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Biomedical subjects

Q Wei

Publications and source records attributed to Q Wei.

At least 73 records · Page 4Linked to original sources

DNA repair capacity in healthy medical students during and after exam stress.

There has been extensive research into the effects of stress on immune function but little on the effects of stress on DNA repair capacity (DRC), a process central to maintaining a normal cell cycle. Defective DRC is one of the factors responsible for carcinogenesis. In the present study we assessed DRC in healthy medical students during times of high and low stress. Sixteen medical students were evaluated during the third day of a 5-day exam period and then again 3 weeks later, after vacation. At both time points, participants underwent a brief physical examination, had venous blood drawn, and completed questionnaires to identify subjective stress levels. The DRC was assessed by the host-cell reaction assay, which measures nucleotide excision repair capacity. Participants reported significantly higher levels of subjective stress during the exam period than after vacation. DRC was also significantly higher during the exam period than after vacation, suggesting a positive association between subject stress levels and DRC. The results are discussed in relation to previous findings and implications for cancer research.

Adult↗

Human mu-opioid receptor overexpressed in Sf9 insect cells functionally coupled to endogenous Gi/o proteins.

Human mu-opioid receptor (HmuOR) with a tag of six consecutive histidines at its carboxyl terminus had been expressed in recombinant baculovirus infected Sf9 insect cells. The maximal binding capacity for the [3H] diprenorphine and [3H]ohmefentanyl (Ohm) were 9.1 +/- 0.7 and 6.52 +/- 0.23 nmol/g protein, respectively. The [3H] diprenorphine or [3H] Ohm binding to the receptor expressed in Sf9 cells was strongly inhibited by mu-selective agonists [D-Ala2, N-methyl-Phe4, glyol5]enkephalin (DAGO), Ohm, and morphine, but neither by delta nor by kappa selective agonist. Na+ (100 mM) and GTP (50 microM) could reduce HmuOR agonists etorphine and Ohm affinity binding to the overexpressed HmuOR. mu-selective agonists DAGO and Ohm effectively stimulated [35S]GTP-gammaS binding (EC50 = 2.7 nM and 6.9 nM) and inhibited forskolin- stimulated cAMP accumulation (IC50 = 0.9 nM and 0.3 nM). The agonist-dependent effects could be blocked by opioid antagonist naloxone or by pretreatment of cells with pertussis toxin (PTX). These results demonstrated that HmuOR overexpressed in Sf9 insect cells functionally coupled to endogenous G(i/o) proteins.

Animals↗

Protean clinical manifestations in children with leukemias containing MLL-AF10 fusion.

Translocations involving the MLL gene on chromosome 11q23 occur in 5-10% of human leukemias, and involve fusion with more than 30 different partner genes. The MLL-AF10 fusion produced by the t(10;11)(p12;q23) or ins(10;11)(p12;q23q13) occurs in a small percentage of acute leukemias, most commonly acute myelogenous leukemia (AML) of the M5 FAB subtype. We report two cases of AML (M5a and M0) and one case of acute lymphoblastic leukemia containing MLL-AF10 fusion. Each case had varied clinical characteristics, despite expressing similar MLL-AF10 fusion transcripts. Including the three cases described in this report, we identified a total of 38 cases of leukemia with MLL-AF10 fusion. Approximately one-third of these are not M5 AML. Taken together, these findings emphasize that while the sentinel molecular event may be identical in a disease, the clinical presentation and outcome can vary widely.

Artificial Gene Fusion↗

Conditional expression of a truncated fragment of nonmuscle myosin II-A alters cell shape but not cytokinesis in HeLa cells.

