PGM1 and ENO1 Promote the Malignant Progression of Bladder Cancer via Comprehensive Analysis of the m6A Signature and Tumor Immune Infiltration.

BACKGROUND: While N6-methyladenosine (m6A) modification of RNA and the tumor immune microenvironment both influence the progression of cancer, little attention has been paid to interactions between these two factors. Thus, we systematically explored potential biomarkers in the malignant progression of bladder urothelial carcinoma (BLCA) via combining expression of m6A methylation regulators with tumor immune infiltration. METHODS: We extracted m6A regulators from published literature, downloaded BLCA RNA-seq and clinical information from the Cancer Genome Atlas database, and integrated three main bioinformatic methods and qPCR to explore the biological variations in the malignant progression of BLCA. RESULTS: FTO, IGF2BP3, and YTHDC1 have a significant difference in bladder cancer and prognosis. Two subgroups (clusters 1 and 2) were identified according to three key m6A regulators; cluster 1 was preferentially associated with poor prognosis and immune infiltration relative to cluster 2 significantly. We further identified PGM1 and ENO1 as potential prognostic biomarkers, as they were correlated with FTO and IGF2BP3 positively but with YTHDC1, negatively. M2 macrophage and TFH cells were highly infiltrated in BLCA and were associated with BLCA prognosis. Finally, PGM1 and ENO1 were correlated with M2 macrophage and TFH cells and their surface markers CD163and CXCR5. CONCLUSIONS: PGM1 and ENO1 are highly correlated with the malignant progression of BLCA, and the expression of these genes may be new indicators for the diagnosis and prognosis of BLCA.

Clinical application, evaluation and analysis of influencing factors of on tuberculosis γ-Interferon enzyme-linked immunosorbent assay.

OBJECTIVE: To assess sensitivity and specificity of the interferon gamma release assay test, and to pinpoint the influencing factors that should be taken care of in clinical application. METHODS: This study was conducted at the First People's Hospital in the Yunnan Province of China from October 2018 to March 2019, and comprised samples collected from outpatient and inpatients. To detect mycobacterium tuberculosis, acid-fast staining on sputum smear was performed on relevant tissues suspected of extrapulmonary tuberculosis. Tuberculosis interferon gamma release assay test and pathology samples were examined. Tuberculosis-specific cell reaction assay kit was used for sampling. SysmexXN-2000 haematology analyser, VACUETTE SRS100/II and Beckman Coulter AU5800 were used to perform various analyses. Data was grouped and analysed using R statistical software. RESULTS: Of the 960 samples, 516(53.75%) cases tested positive for tuberculosis infection and 444(46.25%) tested negative. The sensitivity of the pathological results was 86% and the specificity was 96%. The sensitivity of the interferon gamma release assay test was 95% and specificity 82%. Interferon release test, pathological results and final diagnosis showed significant comparisons (p<0.05). Significant relationships were also established for factors, such as age, interferon release quantity, lymphocyte, C-reactive protein and counts of mono-nuclear cell (p<0.05). CONCLUSION: Interferon gamma release assay test was found to have high consistency with pathological results and final diagnosis and can be used as a subsidiary to traditional clinical imaging and pathological judgment.

A new contrast-to-noise ratio for image quality characterization of a coded-aperture γ camera.

The gamma-ray imaging technique was developed and is widely used in several nuclear engineering fields. Specifically, compared with the traditional point-by-point radiation detector, the coded-aperture gamma camera has advantages of a wide field of view, high angular resolution, and high efficiency. Several methods for characterizing image quality, including the figure of merit (FOM) method and the contrast-to-noise ratio (CNR) method, were assessed and developed. These methods have their respective drawbacks depending on the circumstances. The FOM lacks reliability in exhibiting the impact of background noise fluctuation on the purity of a real image. The CNR characterizes image quality with inconsistent sensitivity while the source moves along the X and Y directions. Therefore, a new CNR method was proposed to achieve better performance and greater consistency in real imaging. With our coded-aperture imaging system developed in the laboratory, we performed simulations within the MATLAB and Geant4 platforms and real imaging experiments to analyze and compare images and the results of these three characterization methods. The results show that the new CNR method is reliable and practical in regard to real imaging performance.

