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Biomedical subjects

R Hu

Publications and source records attributed to R Hu.

At least 73 records · Page 4Linked to original sources
PubMed1997

MMAC1/PTEN mutations in primary tumor specimens and tumor cell lines.

A candidate tumor suppressor gene, MMAC1/PTEN, located in human chromosome band 10q23, was recently identified based on sequence alterations observed in several glioma, breast, prostate, and kidney tumor specimens or cell lines. To further investigate the mutational profile of this gene in human cancers, we examined a large set of human tumor specimens and cancer cell lines of many types for 10q23 allelic losses and MMAC1 sequence alterations. Loss of heterozygosity (LOH) at the MMAC1 locus was observed in approximately one-half of the samples examined, consistent with the high frequency of 10q allelic loss reported for many cancers. Of 124 tumor specimens exhibiting LOH that have been screened for MMAC1 alterations to date, we have detected variants in 13 (approximately 10%) of these primary tumors; the highest frequency of variants was found in glioblastoma specimens (approximately 23%). Novel alterations identified in this gene include a missense variant in a melanoma sample and a splicing variant and a nonsense mutation in pediatric glioblastomas. Of 76 tumor cell lines prescreened for probable LOH, microsequence alterations of MMAC1 were detected in 12 (approximately 16%) of the lines, including those derived from astrocytoma, leukemia, and melanoma tumors, as well as bladder, breast, lung, prostate, submaxillary gland, and testis carcinomas. In addition, in this set of tumor cell lines, we detected 11 (approximately 14%) homozygous deletions that eliminated coding portions of MMAC1, a class of abnormality not detected by our methods in primary tumors. These data support the occurrence of inactivating MMAC1 alterations in multiple human cancer types. In addition, we report the discovery of a putative pseudogene of MMAC1 localized on chromosome 9.

Brain Neoplasms↗
PubMed1997

Human mitogen-activated protein kinase kinase 4 as a candidate tumor suppressor.

Mitogen-activated protein kinases function in signal transduction pathways that are involved in controlling key cellular processes in many organisms. A mammalian member of this kinase family, MKK4/JNKK1/SEK1, has been reported to link upstream MEKK1 to downstream stress-activated protein kinase/JNK1 and p38 mitogen-activated protein kinase. This mitogen-activated protein kinase pathway has been implicated in the signal transduction of cytokine- and stress-induced apoptosis in a variety of cell types. Here, we report that two human tumor cell lines, derived from pancreatic carcinoma and lung carcinoma, harbor homozygous deletions that eliminate coding portions of the MKK4 locus at 17p, located approximately 10 cM centromeric of p53. In addition, in a set of 88 human cancer cell lines prescreened for loss of heterozygosity, we detected two nonsense and three missense sequence variants of MKK4 in cancer cell lines derived from human pancreatic, breast, colon, and testis cells. In vitro biochemical assays revealed that, when stimulated by MEKK1, four of the five altered MKK4 proteins lacked the ability to phosphorylate stress-activated protein kinase. Thus, the incidence of coding mutations of MKK4 in the set of cell lines is 6 of 213 (approximately 3%). These findings suggest that MKK4 may function as a suppressor of tumorigenesis or metastasis in certain types of cells.

DNA, Neoplasm↗
PubMed1997

Neuronal and glial apoptosis after traumatic spinal cord injury.

Cell death was examined by studying the spinal cords of rats subjected to traumatic insults of mild to moderate severity. Within minutes after mild weight drop impact (a 10 gm weight falling 6.25 mm), neurons in the immediate impact area showed a loss of cytoplasmic Nissl substances. Over the next 7 d, this lesion area expanded and cavitated. Terminal deoxynucleotidyl transferase (TdT)-mediated deoxyuridine triphosphate-biotin nick end labeling (TUNEL)-positive neurons were noted primarily restricted to the gross lesion area 4-24 hr after injury, with a maximum presence at 8 hr after injury. TUNEL-positive glia were present at all stages studied between 4 hr and 14 d, with a maximum presence within the lesion area 24 hr after injury. However 7 d after injury, a second wave of TUNEL-positive glial cells was noted in the white matter peripheral to the lesion and extending at least several millimeters away from the lesion center. The suggestion of apoptosis was supported by electron microscopy, as well as by nuclear staining with Hoechst 33342 dye, and by examination of DNA prepared from the lesion site. Furthermore, repeated intraperitoneal injections of cycloheximide, beginning immediately after a 12.5 mm weight drop insult, produced a substantial reduction in histological evidence of cord damage and in motor dysfunction assessed 4 weeks later. Present data support the hypothesis that apoptosis dependent on active protein synthesis contributes to the neuronal and glial cell death, as well as to the neurological dysfunction, induced by mild-to-moderate severity traumatic insults to the rat spinal cord.