A truncated fragment of the nonmuscle myosin II-A heavy chain (NMHC II-A) lacking amino acids 1-591, delta N592, was used to examine the cellular functions of this protein. Green fluorescent protein (GFP) was fused to the amino terminus of full-length human NMHC II-A, NMHC II-B, and delta N592 and the fusion proteins were stably expressed in HeLa cells by using a conditional expression system requiring absence of doxycycline. The HeLa cell line studied normally expressed only NMHC II-A and not NMHC II-B protein. Confocal microscopy indicated that the GFP fusion proteins of full-length NMHC II-A, II-B, and delta N592 were localized to stress fibers. However, in vitro assays showed that baculovirus-expressed delta N592 did not bind to actin, suggesting that delta N592 was localized to actin stress fibers through incorporation into endogenous myosin filaments. There was no evidence for the formation of heterodimers between the full-length endogenous nonmuscle myosin and truncated nonmuscle MHCs. Expression of delta N592, but not full-length NMHC II-A or NMHC II-B, induced cell rounding with rearrangement of actin filaments and disappearance of focal adhesions. These cells returned to their normal morphology when expression of delta N592 was repressed by addition of doxycycline. We also show that GFP-tagged full-length NMHC II-A or II-B, but not delta N592, were localized to the cytokinetic ring during mitosis, indicating that, in vertebrates, the amino-terminus part of mammalian nonmuscle myosin II may be necessary for localization to the cytokinetic ring.

Actin Cytoskeleton↗

XPD/ERCC2 polymorphisms and risk of head and neck cancer: a case-control analysis.

DNA repair capacity is central in maintaining normal cellular functions. Variants of several DNA repair genes,including the nucleotide excision repair gene XPD, have been described recently. Because we previously reported that patients with squamous cell carcinoma of the head and neck (SCCHN) had lower DNA repair capacity than healthy controls, we hypothesized that inherited polymorphisms of XPD may contribute to genetic susceptibility to SCCHN, a tobacco-related cancer. To test this hypothesis, we conducted a hospital-based case-control study of 189 SCCHN patients and 496 cancer-free controls who were frequency-matched on age, gender and smoking status. All subjects were non-Hispanic whites. Two XPD polymorphisms (C22541A and A35931C) were typed using the restriction enzymes TfiI and PstI, respectively. Multivariate logistic regression analysis was performed to calculate adjusted odds ratios (ORs) and 95% confidence intervals (CIs). In the controls, the frequencies of the variant 22541A and 35931C alleles were 44.7% and 33.8%, respectively. The frequency of the 22541A homozygous genotype (22541AA) was lower in cases (15.9%) than in controls (20.4%) but was not associated with risk (adjusted OR = 0.90; 95% CI = 0.52-1. 56) for SCCHN. The frequency of the 35931C homozygous genotype (35931CC) was higher in cases (16.4%) than in controls (11.5%) and associated with a borderline increased risk (adjusted OR = 1.55; 95% CI = 0.96-2.52) for SCCHN. The risk was higher in older subjects (OR = 2.22; 95% CI = 1.03-4.80), current smokers (OR = 1.83; 95% CI = 0.79-4.27) and current drinkers (OR = 2.59; 95% CI = 1.25-5.34) in the stratification analysis. These results suggest a gene-environment interaction, but this did not reach statistical significance. The findings are limited due to the relatively small numbers in the subgroups and need to be verified by further investigations.

Adult↗

Transfusion-acquired, autochthonous human babesiosis in Japan: isolation of Babesia microti-like parasites with hu-RBC-SCID mice.

We have isolated piroplasms from a patient who developed the first case of human babesiosis in Japan by using NOD/shi-scid mice whose circulating erythrocytes (RBCs) had been replaced with human RBCs (hu-RBC-SCID mice). Following inoculation of the patient's blood specimen into hu-RBC-SCID mice, parasites proliferated within the human RBCs in the mice, resulting in a high level of parasitemia. Parasite DNA was prepared from blood samples of the patient and the mice, and the nuclear small-subunit rRNA gene (rDNA) was amplified and sequenced. Both DNA samples gave rise to identical sequences which showed the highest degree of homology (99.2%) with the Babesia microti rDNA. Because the patient had received a blood transfusion before the onset of babesiosis, we investigated the eight donors who were involved. Their archived blood samples were analyzed for specific antibody and parasite DNA; only a single donor was found to be positive by both tests, and the parasite rDNA sequence from the donor coincided with that derived from the patient. The donor's serum exhibited a high antibody titer against the isolate from the patient, whereas it exhibited only a weak cross-reaction against B. microti strains isolated in the United States. We conclude that the first Japanese babesiosis case occurred due to a blood transfusion and that the etiological agent is an indigenous Japanese parasite which may be a geographical variant of B. microti. Our results also demonstrated the usefulness of hu-RBC-SCID mice for isolation of parasites from humans and for maintenance of the parasite infectivity for human RBCs.

