Cord blood T cells were enriched by nylon wool colomn, and effects of PHA-stimulated T cell supernatant collected from 18 h to 7 days on the proliferation of CFU-c were studied. The results showed that the supernatant collected at 18 h (PHA-TCM) could significantly inhibit the growth of CFU-c and the inhibition was PHA-TCM dose dependent, suggesting there is a CFU-c inhibitory activity in PHA-TCM. Kinetic studies demonstrated that the activity was decreased in the supernatant collected at 48 h and disappeared at 7 days. On the other hand, unstimulated T cell supernatant and PHA alone had no inhibitory effect on CFU-c growth. Indomethacin did not affect the production of the inhibitory activity and no interferon activity could be detected in PHA-TCM. These suggested that the inhibition was mediated by a non-interferon, non-prostaglandin suppressor. Further studies revealed that the suppressor was a protein stable at 56 degrees C and lost in pH 2 and pH 11 for 3 h, its molecular weight was large than 10,000 dolton.

The effect of bleomycin A5 (BLM) alone and along with calmodulin inhibitor N-(4-aminobutyl)-5-chloro-2-naphthalene sulfonamide (W-13) on the proliferation of S-180 cells in vitro were studied. IC50 of BLM alone to the cells was about 2.63 micrograms/ml, which was decreased to 1/3.8 and 1/9.5 of 2.63 micrograms/ml when plus W-13 1, 5 micrograms/ml respectively. The results indicated that nontoxic doses of W-13 enhanced the inhibition of cell proliferation under the condition of BLM 0.5-2.5 micrograms/ml. In colony forming test, the survival fraction of S-180 cells treated with BLM plus W-13 was decreased to 1/87-240 of the cells treated with BLM alone. The results suggest that W-13 can enhance antitumor activity of BLM in vitro and may be used as an enhancer of BLM in vivo.

Several factors which may influence the colony-forming efficiency of mouse CFU-f, including media, culture dishes, mouse strains, and O2 concentrations in the incubator were tested. Among them, the O2 concentration was found to be the most important factor, when a mixed gas consisting of 5% O2, 5% CO2 and 90% N2 was used to replace the usual 5% CO2 in air, the colony-forming efficiency of CFU-f increased markedly and the influence of other factors on cell growth diminished. Although the use of a lit candle to consume O2 in the air provided good conditions for colony formation of several tumor cell lines in soft agar, it is not suitable for the growth of CFU-f.

Twenty-nine guinea pigs were divided into three groups. The first group received distilled water 2 ml/kg ip daily; the second group received DDP 2 mg/kg ip daily; and the third group DDP 4 mg/kg ip daily. ABR hearing threshold was tested before injection. Twenty-four hours after systemic administration, ABR hearing threshold was tested again. The animals were killed and the cochlear specimens were observed under light, scanning and transmission electron microscopes. No abnormality regarding ABR hearing threshold and cochlear morphology was observed in the first group. In the second group, ABR hearing threshold was elevated, inner and outer hair cells, supporting cells, striae vascularis and spiral ganglions were damaged. In the third group, ABR hearing threshold elevated markedly. All above-mentioned cells were severely damaged. The experiment showed that the cochlear damage induced by DDP was dose related. The severest damage was seen in the second turn of the cochlea. The damage to the Deiters' cells was earliest and severest. The outer hair cells were more vulnerable to the damage than the inner hair cells did.

By scanning and transmission electron microscopy, leukemic cells were observed after CFU-Mix culture. Even though granulocytic growth factor, erythropoietin and lymphocytic growth factor were added in vitro, acute leukemic cells still showed defective differentiation and maturation. These were characterized by abnormal colony which consisted of smooth cells, bizarre shape, nuclear-cytoplasmic asynchrony in development, and appearance of nuclear bleb. However, chronic myelogenous leukemic cells were more mature than the acute ones, manifesting in normal colony with finger-like projections and ruffled membrane. Macrophages and eosinophils could be observed. It is suggested that there is a difference in differentiation between acute and chronic leukemic cells.

