-E and BFU-E in Yang deficiency group was significantly higher than that in the other two groups (P less than 0.01 and P less than 0.05). It also showed that the sensitivity of progenitor cells to androgenic hormones of Yang deficiency group was preferential to all (P less than 0.005 and P less than 0.05). The percentage of immunosuppressive type of aplastic anemia in Yin deficiency group was much higher than those in the other two groups (P less than 0.005). These observations suggested that TCM classification for aplastic anemia in this paper has objective material foundation.