A rapid and accurate method for the detection of four aminoglycoside modifying enzyme drug resistance gene in clinical strains of Escherichia coli by a multiplex polymerase chain reaction.

BACKGROUND: Antibiotics are highly effective drugs used in the treatment of infectious diseases. Aminoglycoside antibiotics are one of the most common antibiotics in the treatment of bacterial infections. However, the development of drug resistance against those medicines is becoming a serious concern. AIM: This study aimed to develop an efficient, rapid, accurate, and sensitive detection method that is applicable for routine clinical use. METHODS: Escherichia coli was used as a model organism to develop a rapid, accurate, and reliable multiplex polymerase chain reaction (M-PCR) for the detection of four aminoglycoside modifying enzyme (AME) resistance genes Aac(6')-Ib, Aac(3)-II, Ant(3″)-Ia, and Aph(3')-Ia. M-PCR was used to detect the distribution of AME resistance genes in 237 clinical strains of E. coli. The results were verified by simplex polymerase chain reaction (S-PCR). RESULTS: Results of M-PCR and S-PCR showed that the detection rates of Aac(6')-Ib, Aac(3)-II, Ant(3″)-Ia, and Aph(3')-Ia were 32.7%, 59.2%, 23.5%, and 16.8%, respectively, in 237 clinical strains of E. coli. Compared with the traditional methods for detection and identification, the rapid and accurate M-PCR detection method was established to detect AME drug resistance genes. This technique can be used for the clinical detection as well as the surveillance and monitoring of the spread of those specific antibiotic resistance genes.

Loop-Mediated Isothermal Amplification Assay for Rapid Detection of Hypothetical Protein Gene in Escherichia Coli Clinical Isolates.

BACKGROUND: Escherichia coli is the most common pathogenic bacteria that frequently causes life-threatening opportunistic human infections, diarrhea, and septicemia in immunocompromised hosts. METHODS: This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of a hypothetical protein from an E. coli-specific gene (GenBank ID: 13702648). The gene was obtained through local and online BLAST, and specific primers were designed for this gene. Reaction conditions were optimized at 65ºC for 30 minutes and 80ºC for 2 minutes, whereas the reaction system contained 5.2 mM Mg2+, 8 U of Bst 2.0 DNA polymerase, 1.4 mM deoxyribonucleotide, and 0.2 and 1.6 µM of the outer and inner primers, respectively. The LAMP method was evaluated using 240 strains of E. coli and 150 strains of non-E. coli. RESULTS: Positive reactions were observed on all 240 strains of E. coli while all non-E. coli strains were negative. Plasmids with the specific gene and mice blood with E. coli were used for sensitivity analysis. The detection limit of LAMP was 100 bacterium/reaction. CONCLUSIONS: Results showed that the LAMP targeted to the hypothetical protein (GenBank ID: 13702648) is a fast, specific, sensitive, inexpensive, and suitable method for the detection of E. coli.

Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method.

Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse blood with K. pneumoniae were used for sensitivity analysis. The detection limit of the LAMP was 1 bacterium/reaction. The results showed that the LAMP targeted to ureR_1 is a fast, specific, sensitive, inexpensive, and suitable method for the detection of K. pneumoniae.

Establishment of loop-mediated isothermal amplification for rapid detection of Pseudomonas aeruginosa.

Pseudomonas aeruginosa is one of the three most pathogenic bacteria that frequently cause life-threatening opportunistic human infections, pneumonia, and lower respiratory tract infections in immunocompromised hosts, particularly in the burns ward. The present study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of P. aeruginosa-specific gene hypothetical protein (GenBank ID: 882161). The gene was obtained through local and online BLAST, and specific primers were designed for this gene. Reaction conditions were optimized at 65°C for 30 min and 80°C for 2 min, whereas the reaction system contained 5.2 mM Mg2+, 8 U Bst 2.0 DNA polymerase, 1.4 mM deoxyribonucleotide and 0.2 and 1.6 µM of the outer and inner primers, respectively. The LAMP method was evaluated using 150 P. aeruginosa and 170 non-P. aeruginosa strains. Positive reactions were observed on 150 P. aeruginosa strains, whereas all non-P. aeruginosa strains exhibited negative results. Plasmids with the specific gene and mouse blood with P. aeruginosa were used for sensitivity assay. The detection limit of LAMP was 1 bacterium/reaction. Results indicated that the LAMP method targeted to hypothetical protein is a fast, specific, sensitive, inexpensive and suitable method for detection of P. aeruginosa.