Fresh fetal liver obtained from 3- to 6-month fetus was prepared. Fetal liver cell suspension (FLC) or fetal liver cell-free suspension (FLCF) were then transfused into two groups of patient of aplastic anemia. 15 of 21 patients of aplastic anemia treated with FLC showed reconstitution of haemopoietic function or improvement of peripheral blood pictures, while 27 of 30 patients treated with FLCF showed reconstitution or improvement. It is verified that there is a stimulating factor for CFU-CM, BFU-E, and CFU-E and also a immunologic stimulant for improving the nonspecific immunologic function of the organism as shown by clinical analysis and experimental study. It is obvious that the therapeutic effect of FLCF is much better than that of the FLC.

Chronic myelomonocytic leukemia (CMML) is a polymorphous malignant hematological stem cell disorder, characterized by abnormal hyperplasia of mature or immature cells of both monocytic and granulocytic series and with abnormal cellular morphology. It is an independent entity of chronic leukemia, as its prestage course is manifested by refractory anemia with monocytosis and the disease gradually evolves to CMML. In some cases, it finally becomes acute leukemia. In this study, the average white cell count of the patients was 29.3 x 10(9)/L.14 cases had leucocytosis, 7 leucopenia and 5 normal count. The absolute value of monocytes was 19 x 10(9)/L and the proportion of monocytes was 10-87%, with an average of 49%. In the leukopenic group with white cell count less than 4 x 10(9)/L, the absolute value of monocytes was less than 1 x 10(9)/L in 5 of the 7 cases. However, it was noticed that all the 5 cases had a proportion of monocytes greater than or equal to 10%. The authors would like to take this percentage as the diagnostic criteria for CMML, which is different from that adopted in FAB classification of 1982 as well as in the literatures. Statistics showed that P value of M/E, Mo/E, Mo/M were of apparent significance in the differentiation of CMML from normal controls and patients with other hematological diseases such as RA, RAEB, CML, CNL, M4 and M5.

By means of soft agar colonization and limiting dilution, 5 clonal cells with different metastatic capacity, [one with high metastatic rate (78.6-89.4%), three moderate (33.3-66.6%) and one low (15-15.4%)], were separated from Hca-F25/L cell line. Their common features were: All clones metastasize to lymphatic system except a small percentage (1.88%) associated with lung metastasis; five clones as well as their parents all reacted with Keratin monoclonal antibody (HK2), which indicated the epithelial character with poor differentiation; Cells of each clone have highly identical karyotype. Five clone lines have similar marker chromosomes (M1, M2, M3, M4) with different stem line karyotypes. We also found different features of five cell lines in pathomorphological indicators and cell electrophoresis rate. Our results indicated that the Hca-F25/L strain is heterogeneous in phenotype and karyotype.

Pingyangmycin (PYM), produced by Streptomyces pingyangensis n. sp., was found to be identical to bleomycin A5. In the present study, a comparative observation was carried out in 10 human cancer cell lines. As determined by a colony-forming assay, the dose-response curves obtained from cells exposed to PYM for 1 h were of one type only: biphasic exponential. The sensitivities of these cells derived from different types of tumors, however, varied with a broad range of ID50 values (0.03-0.82 microgram/ml). A hepatoma cell line (BEL-7402) and three lines derived from squamous carcinomas of the esophagus (Eca109 and CaEs17) or the nasopharynx (CNE) were relatively sensitive (ID50 less than 0.20 microgram/ml) to PYM which is known to have clinical activity against these diseases. Two gastric adenocarcinoma cell lines (MGc80-3 and BGC-823) and a pulmonary adenocarcinoma cell line (SPC-A-1) appeared to be less sensitive to the drug, with ID50 values of 0.21-0.47 microgram/ml. PYM was 7-fold more effective against LTEP-78 cells derived from pulmonary squamous carcinoma as opposed to SPC-A-1 cells, resulting in a low ID50 value of 0.04 microgram/ml. However, PYM as a single agent has not yet received full evaluation in relation to this type of lung cancer. In contrast with other cell lines of squamous cancer origin, HeLa and CC-801 cells derived from uterine cervix carcinomas which have been evaluated as highly responsive to PYM had the highest ID50 values (greater than 0.70 microgram/ml).

