Use of defective herpes-derived plasmid vectors.
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Biomedical subjects
Publications and source records attributed to L Yu.
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Nephrotoxicity is the major adverse effect of conventional amphotericin B (AMB/D), often limiting administration of full dosage. The new liposomal amphotericin B seems to be less toxic. The new liposomal amphotericin B seems to be less toxic. In this study, it is proposed that solubilizing the standard AMB/D preparation with 10% lipid emulsion will attenuate nephrotoxicity. Rats were injected with either AMB/D (Fungizone), AMB, AMB/D plus lipid emulsion (AMB/D/LE), or sodium deoxycholate (D). Renal function studies were performed on day 5. To assess a direct tubular toxic effect, isolated rat proximal tubule suspensions and inner medullary collecting duct cells in culture were exposed to AMB/D, AMB, AMB/D/LE, liposomal amphotericin B, and D for 60 min in normoxia. Lactate dehydrogenase (LDH) release was assessed as an index of cell injury. Creatinine clearance (ml/min per 100 g) averaged 0.79 +/- 0.04 in control rats, 0.29 +/- 0.09 in AMB rats (P < 0.001 versus control), 0.38 +/- 0.04 in AMB/D rats, 0.46 +/- 0.05 in D rats, and 0.78 +/- 0.03 in AMB/LE rats. Renal blood flow (ml/min per 100 g) was 3.45 +/- 0.31 in control, 1.29 +/- 0.28 in AMB, 1.42 +/- 0.23 in AMB/D, 3.03 +/- 0.39 in D, and 2.71 +/- 0.21 in AMB/D/LE rats. The fractional excretion of potassium (%) was 27.3 +/- 1.18 in control rats, 61.6 +/- 7.00 in AMB/D rats, 58.4 +/- 15.32 in AMB rats, and 37.9 +/- 2.06 in AMB/D/LE rats. LDH release (%) in proximal tubules incubated with AMB/D and D was 43.6 +/- 3.39 and 58.6 +/- 4.20, respectively. Addition of lipid emulsion decreased LDH release: 21.6 +/- 1.22 for AMB/D/LE and 26.4 +/- 3.03 for deoxycholate plus lipid emulsion. AMB did not demonstrate any toxic effect in proximal tubule suspensions. D was not toxic to inner medullary collecting duct cells at 0.16 mg/ml, whereas D at a higher dose and AMB induced a significant LDH release. Addition of lipid emulsion did not affect the antifungal activity as assessed by the Etest method. In conclusion, an alternative way of administering standard AMB with reduced nephrotoxicity is proposed.
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OBJECTIVE: To determine whether autoimmunity is a prominent feature of NIDDM among diabetic members in families with a strong history of NIDDM or in families with a mixture of NIDDM and IDDM. RESEARCH DESIGN AND METHODS: We determined GAD and islet cell (ICA512) autoantibodies from 215 NIDDM individuals and from 14 individuals with impaired glucose tolerance (IGT) of 68 families, including 1 family with maturity-onset diabetes of the young (MODY) and 3 families ascertained specifically for a mixture of NIDDM and IDDM. We tested 2 control populations: 50 unrelated spouses form Utah families, including 29 spouses with either IGT or NIDDM and 198 random nondiabetic white individuals from Colorado. RESULTS: We detected either GAD or ICA512 autoantibodies in 11 members of seven families and in one spouse used as a control subject. In two families, two affected individuals showed evidence of autoimmunity, but NIDDM individuals in each of the seven families showed no evidence of autoimmunity. Among the five families with both IDDM and NIDDM individuals (three families ascertained for a mixture and two families ascertained with an incidental IDDM child), antibodies were detected in members of only one family. Antibody-positive individuals were significantly younger at diabetes onset and had low waist-to-hip ratios, but were not more likely to be insulin treated. CONCLUSIONS: Autoimmunity is an important cause of apparent NIDDM, even among families with a strong history of NIDDM. However, autoimmunity among affected family members appeared to be a chance event and not the manifestation of a different genetic cause of diabetes.
Nitric oxide synthase has been identified in several epithelial cells in the kidney, including proximal tubular cells, thick ascending limb, inner medullary collecting duct, and interstitial cells. Nitric oxide (NO) plays an important role in renal hemodynamics and sodium tubular transport. We have demonstrated that NO participates in hypoxia/reoxygenation (H/R) injury in isolated rat proximal tubules (PT) suspensions. In this in vitro model L-arginine addition enhanced H/R injury while L-NAME almost completely prevented injury. These effects were less intense in chronic supplemented rats with L-arginine and L-NAME, suggesting that NO synthase manipulation had interfered with PT susceptibility to H/R injury. In contrast, L-arginine protected IMCD cells in culture from hypercholesterolemic rats against hypoxia. Moreover, acute infusion of L-arginine before bilateral renal artery clamping was protective while L-arginine chronic administration and L-NAME were deleterious in this ARF model. The L-arginine protection was not observed in unilateral renal clamping plus contralateral nephrectomy in normal rats, but L-arginine was protective in hypercholesterolemic rats. Taken together, these results suggest that the net effect of NO stimulation is variable, and that it is the result of a balance between beneficial hemodynamic effects and cytotoxicity.
