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Biomedical subjects

X J Fan

Publications and source records attributed to X J Fan.

At least 19 recordsLinked to original sources

A serum proteomics approach to the diagnosis of ectopic pregnancy.

An ectopic pregnancy (EP) occurs when implantation of the embryo occurs outside of the uterus. If left untreated, the developing fetus will continue to grow, leading to life-threatening consequences for the mother. A major difficulty with the diagnosis of ectopic pregnancy is that methods of detection are limited, and some, such as ultrasound, are not very reliable in the earliest days of gestation. Currently, no effective serum test exists to distinguish an ectopic pregnancy from a normal intrauterine pregnancy. The incidence of ectopic pregnancy is increasing and has doubled in the last 20 years. It is now the second most common cause of maternal death in the first trimester of pregnancy. To address this issue, we initiated a project to identify serum markers of ectopic pregnancy. The subjects for these studies presented at the Hospital of the University of Pennsylvania. We obtained over 140 serum samples from women with suspected ectopic pregnancy: women presenting with pain and/or bleeding in the first trimester of pregnancy. The approximate racial breakdown of the subjects is as follows: African American, 36%; Caucasian, 3%; Asian, 2%; Hispanic, 1%; unknown, 58%. Serum samples from 139 women (62 with ectopic pregnancy and 77 with a normal intrauterine pregnancy) were applied to WCX2 (weak ion exchange) protein chip surfaces and analyzed for serum markers using surface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). Several proteins in the 7500-18,000 Da mass range were identified that may discriminate an ectopic pregnancy from an intrauterine pregnancy. The most promising markers were analyzed using classification and regression tree analysis (CART) with and without clinical variables (serum hCG value, length of amenorrhea). Two different algorithms were developed that classify the patients on the basis of sensitivity (number of EPs who screen positive/# of EPs) or specificity (# of healthy patients who screen negative/# of healthy). Our current approach is to refine these two "rule sets" to segregate patients into three groups: those who need immediate intervention for a probable ectopic pregnancy, those who appear to have a normal pregnancy, and those who need further monitoring for diagnosis.

Biomarkers↗

A new method for characterizing denitrifying phosphorus removal bacteria by using three different types of electron acceptors.

This study investigated the characteristics of denitrifying phosphorus removal bacteria by using three different types of electron acceptors as well as the positive role of nitrite in phosphorus removal process. Denitrifying phosphorous removal bacteria was enriched under anaerobic-anoxic (A/A) condition. To understand A/A sludge better, sludge from two other sources were also studied. These include sludges obtained from a lab-scale anaerobic-anoxic-aerobic (A/A/O) system and a local sewage treatment plant. Three types of possible electron acceptors (oxygen, nitrate and nitrite) were examined for their roles in phosphorus uptake. The results obtained indicated that oxygen, nitrate and nitrite were able to act as electron acceptors successfully. This observation suggested that in addition to the two well-accepted groups of phosphorus removal bacteria (one can only utilize oxygen to take up phosphorus, P(O), while the other can use both oxygen and nitrate, P(ON)), a new group of phosphorus removal bacteria, P(ON(n)), which could use oxygen, nitrate or nitrite to take up phosphorus was identified. The relative population of these three types of bacteria could be calculated from results obtainable from phosphorus uptake batch experiments with either oxygen or nitrate or nitrite as electron acceptor. The results obtained in this study showed that A/A sludge had similar phosphorus removal performance as the A/A/O sludge. However, it has better denitrifying phosphorus removal capability, which was demonstrated by the relative population of the three groups of bacteria. The results also suggested that nitrite was not an inhibitor to phosphorus removal process. Instead, it is an alternative electron acceptor to oxygen or nitrate.

Bacteria↗

[The analysis of rice total RNA by capillary electrophoresis].

When the RNase was eliminated through backing at 200 degrees C or treated by diethylpyrocarbonate (DEPC), the total RNA of rice could be separated within 15 min using 1.0%T, 0%C linear polyacrylamide as sieving matrix and 7 mol/L urea as denaturant. The tRNA of rice can be separated into two classes and about nine peaks when high concentrated polyacrylamide sieving matrix (5.0%T, 0%C) was used. This technique could provide one of the rapid and accurate methods for the determination of RNA in plants.

English Abstract↗

[Determination of human PDGF-B promoter binding nuclear protein by capillary electrophoresis].