Animals↗
PubMed1997

Characterization of immobilization of an enzyme in a modified Y zeolite matrix and its application to an amperometric glucose biosensor.

A new approach to construct an amperometric biosensor is described. Without using bovine serum albumin-glutaraldehyde, glucose oxidase (GOx) was immobilized on a dealuminized Y zeolite (DAY)-modified platinum electrode to construct a glucose sensor. The large specific surface area of the zeolite substrate resulted in high enzyme loading. The immobilized GOx in this manner was stable and could maintain its high activity for at least 3 months. The interactions between the zeolite and the enzyme were investigated by means of Fourier transform infrared spectra, and the pore distribution and the surface acid property of DAY were preliminarily studied. The results showed that the hydrophilic property and the existing mesopores of DAY played important roles in the enzyme immobilization. This resulting biosensor exhibited good reproducibility and selectivity, owing to the uniform pore structure and unique ion-exchange property of the zeolite. The biosensor responded rapidly to glucose in the linear range from 2.0 x 10(-6) to 3.0 x 10(-3) M, with a detection limit of 0.5 microM.

Biosensing Techniques↗
PubMed1997

Assessment of vertebral body motion during spine surgery.

STUDY DESIGN: In vitro and in vivo assessment of the accuracy of devices proposed for tracking spine motion during surgery; in vivo assessment of vertebral motion during spine surgery. OBJECTIVES: 1) To quantify the accuracy of newly designed vertebral body trackers; 2) to demonstrate the feasibility of tracking vertebral motion in a cadaveric model; and 3) to quantify the vertebral motion that occurs during spinal surgery. SUMMARY OF BACKGROUND DATA: Computer techniques are beginning to be applied to spine surgery. Validation of accuracy of methods of spinal tracking has not been reported. No information exists on the amount of vertebral motion that occurs during surgery. Because the new techniques require accurate positional information for the vertebral body, it is important to understand and evaluate methods of tracking vertebrae. METHODS: An optical tracking system (Northern Digital, Waterloo, Ontario, Canada) was used to track custom-designed trackers. The reliability and accuracy of the trackers were evaluated in vitro. The proposed tracking methodology for human testing was performed using a cadaveric model, and after successful completion, human testing was done in the operating room to evaluate the motion of two vertebral bodies during exposure for instrumentation of the lumbar spine. This technique was used to evaluate the custom designed trackers effectiveness for tracking vertebral bodies for pedicle screw insertion. RESULTS: The trackers developed were accurate and capable of tracking the motion of the spine. Measured motion of L3 and L4 during breathing was 1.3 mm, peak to peak. Maximal intraoperative motion of the vertebral bodies was 12.3 mm during maneuvers simulating dissection of soft tissue and targeting of spinal pedicles. CONCLUSIONS: Significant motion occurs in lumbar vertebral bodies during surgery. Breathing motion alone is up to 1.3 mm, and surgeon-induced motion up to 10 times greater. Vertebral body trackers for use with an optical position sensor were capable of measuring this motion.

Adult↗
PubMed1997

Identification of a candidate tumour suppressor gene, MMAC1, at chromosome 10q23.3 that is mutated in multiple advanced cancers.

Deletions involving regions of chromosome 10 occur in the vast majority (> 90%) of human glioblastoma multiformes. A region at chromosome 10q23-24 was implicated to contain a tumour suppressor gene and the identification of homozygous deletions in four glioma cell lines further refined the location. We have identified a gene, designated MMAC1, that spans these deletions and encodes a widely expressed 5.5-kb mRNA. The predicted MMAC1 protein contains sequence motifs with significant homology to the catalytic domain of protein phosphatases and to the cytoskeletal proteins, tensin and auxilin. MMAC1 coding-region mutations were observed in a number of glioma, prostate, kidney and breast carcinoma cell lines or tumour specimens. Our results identify a strong candidate tumour suppressor gene at chromosome 10q23.3, whose loss of function appears to be associated with the oncogenesis of multiple human cancers.

Amino Acid Sequence↗
PubMed1997

The role of MMAC1 mutations in early-onset breast cancer: causative in association with Cowden syndrome and excluded in BRCA1-negative cases.

