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With the opening of the second-child policy and the increase of cesarean section rate,the incidence of surgical site infection after cesarean section also increases,which seriously affects the physical recovery after delivery.In this paper,the types of infection bacteria,high risk factors,prevention,diagnosis and treatment measures are comprehensively expounded.It is emphasized that early identification of risk factors and establishment of effective prevention and treatment measures are essential to reducing the incidence and mortality of incision infection in cesarean section.
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<p><b>OBJECTIVE</b>To express and purify the mouse endothelial cell-targeted recombinant Notch ligand protein mD1R, and to investigate its effect on hematopoiesis after carbon tetrachloride damage.</p><p><b>METHODS</b>PCR was performed to clone and construct the expression vector pET22b(+)-mD1R. The mD1R successfully transformed into E. coli was induced by IPTG, and purified with Ni-beads affinity chromatography. The target protein was detected by SDS-PAGE. The fluorescence-activated cell sorting analysis (FACS), cell adhesion test, immunofluorescence staining and quantitative real-time PCR were employed to detect the endothelial cell-targeted and Notch signaling-activated biological characteristics of mD1R. The carbon tetrachloride mouse model was established to observe the effects of mD1R on the hematopoietic stem cell (HSC), myeloid cells and lymphoid cells by flow cytometry. The LinScal-1c-Kit cells were sorted by magnetic bead, FACS was performed to analyze the cell cycle, and RT-PCR was employed to observe the expression of interleukin (IL)-10.</p><p><b>RESULTS</b>The prokaryotic expression vector was successfully cloned and constructed. The purity and the activity were confirmed in mD1R recombinant protein. The purified mD1R activated the Notch signaling pathway of hematopoietic stem cells in carbon tetrachloride damaged mouse, and internally elevated the number of HSC and long-term HSC to 2.96-fold and 6.18-fold. In addition, mD1R improved the amplification of the myeloid progenitor cells and the myeloid-derived suppressor cells, particularly the granulocyte/monocyte into blood. Mechanistically, the further analyses suggested that Notch pathway could increase the proliferation of HSC and enhance expression of IL-10 after stress injury.</p><p><b>CONCLUSIONS</b>A new and activated recombinant Notch ligand protein has been obtained successfully to communicate hematopoietic stem cells and hematopoietic microenvironment. The Notch- mediated intrinsic hematopoiesis has been regulated by the anti-inflammatory factor after stress injury.</p>
Subject(s)
Animals , Mice , Carbon Tetrachloride , Escherichia coli , Hematopoiesis , Hematopoietic Stem Cells , Ligands , Receptors, Notch , Signal TransductionABSTRACT
Objective:To investigate the value of autoantibodies and serum levels of IgG4 and CA19-9 in the diagnosis of IgG4 associated cholangitis (IgG4-SC).Methods:Detect the serum IgG4 and CA19-9 of 41 clinical cases of IgG4-SC patients,162 clinical cases of non IgG4-SC patients and 40 healthy human serum samples by immunoassay and direct chemiluminescence methods, also detect the antinuclear antibodies (ANA),anti neutrophil antibody (ANCA),anti smooth muscle antibody (SMA) and anti mitochondrial antibody (AMA) of the above serum samples by indirect immunofluorescence and analyze the detection results.Results:①The positive rates of ANA,ANCA,SMA and AMA in patients with IgG4-SC were 41.46%,7.32%,0 and 2.44%.Among them,the positive rate of ANA was significantly different from that of the normal control group(P<0.01),and the positive rate of SMA and AMA was significantly different from that of non IgG4-SC group(P<0.01),and so as the positive rate of ANCA do with that of PSC group.②The number of serum IgG4 and CA19-9 increased samples were significantly compared with the normal control group (P<0.01);the area under the ROC curve (AUC) was 0.979 and 0.646,respectively,and P<0.05.Conclusion:The high level of serum IgG4 and CA19-9 and autoantibody detection are of great accuracy and important clinical value in the differential diagnosis of IgG4-SC.