With the use of 1.0% T, 0% C linear polyacrylamide as sieving matrix, 0.25 x TBE(Tris 89 mmol/L, boric acid 89 mmol/L, EDTA 2 mmol/L) as running buffer and 15 degrees C as column temperature, the human PDGF-B promoter binding nuclear protein can be determined within 50 min with good resolution. The results proved that there are two proteins having strong ability binding human PDGF-B promoter, which similar to that in slab gel electrophoresis. This technique can provide one of the rapid and accurate separation methods in the studying of the formation and repression behavior of DNA binding protein based on PDGF gene as the target.

DNA-Binding Proteins↗

Molecular determinants of the caspase-promoting activity of Smac/DIABLO and its role in the death receptor pathway.

Smac/DIABLO is a mitochondrial protein that is released along with cytochrome c during apoptosis and promotes cytochrome c-dependent caspase activation by neutralizing inhibitor of apoptosis proteins (IAPs). We provide evidence that Smac/DIABLO functions at the levels of both the Apaf-1-caspase-9 apoptosome and effector caspases. The N terminus of Smac/DIABLO is absolutely required for its ability to interact with the baculovirus IAP repeat (BIR3) of XIAP and to promote cytochrome c-dependent caspase activation. However, it is less critical for its ability to interact with BIR1/BIR2 of XIAP and to promote the activity of the effector caspases. Consistent with the ability of Smac/DIABLO to function at the level of the effector caspases, expression of a cytosolic Smac/DIABLO in Type II cells allowed TRAIL to bypass Bcl-xL inhibition of death receptor-induced apoptosis. Combined, these data suggest that Smac/DIABLO plays a critical role in neutralizing IAP inhibition of the effector caspases in the death receptor pathway of Type II cells.

Amino Acid Sequence↗

Helicobacter pylori increases proliferation of gastric epithelial cells.

The direct and indirect effects of helicobacter pylori on cell kinetics of gastric epithelial cell line AGS were investigated by flow cytometric analysis of Ki-67 positive cells and by MTT assay. Flow cytometric analysis of Ki-67 positivity permits detection of cells that are in S-phase, whereas the MTT assay is a colometric measure of the number of viable cells. In the absence of added stimulants, 23.06 (4.88)% mean (SD) of AGS cells were Ki-67 positive. When cells were preincubated in the presence of H pylori, there was a significant increase in Ki-67 positivity (66.20 (7.89)%, p < 0.001). This increase was not seen in cells cultured in the presence of Campylobacter jejuni (24.63 (8.11)% or Escherichia coli (21.66 (9.78)%). Pre-incubation of AGS cells with supernatants from both H pylori and mitogen activated peripheral blood lymphocytes also increased the per cent of cells that were Ki-67 positive (72.93 (8.68) and 69.96 (12.35)%; p, 0.001) respectively. Similar results were also found in MTT assay. These data show that both H pylori directly and the immune/inflammatory response to H pylori indirectly can influence the rate of epithelial cell proliferation, suggesting this bacterium may be an initiating step in gastric carcinogenesis and an important co-carcinogenic factor in H pylori positive subjects.

Antibodies, Monoclonal↗

Effect of IL-4 on peripheral blood lymphocyte proliferation: implication in immunopathogenesis of H. pylori infection.

Interleukin (IL) 4 is a type 2 cytokine which has a negative immunoregulatory role in human infection. IL-4 suppresses the production of interferon-gamma and enhances IL-10 synthesis. However, the effect of IL-4 on proliferative response of lymphocytes remains to be elucidated. We have previously reported an increase in production of IL-4 in subjects with Helicobacter pylori (H. pylori) infection. To evaluate whether the increased IL-4 is responsible for the down-regulation of immune responses in H. pylori infection, we observed the proliferative response of peripheral blood lymphocytes (PBL) co-cultured with phythaemagglutinin (PHA) or H. pylori in the presence and absence of added IL-4. As we have previously shown, PHA and H. pylori may increase PBL proliferation (P < 0.001). An increase in PBL proliferation was observed when PBL were co-cultured with PHA (P < 0.001) or H. pylori (P < 0.001) in the presence of IL-4 compared to that in the absence of IL-4. The optimal dose of IL-4 to give maximal lymphocyte proliferation is 50 pg/ml for the PHA-stimulated group or 100 pg/ml for the H. pylori-stimulated group. The data suggest that the increased IL-4 does not directly contribute to suppressed lymphocyte proliferation in H. pylori infection. Further studies will be required to determine the role of IL-4 in other aspects of down-regulation of immune responses in H. pylori infection.

Cells, Cultured↗

Up-regulation of CD44 and ICAM-1 expression on gastric epithelial cells by H. pylori.

