[Safety and efficacy analysis of endovascular thrombectomy in patients with large vascular occlusion with low Alberta Stroke Program Early Computed Tomography Score].

Objective: To evaluate the safety and efficacy of endovascular thrombectomy (EVT) in acute anterior circulation large vessel occlusive stroke (ALVOS) and explore the related influencing factors for prognoses in patients with low Alberta Stroke Program Early Computed Tomography Score (ASPECT). Methods: Patients with acute ALVOS who underwent EVT in Yijishan Hospital of Wannan Medical College from January 2019 to June 2022 were sequentially enrolled. (1) Patients were divided into a low ASPECT group (0-5) and a non-low ASPECT group (6-10), and the differences between the two groups were compared with respect to incidence of perioperative complications and good prognosis rate [modified Rankin scale (mRS) score≤2] 90 days after onset. (2) According to the prognoses 90 days after onset, the low ASPECT group was divided into the good prognosis (mRS score≤2) and poor prognosis (mRS score>2) subgroup. Univariate analysis and multivariate logistic regression analysis were used to investigate the independent risk factors for prognoses of the low ASPECT patients after EVT. Results: A total of 582 patients [age 26-94(69±11) years, 345 male patients (59.3%)] were enrolled for analysis. The baseline ASPECT score was 8 (7, 10), and the baseline NIHSS score was 14 (11, 18). Among them, 102 (17.5%) patients were in the low ASPECT score group and 480 (82.5%) patients were in the non-low ASPECT score group. In the total cohort, patients in the low ASPECT score group had a higher incidence of symptomatic intracranial hemorrhage, lower 90-day good prognosis rate, and higher 90-day mortality rate. Further, propensity score matching statistical analysis showed that patients in the low ASPECT score group had a significantly higher incidence of malignant brain edema after EVT treatment (40.0% vs. 17.6%, χ2=9.13, P=0.003), and a significantly lower 90-day good prognosis rate (24.7% vs. 41.6%, χ2=4.96, P=0.026), but there was no significant difference in the incidence of symptomatic intracranial hemorrhage and 90-day mortality between the two groups (40.3% vs. 26.0%, χ2=3.55, P=0.060). Among 102 patients with low ASPECT score, 22 (21.6%) patients had good prognosis and 80 (78.4%) had poor prognosis. Multivariate logistic regression analysis showed that history of atrial fibrillation (OR=4.478, 95%CI 1.186-16.913, P=0.027) was an independent risk factor for poor prognosis of EVT in patients with low ASPECT score, while good collateral circulation (grade 2 vs. grade 0: OR=0.206, 95%CI 0.051-0.842, P=0.028) was a protective factor for good prognosis of EVT in patients with low ASPECT score. Conclusions: Although the 90-day good prognosis rate of EVT treatment for patients with low ASPECT score was lower than that of the non-low ASPECT group, 21.6% patients still benefitted from EVT treatment, especially patients with non-atrial fibrillation and good collateral circulation. Future studies involving more patients are needed to validate our observations.

[Therapeutic effect of IGF-1R-targeting inhibitor (TAE226) on malignant pleural effusion in nude mice].