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Objective:To explore the clinical value of detecting serum 14-3-3η and auto-antibodies in rheumatoid arthritis (RA),and compare their performance in RA diagnosis.Methods: Serum samples of 134 RA patients,90 non-RA inflammatory arthropathy patients,70 of whom with osteoarthritis(OA)and 20 with ankylosing spondylitis(AS),and 40 healthy controls from the second affiliated hospital of Nanchang University were collected.Concentrations of 14-3-3η,anti-CCP,anti-RA33,anti-Sa were detected with enzyme linked immunosorbent assay(ELISA),with RF detected by immunonephelometry.Diagnostic utilities of them for RA were evaluated and compared then.Results:① Serum levels of 14-3-3η,anti-CCP,anti-RA33,anti-Sa and RF were significantly higher in patients with RA than non-RA inflammatory arthropathy patients and healthy controls,the differences between groups were statistically significant;② ROC curves were conducted according to the serum levels detected.The AUC of 14-3-3η,anti-CCP,anti-RA33,anti-Sa and RF were 0.831(95% CI:0.782-0.881),0.852(95% CI:0.802-0.901),0.615(95% CI:0.546-0.684),0.706(95% CI:0.643-0.770)and 0.739(95% CI:0.676-0.802)respectively,with P values<0.01.Among all index,only anti-CCP and 14-3-3η were of moderate diagnostic value,at the threshold of 24.10 U/ml and 2.59 ng/ml individually;③anti-Sa was of highest specificity and RF was of highest sensitivity among all indexes detected;the specificity of 14-3-3η was merely moderately inferior to anti-Sa and anti-RA33,but its sensitivity was superior to them both.Conclusion:Serume14-3-3η,anti-CCP,anti-RA33,anti-Sa and RF levels increased remarkably in patients with RA,and contributed to RA diagnosis.Meanwhile,14-3-3η was advantageous,to some extent,in the sensitivity and specificity over auto-antibodies,and can be utilized as a reference index in diagnosing RA.
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Objective:To investigate the clinical significance of bone metabolism indexes and human leukocyte antigen B27 (HLA-B27)in ankylosing spondylitis(AS)and analyze their diagnostic value. Methods:The subjects were 94 cases of AS patients,239 cases with other diseases and 80 healthy controls, and the results were retrospectively surveyed. Results:In the AS group, the concentration of the β-collagen specific sequence (β-CTX ) was higher ( P<0. 05 ) while the concentrations of osteocalcin ( OC ) , 25-hydroxyvitamin D[25-(OH)D] and parathyroid hormone(PTH)were lower than those in the control group. In the AS group,there was a positive correlation between the concentration of C-reactive protein(CRP)and β-CTX(P<0. 01),and the concentration of CRP was negatively correlated with the concentration of 25-(OH)D(P<0. 05). However,the concentrations of other bone metabolic indexes had no correlation with the concentration of CRP(P<0. 05). The positive rate of HLA-B27 in the AS group was significantly higher than that in other groups(P<0. 05),and HLA-B27 had highly sensitive and specific in the diagnosis of AS. In the AS group,the sensitivity of the concentration of 25-( OH) D was higher than β-CTX,while its specificity was lower than β-CTX. Conclusion:Bone metabolic indexes have great value in early screening and clinical diagnosis of AS,especially β-CTX and 25-( OH) D were more obvious.
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Objective:To investigate the clinical significance of bone metabolism indexes and human leukocyte antigen B27 (HLA-B27)in ankylosing spondylitis(AS)and analyze their diagnostic value. Methods:The subjects were 94 cases of AS patients,239 cases with other diseases and 80 healthy controls, and the results were retrospectively surveyed. Results:In the AS group, the concentration of the β-collagen specific sequence (β-CTX ) was higher ( P<0. 05 ) while the concentrations of osteocalcin ( OC ) , 25-hydroxyvitamin D[25-(OH)D] and parathyroid hormone(PTH)were lower than those in the control group. In the AS group,there was a positive correlation between the concentration of C-reactive protein(CRP)and β-CTX(P<0. 01),and the concentration of CRP was negatively correlated with the concentration of 25-(OH)D(P<0. 05). However,the concentrations of other bone metabolic indexes had no correlation with the concentration of CRP(P<0. 05). The positive rate of HLA-B27 in the AS group was significantly higher than that in other groups(P<0. 05),and HLA-B27 had highly sensitive and specific in the diagnosis of AS. In the AS group,the sensitivity of the concentration of 25-( OH) D was higher than β-CTX,while its specificity was lower than β-CTX. Conclusion:Bone metabolic indexes have great value in early screening and clinical diagnosis of AS,especially β-CTX and 25-( OH) D were more obvious.