Interaction between lymphocytes and epithelial cells may play a key role in Helicobacter pylori (H. pylori)-associated gastric mucosal inflammation. This interaction process is at least partially mediated by various cell adhesion molecules. The aims of the present study were to assess using flow cytometric analysis whether H. pylori directly or supernatants from H. pylori-activated peripheral blood mononuclear cells (PBMC) can affect the expression of adhesion molecules on the gastric epithelial cell line AGS in vitro. The results showed that resting AGS cells expressed CD44 and ICAM-1. Co-culture of AGS with H. pylori or cytokine-rich supernatants from H. pylori-activated PBMC resulted in up-regulation of expression of CD44 and ICAM-1 on AGS cells. These data suggest that H. pylori directly and indirectly through inflammatory cytokines may contribute to alternations in adhesion molecule expression on gastric epithelial cells. This may be of pathological significance in H. pylori-associated gastric mucosal inflammation and carcinogenesis.

Epithelium↗

Increased gastric production of interleukin-8 and tumour necrosis factor in patients with Helicobacter pylori infection.

AIMS: To investigate the role of interleukin-8 (IL-8) and tumour necrosis factor (TNF) in patients infected with Helicobacter pylori. METHODS: The study population comprised 52 patients with dyspepsia attending for upper gastrointestinal endoscopy. Of these patients, 35 were infected with H pylori. IL-8 and TNF concentrations in plasma, gastric juice, and gastric biopsy homogenate supernatant fluid were measured by radioimmunoassay and L929 cell bioassay, respectively. RESULTS: The concentrations of IL-8 and TNF in gastric juice and gastric biopsy homogenates were substantially greater in patients infected with H pylori. In H pylori positive patients IL-8 concentrations in gastric juice and gastric biopsy homogenates were higher in those with moderate gastritis than in those with mild gastritis. There was a positive correlation between IL-8 and TNF concentrations in gastric juice and gastric biopsy homogenate supernatant fluid from H pylori positive patients. There were no significant differences between H pylori positive and negative patients with respect to IL-8 and TNF plasma concentrations. CONCLUSION: This study suggests that increased gastric production of IL-8 and TNF may be implicated in the pathogenesis of H pylori associated gastroduodenal disease.

Adult↗

Helicobacter pylori-induced lipid peroxidation in peripheral blood lymphocytes.

Several lines of evidence implicate Helicobacter pylori infection in the pathogenesis of gastritis and peptic ulceration. To investigate whether H. pylori can cause lipid peroxidation in lymphocytes in vitro and to look for experimental evidence of lipid peroxidation induced by H. pylori, the lipid peroxide (LPO) level in peripheral blood lymphocytes was measured using the thiobarbituric acid fluorescence method. In the absence of added H. pylori, the LPO level in lymphocytes was 0.133 +/- 0.033 nmol/10(6) cells, and in the co-culture of H. pylori with peripheral blood mononuclear cells 0.340 +/- 0.097 nmol/10(6) cells. A significant difference was found between the two groups (p < 0.001). Antioxidants, either superoxide dismutase or catalase, could inhibit LPO production in lymphocytes. The present data provide further evidence that H. pylori can induce lipid peroxidation, which may be responsible for the pathogenesis of H. pylori-associated mucosal damage.

Catalase↗

Gastric T lymphocyte responses to Helicobacter pylori in patients with H pylori colonisation.

Helicobacter pylori has been identified as a dominant factor in the pathogenesis of duodenal ulcer. The aim of this study was to examine peripheral blood and gastric lymphocyte proliferation and cytokine production in patients with H pylori colonisation. Sixty five dyspeptic patients attending for endoscopy were studied; 35 of these were H pylori positive and 30 H pylori negative as assessed by culture, histology, and rapid urease test. H pylori antigen was capable of stimulating peripheral blood lymphocyte proliferative responses even in H pylori negative patients. Peripheral blood lymphocyte proliferative responses to H pylori (but not to purified protein derivative or phythaemagglutinin) were significantly lower in H pylori positive than H pylori negative patients. Similarly, antigen specific proliferative responses and interferon gamma production by gastric lamina propria lymphocytes were also depressed in H pylori positive patients compared with H pylori negative patients. CD8 and CD22 positive lamina propria lymphocytes were increased in H pylori positive patients. These data show that antigen specific responses to H pylori are significantly lower in H pylori positive patients and could indicate activation of antigen specific suppression.

Adolescent↗

Interferon-gamma and tumour necrosis factor production in patients with Helicobacter pylori infection.