Rapid, specific, and sensitive detection of the ureR_1 gene in Klebsiella pneumoniae by loop-mediated isothermal amplification method

Klebsiella pneumoniae is one of the main pathogenic bacteria that causes nosocomial infections, such as pneumonia, urinary tract infection, and sepsis. Therefore, the rapid and accurate detection of K. pneumoniae is important for the timely treatment of infectious patients. This study aimed to establish a loop-mediated isothermal amplification (LAMP) method for the rapid and sensitive detection of K. pneumoniae-specific gene ureR_1 (Gene ID: 11847803). The ureR_1 gene was obtained through local and online BLAST, and the specific primers were designed for its detection. Positive reactions were observed on all 140 K. pneumoniae clinical isolates while all the 82 non-K. pneumoniae clinical isolates were negative. Plasmids with the specific gene and the mouse blood with K. pneumoniae were used for sensitivity analysis. The detection limit of the LAMP was 1 bacterium/reaction. The results showed that the LAMP targeted to ureR_1 is a fast, specific, sensitive, inexpensive, and suitable method for the detection of K. pneumoniae.

The Role of Thioredoxin-1 in Suppression Sepsis Through Inhibiting Mitochondrial-Induced Apoptosis in Spleen.

Sepsis is a serious public health issue and the leading cause of death in critically ill patients in intensive care units. Thioredoxin-1 (Trx-1) is a protein of regulating redox, as well as a modulator of inflammation and apoptosis. Our previous study reported that Trx-1 decreased endoplasmic reticulum-mediated inflammation involved in lung in a model of experimental sepsis. However, its effect on mitochondrial-mediated apoptosis in spleen has not been reported. We studied whether Trx-1 could prevent spleen cells apoptosis in sepsis. In the present study, we showed that the apoptosis in spleen was decreased in sepsis induced by cecal ligation and puncture (CLP) in Trx-1 overexpression transgenic (Tg) mice compared with wild-type mice. Colony forming units in the peritoneal cavity and the level of procalcitonin in plasma were significantly decreased in Trx-1 Tg mice 12 h after CLP. The expressions of c-jun-N-terminal kinase, Bax, caspase-9, and caspase-3 were increased in spleen, which were suppressed in Trx-1 Tg mice. However, the decreased Bcl-2 expression in sepsis was recovered in Trx-1 Tg mice. Our results suggest that overexpression of Trx-1 provides protection against sepsis through suppressing mitochondria-induced apoptosis pathway in spleen. This study may provide a new target for clinical intervention, as well potential strategies for treatment of sepsis.

Haematological and electrophoretic characterisation of ß-thalassaemia in Yunnan province of Southwestern China.

OBJECTIVES: ß-Thalassaemia is widely found in Southwestern China. Characterisation of ß-thalassaemia can improve screening and prenatal diagnosis for at-risk populations. DESIGN: A retrospective study. METHODS: In this study, the levels of haemoglobin alpha 2 (HbA2) and haemoglobin alpha (HbA) were analysed by gender for a total of 15 067 subjects screened by capillary electrophoresis. The cut-off value with the highest accuracy was established to identify ß-thalassaemia in 723 patients suspected to have this disease. Haematological and electrophoretic characterisation of eight common types of ß-thalassaemia were analysed in 486 ß-thalassaemia subjects. RESULTS: HbA levels were significantly higher in men than in women, but there was no significant difference on HbA2 levels. A new cut-off value for the diagnosis of ß-thalassaemia (HbA2≥4.0%) with the highest accuracy was proposed for the studied populations. Haemoglobin (Hb) was significantly higher in men compared with women (p<0.05), whereas no statistically significant differences were found for mean cell volume (MCV), mean cell haemoglobin (MCH), HbA and HbA2. The haemoglobin E (HbE) group showed comparatively higher values for haematological indices (Hb, MCV and MCH) than the other genotypes in heterozygous ß-thalassaemia groups (p<0.05), and -28 (A>G) (HBB (ß-globin):c.-78A>C) had significantly higher HbA2 values compared with other ß-thalassaemia. CONCLUSIONS: Ethnic groups have diversified ß-globin gene mutations and considerable haematological variations. Our study will lay the foundation for screening programmes and clinical management of thalassaemia in Southwestern China.