OBJECTIVE: To study the therapeutic effect of IGF-1R inhibitor TAE226 on malignant pleural effusion (MPE) in nude mice. METHODS: Human lung carcinoma A549 cells were injected into the pleural cavity of nude mice to establish MPE model. The mice were randomly divided into model group and treatment group, and were orally administered with distilled water and TAE226 (20 mg/kg) in the same volume, respectively. The volume of pleural effusion and tumor weight of the two groups were observed. HE staining was used to reveal the histological changes and enzyme-linked immunosorbent assay (ELISA) was used to detect the IGF-1R protein expression. IGF-1R mRNA level in the tumor tissue was determined by RT-PCR. Microvessel density (MVD) and cell proliferation index (PI) were assessed by immunohistochemical analysis. The protein expression levels of IGF-1R, p-IGF-1R, PI3K and p-PI3K in the tumor tissue were determined by Western blotting. RESULTS: The volumes of pleural effusion were (241.4±89.7) µl and (121.7±78.8) µl in the model and treatment groups, respectively (P<0.05). The tumor weight of treatment group was (316.7±186.3) mg, significantly lower than that of the model group (671.4±281.4) mg (P<0.05). RT-PCR analysis showed that IGF-1R mRNA level was 0.914±0.029 in the treatment group, significantly lower than that of the model group (1.152±0.037, P<0.01). The ELISA data revealed that IGF-1R protein expression level of the model group was significantly higher than that of the treatment group [(41.0±4.7) µg/L vs. (24.0±3.1) µg/L, P<0.01]. Immunohistochemical analysis showed that there were significant differences between MVD and PI in the model and treatment groups [MVD, 34.75±3.49 vs. 22.25±3.63; PI, (75.25±7.15)% vs. (45.75±5.12)%; P<0.01 for both). Western blot data showed that IGF-1R and PI3K protein expression levels were not significantly different between the model and treatment groups (1.03±0.33 vs. 0.98±0.37 and 1.05±0.28 vs. 0.98±0.19), respectively (P>0.05), but p-IGF-1R and p-PI3K protein expression levels had significant differences between the two groups (1.08±0.10 vs. 0.51±0.08 and 1.12±0.09 vs. 0.86±0.09), respectively (P<0.01 for both). CONCLUSIONS: The IGF-1R inhibitor can effectively inhibit the formation of malignant pleural effusion. Its mechanism may be related to the suppression of tumor cell proliferation, invasion and angiogenesis through inhibition of PI3K signaling. TAE226 treatment may be a potential therapeutic regimen of treating malignant pleural effusion.

Gene transfer of pro-IL-18 and IL-1beta converting enzyme cDNA induces potent antitumor effects in L1210 cells.

We report in a murine model of acute lymphoid leukemia L1210 the potent antitumor efficiency of a combinatorial delivery of pro-IL-18 gene modified L1210 (Lp18) and IL-1beta converting enzyme (ICE) gene modified L1210 (LpICE). Live leukemia cells Lp18 or Lp18 plus LpICE showed apparently reduced leukemogenicity with a survival rate of 40 or 50% at 50 days after intraperitoneal (i.p.) inoculation of a lethal dose of cells, respectively. Combination of Lp18 and LpICE was capable of inhibiting accumulation of bloody ascites, synergistically superior to Lp18 or LpICE alone. All surviving mice were rechallenged with parental L1210 cells at day 50, and all survived up to day 80, suggesting that gene-modified cells induced immune protection. Moreover, NK cytotoxicity and CTL activity were both enhanced in mice injected with Lp18, especially Lp18 plus LpICE. Levels of IFN-gamma were not altered significantly by inoculation of Lp18 or Lp18 plus LpICE. Our results demonstrate that IL-18 is a useful candidate gene in gene therapy of lymphoma or lymphoid leukemia, and ex vivo combinatorial delivery of Lp18 plus LpICE either as a single approach or as an adjunct to concomitant radiotherapy or chemotherapy, may be more efficient in a situation of minimal residual disease.

Enhancement of J6-1 human leukemic cell proliferation by cell-cell contact: role of an M-CSF-like membrane-associated growth factor MAF-J6-1.

Density-dependent cell proliferation and cluster formation are growth phenotypes frequently associated with leukemia cells. The secretion of autocrine growth factor, such as granulocyte-macrophage colony stimulating factor (GM-CSF) and interleukin 1 (IL-1), has been implicated as one possible mechanism in leukemogenesis. In many cases, however, leukemia cells do not appear to produce autocrine growth stimulators. J6-1 is an established human myeloid leukemia cell line that exhibits both density-dependent and cluster-forming growth characteristics. The effect of direct cell-cell contact on J6-1 cell proliferation was investigated. We have isolated from J6-1 cells a membrane-bound factor (designated as MAF-J6-1) that promoted the colony formation by both J6-1 cells and mouse bone marrow CFU-GM. The growth-promoting activity of MAF-J6-1 can be neutralized by either anti-macrophage-CSF (M-CSF or CSF-1) or anti-MAF-J6-1 monoclonal antibodies (MAb), suggesting that MAF-J6-1 is related to M-CSF. Using an immunoblot analysis with anti-MAF-J6-1 MAb, the MW of this membrane-associated factor was estimated to be 80 kDa. Both antibodies also induced a modest growth inhibition on J6-1 cells in vitro. Similarly, addition of exogenous recombinant human M-CSF augmented the colony formation by J6-1 cells, an effect also neutralized by both antibodies. Using an in situ hybridization technique, J6-1 cells were found to express a high level of c-fms proto-oncogene, which encodes the receptor for the M-CSF. Taken together, our results suggest that the membrane-bound MAF-J6-1 promote J6-1 cell proliferation and cluster formation through a 'juxtacrine' mechanism.