The production of the cytokines, interferon-gamma and tumour necrosis factor by human antral mucosa cells and stimulated peripheral blood mononuclear cells were determined by enzyme linked immunosorbent assay and L929 bioassay respectively. Tumour necrosis factor production by peripheral blood mononuclear cells in response to Helicobacter pylori stimulation was depressed in Helicobacter pylori positive individuals, compared to Helicobacter pylori negative individuals (P < 0.05). There was no difference in tumour necrosis factor production by peripheral blood mononuclear cells in response to purified protein derivative. However, tumour necrosis factor production by cells isolated from gastric mucosa during short term culture was significantly higher in Helicobacter pylori positive patients (P < 0.05) than negative patients, indicating a probable macrophage response. Levels of interferon-gamma did not differ significantly in the gastric explant culture from the two groups. The results show that Helicobacter pylori negative patients have a stronger peripheral cellular immune response to Helicobacter pylori infection. The higher levels of tumour necrosis factor production by antral mucosa cells in Helicobacter pylori positive patients may reflect the infiltration of T lymphocytes and macrophages within the local mucosa.

Adult↗

[In vitro effect of catechin on cell growth].

Experimental study on the effect of catechin, an extract of Chinese green tea, on cell growth of HeLa, KB and 3T3 cell lines and its possible mechanism are reported. The proliferative survival curves showed that HeLa and KB were sensitive to catechin, while 3T3 was less so. The survival curve of HeLa cells treated with catechin varied with the dose and duration of exposure. 3H-TdR incorporation rate into HeLa cells was inhibited by catechin and the inhibition rate was also dose and exposure dependent. It suggests that one of the mechanisms of cell growth inhibition by catechin may probably due to inhibition of DNA synthesis.

3T3 Cells↗

Levels of O6-methylguanine acceptor protein in extracts of human breast tumor tissues.

We have measured the abilities of extracts of tissues from human breast tumors to demethylate adducts of O6-meG in exogenous DNA by transfer of the methyl group to an acceptor protein. The results have shown that all 21 specimens examined (including 5 non-neoplastic, 11 malignant tumors and 5 benign growth) contained significant amounts of O6-meG acceptor activity, removing on average 221.1 +/- 2.1 (SEM) fmol O6-meG per mg protein or 10.07 +/- 0.98 (SEM) fmol O6-megG per microgram DNA in the extracts. There were also wide interindividual variations, which were not age-dependent, and there were no significant differences between the non-neoplastic and neoplastic tissues obtained from individuals with benign or with malignant disease. It was estimated that the average number of O6-meG acceptor molecules per cell in normal human breast tissues was calculated as 46,000 +/- 7000 (SEM).

Adult↗

Study on enhancement of tumor-killing effect and formation of new fluorophore in tumor cells.

Hyperthermia (42-44 degrees C) and photosensitizing therapy can destroy S180 tumor cells, reduce malignant ascites and prolong the survival times of mice with carcinomas. The highest curative effect was observed when using a combination of the two treatments. Heating to 44 degrees C has a greater destructive effect on tumor cells than has heating to 42 degrees C. The results show that this is due to a synergistic interaction between these two treatments. The fluorescence spectrum of S180 cells was determined before and after treatment, and the indication was that the synergistic effect is probably related to a new fluorescence product; the greater the intensity of the new fluorescence, the more marked the synergy of hyperthermia and photosensitizing therapy. The maximum emission wavelength was 460nm (excitation wavelength 370nm).

Animals↗

[Experimental studies on the hyperthermia-photodynamic effect of tumor cells photosensitized with Y-HpD].

Experimental studies were made on the hyperthermia-photodynamic effect of tumor cells photosensitized with Y-HpD (China-made). Fluorescence spectra were determined in different animal and human cancer tissues before and after treatment. The results showed: Hyperthermia (42-44 degrees C) significantly increased the efficacy of photodynamic therapy (PDT) with a prolongation of survival time of mice and significant difference between PDT groups with and without hyperthermia (P less than 0.05). However, this difference was not present with mild hyperthermia (39-40 degrees C); A new fluorescence band was first observed in animal and human cancer cells after PDT treatment (with or without hyperthermia), with a maximum emission wave-length of 460 nm (excitation wave-length, 370 nm). The intensity of the band being correlated with cancer cell destruction, and the highest fluorescence intensity together with the strongest cytocidal effect were seen in the PDT-plus-heat group. Most Y-HpD was found the cytoplasm after treatment, with little Y-HpD left in the nucleus. The amount of Y-HpD in the cells decreased with the formation of fluorophores in the cytoplasm as a product of photochemical reactions. The authors suggest that hyperthermia may activate Y-HpD, and the increase of cellular Y-HpD then enhances the photochemical reaction.

Animals↗