Animals↗

Effect of vanadyl ions on calcineurin and its A subunit.

Calcineurin (CN) exhibits a bimodal regulation by different concentrations of vanadyl ions (VO2+) in the presence of Mn2+. Low concentrations of VO2+ inhibit the enzyme, with 50 microM VO2+ completely inhibiting CN activity, while high concentrations, up to 500 microM VO2+, stimulate the CN activity. A similar bimodal regulation of CN was not observed with either calcium or vanadate under the same conditions. X-band electron spin resonance spectroscopy, used to study the binding of VO2+ to the catalytic subunit A of calcineurin, show that there are two kinds of binding sites in the A subunit.

Animals↗

Gender difference in smoking effect on chromosome sensitivity to gamma radiation in a healthy population.

In the general population, there is variation in radiosensitivity associated with cancer risk. However, data on the role of epigenetic factors in the variation of radiosensitivity are scarce. Thus we investigated the effects of smoking and age on the radiosensitivity of human lymphocytes by measuring the frequency of chromosome aberrations after in vitro exposure to gamma rays in peripheral lymphocytes from 441 healthy subjects (18-95 years old). We analyzed the frequency of both spontaneous (baseline) and in vitro gamma-ray-induced (1.5 Gy) chromatid breaks in 50 well-spread metaphases per subject. The overall mean frequencies of spontaneous and induced breaks were 0.02 and 0.45 per cell, respectively. The mean frequency of induced breaks was significantly higher in men than in women (P = 0.03) but did not differ by age or ethnicity. Donors who had ever smoked showed a small but significantly increased frequency of induced breaks (mean = 0.47) compared to nonsmokers (mean = 0.41; P = 0.005). Further stratification and multivariate analyses revealed that the smoking effect was more pronounced in men than in women. These findings support a smoking effect on radiosensitivity in a healthy population, particularly in men. Therefore, when evaluating the association between radiosensitivity and susceptibility to smoking-related cancers, the effect of smoking should be taken into account.

Adult↗

The value of serum GPDA-F for the diagnosis of primary hepatocellular carcinoma.

OBJECTIVE: To investigate the role of glycylproline dipeptidyl aminopeptidase isoenzymes in the diagnosis of primary hepatocellular carcinoma (PHC). METHODS: We developed a stage gradient polyacrylamide gel electrophoresis system to separate serum GPDA isoenzymes. Total GPDA activities, alpha-fetoprotein, the sizes of tumors and alanine aminotransferase (ALT) activities were also measured simultaneously and the correlation between GPDA-F and these indices was analyzed. RESULTS: Serum GPDA was separated into two bands, namely fast band (GPDA-F) and slow band (GPDA-S). GPDA-F was negative in all healthy persons as well as in the patients with benign liver filling defects, while it was positive in 85.3% cases of PHC. Liver cirrhosis, chronic hepatitis, extrahepatic carcinoma and metastatic liver carcinoma had low positive rates. GPDA-F was positively correlated with serum total GPDA activities, but had no correlation with AFP and size of the tumors. There was the correlation between GPDA-F and ALT in benign liver diseases, but no correlation between GPDA-F and ALT in PHC. Serial measurements of serum GPDA-F showed that GPDA-F was persistently positive in PHC but might change into negative in benign liver diseases. Dynamic determination of GPDA-F might be helpful to differentiate true positive of PHC from false positive of benign liver diseases. CONCLUSION: GPDA-F is a new serum marker of PHC. Measurement of serum GPDA-F is of value for the diagnosis of PHC, especially for those at early stage or with negative AFP.

Alanine Transaminase↗

Reduced expression levels of nucleotide excision repair genes in lung cancer: a case-control analysis.

