Scutellarin ameliorates diabetic nephropathy via TGF-ß1 signaling pathway.

Breviscapine, a natural flavonoid mixture derived from the traditional Chinese herb Erigeron breviscapus (Vant.) Hand-Mazz, has demonstrated a promising potential in improving diabetic nephropathy (DN). However, the specific active constituent(s) responsible for its therapeutic effects and the underlying pharmacological mechanisms remain unclear. In this study, we aimed to investigate the impact of scutellarin, a constituent of breviscapine, on streptozotocin-induced diabetic nephropathy and elucidate its pharmacological mechanism(s). Our findings demonstrate that scutellarin effectively ameliorates various features of DN in vivo, including proteinuria, glomerular expansion, mesangial matrix accumulation, renal fibrosis, and podocyte injury. Mechanistically, scutellarin appears to exert its beneficial effects through modulation of the TGF-ß1 signaling pathway, as well as its interaction with the Erk and Wnt/ß-catenin pathways.

The Concordant Disruption of B7/CD28 Immune Regulators Predicts the Prognosis of Oral Carcinomas.

Immune modulation is a critical factor in determining the survival of patients with malignancies, including those with oral squamous cell carcinoma (OSCC) and head and neck SCC (HNSCC). Immune escape or stimulation may be driven by the B7/CD28 family and other checkpoint molecules, forming ligand-receptor complexes with immune cells in the tumor microenvironment. Since the members of B7/CD28 can functionally compensate for or counteract each other, the concomitant disruption of multiple members of B7/CD28 in OSCC or HNSCC pathogenesis remains elusive. Transcriptome analysis was performed on 54 OSCC tumors and 28 paired normal oral tissue samples. Upregulation of CD80, CD86, PD-L1, PD-L2, CD276, VTCN1, and CTLA4 and downregulation of L-ICOS in OSCC relative to the control were noted. Concordance in the expression of CD80, CD86, PD-L1, PD-L2, and L-ICOS with CD28 members was observed across tumors. Lower ICOS expression indicated a worse prognosis in late-stage tumors. Moreover, tumors harboring higher PD-L1/ICOS, PD-L2/ICOS, or CD276/ICOS expression ratios had a worse prognosis. The survival of node-positive patients was further worsened in tumors exhibiting higher ratios between PD-L1, PD-L2, or CD276 and ICOS. Alterations in T cell, macrophage, myeloid dendritic cell, and mast cell populations in tumors relative to controls were found. Decreased memory B cells, CD8+ T cells, and Tregs, together with increased resting NK cells and M0 macrophages, occurred in tumors with a worse prognosis. This study confirmed frequent upregulation and eminent co-disruption of B7/CD28 members in OSCC tumors. The ratio between PD-L2 and ICOS is a promising survival predictor in node-positive HNSCC patients.

Obesity is negatively associated with dental caries among children and adolescents in Huizhou: a cross-sectional study.

BACKGROUND: Obesity and dental caries among children and adolescents are growing worldwide public health problems. They share some common and modifiable influences. The objective of this study was to evaluate the prevalence of obesity and dental caries among children and adolescents in Huizhou and explore the association between Body Mass Index (BMI) category and dental caries. METHODS: This cross-sectional study enrolled 105,181 students (55,500 males and 49,681 females) from 87 schools in Huizhou. Height and weight were measured, and BMI was calculated. Based on Chinese BMI standards, students were classified into underweight, normal weight, overweight, and obesity groups. Dental caries was diagnosed according to criteria recommended by World Health Organization (WHO). We used the Chi-square test to compare proportions of groups and performed Association Rules Mining to identify patterns and combinations of BMI categories and dental caries. Finally, a multilevel logistic regression model was applied to analyze the association between BMI category and dental caries when confounders were considered. RESULTS: The prevalence of underweight, overweight, and obesity among children and adolescents was 7.56%, 8.85%, and 2.95%, respectively. The overall prevalence of dental caries was 58.10%, with a lower prevalence among boys than girls. Students from primary schools and remote towns more easily suffer from dental caries. Caries prevalence of students belonged to underweight, normal, overweight, and obesity was 65.6%, 58.8%, 49.6%, and 46.1% individually. With increasing BMI levels, the prevalence of dental caries decreased. Further, this trend still exists in each subgroup of gender, educational stage, school type, and area. Association rules indicate underweight has a positive effect on the occurrence of dental caries, while overweight or obesity has a negative impact on the occurrence of dental caries. The three-level logistic regression model results show that BMI category is inversely associated with dental caries after adjusting confounders. CONCLUSION: Obesity is negatively associated with dental caries among children and adolescents in Huizhou. Further research is required to investigate how dietary habits, oral hygiene habits, and parental socioeconomic status mediate the association between BMI and dental caries.

