YM155 inhibits tumor growth and enhances chemosensitivity to cisplatin in osteosarcoma.

OBJECTIVE: Chemoresistance is the principal reason for poor survival and disease recurrence in osteosarcoma patients. Survivin, a family member of the inhibitor of apoptosis proteins, plays an important role in inhibition of apoptosis. Survivin is expressed in a vast majority of human cancers, which is often correlated with poor prognosis in a wide variety of cancer patients. Furthermore, survivin expression is often related with chemoresistance in cancer cells, including osteosarcoma (OS). Here, we evaluated the therapeutic potential of YM155, a selective survivin suppressant alone and in combination with cisplatin using human OS models. MATERIALS AND METHODS: U-2 OS, SW1353, MG-63 cells were treated with YM155, and/or cisplatin, and cell viability, apoptosis, survivin protein expression levels were then evaluated. Furthermore, the efficacy of YM155 combined with cisplatin was further examined in established xenograft models. RESULTS: YM155 was sufficient to induce spontaneous apoptosis of OS cells. Combination with YM155 significantly augmented the cytotoxicity of cisplatin in OS cells. Combination treatment of YM155 and cisplatin showed antiproliferative effects and induced a greater rate of apoptosis than the sum of the single-treatment rates and promoted tumor regression in established OS xenograft models. CONCLUSIONS: Our findings provide evidence that YM155 could act as a survivin inhibitor on OS cells. Chemotherapeutic approaches using YM155 might enhance the benefit of the cisplatin in the treatment of OS cells. YM155 could be further developed as a potential therapeutic agent for the treatment of OS.

Effects of VEGF/VEGFR/K-ras signaling pathways on miRNA21 levels in hepatocellular carcinoma tissues in rats.

The aim of this study was to investigate the effects of the vascular endothelial growth factor (VEGF)/VEGF receptor (VEGFR)/K-ras signaling pathways on miRNA21 levels in hepatocellular carcinoma tissues in rats. Eighteen male Sprague-Dawley rats were randomly divided into normal control, model, and VEGF blocking agent groups (N = 6/group). The expression of VEGF mRNA, K-ras protein, and miRNA21 increased significantly (P < 0.05) in the model group compared with the normal control group, and decreased dramatically in the VEGF blocking agent group compared to the model group. The expression of VEGFR mRNA in the model group was higher than that of the control group, and the expression of VEGFR mRNA in the VEGF blocking agent group was significantly higher than that of the control group (P < 0.05). Statistically, there was no difference between the expression of VEGFR mRNA for the VEGF blocking agent group and the model group (P > 0.05). Finally, the expression of the miRNA21 gene in the VEGF blocking agent group was higher than in the control group, and there was a significant statistical difference noted; Pearson's correlation analysis demonstrated that the expression of K-ras protein was positively correlated with miRNA21 in the experimental groups (P = 0.001). The above results showed that the VEGF/VEGFR/K-ras signaling pathway might promote the occurrence and development of hepatocellular carcinoma cells through regulating expression of miRNA21, which has potential clinical value for the development of therapies against biological targets and determining prognosis for patients with hepatocellular carcinoma.

Matrine promotes G0/G1 arrest and down-regulates cyclin D1 expression in human rhabdomyosarcoma cells.

Matrine has a broad-spectrum of anti-cancer effects and is efficient in the inhibition of proliferation of hepatoma cells, leukemia cells and neuroblastoma cell. However, its efficacy and tentative mechanisms in rhabdomyosarcoma have not been addressed before. This study aimed to investigate the effects of Matrine on cell cycle and expression of cyclin D1 in human rhabdomyosarcoma cells (RD cell line). RD cell line was treated with different concentrations (0, 0.5, 1.0, and 1.5 mg/mL) of Matrine, and cell proliferation and cell cycle were evaluated using, respectively, MTT assay and flow cytometry. The effect of Matrine on cyclin D1 mRNA levels was measured by RT-PCR. There was a dose-dependent inhibition of proliferation in the matrine-treated group (inhibition of proliferation rate in control cells 12.70 ± 0.35%; Matrine-treated cells [0.5, 1.0, and 1.5 mg/mL]: 31.16 ± 0.11%, 42.96 ± 0.9%, and 57.26 ± 0.8%). The G0 / G1 ratio in study groups were, respectively, 58.44 ± 3.57%, 64.79 ± 2.03%, 69.97 ± 2.89% and 75.03 ± 1.23%.Cyclin D1 mRNA levels progressively diminished (control group ratio of cyclin D1 / ß-actin: 0.59 ± 0.06; Matrine: 0.35 ± 0.05, 0.27 ± 0.02 and 0.04 ± 0.03). All aforementioned changes were significant (P<0.05). In conclusion, Matrine markedly suppresses cell proliferation in RD cells by decreasing expression of cyclin D1 mRNA and blocking the cell cycle at the G0 / G1 stage.