ATP binding cassette subfamily A member 7 rs3764650 polymorphism and the risk of Alzheimer's disease.

We did a meta-analysis to assess the association between ABCA7 rs3764650 polymorphism and the risk of Alzheimer's Disease (AD). A total of 10 eligible studies with 20511 patients and 40503 controls met the inclusion criteria. ABCA7 rs3764650 polymorphism was significantly associated with AD risk (OR=1.21, 95% CI 1.16-1.26, P<0.00001; I2=5%). In the subgroup analysis by race, statistically significant associations were found in Asians (OR=1.09, 95% CI 1.01-1.18, P =0.03; I2=0%) and in Caucasians (OR=1.25, 95% CI 1.19-1.31, P<0.00001; I2=0%). In conclusion, this meta-analysis suggested that ABCA7 rs3764650 polymorphism is significantly associated with AD risk.

S-propranolol protected H9C2 cells from ischemia/reperfusion-induced apoptosis via downregultion of RACK1 Gene.

Ischemia-reperfusion (I/R) injury can lead to apoptotic death of heart cells and subsequently heart failure. Propranolol is widely used in the management of cardiovascular disorders, but the mechanism is still unclear. Our previous studies showed that activated protein kinase C1 (RACK1) was significantly down-regulated in human umbilical vein endothelial cells by S-propranolol. RACK1 may be a target protein of S-propranolol during I/R. At present, we constructed a lentiviral expression vector for RNA interference (RNAi) of RACK1. The interference efficiency of the lentivirus was confirmed by RT-PCR and western blot. H9C2 cells infected with Lv-RACK1-shRNA or control were subjected to simulate I/R in the presence and absence of S-propranolol. The release of cytokines and chemokines was determined by ELISA assay. Flow cytometry was employed to determine mitochondrial membrane potential (MMP), Ca(2+) concentration, reactive oxygen species (ROS) production, and cell apoptosis. We found that RACK1 RNAi and S-propranolol treatment remarkably protected I/R injured cells from apoptosis via attenuating the release of cytokines and chemokines, Ca(2+) overload, ROS concentration, and MMP. Furthermore, RACK1 RNAi and S-propranolol, separately and in combination, significantly reduced caspase-3 activity, cytochrome c release and JNK activation. RACK 1 can be considered as a target for drug development.

Advanced light-trapping effect of thin-film solar cell with dual photonic crystals.

A thin-film solar cell with dual photonic crystals has been proposed, which shows an advanced light-trapping effect and superior performance in ultimate conversion efficiency (UCE). The shapes of nanocones have been optimized and discussed in detail by self-definition. The optimized shape of nanocone arrays (NCs) is a parabolic shape with a nearly linearly graded refractive index (GRI) profile from the air to Si, and the corresponding UCE is 30.3% for the NCs with a period of 300 nm and a thickness of only 2 µm. The top NCs and bottom NCs of the thin film have been simulated respectively to investigate their optimized shapes, and their separate contributions to the light harvest have also been discussed fully. The height of the top NCs and bottom NCs will also influence the performances of the thin-film solar cell greatly, and the result indicates that the unconformal NCs have better light-trapping ability with an optimal UCE of 32.3% than the conformal NCs with an optimal UCE of 30.3%.

Multi-year ozone concentration and its spectra in Shanghai, China.