Composition and Antithermal Quenching of Noninteger Stoichiometric Eu2+-Doped Na-ß-Alumina with Cyan Emission for Near-UV WLEDs.

Phosphors with high quantum efficiency and thermal stability play a key role in improving the performance of phosphor-converted white light-emitting diodes (pc-WLEDs). A near-UV-pumped LED shows a great advantage due to its reduction of the negative effect of blue light on human health. In this work, we propose a series of near-UV excitable cyan-emitting Eu2+-activated phosphors with a nominal composition of Na2-2xAl11O17+a:xEu2+ (x = 0.01-0.40), which crystallize in a sodium ß-alumina phase with a composition close to Na1.22Al11O17.11. An excess amount of the sodium carbonate raw material makes up the volatile Na during the high-temperature process. The noninteger stoichiometric composition promotes the rigidity of the crystal structure with a slight excess of Na insertion into layers between spinel blocks of the NaAl11O17 matrix. The nonequivalent substitution of Na+ by Eu2+ generates intrinsic defects acting as carrier traps. As a result, the phosphor with an optimal nominal composition Na1.6Al11O17+a:0.20Eu2+, under the excitation at 365 nm, shows an asymmetric cyan emission band at 468 nm with internal and external quantum efficiencies of 81.3 and 56.9%, respectively. Remarkably, the phosphor exhibits antithermal quenching within 200 °C. A pc-WLED with a high color rendering index (87.2) suggests great potential of the phosphor in pc-WLEDs. Therefore, a combination of a rigid structure and deep trap level is an effective way in exploring new phosphors with high quantum efficiency and thermal stability.

LncRNA MIR31HG Drives Oncogenicity by Inhibiting the Limb-Bud and Heart Development Gene (LBH) during Oral Carcinoma.

The miR-31 host gene (MIR31HG) encodes a long non-coding RNA (LncRNA) that harbors miR-31 in its intron 2; miR-31 promotes malignant neoplastic progression. Overexpression of MIR31HG and of miR-31 occurs during oral squamous cell carcinoma (OSCC). However, the downstream effectors modulated by MIR31HG during OSCC pathogenesis remain unclear. The present study identifies up-regulation of MIR31HG expression during the potentially premalignant disorder stage of oral carcinogenesis. The potential of MIR31HG to enhance oncogenicity and to activate Wnt and FAK was identified when there was exogenous MIR31HG expression in OSCC cells. Furthermore, OSCC cell subclones with MIR31HG deleted were established using a Crispr/Cas9 strategy. RNA sequencing data obtained from cells expressing MIR31HG, cells with MIR31HG deleted and cells with miR-31 deleted identified 17 candidate genes that seem to be modulated by MIR31HG in OSCC cells. A TCGA database algorithm pinpointed MMP1, BMP2 and Limb-Bud and Heart development (LBH) as effector genes controlled by MIR31HG during OSCC. Exogenous LBH expression decreases tumor cell invasiveness, while knockdown of LBH reverses the oncogenic suppression present in MIR31HG deletion subclones. The study provides novel insights demonstrating the contribution of the MIR31HG-LBH cascade to oral carcinogenesis.

Photoinduced NO and HNO Production from Mononuclear {FeNO}6 Complex Bearing a Pendant Thiol.