Cowden syndrome (CS) is an autosomal dominant disorder associated with the development of hamartomas and benign tumors in a variety of tissues, including the skin, thyroid, breast, endometrium, and brain. It has been suggested that women with CS are at increased risk for breast cancer. A locus for CS was recently defined on chromosome 10 in 12 families, resulting in the identification of the CS critical interval, between the markers D10S215 and D10S541. More recently, affected individuals in four families with CS have been shown to have germ-line mutations in a gene known as "PTEN," or "MMAC1," which is located in the CS critical interval on chromosome 10. In this study, we report three novel MMAC1 mutations in CS and demonstrate that MMAC1 mutations are associated with CS and breast cancer. Furthermore, we also show that certain families and individuals with CS do not have mutations in the coding sequence of MMAC1. Finally, we did not detect MMAC1 mutations in a subpopulation of individuals with early-onset breast cancer, suggesting that germ-line mutations in this gene do not appear to be common in this group.

Breast Neoplasms↗
PubMed1997

Cytokine pattern in relation to disease progression in human immunodeficiency virus-infected children.

Cytokine mRNA expression and stimulus-induced cytokines were examined in peripheral blood mononuclear cells in 62 human immunodeficiency virus (HIV)-infected children and uninfected controls. Compared with that in controls, constitutive mRNA expression in patients was increased for tumor necrosis factor (TNF)-alpha, interferon (IFN)-gamma, and interleukin (IL)-10 and decreased for IL-12; it was undetectable for IL-2 and IL-4 in both patients and controls. Stimulus-induced secretion of TNF-alpha, IFN-gamma, IL-12, and IL-4 was less than that in controls; IL-10 secretion was similar. There was no increase in stimulus-induced or constitutive IL-4 or IL-10 in children with severe immunologic deficit compared with controls. A higher stimulus-induced IL-10 secretion and a lower constitutive TNF-alpha mRNA were associated with a slower rate of disease progression, and TNF-alpha mRNA expression correlated with lower plasma HIV RNA. Thus, constitutive cytokine mRNA expression differs from stimulus-induced cytokine responses. The dominant defect in HIV-infected children appears to be one of reduced type 1 cytokines, predominantly IL-2.

Acquired Immunodeficiency Syndrome↗
PubMed1997

Effects of Babesia microti infection on feeding pattern, engorged body weight, and molting rate of immature Ixodes scapularis (Acari: Ixodidae).

Effects of Babesia microti Franca on the pattern of feeding time, the body weight of engorged ticks, and the molting rate of the blacklegged tick, Ixodes scapularis Say, were determined. Using the Syrian golden hamster, Mesocricetus auratus (Waterhouse), as an animal model, we found no significant differences in patterns of feeding time determined for both larvae and nymphs that fed on B. microti-infected and uninfected hamsters. However, an infection of B. microti in hamsters delayed engorgement of the ticks. The presence of B. microti had no adverse effects on the body weight of engorged ticks. The mean body weight of groups of 5 engorged larvae fed on infected hamsters was not significantly different from that on uninfected hamsters. The mean body weight of individual nymphs that fed on infected hosts was significantly higher than that on uninfected hosts. Larvae fed on infected hamsters molted in greater numbers than those fed on uninfected hamsters. However, there was no significant difference in molting rates of nymphs derived from infected and uninfected hamsters. In addition, greater body weights and higher molting rates were observed in both larvae and nymphs that fed on uninfected white-footed mice, Peromyscus leucopus Rafinesque, compared with those on uninfected hamsters. Nymphs fed on mice spent a significantly longer time achieving repletion than those on uninfected hamsters. We suggest a mutualistic relationship in the interactions between B. microti and I. scapularis. This mutualism may potentially enhance the long-term coexistence and survival of both species. Furthermore, we suggest that the pattern of feeding time, the body weight of engorged ticks, and the molting rate of immature I. scapularis are host-dependent.

Animals↗
PubMed1997

Neurologic evaluation of infant and adult rats before and after sciatic nerve blockade.