Clinical study of Lipo PGE1-inhibiting platelet activation in acute rejection after kidney transplantation.

To observe the efficacy of the platelet activation inhibitor Lipo PGE1 therapy in the recovery of graft function after an acute rejection episode after kidney transplantation. Forty patients with acute rejection after kidney transplantation were randomly assigned into groups treated with or without Lipo PGE1. The expression levels of CD61, CD63, and PAC-1 on platelet surfaces were assayed by flow cytometry. The recovery time for graft function and 1-year patient and graft survival rates were recorded. Compared with controls, the expression levels of CD61, CD63, and PAC-1 were lower among acute rejection patients who received Lipo PGE1 therapy. The recovery time for graft function was shorter and the 1-year patient and graft survival rates higher. Lipo PGE1 therapy in patients with acute rejection episodes may inhibit platelet activation thereby benefiting graft functional recovery. The 1-year survival rates of patients and grafts might be increased if the expression levels of CD61, CD63, and PAC-1 on the platelet surfaces was decreased by Lipo PGE1 therapy.

Relationship between the expression levels of CD61, CD63, and PAC-1 on platelet surface in peripheral blood and the transplanted kidney function.

OBJECTIVE: To analyze the relationships between the expression levels of CD61, CD63, and PAC-1 on the platelet surface and the incidences of acute rejection and tubular necrosis as well as the recovery of graft function after renal transplantation. METHODS: The expression levels of CD61, CD63, and PAC-1 on platelet surfaces were assayed by flow cytometry in 86 patients with different stages of uremia before and after transplantation. Patients were divided into three groups: 29 patients with normal graft function, 30 with acute rejection, and 27 with acute tubular necrosis. Patients with acute rejection were randomly assigned into groups treated with or without anticoagulants. RESULTS: The expression levels of CD61, CD63, and PAC-1 on platelet surfaces significantly increased (P <.05) among patients with acute rejection, as compared with those with normal graft function or acute tubular necrosis. Compared with controls, the expression levels of CD61, CD63, and PAC-1 were lower among acute rejection patients who, received anticoagulant therapy. The recovery time for graft function shorter and, the 1-year patients and graft survival rates higher. CONCLUSIONS: The pretransplant expression levels of CD61, CD63, and PAC-1 on platelet surface were significantly higher among patients with acute rejection, suggesting that this complication rather than acute tubular necrosis may be related to platelet activation. Patients with acute rejection displayed benefit from anticoagulant therapy.

Isolation and regional mapping of NotI and EagI clones from human chromosome 21.

NotI and EagI boundary libraries were constructed for human chromosome 21. One hundred forty-seven clones were isolated from the somatic cell hybrid 72532X-6 and localized using a hybrid mapping panel. After identification of those clones, which were isolated more than once, as well as those probes derived from a previously unrecognized integrated non-chromosome-21 fragment, 58 individual boundary clones (plus 2 additional NotI-EcoRI clones isolated from a flow-sorted library) were localized to 11 separate regions. The distribution of these probes is highly nonrandom, with 50% of the clones located in the distal band 21q22.3. Two probes, Not50 and Eag101, map to regions in the very proximal long arm which may contain the gene responsible for familial Alzheimer's disease (AD1), and Not50 would appear to be more proximal than D21S16 (E9). Twenty-eight probes map to the region between superoxide dismutase (SOD1) and the ETS2 oncogene, which appears to contain genes responsible for many of the phenotypic features of Down syndrome. Twenty clones contain (GT)n repeats, as determined by hybridization to a CA polymer, and should provide additional highly polymorphic probes. Closure of gaps in the physical linkage map of chromosome 21 should be facilitated by the isolation of these probes, as they identify many of the unmethylated CpG-rich islands that have hindered pulsed-field gel analysis. They will also be useful in identifying a set of genes in proximity to NotI and EagI restriction sites, as well as conserved DNA sequences for comparative mapping studies.

[Experimental study on healing of old tear in the avascular portion of menisci in dogs].

32 adult mongrel dogs (male 14, female 18, average weight 16.7 kg) were used to study the healing process of old tear in the avascular portion of the menisci. They were divided into two groups--Control group and repair group which were divided into two subgroups, that is, debrided and undebrided groups. Dogs were sacrificed 2.4 weeks and 4.5 months and one year after repair respectively. Finally results were analysed by means of light and electron microscopy. The following conclusions were drawn: (1) A meniscal tear in the avascular portion would not heal by itself despite some cellular response on the edge of the tear. (2) Tear would not heal after simple vascular access channel procedure. (3)Tear healed with fibrocartilage when vascular access channel with synovial flap implantation was used. Therefore this technique can be considered as an alternative to meniscectomy in the management of torn meniscus clinically. (4) Debridement has a bad effect on the healing process because it makes the tear wider and longer and remove the activated cells. (5) The vascular access channel about one half of the thickness of the peripheral edge of the meniscus does not change the shape of the meniscus.