84 cases observed were divided into 4 types according to principles of diagnosis and treatment based on an overall analysis of symptoms and signs, namely: Yang-deficiency of the Spleen and Kidney (62 cases, 73.8%), Yin-deficiency leads to internal heat (6 cases), deficiency of both the Heart and the Spleen (7 cases), and prostration of Qi after loss of blood (9 cases). The patients were treated with the method of warming and tonifying the Spleen and Kidney by using Er-Xian decoction of warming the Kidney. The three other types were also treated with the method after being relieved with the methods of tonifying the Heart and the Spleen, of nourishing Yin to relieve internal heat and cooling blood, and of strengthening Yang to stop chronic hemorrhage respectively so that function of the blood and Qi would promote each other and be improved. The total effective rate was 84.5%, and remission rate was 47.6% among 84 cases, but 91.9% and 50.0% in the type of Yang-deficiency of the Spleen and Kidney alone. Among the effective cases there was a remarkable improvement in the hemogram after treatment (P less than 0.001, less than 0.01), and the long-termed curative effect was also exciting. The mechanism of Er-Xian decoction of warming the Kidney was investigated through the nourishment of the hemopoiesis stem cell. The experiment showed that the decoction could increase CFU-S and GM-CFU in the bone marrow under the lower hemopoietic function of the bone marrow, and increase GM-CFU and CFU-E in the bone marrow of normal rats (P less than 0.001, less than 0.01, less than 0.05). The histological findings showed that there was a reduction in hemorrhage and hyperemia in the bone marrow between the decoction group and the control group, and the recovery of hemopoietic function was better than the latter.

Murine lymphokine-activated killer (LAK) cells were generated from spleen cells of C57/BL6 mice by culture of spleen cells in vitro for 72 hours in medium containing 500 units/ml recombinant human interleukin 2 (IL-2), and effects of these LAK cells on proliferation of syngenic myeloid progenitor cells (CFU-GM) were observed. After 3 days culture, LAK cells were assayed for their cytotoxicity in a 4 hours 51Cr-release test. Either natural killer (NK) cell sensitive YAC-1 lymphoma cells or NK cell resistant LP-3 and WEHI-164 fibrosarcoma cells were efficiently lysed by murine LAK cells. When LAK cells were added into culture system in a final concentration of 5 x 10(4)/ml, 2 x 10(5)/ml, 8 x 10(5)/ml, CFU-GM were increased by 55.2%, 165.5%, and 194.4% of control respectively. LAK-CM also showed augmentative effect on CFU-GM growth. When 10% (v/v) of LAK-CM were added into culture system, CFU-GM were increased by 51.4% of control, but LAK-CM alone could not stimulate CFU-GM growth. Again, effects of LAK-BMC interaction on CFU-GM formation were investigated. CFU-GM were inhibited to 27.6% of control when 1 x 10(5) BMC were mixed with 8 x 10(5) LAK cells and incubated for 4 hours prior to CFU-GM culture. These data suggest that (1) LAK cells may secrete co-CSF which showed synergistic effect with CSF on CFU-GM proliferation: (2) When LAK cells contact with BMC, they showed significant cytotoxicity to myeloid progenitor cells which mediated decrease of CFU-GM formation.

