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Biomedical subjects

Xiao-Yang Wang

Publications and source records attributed to Xiao-Yang Wang.

9 recordsLinked to original sources

[Expression of p53 in neonatal mice following hypoxia-ischemia and effects of its inhibitor on neonatal brain injury].

OBJECTIVE: p53-induced apoptosis is crucial in the development of hypoxic-ischemia (HI) brain damage and neurodegenerative disorders. Some experimental research has shown that a synthetic inhibitor of p53 can protect neurons against apoptosis. This study aimed to explore the expression of p53 in neonatal mice following HI brain damage and the effect of p53 inhibitor (pifithrin-alpha, PFT-alpha) on brain damage. METHODS: HI was induced in 9-day-old mice pups by ligation of left carotid artery and 10% oxygen exposure for 55 minutes. The pups were sacrificed and the brains were taken out at 3, 8, 24, and 72 hrs post-HI. The brains were sectioned and stained with antibody against p53 and microtubule-associated protein 2 (MAP-2). PFT-alpha was injected intraperitoneally: in experiment 1, immediately after HI with different dosages (1, 2 and 8 mg/kg); in experiment 2, 2 mg/kg at different HI times (1 hr before HI, and immediately and 1 hr after HI). Control animals without HI received injections of 0.5% dimethyl sulfoxide. Brain damage was evaluated by gross morphology scoring at 72 hrs after HI. RESULTS: The number of p53 positive cells in the cortex, hippocampus and striatum of the ipsilateral hemisphere increased significantly and peaked at 3-8 hrs post-HI when compared with those of contralateral hemisphere as well as normal controls. The positive cells distributed mainly in the MAP-2 negative area. Both different dosages and different injection time PFT-alpha treatment did not reduce the extent of brain damage. CONCLUSIONS: The immunoactivity of p53 increased significantly as early as 3 hrs post-HI. The distribution area of p53 expression was consistent with that of brain damage. The p53 inhibitor PFT-alpha has no protective effects against HI brain damage in neonatal mice.

Animals↗

[Neuroprotective effect of hypothermia on hypoxic-ischemic brain injury in mice].

OBJECTIVE: The study was to investigate the effect of different temperatures during hypoxia on brain injury in mice of different ages. METHODS: Newborn C57/BL6 mice at 7 days or 21 days of life were subjected to left carotid artery ligation followed by exposure with 10% oxygen. The mice were kept in a incubator with a predetermined, constant temperature, either 34 degrees centigrade (Hypothermia group) or 36 degrees centigrade (Normothermia group). Brain injury was evaluated 7 days after hypoxia-ischemia (HI). Active caspase-3 and apoptosis-inducing factor (AIF) expressions in the brain tissue were detected by immunohistochemistry and Western Blot was used to evaluate the phosphor-Akt (P-Akt) expression in the brain tissue at 24 hrs post-HI. RESULTS: Brain injuries, including the cortex, hippocampus, striatum and thalamus injuries, occurred in the Normothermia group at 7 days post-HI. The brain cortex showed cystic cavitation in the postnatal day (P)7 pups mice and laminar infarct of the brain cortex was observed in P21 mice. In the Hypothermia group, the P7 mice did not present with laminar infarct of the cortex and had lower scores of neuropathological lesions in cortex, hippocampus, striatum and thalamus than P7 mice from the Normothermia group (P < 0.01); the cortex injuries were significantly relieved but the injuries of hippocampus, striatum and thalamus in P21 mice were similar to those from the Normothermia group. Active caspase-3 (7.0 +/- 5.6) and AIF positive cells (3.7 +/- 6.2) in the cortex of P7 mice from the Hypothermia group were significantly lower than those of the Normothermia group (51.5 +/- 23.2 and 31.8 +/- 22.4) at 24 hrs post-HI (P < 0.01). Wetstern Blot showed the P-Akt expression was obviously decreased in the ipsilateral hemisphere to the occlusion compared with that of the contralateral hemisphere after HI in the Normothermia group (P < 0.05), while in the Hypothermia group the P-Akt expression was not significantly different between the two hemispheres. CONCLUSIONS: Hypothermia has protective effects against HI insults. The protection was more pronounced for the immature brain than the mature brain.