[Designation, classification and treatment of endometrial stromal sarcoma--a report of 9 cases].

From 1958 to 1984, 13 cases were diagnosed as endometrial stromal sarcoma in our hospital, 9 of whom having complete data and been followed for over 5 years were analysed. The results indicated that degree of cellular atypia was a major factor related to prognosis. According to the degree of cellular atypia with consideration of cellular mitotic rate, the tumors were divided into two types: low and high grade malignancies. The former was likely to develop recurrence in the pelvis whereas the latter was prone to distant metastasis. There was no definite relation between the time and initial site of recurrence or metastasis and the stage of the lesion. Operation is the treatment of choice for primary as well as recurrent lesions. The operation should be total hysterectomy and bilateral salpingo-oophorectomy with removal of all visible tumor tissues. In this series, 5 patients received prophylactic postoperative pelvic radiation, none developed local recurrence and 3 of them have survived for 13-19 years free of tumor. Two of 3 treated with postoperative chemotherapy developed recurrence and died 3 months and 4 years after chemotherapy, respectively.

A survey of accessory auricle anomaly. Pedigree analysis of seven cases.

We performed an investigation of 50 accessory auricles on subjects encountered during 1983 through 1985. Fifteen of the families were found to have the same accessory auricular deformity among the probands' family members. From these 15 families, we selected seven pedigrees that were not accompanied by auricular deformities who had filial generations for analysis. Pedigrees 1 through 6 showed autosomal dominant inheritance; two also had irregular dominant properties (pedigrees 4 and 7), and one (pedigree 7) could not be excluded from the possibility of having an X-linked recessive inheritance. During the investigation, there were latent accessory auricles in which the cartilages were seen to be subcutaneous only or protruding slightly. Genetically, the pathogenesis of an accessory auricle should be related to the accessory auricular gene.

A nuclear protein associated with human cancer cells binds preferentially to a human repetitive DNA sequence.

A protein (Rp66) of 66 kDa was shown by DNA-binding protein blot assay to bind to a human repetitive DNA sequence (low-repeat sequence; LRS) in each of 10 transformed human cell lines examined. This protein-DNA interaction was not observed in 11 normal human cell cultures or in the Chinese hamster cell line CHO-K1. Gel retardation assay confirmed the specificity of the protein-DNA binding between Rp66 and LRS. In a histiocytic lymphoma human cell line, U937, that can be induced to differentiate in the presence of phorbol ester, this binding disappeared after cell differentiation. These together with other results cited suggest a regulatory role for these repetitive sequences in the human genome, with particular application to cancer.

[Relationship between central recurrence and time-dose factors in patients with carcinoma of uterine cervix after radiotherapy].

Time-dose factor (TDF) value at point A was evaluated in 75 patients with central recurrence and 100 patients free of cancer for more than 5 years after radiotherapy serving as control. The results showed that the mean TDF at point A was 131 +/- 16 and, 139 +/- 15 in patients with Stage II lesions in central recurrence and control groups and 134 +/- 18 and 144 + 14 in patients with Stage III lesions in both groups, respectively. The TDF was all lower in the recurrence groups than those in the control groups. There was significant difference between the two groups (P less than 0.02 and P less than 0.01). The results also showed that the TDF at point A was significantly different between the recurrent and control groups (P less than 0.05) when it was more than 135 and 140 in Stage II and III lesions. It is pointed out that, even with same dose delivered to point A, the lower TDF value at point A is possibly the reason of central failure in cervical cancer treatment. The authors suggest that total TDF value at point A in Stage II and III lesions be no less than 135 and 140.

[Carcinoma of the uterine cervix associated with pregnancy--analysis of 458 cases].

From 1958-1978, 458 patients with carcinoma of the cervix complicated with pregnancy were treated in our hospital. It comprised 4.3% of the total number of cervical carcinoma admitted for radiotherapy during the same period. In these patients, 112 cases were antenatal and 346 cases were postnatal. The five year survival rates were 56.2% and 44.7%, respectively, both being lower than that (69.4%) of nonpregnant patients. Statistics showed: 1. the earlier the period of pregnancy, the better the prognosis. 2. within six months postpartum, the earlier the treatment, the higher the survival rate. These results suggest that pregnancy has an unfavorable effect on cervical carcinoma. Irradiation treatment was essentially the same for patients who were pregnant or not. Termination of pregnancy, however, was a prerequisite. In the first trimester of pregnancy, spontaneous abortion with radiation was advocated. In the second and third trimesters of pregnancy, hysterotomy and removal of foetus for proper irradiation was done. Intracavitary irradiation was recommended before termination of pregnancy and postabortal or postpartum external radiation was started as early as possible in order to achieve more favorable results.