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<p><b>OBJECTIVE</b>To investigate the difference in urinary proteome between patients with bladder urothelial carcinoma (BUC) and healthy volunteers and to provide a basis for the early diagnosis of BUC.</p><p><b>METHODS</b>The urine samples from BUC patients and healthy volunteers (controls) were treated by 25% ethanol precipitation and two-dimensional gel electrophoresis (2-DE), and the obtained urinary proteins were subjected to Coomassie brilliant blue staining and analysis by PDQuest 8.0 (2-DE image analysis software); the differentially expressed proteins were sequenced by matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry and identified using the Swiss-Prot database; the differential expression of these proteins was verified by western blot.</p><p><b>RESULTS</b>High-resolution and high-reproducibility 2-DE images were obtained from the urine samples of BUC patients and controls, with 789 ± 18 and 762 ± 14 protein spots, respectively. Compared with the control group, the BUC grouP had significantly decreased expression of 6 protein spots and significantly increased expression of 11 protein spots. The mass spectrometry revealed five proteins with increased expression in the BUC group, including fibrinogen, lactate dehydrogenase B, apolipoprotein A1, clusterin, and haptoglobin, and the results were confirmed by western blot.</p><p><b>CONCLUSION</b>There is significant difference in urinary proteome between BUC patients and healthy volunteers; the identification of differentially expressed proteins in urine lays the foundation for identifying potential molecular markers in early diagnosis of BUC.</p>
Subject(s)
Aged , Female , Humans , Male , Middle Aged , Case-Control Studies , Early Detection of Cancer , Proteomics , Methods , Urinary Bladder Neoplasms , Diagnosis , UrineABSTRACT
<p><b>OBJECTIVE</b>To observe the effects of complement fragment C3f on expression and secretion of collagen I, III and transforming growth factor( TGF)-beta1 in human embryonic lung fibroblast (MRC-5) cells.</p><p><b>METHODS</b>MRC-5 cells were cultured with C3f (the synthetic 17 peptides fragments of complement C3). The extracellular and intracellular expression levels of type I, III collagens and TGF-beta1 in MRC-5 cultures were detected by ELISA and immunohistochemistry, respectively.</p><p><b>RESULTS</b>The expression levels of type I, III collagen and TGF-beta1 in the supernatant of MRC-5 cultures decreased significantly with the concentrations of C3f as compared with controls (P < 0.05). Also the expression level of TGF-beta1 in MRC-5 cytoplasm reduced significantly as compared with controls (P < 0.05).</p><p><b>CONCLUSION</b>The results of present in vitro study showed that the complement fragment C3f could reduce the formation of TGF-beta1 and type I, III collagens in MRC-5 cells, and inhibit the lung tissue fibrosis.</p>
Subject(s)
Humans , Cell Line , Collagen Type I , Metabolism , Collagen Type III , Metabolism , Complement C3b , Pharmacology , Fibroblasts , Metabolism , Lung , Cell Biology , Embryology , Transforming Growth Factor beta1 , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To explore the correlation between the exposure levels and serum protein fingerprints in population exposed to silica.</p><p><b>METHODS</b>Liquid chip time-of-flight mass spectrometry technology was used to investigate the serum profiles in control group (30 cases), group exposed to silica (30 cases), silicosis group (I stage, 25 cases) and suspected silicosis group (30 cases), and screen the differential expression proteins. The correlation between the levels of the differential expression proteins and the exposure levels was performed.</p><p><b>RESULTS</b>Five differential expression proteins were found among 4 groups, the expression of 5081 and 5066 proteins was upregulated, and the expression of 3954, 2021 and 1777 proteins was downregulated. There was no the correlation between the exposure levels and the peak with M/Z among those proteins.</p><p><b>CONCLUSION</b>the results of present investigation indicated there was no correlation between the exposure levels and protein/peptide peak.</p>
Subject(s)
Adult , Female , Humans , Male , Middle Aged , Blood Proteins , Case-Control Studies , Dust , Mass Spectrometry , Occupational Exposure , Peptide Mapping , Proteomics , Silicon Dioxide , Toxicity , Silicosis , BloodABSTRACT