Epidemiological studies have indicated that reduced DNA repair capacity and increased DNA adduct levels are associated with increased risk of lung cancer. Nucleotide excision repair (NER) is the major pathway in humans for repairing DNA adducts induced by smoking-related carcinogens, such as benzo[a]pyrene diol epoxide. We hypothesized that genetically determined baseline expression level of genes involved in NER is associated with risk of lung cancer. In a pilot case-control study, we measured the relative expression levels of five NER genes [ERCC1, XPB/ERCC3, XPG/ERCC5, CSB/ERCC6 and XPC (ERCC, excision repair cross-complementing; CSB, Cockayne's syndrome complementary group B)] in phytohemagglutinin-stimulated peripheral lymphocytes obtained from 75 lung cancer patients and 95 controls using a newly developed multiplex RT-PCR assay. Cases and controls were matched on age, sex, ethnicity and tobacco use. The expression level of the beta-actin gene was used as an internal control for the relative quantitation. We observed a 12.2 and 12.5% decrease in the baseline expression levels of XPG/ERCC5 and CSB/ERCC6, respectively, in cases compared with controls. These differences were statistically significant (P < 0.01) when the median expression level in the controls was used as the cut-off point, the lung cancer patients were significantly more likely than the controls to have reduced expression levels of XPG/ERCC5 [odds ratio (OR), 2.32; 95% confidence interval (CI), 1.22-4.43] and CSB/ERCC6 (OR, 2.49; 95% CI, 1.28-4.84). There was also a dose-response relationship between reduced expression levels and increased lung cancer risk (trend test: P < 0.01). Our results suggest that individuals whose expression levels of XPG/ERCC5 and CSB/ERCC6 are reduced may be at higher risk of lung cancer.

Actins↗

[In vitro study of C3a des Arg produced by plasma incubation with blood purification materials].

This study was performed to evaluate the degree of complement C3 activation of five biomedical polymers (cellulose acetate (CA), polymethylmethacrylate (PMMA), polysulphone (PS), polyethersulphone (PES), sulphonated polyethersulphone(PES-SO3) which are used to make hemofilters. The concentration of the activation derivative C3a des Arg was measured by radioimmunoassay (RIA) in human plasma after incubation with these five polymers for 30, 60, 90 and 120 minutes. The experimental results indicated that the presence of polymers caused an increase in C3a des Arg. The potential to activate complement C3 decreased in sequence of CA, PMMA, PES, PS and PES-SO3. CA activated the most complement C3. PES-SO3 activated the least complement C3, and the concentration of C3a des Arg decreased with the increase in sulphonation degree. The investigation demonstrates that with good biocompatibility PES is a promising material to make biocompatible hemofilters. Furthermore, it indicates that PES containing sulphonate groups can achieve a marked reduction in complement activation and supports the sulphonation as a means of improving the biocompatibility of PES for blood purification application. RIA is a reliable and precise quantitative determination of human C3a des Arg and is suitable for testing the activation of complement of various synthetic materials that are utilized or to be utilized in the medical field.

Complement Activation↗

[The xenoantigenicity of Chinese inbred-line pigs].

OBJECTIVE: The major obstacle in pig to human transplantation is acute and hyperacute rejection (HAR) triggered mainly by alpha-galactosyl residues(alpha-Gal) in donor. Since the inbred-line Banna pig(IBNP) and Wuzhishan pig (IWZSP) are highly inbred and may be the potential donor for xenotransplantation, it is important to investigate the reaction between human serum and inbred-line pig tissues as well as the distribution of alpha-Gal in these tissues. METHODS: Samples from heart, liver, spleen, lung, kidney, pancreas, small intestine, thymus, skin, lymph node and blood vessels at all levels were collected from four 8 to 11-month-old male IBNPs and one IWZSP. Affinity-immunohistochemistry assays were conducted following routine procedures on paraffin sections with normal human sera of blood type A, B, O, AB and BSI-B4(alpha-Gal specific binding lectin) as the primary antibodies or affinity reagents. Sections digested by alpha-galactosidase were also examined as control. RESULTS: Parallel results were obtained from these pig tissues stained against human sera and BSI-B4. There was no significant difference both in the antigens recognized by sera of different blood types or BSI-B4 and in the distribution of alpha-Gal. The strongest alpha-Gal positive staining was appeared in vascular endothelial cells at all levels and partial parenchyma cells. However, tissues of cartilage, peripheral nerve and muscle were negative. After digested by alpha-Galactosidase, all samples were negative against BSI-B4 and human sera except few positions that showed different staining. CONCLUSION: The distribution of target antigen is similar in various tissues of the two kinds of pigs. Though alpha-Gal is the major xenoantigen in IBNP and IWZSP, there may be some unknown antigens related to pig to human transplantation. Possibly the level and distribution of antigen expression in pig tissues are not the first affair to be considered, and these pigs should be genetically modified in order to eliminate rejection in pig to human xenotransplantation.