The periodic properties of surface ozone variation were studied at five stations with different environmental conditions in Shanghai based on multi-year observations of ozone concentration and UV radiation using spectral decomposition methods. The spectra of surface ozone have distinct peaks at semi-diurnal, diurnal, intraseasonal, semiannual, annual, and quasi-biennial periods. The spectra for the frequency band larger than the semi-diurnal follow a -5/3 power law at all the stations. The diurnal peak values for all stations in different years are similar to each other, while the semi-diurnal peak values are somewhat different among the stations. The peak value of semi-diurnal cycle at the station Dongtan (ecological environment area) is smaller than that at the other stations. The monthly mean of surface ozone has a significant seasonal variation with a maximum in May, a secondary maximum in fall, a lower value in summer (July and August), and a minimum in December or January. However the seasonal variation of UV radiation monthly mean shows a relatively higher value in summer (July and August), and for other months it is closely related to the ozone monthly mean. These secondary peaks of the ozone monthly mean in fall might be caused by the UV radiation coming back to its relevant value after falling off during the Asia summer monsoon; it was not related to biomass burning. The intraseasonal cycling of ozone might be related to the MJO (Madden-Julian Oscillation). Further studies are needed to understand the relationship between the local ozone intraseasonal variation and the MJO. The quasi-biennial variation of ozone in Shanghai might be a local reflection of climate change and could be associated with ENSO (El-Nino Southern Oscillation). Further studies will be needed to understand the relationship of the quasi-biennial variation of ozone to ENSO.

[Effective antiviral therapy with entecavir in chronic hepatitis B virus carriers].

OBJECTIVE: To evaluate the short-term effect and safety of entecavir for the treatment of chronic hepatitis B (CHB) virus carriers. METHODS: Ninety-three cases of CHB virus infection (hepatitis B surface antigen (HBsAg)-positive, hepatitis B e antigen (HBeAg)-positive, hepatitis B core antibody (HBcAb)-positive, HBV DNA≥1x10(5) copies/mL) were divided into two groups: CHB virus carrier (47 cases) and CHB (46 cases). All of the 93 cases were given 0.5 mg entecavir orally once a day for 48 weeks. Virology, serology and biochemistry tests were perrmed at treatment weeks 0, 4, 12, 24 and 48. Side effects of entecavir and the incidence of liver cirrhosis and hepatocellular carcinoma were recorded. RESULTS: The CHB virus carrier and CHB group had complete virological response rates of 14.9% and 17.4% at week 4, 51.1% and 63.0% at week 12, 76.6% and 89.1% at week 24, and 97.9% and 100% at week 48, respectively; there was no significant difference between the two groups (P>0.05). The CHB virus carrier and CHB group had partial virological response rates of 42.6% and 47.8% at week 4, 57.44% and 65.2% at week 12, 85.0% and 89.1% at week 24, and 100% and 100% at week 48, respectively; there was no significant difference between the two groups (P>0.05). No cases in either group experienced virologic breakthrough during the treatment course. The CHB virus carrier and CHB group had serological response (HBeAg-negative) rates of 0 and 4.3% at week 4, 2.1% and 8.7% at week 12, 4.3% and 13.0% at week 24, and 8.5% and 21.7% at week 48, respectively; there was no significant difference between the two groups (P>0.05). The CHB virus carrier and CHB group had HBeAg seroconversion rates of 0 and 0 at week 4, 0 and 4.4% at week 12, 2.1% and 10.9% at week 24, and 6.4% and 17.4% at week 48, respectively; there was no significant difference between the two groups (P>0.05). No case in either group showed HBsAg-negativity and seroconversion during the treatment course. The CHB group had a biochemical response (alanine aminotransferase normalization) rate of 26.1% at week 4, 65.2% at week 12, 91.3% at week 24, and 97.8% at week 48.No case in either group showed biochemical breakthrough during the treatment course. There were no cases of liver cirrhosis or hepatocellular carcinoma in either group. There were no side effects of the entecavir treatment experienced in either group. CONCLUSION: Antiviral therapy with entecavir is effective, safe and well tolerated in CHB virus carriers.

Enantioselective pharmacodynamics of propranolol in HUVEC cells: a study using chiral 2D gel electrophoresis and mass spectrometry.

Propranolol (PRO), a nonselective ß-adrenergic receptor (ß-AR) antagonist, has two enantiomers, R(+)-PRO and S(-)-PRO, which have diverse biological effects. For example, S(-)-PRO blocks the ß-receptor ~100 times more strongly than R(+)-PRO. However, the signaling pathway that causes this difference remains unclear. This pathway may affect the expression of numerous proteins, some of which play key roles during the drug action process. Therefore, we treated human umbilical vein endothelial cells (HUVECs) with R(+)-PRO and S(-)-PRO in order to identify differentially expressed proteins and to determine their functions in the drug action process. Of the 22 differentially expressed protein spots investigated, 14 demonstrated higher expression levels in the R(+)-PRO-treated cells, while 8 demonstrated lower expression levels in the same cells. Mass spectrometry identified 10 of the differentially expressed proteins: 4 signaling molecules, 2 metabolic enzymes, 3 heat shock proteins and 1 cytoskeleton protein. Our results suggest that these differentially expressed proteins, particularly guanine nucleotide-binding protein subunit ß-2-like 1 (GBLP), are the key biomacromolecules underlying the mechanism by which PRO enantiomers induce stereoselective cellular responses. The results aid in clarifying the role of PRO in the treatment of arrhythmia and angina.