In this paper, the morphological and histological characters of crude drugs derived from Ephedra likiangensis Florin, E. likiangensis f. mairei (Florin) C. Y. Cheng, E. saxatilis Royle ex Florin, E. gerardiana Wall, E. gerardiana var. congesta C. Y. Cheng, E. minuta Florin, E. minuta var. dioeca C. Y. Cheng, E. lepidosperma C. Y. Cheng, E. lomatolepis Schrenk and E. intermedia var. tibetica Stapf are described, compared and illustrated with line drawings. It was found that all the Ephedra spp. growing in southwester nChina possess the following features in common, viz. (1) numerous distinct longitudinal ridges are present on the stem; (2) xylem fibres and tracheids possess steep, tertiary spiral thickenings; (3) stone cells (sclereids) are absent from the nodes. According to the above characters, the drugs produced in southwestern China can be differentiated from those produced in northern China. The results also show that the histological characters of E. intermedia var tibetica Stapf produced in southeast ern Xizang (Tibet) are different from those produced in north western Xizang. The former possesses the xylem fibres and tracheids with steep, tertiary spiral thickenings, while the latter possesses sclereids at stem-nodes. It may be related to their habitat factors and interspecies hybridization. A key for identification of all the crude drugs derived from Chinese Ephedra spp. studied in this series of paper, including thirteen species, three varieties and one form is presented.

The total alkaloids of T. glandulosissimum and its main component hernandezine were found to be effective for treatment of mice bearing P388 leukemia, S180 ascites and C26 colon cancer. Although hernandezine inhibited the growth of mouse L1210 cells and human oral cancer KB cells in vitro markedly, its inhibitory effect on normal hemopoietic progenitor cells (CFU-GM) in mice was relatively low. Preliminary results showed that hernandezine blocked cell-cycle transfer from G1 to S phase, and its cytocidal action might be cell cycle specific. In addition, two other components, thalidezine and isothalidezine, isolated from the same plant exerted similar inhibitory effect on L1210 cells.

During autologous bone marrow graft in treatment of malignant diseases, it is critical to purge malignant cells from the marrow. In the present study, the sensitivity to photodynamic inactivation of 3 leukemic cell lines was compared with their counterpart normal hematopoietic cells. After mouse leukemic L1210 cells were treated with a preparation of hematoporphyrin derivatives, YHpD, 10 micrograms/ml for 1 h and irradiated with black light (peak wavelength 395 nm, light intensity 0.6 mW/cm2) for 5 min, the survival rate of clonogenic cells decreased to less than 10%, while that of bone marrow granulocyte-macrophage progenitor cells (CFU-GM) in DBA/2 mice remained at nearly normal level (greater than 80%). Similar results were obtained when human leukemic HL-60 cells were compared to human CFU-GM and mouse leukemic L615 cells to CFU-GM in 615 strain mice. It is suggested that hematoporphyrin photoradiation may be useful for selectivity killing leukemic cells in bone marrow.

The killing effects of 514.5 nm argon laser and 630 nm dye laser radiation on the murine SP2/0 myeloma cells cultured in thin layer containing HPD were measured by clonogenic assay. It was found that the former was 3.34 fold higher than the latter. Improved argon laser photodynamic therapy was used in the treatment of 104 superficial bladder tumors in 40 patients. Firstly, high output (6-7 W/cm2) argon laser contact and interstitial radiation were used to eradicate visible tumors, and then cylindrical optic fiber was used to deliver argon laser (2.1-3.47 W/cm2) for whole bladder mucosal scattered irradiation to destroy small multi-focal tumors and reduce recurrence. In this series, the follow-up was 7-34 months. All patients achieved complete response and only 7 (17.5%) recurred.

Bone marrow cells from 33 patients with AML were cultured in vitro using PHA-LCM. The test consisted of two phases: an initial liquid phase and then a semi-liquid phase as described by Dicke et al. Drug sensitivity test for homoharringtonine (H) and cytarabine (A) were performed with clonogenic assay. All patients were treated with these drugs. The results showed that PHA-induced leukemic colonies (CFU-AML) varied from 0 to 812/2 x 10(5) cells (median 175). Three patterns of cell growth were recognized in analysis: high degree (7 patients) with more than 250 CFU-AML colonies, median degree (17 patients) with 50-250 and low, degree with 0 to 49 colonies (9 patients). Drug sensitivity was closely related to the cell growth patterns as evaluated with the clinical outcome. Patients with "high" growth pattern needed more sensitive drugs. When clonogenic assay showed "low" growth pattern, all patients responded to H and A very well regardless the degree of drug sensitivity. Most patients with M3 had high or median growth patterns and relatively low sensitivity to HA. Only one of the six patients with M3 got remission.