Animals↗

[Expression of matrix metalloproteinases and tissue inhibitor of metalloproteinases in colorectal neoplasm].

OBJECTIVES: To explore the relationship of matrix metalloproteinases (MMPs) and tissue inhibitor of metalloproteinases (TIMPs) to the oncogenesis and development of colorectal neoplasm. METHODS: RT-PCR was used to assay the level of MMP-2, MT1-MMP, MMP-7, TIMP-2, TIMP-3 mRNA in 28 cases of colorectal cancer, including tumor tissue and surrounding normal tissue. RESULTS: MMP-2, MT1-MMP, TIMP-2 and TIMP-3 were over-expressed in tumor and normal tissues, and MMP-7 was strong-expressed in tumor tissue but was weak-expressed only in one case of normal tissue. The expression of MMP-7 in tumor tissue was correlated to Dukes stage (P < 0.01). The expression of TIMP-2 in tumor tissue with positive-node was much higher than that with negative-node (P < 0.01). The expression of TIMP-3 was decreased with the Dukes stage and the depth of invasion (P < 0.01). There were no correlations between MMPs and TIMPs (P > 0.1). CONCLUSIONS: The expression of MMP-7 mRNA has a high specificity in colorectal cancer. MMP-7 may become a sensitive tumor marker. Inducing TIMP-2, TIMP-3 or suppressing MMP-2, MT1-MMP, MMP-7 mRNA's expressions may inhibit the invasion and metastasis of cancer.

Adult↗

[A study on apoptosis and activity of SAPK/JNK in adrenocortical cells induced by cadmium chloride].

OBJECTIVE: To study apoptosis induced by cadmium chloride and the alteration in activity of stress activated protein kinase (SAPK) in adrenocortical cells. METHODS: Fasciculata-glomerulosa (FG) cells of male guinea pigs were dispersed and primarily cultured in vitro. Features of apoptotic cells were observed using combined labeling with annexin-V and propidium iodide (PI) and flow cytometry, and activity of SAPK was determined with immunoprecipitation and chemiluminescence assay. RESULTS: Apoptosis rate of FG cells increased with dose of cadmium chloride (CdCl(2)) two hours after treatment with 6.25 - 200.00 micromol/L of it. The average percentages of apoptotic cells ranged from 9.90% to 82.35%, with significant difference in the groups treated with 25, 50, 100 and 200 micromol/L of CdCl(2), as compared with the control group (P < 0.01). Regression analysis showed that occurrence of apoptosis correlated with the dose of cadmium chloride in a dose-response pattern. In the meanwhile, there were obviously elevated percentages of apoptotic cells as the increase in duration of incubation, ranging from 5.58% to 73.08% for incubating cells with 50.00 micromol/L of CdCl(2), from 15 minutes to four hours. Duration of incubating cells with cadmium chloride correlated to occurrence of apoptosis in a time-effect manner. There were no remarkable changes in the activity of SAPK in adrenocortical cells two hours after incubation with 25.00 - 100.00 micromol/L of CdCl(2). CONCLUSIONS: Apoptosis can be induced by cadmium chloride in adrenocortical cells and its mechanism should be studied further.

Adrenal Cortex↗

[Preparation of immobilized metal-chelated affinity membrane and its application to purification of Cu/Zn-superoxide dismutase].

Immobilized metal-chelated affinity membranes were prepared using macropore cellulose filter paper as matrix. The matrix was treated with alkaline, activated with epichlorohydrin and coupled with iminodiacetate sodium, and then Cu2+ was immobilized. Cu/Zn-superoxide dismutase(SOD) was purified with prepared affinity membranes. The special activity of the purified Cu/Zn-SOD was increased 10.7 times from 645 U/mL to 6,882 U/mL. The protein recovery and the activity recovery were 92.3% and 985%, respectively. A new method to prevent the leaking of metal ion was developed and Cu2+ concentration in the effluent was lowered to 86 micrograms/L.

Chelating Agents↗