Depression and survival in Chinese patients with gastric cancer: a prospective study.

AIM: Depression is thought to be a predictor of poor survival among cancer patients. In our study, we aimed to investigate the association between depression and survival in patients with gastric cancer. METHODS: The subjects were a total of 300 patients aged 20-75 years who had histological confirmed diagnosis of gastric cancer from January 2004 to May 2006. Three months after patients diagnosis, depression was scored using by the Depression Status Inventory (DSI) designed by Willian WK Zung. The follow-up period consisted of a total of 13,643 person-months. A Cox's regression analysis was used to assess the association between depression and survival. RESULTS: The percentage of subjects with depression according to the DSI depression criteria was 31%. Tumor stage and treatment methods were significantly associated with depression of patients. Age (60 years or older), annual income, tumor stage, lymph nodes metastasis and treatment were significantly associated with increased hazard ratio (HR) for gastric cancer survival. The adjusted HR for mortality risk in gastric cancer patients with depression tended to be high (HR=3.34, 95% CI=1.23-5.49) and a significant trend was found (P<0.05). CONCLUSION: The data obtained in this prospective study in Chinese support the hypothesis that depression is associated with poor survival among gastric cancer patients. Further studies with a large sample and longer term follow-up period are needed.

Green tea drinking habits and gastric cancer in southwest China.

AIM: The purpose of this study was to investigate relationships between green tea consumption and gastric cancer development. METHODS: A population-based case-control study including 200 cases and 200 controls was conducted in the southwest area of China from May 2010 to February 2011. A self-designed questionnaire was used to collect data on factors influencing gastric cancer development, including tea drinking, conditional logistic regression being used to calculate odds ratios (ORs) and corresponding 95% confidence intervals (95% CIs). RESULTS: Cases with higher economic status had a reduced risk of gastric cancer, while those with cancer family history, drinking and smoking showed increased risk. Hot and very hot tea temperature was significantly related to high risk of gastric cancer with ORs (95%Cl) of 1.82 (1.03-3.52) and 3.07 (1.78-7.36), respectively. Further analysis indicated elevated risk of gastric cancer in former drinkers, former smokers and current drinkers when the measured tea temperature was hot. CONCLUSION: Drinking tea at high temperature increases the gastric cancer risk, especially in drinkers and smokers.

ICAT as a potential enhancer of monocytic differentiation: implications from the comparative proteome analysis of the HL60 cell line stimulated by all-trans retinoic acid and NSC67657.

A novel sterol mesylate compound (NSC67657) was recently identified and reported by National Cancer Institute that could efficiently induce the differentiation of HL60 cells into monocytes in vitro and in vivo. The expression of many proteins would have been changed during the differentiation process, and some proteins may have played key roles in the differentiation of HL60 cell line induced by this drug. Therefore, we treated HL60 cells with NSC67657 and all-trans retinoic acid (ATRA) to identify the differentially expressed proteins and determine their functions in cellular differentiation. Of the 45 differentially expressed protein spots investigated, 24 were either elevated or decreased in both the monocytic and granulocytic differentiating HL60 cells, 8 showed significant changes only when induced by NSC67657, and 13 showed significant changes only when induced by ATRA. After verification by RT-PCR, Western blotting, and immunocytochemistry, only the protein ICAT was found to be elevated by NSC67657 treatment alone. Although the over-expression of ICAT is not sufficient to induce the differentiation of HL60 cells into monocytes, it did increase the proportion of CD14+ cells in cells pretreated with NSC67657. Successful application of multiple techniques including two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization time-of-flight mass spectrometry, Western blotting, and eukaryotic electroporation revealed that proteomic and molecular biological analyses provide valuable tools in drug development research.