BACKGROUND: Only limited data exist comparing differences in sensory function and responses to neural blockade in infant and adult rats. Therefore, the authors sought (1) to compare baseline thermal, proprioceptive, and postural responses in infant, adolescent, and adult rats; and (2) to compare the effects of sciatic nerve blockade on thermal, proprioceptive, and postural responses in infant, adolescent, and adult rats. METHODS: Infant, adolescent, and adult rats were evaluated for proprioceptive, thermal, and mechanical nociceptive and motor function before and after sciatic blockade using a detailed neurologic examination. RESULTS: Mechanical and thermal nociception were present in all rats, starting from age 1 day. The withdrawal reflex latency to pinch was rapid at all ages, whereas that reaction to thermal stimulus depended on both age and temperature. In contrast, the tactile placing response and hopping response were absent at birth and developed completely during the first 10 days of life. The extensor postural thrust was absent in the first 2 weeks of life and developed variably during the first 50 days of life. Sciatic blockade duration is shorter in infant rats than in adult rats receiving the same dose per kilogram. A brief halothane general anesthetic at the time of sciatic injection in infant or adult rats does not alter the duration of blockade. CONCLUSIONS: Infant rats show increased sensitivity to noxious thermal stimuli and similar response to deep mechanical stimuli compared with adult rats. Their proprioceptive and motor responses develop during the first 2 weeks of life. When doses are scaled by body weight, block duration is shorter in infant than in adult rats.

Age Factors↗
PubMed1997

Clonal origin of multiple lung cancers: K-ras and p53 mutations determined by nonradioisotopic single-strand conformation polymorphism analysis.

Disease stage is the most important factor in determining prognosis and treatment of lung cancer. Staging of lung cancer is complicated by presentation of multiple pulmonary malignant lesions with a similar histology. It is a dilemma to decide if these lesions are synchronous primaries arising from different malignant clones or metastases from a single clone. Lung cancer is associated with multiple genetic abnormalities including mutations of K-ras and p53, which are believed to occur prior to onset of metastasis. To determine the clonal origin of multiple pulmonary malginant nodules, we analyzed point-mutations of K-ras and p53 by microdissection, polymerase chain reactions (PCR), nonradioisotopic single-strand conformation polymorphism (SSCP) analysis, and DNA sequencing. Each pulmonary lesion was microdissected from paraffin slides. Genomic DNA was amplified by two sequential PCRs followed by electrophoresis in a minigel and silver staining. Deoxyribonucleic acid sequencing was performed if necessary to confirm a mutation found upon SSCP analysis. Applying this molecular approach, we were able to differentiate the clonal origins of multiple malignant nodules of the lung as exemplified by the two cases presented.

Adenocarcinoma↗
PubMed1997

Cell death suggestive of apoptosis after spinal cord ischemia in rabbits.

BACKGROUND AND PURPOSE: After spinal cord ischemia, some neurons remain viable after an ischemic insult but may be at risk of dying during reperfusion. We searched for morphological and biochemical features of apoptosis, which is a mechanism of delayed neuronal death, in a rabbit model of spinal cord ischemia. METHODS: The infrarenal aorta of White New Zealand rabbits (n = 24) was occluded for 40 minutes using a loop tourniquet. Rabbits were killed after 12, 24, or 48 hours (n = 8 per group). The loop was placed but never tightened in sham-operated rabbits (n = 6). The lumbar segment of the spinal cord (L5 to L7) was used for morphological studies, including hematoxylin and eosin staining and a modified terminal deoxynucleotidyltransferase-mediated dUTP-biotin nick end-labeling (TUNEL) staining method. Electron microscopy was used to examine ultrastructural morphology. In addition, lumbar tissue was used for biochemical investigation of DNA laddering by agarose gel electrophoresis. RESULTS: After ischemia, the affected areas contained neurons with positive TUNEL staining. Positive neurons were located in laminae III to IX, although most were concentrated in the intermediate and ventral areas. Adjacent sections stained with hematoxylin and eosin exhibited ischemic cell changes (red and ghost neurons), while apoptotic bodies were also apparent. In addition, electron microscopy of ischemic tissue samples exhibited ultrastructural characteristics of apoptosis, including nuclear condensation and relatively normal organelle morphology. Finally, isolated DNA revealed a ladder on agarose gel electrophoresis, indicating DNA fragmentation into approximately 180 multiples of base pairs. CONCLUSIONS: Spinal cord ischemia in rabbits induces morphological and biochemical changes suggestive of apoptosis. These data raise the possibility that apoptosis contributes to neuronal cell death after spinal cord ischemia.

Animals↗
PubMed1997

[Relationship of C-erbB-2 oncogene overexpression to estrogen progesterone receptors in brease cancer and its prognostic significance].

In order to understand the relationship of C-erbB-2 oncogene overexpression to ER and PR in breast cancer and its prognostic significance, we examined overexpression of C-erbB-2 oncogene in 106 breast carcinomas by using immunohistochemical techniques (LSAB). The results showed that the positive rate of C-erbB-2 overexpression was 63.21% (67/106). The overexpression of C-erbB-2 oncogene related negatively with survival. 81.63% of the cases with overexpression of C-erbB-2 oncogene survived < or = 5 years and 34.29% survived > 10 years. There were significant associations of C-erbB-2 overexpression with advance clinical stage, high histological grade, and positive axillary node status in breast cancers. Negative relationship between hormone receptors and C-erbB-2 oncogene. All of these findings suggested that overexpression of C-erbB-2 oncogene might be an important prognostic factor and the detection of C-erbB-2 oncogene might be arranged as a regular pathological examination in the cases of breast cancer.