<p><b>OBJECTIVE</b>To analyze the early expression differences of lung tissue proteins in rats exposed to silica using comparative proteomics method, to explore the effects of Chinese traditional medicine (Gymnadenia conopse alcohol extract, GcAE) on silicosis (50 mg/ml).</p><p><b>METHODS</b>Adult male Wistar rats were randomly divided into silica-treated group and GcAE-treated group, four rats a group. The rats were exposed to silica by intratracheal (IT) instillation of 1 ml silica suspension for 24 h. After exposure, the rats in GcAE-treated group were intragastric administration with 0.8 ml GcAE (0.8 ml/100 g a day) and the rats in silica-treated group were intragastric administration with 2 ml sterilized saline a day for 14 days. Then all rats were sacrificed and lung tissues were collected. The total proteins were separated by means of two-dimensional gel electrophoresis (2-DE) and the differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blotting was used to validate the expression of certain candidate proteins in lung tissues.</p><p><b>RESULTS</b>Obvious pathological changes of lung could be observed in silica-treated group, such as the thicken of interalveolar septum, which was infiltrated with lymphocytes, macrophages and a few neutrophils with the proliferation of fibroblasts and smooth muscle cells. The accumulation of collagen, the destruction of alveolus structure and the more dotted fibrosis or granuloma could also be found. However, the pathological changes of lung in GcAE-treated group were lighter than those of silica-treated group. Thirty three differentially expressed proteins were identified, including cathepsin D precursor, peroxiredoxin-1 (Prx-1) and SEC14-like protein 3. Compared with silica-treated group, cathepsin D precursor and Prx-1 were significantly downregulated in GcAE-treated group, and SEC14-like protein 3 was significantly upregulated (P < 0.01). The results of western blot indicated that the expression level of Prx-1 in GcAE-treated group was 0.26 ± 0.02, which was significantly lower than that (0.35 ± 0.04) in silica-treated group (P < 0.01).</p><p><b>CONCLUSION</b>GcAE may inhibit the progress of silicosis in the early period and cathepsin D precursor, SEC14-like protein 3 and Prx-1 may participate in this process.</p>
Subject(s)
Animals , Male , Rats , Lung , Metabolism , Orchidaceae , Plant Extracts , Pharmacology , Proteomics , Methods , Rats, Wistar , Silicosis , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effectiveness and safety of deferasirox (DFX) in the treatment of iron overload in children with β-thalassemia major.</p><p><b>METHODS</b>Twenty-four β-thalassemia major children with iron overload who received regular blood transfusion were randomly enrolled. The serum feritin (SF) levels were measured in the patients after different doses of DFX treatment. The DFX treatment-related adverse events were observed. The values of cardiac MRI T2* and liver MRI T2* were compared between the patients receiving DFX treatment for 5 years and the patients treated with deferoxamine and deferiprone.</p><p><b>RESULTS</b>The patients with iron overload did not respond to DFX at the initial dose of 20-30 mg/kg•d. However, the SF level decreased significantly after the dose of DFX increased to 30-40 mg/kg•d (U=58, P<0.01). Serum liver transaminase elevation was the most common adverse effect, followed by non-progressive elevation in serum creatinine level. The mean SF level was significantly lower (1748±481 ng/mL vs 3462±1744 ng/mL; P<0.05), in contrast, the liver MRI T2* value was significantly higher (8.5±2.9 ms vs 2.7±1.9 ms; P<0.01) in patients receiving DFX treatment for 5 years than in the controls. There were no significant differences in the cardiac MRI T2* value between the two groups.</p><p><b>CONCLUSIONS</b>DFX can reduce SF levels in a dose-dependent manner in children with β-thalassemia major. It can significantly lower liver iron overload but not cardiac overload. Serum liver transaminase elevation and non-progressive elevation in serum creatinine level are major adverse effects in DFX treatment.</p>
Subject(s)