Active Transport, Cell Nucleus↗

[Antineoplastic effect of valproic acid and trichostatin on HL-60 and K562 cells].

The objective of this study was to investigate antineoplastic effects of valproic acid (VPA) and trichostatin (TSA) on HL-60 and K562 cells in vitro, and the synergic effects of VPA or TSA in combination with ATRA. The inhibitory effects of VPA, TSA and ATRA in various concentrations and different combinations on proliferation of HL-60 and K562 cells were observed by cell growth curves, 50% inhibitory concentration (IC(50)), as well as inhibition of leukemia colony growth at different time points. The characteristics of cell differentiation or apoptosis were analyzed by cytochemical staining, differentiation antigen detection, cell cycle assay and A(NBT)/A(MMT) value determination. The results showed that HL-60 cell had a lower IC(50) of VPA and TSA compared with K562 cells. ATRA could significantly enhance the inhibition of VPA, TSA on clonegenicity of HL-60 cells and inhibition of VPA on clonegenicity of K562 cells. HL-60 cells treated with VPA displayed the phenotype of neutrophilic like cells, and showed the increases of NBT reduction rate and CD11b expression. No evidence for K562 differentiation was found. It is concluded that both VPA and TSA inhibit HL-60 cells growth in vitro. VPA induces differentiation of HL-60 cells to granulocyte. VPA and TSA have a moderate anti-proliferative effect on K562 cells. None of these agents induces K562 cell differentiation.

Antineoplastic Agents↗

Fibroblastic, hematopoietic, and hormone responsive epithelial cell lines and culture conditions for elucidation of signal transduction and drug resistance pathways by gene transfer.

Elucidation of signal transduction pathways involved in proliferation, cell cycle progression and the regulation of apoptosis has shown great promise in the treatment of various diseases including neoplastic, inflammatory, autoimmune, immunodeficiency, arthritic and neurodegenerative disorders. By understanding how these signal transduction pathways function, chemotherapeutic targets may be identified which will suppress or eliminate the disease. This information may eventually be translated into therapy, which would either eliminate or safely contain the patient's disease. This chapter will focus on basic tissue culture techniques which are used to elucidate signal transduction pathways. Furthermore, this chapter will provide a general background for understanding how gene transfer techniques can be used to elucidate signal transduction pathways as well as various pitfalls commonly encountered with their usage.

Animals↗

Elucidation of signal transduction pathways by transfection of cells with modified oncogenes.

This chapter will focus on introduction of various wild type (WT) and mutant genes into cells by DNA transfection. Techniques for analysis of the inheritance, expression, and biological effects of the introduced genes will be described. Various strong and weak points about three different techniques of stable gene transfer, including calcium-phosphate DNA precipitation, transfection via liposomes, and transfection via electroporation, will be discussed.

Breast↗

Elucidation of signal transduction pathways by retroviral infection of cells with modified oncogenes.

This chapter will focus on understanding how various wild type (WT), dominant negative (DN), constitutively active (CA), and conditionally active (COND) oncogenes, as well as antisense (AS) genes contained in retroviral vectors may be used to elucidate signal transduction pathways. We will describe methods to introduce these genes into cells and subsequent analysis of inheritance, expression, and biological effects of the genes introduced. Furthermore, we will discuss various strong points about each of these different types of constructs, how they can be used to elucidate signal transduction, apoptotic, and drug resistance pathways as well as various pitfalls commonly encountered with their usage.