Light triggers the formation of HNO from a metal-nitrosyl species, facilitated by an intramolecular pendant thiol proton. Two {FeNO}6 complexes (the Enemark-Felthan notation), [Fe(NO)(TMSPS2)(TMSPS2H)] (1, TMSPS2H2 = 2,2'-dimercapto-3,3'-bis(trimethylsilyl)diphenyl)phenylphosphine; H is a dissociable proton) with a pendant thiol and [Fe(NO)(TMSPS2)(TMSPS2CH3)] (2) bearing a pendant thioether, are spectroscopically and structurally characterized. Both complexes are highly sensitive to visible light. Upon photolysis, complex 2 undergoes NO dissociation to yield a mononuclear Fe(III) complex, [Fe(TMSPS2)(TMSPS2CH3)] (3). In contrast, the pendant SH of 1 can act as a trap for the departing NO radical upon irradiation, resulting in the formation of an intermediate A with an intramolecular [SH···ON-Fe] interaction. As suggested by computational results (density functional theory), the NO stretching frequency (νNO) is sensitive to the intramolecular interaction between the pendant ligand and the iron-bound NO, and a shift of νNO from 1833 (1) to 1823 cm-1 (A) is observed experimentally. Subsequent photolysis of the intermediate A results in HNO production and a thiyl group that then coordinates to the Fe center for the formation of [Fe(TMSPS2)2] (4). In contrast with the common acid-base coupling pathway, the HNO is not voluntarily yielded from 1 but rather is generated by the photopromoted pathway. The photogenerated HNO can further react with [MnIII(TMSPS3)(DABCO)] (TMSPS3H3 = (2,2'2''-trimercapto-3,3',3''-tris(trimethylsilyl)triphenylphosphine; DABCO = 1,4-diazabicyclo[2.2.2]octane) in organic media to yield anionic [Mn(NO)(TMSPS3)]- (5-) with a {MnNO}6 electronic configuration, whereas [MnIII(TMSPS3)(DABCO)] reacts with NO gas for the formation of a {MnNO}5 species, [Mn(NO)(TMSPS3)] (6). Effective differentiation of the formation of HNO from complex 1 with the pendant SH versus NO from 2 with the pendant SMe is achieved by the employment of [MnIII(TMSPS3)(DABCO)].

Scutellarin ameliorates diabetic nephropathy via TGF-ß1 signaling pathway.

Breviscapine, a natural flavonoid mixture derived from the traditional Chinese herb Erigeron breviscapus (Vant.) Hand-Mazz, has demonstrated a promising potential in improving diabetic nephropathy (DN). However, the specific active constituent(s) responsible for its therapeutic effects and the underlying pharmacological mechanisms remain unclear. In this study, we aimed to investigate the impact of scutellarin, a constituent of breviscapine, on streptozotocin-induced diabetic nephropathy and elucidate its pharmacological mechanism(s). Our findings demonstrate that scutellarin effectively ameliorates various features of DN in vivo, including proteinuria, glomerular expansion, mesangial matrix accumulation, renal fibrosis, and podocyte injury. Mechanistically, scutellarin appears to exert its beneficial effects through modulation of the transforming growth factor-ß1 (TGF-ß1) signaling pathway, as well as its interaction with the extracellular signal-regulated kinase (Erk) and Wnt/ß-catenin pathways.

The expression of immune co-stimulators as a prognostic predictor of head and neck squamous cell carcinomas and oral squamous cell carcinomas.

Background/purpose: T cells require second immune checkpoint molecules for activation and immune memory after antigen presentation. We found that inducible co-stimulator (ICOS) has been a favorable prognostic factor amongst B7 immune checkpoint co-stimulators (ICSs) families in head and neck squamous cell carcinoma (HNSCC) and oral SCC (OSCC). Materials and methods: This study analyzed the expression of non-B7 tumor necrosis factor (TNF) superfamily ICSs in the Cancer Genome Atlas (TCGA) HNSCC cohort, our OSCC cohort, and TCGA pan-cancer datasets. The correlation in expression, prognosis, and immune status was assessed. Results: The higher expression of CD27, CD30, CD40L, death domain 3 (DR3), and OX40, presumably on the T cell surface, defined better overall survival of HNSCC patients. Besides, CD27, CD30, CD40L, and OX40 were highly correlated with ICOS expression in tumors. CD27, CD40L, and DR3 expression are higher in HPV+ HNSCC tumors than in HPV- tumors. The combined expression level of CD27/OX40 or CD27/CD40L/OX40 enables the potent survival prediction of small, less nodal involvement, early stage, and HPV + tumor subsets. Tumors expressing high CD27, CD30, CD40L, ICOS, and OX40 exhibited enhanced immune cell infiltration. The high correlation in the expression of these ICSs was also noted in the vast majority of tumor types in TCGA datasets. Conclusion: The findings of this study not only confirm the potential of the concordant stimulation of CD27, CD30, CD40L, ICOS, and OX40 as a crucial strategy in cancer immunotherapy but also inspire further exploration into the field, highlighting the promising future of cancer treatment.

Free Triiodothyronine Is Independently Associated with Nonalcoholic Fatty Liver Disease in Hospitalized Type 2 Diabetes Mellitus Patients.