Enhancement of J6-1 human leukemic cell proliferation by membrane-bound M-CSF through a cell-cell contact mechanism II. Role of an M-CSF receptor-like membrane protein.

We have isolated an M-CSF-like membrane-associated growth factor from human leukemic J6-1 cells that can enhance the growth and colony formation of J6-1 cells in vitro. Indirect evidence suggests that this membrane-associated M-CSF-like growth factor may do so by stimulating a corresponding receptor co-expressed on the adjacent J6-1 cells. The objective of this study is to isolate the putative receptor in J6-1 cells by virtue of its ability to bind and thus "block" the growth of J6-1 cells. Based on this approach, we have isolated from the J6-1 cell membrane an inhibitory activity that can inhibit the clonal growth of J6-1 cells. The activity of this inhibitor can be readily neutralized by either anti-M-CSFR MAb or anti-M-CSFR antiserum, suggesting that it is related to M-CSFR, a product of c-fms proto-oncogene. Judging from Sephadex G-200 gel filtration, the molecular weight (MW) of this putative M-CSFR-like inhibitor was estimated to be approx. 150-180 kDa, comparable with that of M-CSFR. The specificity of M-CSFR-like protein to recognize and block membrane-bound M-CSF also was implicated by its ability to upregulate the steady-state levels of c-fms mRNA in J6-1 cells. Besides its antiproliferative activity in vitro, treatment of J6-1 cells with the putative receptor protein before inoculation effectively blocked the growth and tumor formation in vivo by J6-1 cells in a nude mouse model. These findings suggest that the growth and tumor development by J6-1 leukemic cells may involve a contact-mediated "juxtacrine mechanism".

Autocrine and possible intracrine regulation of HL-60 cell proliferation by macrophage colony-stimulating factor.

The abnormal expression of macrophage colony stimulating factor (M-CSF) isoforms, i.e. membrane bound M-CSF (m-M-CSF) and intracellular M-CSF (c-M-CSF), and their receptor were reported in some leukemia and tumor cells. Furthermore, the nuclear localization of them may be related to poor prognosis and metastasis, while the mechanism is uncertain. We previously reported that m-M-CSF and its receptor played auto-juxtacrine and adhesion molecule role in human leukemia cell line J6-1. In this paper, we show that HL-60 cells highly express M-CSF and its receptor. The localization of positive reactions was mainly in cytoplasma and nuclear in HL-60 cells. In cytoplasma and nuclear, three isoforms of M-CSF were found with molecular weight (MW) of 20, 16 and 14 kDa, while one type of m-CSF receptor (M-CSFR) was discovered with MW of 120 kDa. Immunoprecipitation assay showed that these ligands could exist separately or binding with their receptor. Monoclonal antibody (McAb) against M-CSF and anti-sense oligodeoxynucleotides (ASON) blocking M-CSF expression inhibited the proliferation of HL-60 cells. McAb and ASON regulated the expression of cyclin D1/E, CDK2/4 and p16. Simultaneous administration of both McAb and ASON inhibited the proliferation of HL-60 cells and modulate the expression of cyclins at greater degrees. Our results suggested an autocrine and possible an intracrine loop of M-CSF/M-CSFR in HL-60 cells.

Expression of membrane-associated macrophage colony-stimulating factor (M-CSF) in Hodgkin's disease and other hematologic malignancies.