30 cases of chronic myeloid leukemia (CML) with non-myeloid blast crisis from 1966 to 1986 in PUMC Hospital were investigated. Morphologically 18 cases were lymphoblastic, 4 histiocytic, 3 basophilic, 2 erythro-leukemic, 2 megakaryocytic, and 1 monocytic, the ratio between male and female was 3.3:1, and their age ranged from 16 to 55 years. These results suggest that blast crisis of CML may involve many other cellular derivatives than the myeloid series of the pluripotential stem cells, Spleen was not palpable among half of the patients with lymphoblastic crisis, but all the cases with blast crisis of other morphological types had enlarged lives and spleen, especially those with histiocytic and monocytic crisis of CML. Most of cases of CML with non-myeloid blast crisis had poor prognosis with survival time of less than 6 months. However, cases of CML with lymphoblastic crisis had longer survival duration than those with non-myeloid blast crisis of other types.

The effects of histamine H2-receptor antagonists (cimetidine and ranitidine) on bone marrow hematopoiesis of normal and sublethally irradiated mice were investigated. It was found that non-toxic dosage of cimetidine had no significant effect on CFU-s of the normal mouse, but it inhibited CFU-GM growth with a dosage-dependent relationship and the recovery of CFU-s formation from sublethal gamma-ray irradiation. These results suggest that histamine may play a role in bone marrow regeneration after sublethal irradiation. Histamine H2-receptor antagonists can inhibit hemopoietic reconstruction in regenerating bone marrow.

The cytostatic effects of triptolide on HeLa cells in different proliferation stages and cell cycle phases were studied by colony-forming units assay. An exposure of exponential-phase cells to triptolide 0.02-4.00 micrograms/ml for 0.5 h resulted in a biphasic-exponential dose-survival curve (n = 1, D0 = 0.3 micrograms/ml in the most sensitive population; D0 = 2.8 micrograms/ml in the more resistant population). The plateau-phase cells in the same conditions seemed to have lower sensitivity to the drug. The synchronized cells caused by excess TdR double block and the selective detachment of mitotic cells from monolayer were exposed to triptolide 0.06 micrograms/ml for 0.5 h. The sensitivity of cell cycle phases to the drug ranked as follows: S greater than G2, M greater than G1. The result showed that triptolide is one of the cell cycle phase non-specific agents, but more sensitive to S phase cells.

The proliferation of L 1210 cells ceased rapidly after they were exposed to homoharringtonine (HH) 1 microgram/ml during exponential growth phase. However, 25.3% of the cells were still able to form colonies in soft agar if HH was removed after 24 h of incubation (the colony-forming efficiency for control cells was 62.5%). The clonogenic cells survived from the treatment were still sensitive to HH-continuous exposure. The IC50 of the treated and control cells were 15 and 20 ng/ml, respectively. Yet, the sensitivity of the treated cells to cytarabine decreased enormously. For instance, the survival rate of HH-treated cells remained at 100% level after they were exposed to cytarabine 4-8 micrograms/ml for 1 h, but only 40% control cells survived from the same treatment. When cells were continuously exposed to HH 0.4 micrograms/ml, the colony-forming efficiency decreased exponentially as a function of exposure time. The T1/2 of the clonogenic cells was about 18 h. The DNA contents in L 1210 cells was measured with a flow-cytometer. The results showed that the cell-cycle progress in all cells was interrupted by HH, regardless which phase they belonged to. So the cells seemed to be in a "frozen" state and the histogram unchanged.