Adult↗
PubMed1997

[New method for preparing oridonin by column chromatography].

Compared with the traditional methods, the separation and preparation of oridonin by column chromatography have the advantages of fast speed, harmlessness and non-pollution of solvent, safety, economy and easily obtained, etc. After re-crystallizing the oridonin content is determined by TLC scanning to be as high as 97.03%.

Antineoplastic Agents, Phytogenic↗
PubMed1997

Spatial distribution of larval Ixodes scapularis (Acari:Ixodidae) on Peromyscus leucopus and Microtus pennsylvanicus at two island sites.

Larval blacklegged ticks, Ixodes scapularis, were collected from white-footed mice. Peromyscus leucopus, on Prudence Island (where Microtus pennsylvanicus were not captured) and from meadow voles. M. pennsylvanicus, on Patience Island (where P. leucopus was absent) in Narragansett Bay, Rhode Island from June to October 1992. Ixodes scapularis larvae were also collected by flagging in the vicinity of host captures. On both islands, the relative density of larvae changed from July to September in samples from hosts, but not in flagging samples. Consequently, different sampling techniques can give different assessments of tick populations. Larvae were highly aggregated on both of the host species throughout the sampling period. As the mean relative density of larvae increased in the environment (based on flagging samples), larvae on the hosts became more dense and more crowded. Increased densities of larvae in the environment were not correlated with increased patchiness in the distribution of larvae among host animals on either island. Changes in the spatial distribution of larval I. scapularis on each host species had similar trends as larval densities and distributions within the environment. These results suggest that M. pennsylvanicus can serve as an alternative host for immature I. scapularis in a P. leucopus-free environment and have similar distributional characteristics.

Analysis of Variance↗
PubMed1996

Biomechanical analysis of hemipelvic deformation after corticospongious bone graft harvest from the posterior iliac crest.

STUDY DESIGN: The stiffness of the hemipelvis during simulated physiologic loading and the bone deformation in the remaining posterior ilium after the harvest of increasing sizes of corticospongious bone graft were compared with values in intact bone. OBJECTIVES: To quantify the biomechanical effects of the removal of bone grafts from the posterior ilium and to relate the size of graft removed to the stiffness of the hemipelvis and deformation of the remaining bone. SUMMARY OF BACKGROUND DATA: Bone fractures and pelvic instability have been reported to complicate graft harvest from the posterior iliac crest. There is no quantitative data relating graft size to the mechanical properties of the remaining ilium. METHODS: Seven cadaveric hemipelves were loaded with a materials testing machine through the superior sacrum while supported at the acetabulum and stabilized with a cable fixed to the ilium. Force and displacement histories and deformation in the greater sciatic notch were recorded for the intact bone and after removal of corticospongious bone graft in 1.5-cm increments from the posterior iliac crest. RESULTS: If the length of the removed corticospongious bone graft exceeded 3.0 cm, the stiffness of the posterior pelvic ring decreased, and deformation in the remaining bone increased substantially. CONCLUSIONS: Removal or bone graft in excess of 3 cm from the posterior ilium increases the risk of iatrogenic fatigue fracture.

Bone Transplantation↗
PubMed1996

Additive neuroprotective effects of dextrorphan and cycloheximide in rats subjected to transient focal cerebral ischemia.

Previous studies have implicated both excitotoxicity and apoptosis in the pathogenesis of cerebral infarction induced by focal ischemic insults. Here we tested the possibility that the NMDA antagonist, dextrorphan, and the protein synthesis inhibitor, cycloheximide, would produce additive protective effects in a rodent model of focal ischemia-reperfusion. Transient focal cerebral ischemia was induced by a 90 min period of ligation of the right middle cerebral artery and both common carotid arteries. Administration of either 30 mg/kg dextrorphan or 0.5 mg/kg cycloheximide, given i.p. 15 min before ischemia, reduced infarct volume by about 65%. When optimal concentrations of each drug were given together, infarct volume was reduced by 87% as measured 14 days later. These observations support the idea that both excitotoxicity, and apoptosis dependent on new protein synthesis, contribute to cerebral infarction after transient focal ischemia in the rat.

Animals↗