We have identified a membrane-bound form of M-CSF (m-M-CSF) from an established human leukemic J6-1 cell line. To further understand its biological significance, we studied the expression of this membrane-associated growth factor in the lymph nodes of lymphoma patients and bone marrow smears from patients with hematologic diseases by immunohistochemical staining using anti-M-CSF MAb. We detected a high incidence of m-M-CSF expression in 75% (9/12) of the lymph node sections from patients with Hodgkin's Disease (HD). The antigens were detected primarily in large clusters of mononuclear Hodgkin's cells and the extracellular matrix (EM) surrounding them. In one HD patient with abundant multinucleated Reed-Sternberg (R-S) cells, all of them were intensely stained with anti-M-CSF MAb. In non-Hodgkin's lymphomas (NHL), the incidence (17.6 %) of m-M-CSF expression was lower (3/17). Yet, no m-M-CSF antigens were detected in the lymph nodes from six cases of non-hematologic malignancies and other diseases. A high response also was detected in bone marrow smears obtained from patients with hematologic malignancies, which include myeloid leukemias (32.5%), lymphomas with bone marrow metastasis (50%) and myelodysplastic syndromes (MDS) (37.5 %). By comparison, only 6.8 % of bone marrow smears from non-malignant hematologic diseases and 2.7% of lymphoid leukemias showed positive staining with anti-M-CSF MAb. Our results showed that high expression of m-M-CSF antigens is linked to some types of lymphomas, especially HD. and myeloid leukemias, and may play a role in the development of these hematologic malignancies.

Microskin autograft with pigskin xenograft overlay: a preliminary report of studies on patients.

Split-thickness pigskin graft (STPSG) was used to replace allograft skin for microskin grafting in 16 patients, nine of whom were burn patients, five suffered from traumatic defects and two from diabetic ulcers. The expansion ratios used in these patients ranged from 8:1 to 12:1. The STPSG preparation described was found to be safe for clinical application. The autogenous donor skin was excised from the inguinal area, and the donor site was primarily closed. There were no instances of donor site morbidity. The majority of the STPSG overlays adhered to the wound firmly. Histological examination showed that the microskin grafts proliferated actively immediately beneath the STPSG overlay. The time for the wound to be fully resurfaced varied from 13 to 21 days depending on the expansion ratio employed. There were only two episodes of pseudomonas infection and no further grafting was required in any of the patients. In this study the pigskin xenograft was found to provide a suitable environment for the epithelialization of microskin autografts. When allograft is not available, this is an alternative way of ensuring successful microskin grafting.

Serum lipid concentrations correlate with the progression of chronic renal failure.

OBJECTIVE: To explore the distribution pattern for serum lipid concentrations among patients with different degrees of chronic renal failure; to study the characteristics of abnormal lipid metabolism for chronic renal failure patients when the disease progress further. SETTING: No. 255 Hospital of PLA, Tangshan, Hebei, China; No. 281 Hospital of PLA, Beidanhe, Hebei, China; and the General Hospital of Beijing Military Region, Beijing, China. PRACTICE DESCRIPTION: A total of 240 serum/urine samples from 50 healthy volunteers and from 190 patients with different degrees of chronic renal failure, which fall into four groups according to their glomerular filtration rates, were measured for serum levels of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, high density lipoprotein cholesterol, and for urine albumin concentrations; the levels of these criteria were compared between the control group and diseased groups; the mean concentrations of different lipid variables were paired and subjected to linear regression analysis. MAIN OUTCOME MEASUREMENTS: Glomerular filtration rates were estimated by the iohexol clearance method, in which plasma content of iohexol was measured with high performance liquid chromatography; concentrations of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, high density lipoprotein cholesterol, and albumin were assayed according to standard protocols. RESULTS: Serum levels of triglyceride, lipoprotein(a), lipoprotein(a) cholesterol, total cholesterol, apolipoprotein A1, apolipoprotein B100, low density lipoprotein cholesterol, and urine albumin contents were significantly higher, whereas those of high density lipoprotein cholesterol were lower, in diseased groups than that of the control (p < 0.05, p < 0.01). When the disease progressed, concentrations of these criteria increased or decreased further (p < 0.01, p < 0.05). Significant correlations were found between a few lipid criteria for their mean concentrations in diseased groups. CONCLUSION: The study demonstrates a correlation between abnormalities of lipid metabolism and the degrees of kidney insufficiency, and a correlation within certain kinds of lipid criteria in patients with different degrees of renal damage. The results suggest the existence of multi-correlations in vivo in catabolism and metabolism of lipid, lipoprotein, apolipoprotein, and protein in the patients. The exact mechanism responsible for the association and correlation remains to be clarified.