OBJECTIVE: Free triiodothyronine (FT3) is an independent risk factor for nonalcoholic fatty liver disease (NAFLD) in patients with euthyroid. However, whether FT3 has an independent effect on NAFLD in a population of type 2 diabetes remains unknown. The purpose of this study was to identify the potential role of FT3 in NAFLD with T2DM. DESIGN: A cross-sectional study. Patient. A total of 859 T2DM patients who met the inclusion criteria were included. There were 506 T2DM patients without NAFLD and 353 T2DM patients with NAFLD. METHODS: The independent samples t-test or Wilcoxon rank sum test were used for continuous variables of different distribution types, while the chi-square test was used for categorical variables. Pearson correlation analysis and linear regression were used to analyze the correlation between FT3 and clinical measurements and biochemical indicators. Multivariate logistic regression analysis was used to determine independent predictors. RESULTS: Patients with NAFLD had higher BMI, SBP, and DBP, longer duration of T2DM, and higher islet function index, blood glucose index, liver function index, renal function index, blood lipid index, and FT3. We also found that FT3 was affected by other five indicators, including ALT, CR, GGT, TC, and LDL-C only in the NAFLD group but not in the non-NAFLD group. FT3 was significantly associated with NAFLD in T2DM patients, and the prevalence of NAFLD increased gradually from the lowest FT3 tertile to the highest FT3 tertile (P for trend < 0.001). CONCLUSION: FT3 is independently associated with NAFLD in hospitalized T2DM patients after rigorous adjustment for various metabolic parameters.

Transient receptor potential vanilloid 4 channels as therapeutic targets in diabetes and diabetes-related complications.

With an estimated 425 million diabetes patients worldwide in 2019, type 2 diabetes has reached a pandemic proportion and represents a major unmet medical need. A key determinant of the development and progression of type 2 diabetes is pancreatic -cell dysfunction, including the loss of cell mass, the impairment of insulin biosynthesis and inadequate exocytosis. Recent studies have shown that transient receptor potential vanilloid 4 (TRPV4), a Ca2+ -permeable non-selective cation channel, is involved in -cell replication, insulin production and secretion. TRPV4 agonists have insulinotropic activity in pancreatic -cell lines, but the prolonged activation of TRPV4 leads to -cell dysfunction and death. In addition, TRPV4 is involved in a wide variety of pathophysiological activities, and has been reported to play an important role in diabetes-related complications, such as obesity, cardiovascular diseases, diabetic retinopathy, nephropathy and neuropathy. In a rodent type 2 diabetes model, Trpv4 agonists promote vasodilation and improve cardiovascular function, whereas Trpv4 antagonists reduce high-fat diet-induced obesity, insulin resistance, diabetic nephropathy, retinopathy and neuropathy. These findings raise interest in using TRPV4 as a therapeutic target for type 2 diabetes. In this review, we intend to summarize the latest findings regarding the role of TRPV4 in diabetes as well as diabetes-related conditions, and to evaluate its potential as a therapeutic target for diabetes and diabetes-related diseases.

Research Status of Differentially Expressed Noncoding RNAs in Type 2 Diabetes Patients.

AIMS: Noncoding RNAs (ncRNAs) play an important role in the occurrence and development of type 2 diabetes mellitus (T2DM). This paper summarized the current evidences of the involvement microRNAs, long noncoding RNAs (lncRNAs), and circular RNAs (circRNAs) in the differential expressions and their interaction with each other in T2DM. METHODS: The differentially expressed miRNAs, lncRNAs, and circRNAs in the blood circulation (plasma, serum, whole blood, and peripheral blood mononuclear cells) of patients with T2DM were found in PubMed, GCBI, and other databases. The interactions between ncRNAs were predicted based on the MiRWalk and the DIANA Tools databases. The indirect and direct target genes of lncRNAs and circRNAs were predicted based on the starBase V2.0, DIANA Tools, and LncRNA-Target databases. Then, GO and KEGG analysis on all miRNA, lncRNA, and circRNA target genes was performed using the mirPath and Cluster Profile software package in R language. The lncRNA-miRNA and circRNA-miRNA interaction diagram was constructed with Cytoscape. The aim of this investigation was to construct a mechanism diagram of lncRNA involved in the regulation of target genes on insulin signaling pathways and AGE-RAGE signaling pathways of diabetic complications. RESULTS: A total of 317 RNAs, 283 miRNAs, and 20 lncRNAs and circRNAs were found in the circulation of T2DM. Dysregulated microRNAs and lncRNAs were found to be involved in signals related to metabolic disturbances, insulin signaling, and AGE-RAGE signaling in T2DM. In addition, lncRNAs participate in the regulation of key genes in the insulin signaling and AGE-RAGE signaling pathways through microRNAs, which leads to insulin resistance and diabetic vascular complications. CONCLUSION: Noncoding RNAs participate in the occurrence and development of type 2 diabetes and lead to its vascular complications by regulating different signaling pathways.