Dextran 40 is largely used in clinical medicine as a plasma substitute because of its beneficial effects on the microcirculation and antithrombogenic properties. An unusual adverse reaction of dextran administration is oligoanuric acute renal failure. We report two cases of anuric ARF induced by dextran 40. Diuresis and renal function were quickly resumed after plasma-pheresis treatment. Renal biopsy revealed normal kidneys except for swelling and vacuolation of renal tubules suggestive of osmotic nephrosis.
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A human YAC genomic pool with more than 4,000 clones was constructed by means of "terminal modification" of YAC vector and genomic inserts, as well as optimization of relevant experimental parameters. The transformation rate is approximately 400-500 transformants per microgram DNA. Preliminary characterization of randomly sampled clones demonstrates that YAC-specific inserts, in the average size of 450 kb and ranging 150-750 kb, were detected from 80% of the clones. The vector ligation experiment indicates the efficient reduction of "coligation" by terminal modification.
Using biological activity assay, the production of IL-4 and IL-10 by peripheral blood mononuclear cells (PBMC) was studied in patients with nasopharyngeal carcinoma and lymphoma. The IL-2, IL-4 and IFN gamma levels in both cancer patients were lower than that in normal subjects. In contrast, the IL-10 level was higher in both cancer patients. When rIL-2 was added into the cytokine production system, IL-2, IL-4, and IFN gamma level were upregulated in lymphoma patients. However, in the patients of nasopharyneal carcinoma, only IL-2 level was upregulated. IL-10 activity could not be affected by exogenous IL-2 in both cancer patients. The results of negative IL-4 activity and higher positive percentage (84.6%) of IL-10 activity in ascites from patients with ovarian carcinoma were corresponded to the patients with nasopharyngeal carcinoma and lymphoma. By using in situ hybridization technique, the expression of IL-4 and IL-10 was studied in tumor tissues from patients with gastric, esophageal and breast carcinomas. There was no differences of the percentage of IL-4 and IL-10 expression among three kinds of tumor tissues, the range of positive percentage was 40%-70%. These results indicate that the correction of abnormal upregulation of IL-10 activity should be considered for cancer immunotherapy.
The significance of postoperative chemotherapy was studied in 439 cases of unilateral primary breast cancer which were proved histologically with no axillary lymph node metastasis. The survival rate was analyzed by life table method. The prognosis of node negative breast cancer patients was mainly associated with tumor size. 10 year survival rate of patients with tumor less than 3 cm treated operativel and by operation combined with chemotherapy was 92.60% and 94.13% respectively. If tumor size was larger than 3 cm, the ten year survival rate of the patients treated operatived was 79.89%, and that of the patients with combined theropy 96.02%. The prognosis was statistically different between the two groups. As to age, status of menopause, pathologral type, kind of surgery and status of ER, the prognosis of patients treated by combined therapy was better than that of patients treated by operation alone. The postoperative chemotherapy was not significantly beneficial to breast cancer patients with tumor less than 3 cm.
To evaluate the level of hormones before and after female renal transplantation, we measured pituitary gonadal hormones, estrovite, progestin, and prolactin in 25 renal transplant recipients (RTR) and 25 cases of chronic renal failure (CRF) using engyme immunoassay (EIA). The results indicated that serum RRL, FSH and LH level were reduced in RTR females compared with CRF, whereas E2 and P were normal. Serum PRL levels were elevated in CRF females whereas P levels were significantly lower compared with those of other groups. After clomiphene stimulation test, the plasma levels of LH FSH and E2 elevated, suggesting hypothalamic anovulation. Following successful renal transplantation, uremic hypothalamic disfunction was ameliorated and normal menstrual cycle, fertility was restored. During dialysis, treatment was given using suit therapy rather than trigger the ovulatory. Renal transplantation is the best treatment.
OBJECTIVE: For comparing the photodynamic effect of 49 single laser photosensitizers on two human cancer cell lines. METHODS: After irradiation with a pulsed copper vapour laser, the SW1116 cell line of human colorectal cancer and K562 cell line of erythroleukemia were measured with MTT assay. IC50 and the cell survival rate were observed to evaluate the photodynamic effect of various 49 single photosensitizers. RESULTS: In 49 photosentizers, the structure formula of the leading 9 single photosensitizers belonged to homatoporphyrin group, the average IC50 of which was < or = 1.0 microgram/ml on both cell lines of. The photosensitizers currently used in the clinic, hematoporphyrin-3 (PSD-007), ranked 17th, with IC50 on both cell lines of 1.25 micrograms/ml and 3.75 micrograms/ml, respecively. CONCLUSION: The leading 16 single photosensitizer in ths series studied are better purified and more effective in photodynamic killing of human cancer cell lines in vitro. They can be used in lieu of PSD-007 for cancer treatment.