Neonatal screening for glucose-6-phosphate dehydrogenase deficiency in Taiwan.

Glucose-6-phosphate dehydrogenase (G6PD) deficiency is the most common enzymopathic disease in Taiwan. The mass neonatal screening of G6PD deficiency by fluorometric spot test in Taiwan was started with a pilot program in 1984. The nationwide screening was started on July 1, 1987, and a follow-up system comprising of eighteen referral hospitals, including outlying islands, was organized for confirmatory test, medical care and genetic counseling. From July 1987 to December 1997, 2,971,192 heel blood samples collected on filter paper from 1,143 delivery units were screened by four neonatal screening centers. 46,570 cases were confirmed as G6PD deficiency is estimated to be around 2.1% (male 3.1%, female 0.9%) in Taiwan. The coverage rate of neonatal screening was 99% in 1997. To assess the reliability of the confirmatory test, an external quality assurance (QA) program for G6PD assay was developed. Periodically, 3 or 5 lyophilized quality control materials with different activities of G6PD were sent to each referral hospital by speed post delivery in dry ice. From January 1988 to June 1998, 85 QA services were performed. Two hundred and seven (13.5%) abnormal QA results were found, which were attributed to clerk (11.6%), procedural (16.4%), and instrumental errors (47.3%). In aid to confirm G6PD deficiency, a method to detect the G6PD mutation by using the dried blood samples was developed. The frequencies of the mutant alleles in Taiwan were determined to be 46.8% (1376G > T), 16.2% (1388G > A), 7.9% (95A > G), 6.5% (493A > G), 5.6% (392G >T), 4.6% (1024C > T), 0.5% (487G > A) and 0.5% (519C > G), respectively.

[Significance and modulation of production of tumor growth inhibitor derived from human embryonic myofibroblasts].

In this report, the production and modulation of tumor growth inhibitor (HFDI) derived from human embryonic myofibroblasts were studied. HFDI was obtained from 3/30 cultures of human embryonic myofibroblasts, which was obviously lower than the incidences of Australia Caucasian's myofibroblast (6/6) and adult's skin fibroblast (4/6). It is suggested that the inheritance play certain role in HFDI production. Calf serum had a significant effect on HFDI production, though without any linear relationship. It seems that HFDI is not derived from calf serum but the factors influencing the production of HFDI are in the calf serum. HFDI could be induced by inactivated BCG from non- or low-producing myofibroblasts. It is indicated that the production of HFDI is related to bacterial contamination (in vitro) or infection (in vivo). The hypothesis is proposed that tumor inhibitors be induced by bacterial infection from the fibroblasts. HFDI-like activity was identified in 4 healthy donors' sera suggesting that HFDI could enter the blood.

Establishment of a group of murine leukemic cell lines and investigation of their biological characteristics.

An in vitro-vivo technique for establishment of cell lines on murine leukemia has been developed. Using this method, suppressive T lymphoblastic leukemia L7811-85, L7212-85, non-T, non-B lymphocytic leukemia L1210-86, B lymphocytic leukemia P 388-86 and Friend erythroleukemia FLCL cell lines have been established. Incidence of leukemia with these cell lines was 100%. Along with the increase of generations of cell lines, cell growth accelerated, generation time shortened and cloning efficiencies rose. A following up electron microscopic observation on L7811-85 and L7212-85 showed that the virus particles were "A" particles in original cells. When they became cell lines in vitro, virus particles increased and transformed into typical "C" particles with budding. An inhibitory activity relevant to leukemic cells on proliferation of leukemic cells has been observed in the supernatant of L7811-85 medium and was regarded as an "autocrine".