Use of Low-Dose Recombinant Factor â ¦a for Uncontrolled Perioperative Bleeding.

BACKGROUND: Recombinant activated factor VIIa (rFVIIa) is a prohemostatic agent initially approved for use in hemophilia patients and has also been used for a diverse range of off-label indications in the context of massive uncontrolled blood loss; however, no convincing evidence exists regarding the optimal dose of rFVIIa to treat uncontrolled bleeding in surgical patients. AIM: To evaluate the effects and safety of a very low dose of rFⅦa in patients with uncontrolled perioperative bleeding in the surgical intensive care unit (ICU). METHODS: 55 patients from Beijing Hospital, who received rFⅦa between July 2004 and November 2018 for uncontrolled perioperative bleeding were included. The controls were matched for age, sex, severity, and operation type. The baseline demographics, survival, changes in bleeding and transfusion, coagulation parameters and complications were analyzed. RESULTS: A low dose of rFⅦa (2.0∼3.6 mg, with a median dose of 39.02 µg/kg) appears to be effective in controlling massive hemorrhage (with an effective rate of 74.55%), and can reduce volume of red blood cell transfusion, improve coagulation status, while has a relatively low risk of thromboembolic complications (3.6%). CONCLUSION: In patients with uncontrolled perioperative bleeding, a low dose of rFⅦa could be used when traditional methods are ineffective.

Identification of alternative splicing and lncRNA genes in pathogenesis of small cell lung cancer based on their RNA sequencing.

BACKGROUND: The molecular mechanisms involved in small-cell lung cancer (SCLC) are largely unknown. Recent studies have suggested that long non-coding RNAs (lncRNAs) are likely to play a critical role. OBJECTIVES: There is an urgent need for suitable molecular biomarkers for SCLC diagnosis and for assessing patient prognosis. MATERIAL AND METHODS: In this study, we used public databases to identify mRNA-like candidate lncRNAs. A multi-step computational approach was used to construct a functional SCLC lncRNAs-mediated competing with endogenous RNA (ceRNA) network (LMCN) by integrating genome-wide lncRNAs and mRNA expression profiles, miRNA-target interactions, functional analyses, and clinical survival analyses. RESULTS: The results revealed the significance of lncRNAs interactions with ceRNAs in SCLC, indicating that integration of expression profiles and alternative splicing could be used to identify biomarkers and the underlying pathological changes. The following genes: EPB41L4A-AS1, HOXA-AS2, XIST, DLEU2, FGD5-AS1, ALMS1-IT1, SNHG12, MIR17HG, MIR4720, and SCARNA10 in cluster, as well as shared alternative splicing events, were considered to be critical genes. CONCLUSIONS: Olfactory transduction and endocytosis were the top-enriched pathways in SCLC. The selected cluster, including critical genes, might also be a potential pathway of SCLC pathogenesis. As a result, this research provides the perspective information to explore the potential critical genes and its pathways in SCLC therapy.

Study on the association between vitamin D receptor gene fokI (T/C) polymorphisms and the susceptibility to type 2 diabetic kidney disease of Han nationality in south of China.

AIMS: To investigate the distribution of vitamin D receptor fokI gene polymorphism in Yunnan Han population, and to explore the relationship between SNP of fokI and type 2 diabetic kidney disease. METHODS: We included 276 individuals of Han population of Yunnan in this study: 91 type 2 diabetes patients without kidney disease (DM group), their duration of diabetes is more than 10 years, 89 type 2 diabetes patients with diabetic kidney disease (DKD group), their duration of diabetes is less than 10 years and 96 healthy controls (NC group). We compared the concentration of 25 hydroxy vitamin D in different groups and used taqman probe to detect the genotype and allele of fokI, then analysed the relationship between the polymorphisms of fokI and the susceptibility of diabetic kidney disease. RESULTS: (1) NC group had a significantly higher plasma concentrations of 25 (OH) D than DKD group and DM group (P < 0.01); (2) 25 (OH) D and age, BMI, HbA1c, TG showed a weak negative correlation (P < 0.01); (3) Genotype of fokI showed no differences in DM group and DKD group, same as in DM group and NC group; FF genotype in DKD group is relatively lower than NC group (P < 0.05), and there is no difference in Ff and ff genotype (P > 0.05); In DKD group, f allele was 53.4%, higher than DM group (RR = 1.46, P < 0.05); (4) Logistic regression analysis showed that ff genotype may be a susceptible factor for DKD (P = 0.04, OR = 2.37). CONCLUSION: FokI Ff genotype accounted for a larger proportion of the Han population in Yunnan, and ff genotype may be a susceptible factor for DKD in Yunnan Han population.