OBJECTIVE: To evaluate whether the growth inhibition by retinoic acid and RAR alpha mRNA expression levels were affected by the change of ER expression. METHODS: The ER-negative breast cancer cell line MDA-MB-231 was transfected with the ER gene by stable transfection. RESULTS: In ER-transfected cells not only was the RAR alpha mRNA expression increased, but their growth was inhibited by retinoic acid as well. Estrogen could greatly stimulate the RAR alpha gene expression not only in established ER-positive cell lines but also in ER-transfected MDA-MB-231 cells. CONCLUSION: Our data strongly suggest that ER-mediated enhancement of RAR alpha levels play an important role in RA inhibition of human breast cancer cell growth.
The strategy of isolating the band-specific expression fragments from the probe pool of human chromosome generated by microdissection was reported in present paper. A chromosome 14 q 24.3 band-specific single copy DNA library was constructed based on this probe pool. Using this pool DNA as probe to hybridize the human bone marrow cell cDNA library, 68 primary positive clones were selected from 5 x 10(5) cDNA clones. Of them 32 clones were got in second-round screening and designed as cFD 14-1-32. Finally, 24 bandspecific expression fragments were identified from these 32 positive clones by analysing the results of DNA hybridization. Those band-specific clones can hybridize to both 14 q 24.3 DNA and human genomic DNA, but have no hybridization signal with 17 q 11-12 DNA. Partial sequences of 13 fragments of them were sequenced and were identified as novel cDNA sequences as well as have some homology with known genes in NCBI database. Analysis of expression spectrum of cFD 14-1 suggested that the cDNA fragments thus obtained can be used to isolate the genes not yet be cloned in 14 q 24.3 region.
OBJECTIVES: Some human hepatocellular carcinomas (HCCs) produce thyroxine-binding globulin (TBG). High serum TBG levels in such patients may be associated with increased thyroxine (T4) levels. This study aimed to elucidate the serum TBG and thyroid hormone profile in Japanese patients with HCC, to compare the difference between TBG-producing and -nonproducing HCCs, and to investigate the changes in serum TBG level and the thyroid hormone profile after removal of the tumor. METHODS: The 40 subjects included 20 patients with HCC, 10 healthy controls, and 10 operative controls. Serum TBG, 3,5,3'-triiodothyronine (T3), T4, and free T4 were measured serially for 4 wk after resection of HCC in 16 patients and after control operations in 10 patients. Assay methods were a radioimmunoassay for TBG and enzyme immunoassays for T3, T4, and free T4. RESULTS: Values higher than the mean +/- 2 SD of controls were considered abnormally high. Of patients with HCC, 60% had abnormally high TBG values, 65% had abnormally high T3 values, 39% had abnormally high T4 values, and 6% had abnormally high free T4 values. The mean levels of TBG and T3 were significantly higher than those in healthy controls, but no difference was found for T4 and free T4 levels. There were no significant differences in various clinicopathological factors between patients with high TBG levels and those with normal TBG levels. After resection of HCC, serum TBG decreased significantly in patients with high TBG levels but not in those with normal TBG levels. CONCLUSIONS: This study shows that >50% of HCCs in Japanese patients produce TBG; removal of the tumor reduces serum TBG in such cases.
Tumor necrosis factor alpha (TNF-alpha) is known to induce wasting in humans and animals. This study was undertaken to determine TNF-alpha concentrations in children with sickle cell disease (SCD) and whether high TNF-alpha levels are more likely to be present in children with growth deficits, infection, or pain crisis. Tumor necrosis factor alpha was measured using enzyme immunoassay in 143 blood samples obtained from 101 children. Mean TNF-alpha levels were higher in patients (50 pg/mL) than in 21 control children (19 pg/mL) and in 26 laboratory employees (20 pg/mL). During the follow-up period, 35%, 38%, and 28% of children with SCD had infection, pain crisis, or a blood transfusion, respectively. Mean TNF-alpha concentrations were higher in children who had an infection than in those who did not. No significant effect of pain crisis or blood transfusion was observed. Tumor necrosis factor alpha concentrations were above normal (> 40 pg/mL) in 15% of controls, 34% of children with SCD, and 52% of children with SCD who had an infection and 33% of those who did not. A higher percentage of children who had elevated TNF-alpha levels had weight (46% versus 31%) or height (50% versus 28.6%) deficits than children who had normal TNF-alpha levels. These results indicate that most children with SCD in stable condition have normal TNF-alpha concentrations and that those with high TNF-alpha levels are more likely to have growth deficits.
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