Cell Line↗

Mucosa-associated lymphoid tissue lymphomas with t(11;18)(q21;q21) and mucosa-associated lymphoid tissue lymphomas with aneuploidy develop along different pathogenetic pathways.

t(11;18)(q21;q21) and aneuploidy are recurrent chromosomal aberrations in mucosa-associated lymphoid tissue (MALT) lymphomas. To investigate their relationship and clinical significance, we developed a two-color fluorescence in situ hybridization (FISH) technique to detect t(11;18) and aneuploidy in nuclei isolated from paraffin-embedded tissue. Thirty-seven MALT lymphomas (all previously evaluated for t(11;18) by reverse transcriptase-polymerase chain reaction), 1 large cell lymphoma (LCL) arising subsequent to MALT lymphoma, and 16 controls were tested by FISH using the t(11;18) probe set and multiple centromeric probes. t(11;18)(q21;q21) was present by FISH in 11 of 12 polymerase chain reaction-positive MALT lymphomas (92%). The LCL and its clonally identical antecedent MALT lymphoma both showed t(11;18). The LCL had trisomy 12, and a small subset of MALT lymphoma cells had trisomy 3 and/or 12. Only one other MALT lymphoma with t(11;18) showed aneuploidy (trisomy 3) in a small clone, whereas 15 of 25 t(11;18)-negative MALT lymphomas (60%) showed trisomy of chromosomes 18 (n = 12), 3 (n = 8), 7 (n = 2), and/or 11 (n = 1). t(11;18) and aneuploidy are primarily mutually exclusive events, suggesting different pathogenetic pathways in the development of MALT lymphomas. Both t(11;18) and aneuploidy were seen disproportionately in lung, and both were associated with recurrent disease.

Adult↗

Bioluminescent method for detecting telomerase activity.

BACKGROUND: Telomerase is a promising biomarker in cancer diagnosis and therapy. The elongation of telomeric repeats catalyzed by telomerase is accompanied by release of six PP(i) for each TTAGGG repeat (1 pmol PP(i)/310 pg telomeric repeats). We developed a novel method to measure telomerase activity by use of an enzymatic luminometric PP(i) assay (ELIPA). METHODS: Extracts of cell lines and tissues were incubated with primer at 30 degrees C for 30 min. Released PP(i) was converted to ATP by sulfurylase, and ATP was detected by a luciferase bioluminescence system. The ELIPA results were compared with results obtained with the conventional telomeric repeat amplification (TRAP)-ELISA in 42 lung carcinoma tissues and 27 control tissues without malignancy. RESULTS: The lower detection limits of ELIPA and TRAP-ELISA were 5 and 10 cells, respectively. The within-run imprecision (CV) of ELIPA was < or =12%. When compared with TRAP-ELISA, the correlation coefficient (r) was 0.79. When we used the cutoff value from ROC analysis to distinguish malignant and nonmalignant tissues, the sensitivity and specificity of ELIPA were 83% and 96%, respectively, whereas the sensitivity and specificity of TRAP-ELISA were 71% and 96%, respectively. CONCLUSION: ELIPA is a simple and sensitive homogeneous method to quantify telomerase activity.

Adenosine Triphosphate↗

[Laparoscopic radical hysterectomy and pelvic lymphadenectomy: analysis of 14 cases].

OBJECTIVE: To evaluate the feasibility and safety of surgical removal of the tumors through laparoscopy in patients with early-stage uterine cancer. METHODS: A retrospective analysis of the clinical data of 10 cervical cancer and 4 endometrial cancer cases, in which laparoscopic radical hysterectomy and pelvic lymphadenectomy were performed following the same surgical procedures as in laparotomy. RESULTS: The mean operating time was 302 min, and mean estimated blood loss was 760 ml, with the mean postoperative gastrointestinal recovery time of 30 h and average number of removed lymph nodes of 22. CONCLUSION: Laparoscopic surgery has equivalent curative effect to laparotomy, and laparoscopic radical hysterectomy and pelvic lymphadenectomy in the patients with early-stage uterine cancer is both feasible and efficient.

Adult↗