Interleukin-1ß-mediated suppression of microRNA-101 and upregulation of enhancer of zeste homolog 2 is involved in particle-induced lung cancer.

Lung cancer may be a result of complex factors. Small mineral particle is the well-known inducer of lung cancer. Previous study revealed the high morbidity of lung cancer in Xuan Wei in China, and the main cause of lung cancer is the use of smoky coal there. And it is generally accepted that chronic inflammation induced by small mineral particle may be a cause of lung cancer. But the relationship between chronic lung inflammation and lung cancer is largely unknown. In the present study, we found that silica particle was able to induce the secretion of interleukin-1ß from a Xuan Wei lung cancer cell line, XWLC-05. At the same time, microRNA-101 (mir-101) was found to be downregulated by the treatment of silica particle. Interestingly, the interleukin 1 receptor antagonist and interleukin-1ß antibody can reduce silica particle-induced downregulation of mir-101. Twenty-four Xuan Wei lung tumor tissues were collected to detect the expression level of mir-101 and enhancer of zeste homolog 2 (EZH2), which is the potential target of mir-101. The results showed that mir-101 was down-regulated and EZH2 were upregulated. Subsequently, the roles of mir-101 and EZH2 in tumor growth and progression in vitro were tested. Overexpression of mir-101 mimics was able to suppress the expression of EZH2 in XWLC-05 cells. And this resulted in the inhibited tumor cell growth and attenuated cell migration. The results in the present study showed that particle can induce the secretion of interleukin-1ß. Interleukin-1ß subsequently induces the downregulation of mir-101, which may result in the upregulated level of EZH2, and occurrence of lung cancer. We for the first time proposed the role interleukin-1ß-mir-101-EZH2 axes in the particle-induced lung cancer. Further study may be needed to decipher the detailed mechanism involved.

Identification of low abundance proteins in colorectal cancer tissues.

BACKGROUND: Detection of low abundance proteins always possesses challenges even with the currently available proteomics technologies. OBJECTIVE: We aimed to evaluate the low abundance differentially expressed proteins in colorectal cancerous tissues as compared to colorectal normal tissues. METHODOLOGY: Protein separation was carried out by using the coupling methods of hydroxyapatite chromatography and SDS-PAGE followed by mass spectrometry analysis. RESULTS: Five of the membrane associated low abundance proteins, namely uncharacterized protein C5orf4, coiled-coil domain containing protein 152, DnaJ homolog subfamily C member 22, tumor suppressor candidate 3 and leucine-rich repeat transmembrane neuronal protein 4 that previously only reported at the genome level were identified. CONCLUSIONS: The proteins were isolated from the cancerous tissues as either up-regulated or down-regulated proteins as compared to normal tissues and therefore may play important roles in cancer development.

Analysis of differentially expressed proteins in colorectal cancer using hydroxyapatite column and SDS-PAGE.

Limitation on two dimensional (2D) gel electrophoresis technique causes some proteins to be under presented, especially the extreme acidic, basic, or membrane proteins. To overcome the limitation of 2D electrophoresis, an analysis method was developed for identification of differentially expressed proteins in normal and cancerous colonic tissues using self-pack hydroxyapatite (HA) column. Normal and cancerous colon tissues were homogenized and proteins were extracted using sodium phosphate buffer at pH 6.8. Protein concentration was determined and the proteins were loaded unto the HA column. HA column reduced the complexity of proteins mixture by fractionating the proteins according to their ionic strength. Further protein separation was accomplished by a simple and cost effective sodium dodecyl sulfate-polyacrylamide gel electrophoresis method. The protein bands were subjected to in-gel digestion and protein analysis was performed using electrospray ionization (ESI) ion trap mass spectrometer. There were 17 upregulated proteins and seven downregulated proteins detected with significant differential expression. Some of these proteins were low abundant proteins or proteins with extreme pH that were usually under presented in 2D gel analysis. We have identified brain mitochondrial carrier protein 1, T-cell surface glycoprotein CD1a, SOSS complex subunit B2, and Protein Jade 1 which were previously not detected in 2D gel analysis method.