Adolescent , Adult , Child , Child, Preschool , Female , Humans , Male , Benzoates , Therapeutic Uses , Dose-Response Relationship, Drug , Ferritins , Blood , Iron Chelating Agents , Therapeutic Uses , Iron Overload , Drug Therapy , Transfusion Reaction , Triazoles , Therapeutic Uses , beta-Thalassemia , Blood , TherapeuticsABSTRACT
This study was aimed to investigate the effect of curcumin in combination with bortezomib on the proliferation and apoptosis of human MM cell line H929 in vitro, and to explore its mechanisms. MTT assay was applied to detect the inhibitory effects of curcumin and bortezomib either alone or combined at different concentrations on H929 cells, and flow cytometry was employed to assay the apoptosis rate. In addition, RT-PCR was used to analyze the mRNA expression of gene BCL-2, BAX, cyclin D1. Immunofluorescence technique was performed to study the location changes of NF-κB P65 in different groups. The results showed that both curcumin and bortezomib inhibited the proliferation of MM cell line H929 in dose-dependent manner, and combination of these two drugs displayed synergistical effect. A much higher apoptosis rate was determined by flow cytometry in combinative groups than that in single or control group. And RT-PCR showed, as compared with curcumin or bortezomib group, there was mRNA expression decrease of BCL-2, cyclin D1 but increase of BAX in combined group. The expression of NF-κB P65 in nucleus was downregulated in either the curcumin or bortezomib group, however, distribution of NF-κB P65 in cytoplasm was observed in combined group. It is concluded that the combination of curcumin and bortezomib is much more effective for the inhibiting proliferation and promoting apoptosis of H929 cell line, which may function by inhibiting the transcription of NF-κB and apoptosis-related genes.
Subject(s)
Humans , Apoptosis , Boronic Acids , Pharmacology , Bortezomib , Cell Line, Tumor , Cell Proliferation , Curcumin , Pharmacology , Cyclin D1 , Metabolism , Drug Therapy, Combination , Multiple Myeloma , Metabolism , NF-kappa B , Metabolism , Proto-Oncogene Proteins c-bcl-2 , Metabolism , Pyrazines , Pharmacology , Transcription Factor RelA , Metabolism , bcl-2-Associated X Protein , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To analyze the differences of lung tissue proteins in rats exposed to silica early by using comparative proteomics method and investigate the related mechanism with the occurrence and development of silicosis.</p><p><b>METHODS</b>Adult male Wistar rats were randomly divided into control group and silica-treated group. The animal model was established by intratracheal (IT) instillation with silica suspension. On the 14th day after establishment of animal model, rats were sacrificed and lung tissues were collected. The total proteins were separated by means of two-dimensional gel electrophoresis (2-DE) and the differentially expressed proteins were identified by using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In addition, Western blotting was performed to verify the expression of certain candidate protein.</p><p><b>RESULTS</b>Eleven differential expression protein spots were tested by MALDI-TOF-MS, and six proteins were identified. The levels of cathepsin D precursor, peroxiredoxin-1 (Prx-1), heat shock cognate 71 000 protein (HSP7C), heterogeneous nuclear ribonucleoprotein A3 (hnRNPA3) and fatty acid-binding protein (epidermal, E-FABP) were up-regulated in silica-treated group with the optical density (A) values. These values were 116.50+/-12.56, 148.75+/-22.40; 40.00+/-1.63, 66.00+/-13.93; 51.25+/-7.37, 92.75+/-8.69; 83.00+/-6.48, 122.75+/-24.62; 50.75+/-6.50, 93.50+/-23.10 and 100.25+/-19.99, 142.50+/-21.21 respectively. The statistical difference was observed as compared with control group (t=-2.51, -3.71, -7.28, -3.12, -3.56 and -2.90, P<0.05). However, SEC14-like protein 3 with the A values 153.00+/-11.28, 109.75+/-18.32 was down-regulated (t=4.02, P<0.01). Western blotting showed that in the expression of Prx-1 was higher in silica-treated group (0.61+/-0.05) than that in the control (0.35+/-0.05) (t=-7.24, P<0.01) when calculating the semi-quantification of this protein using ratio of optical density.</p><p><b>CONCLUSION</b>2-DE pattern of lung tissue from rats exposed to silica has been established and six differentially expressed proteins have been identified. Our study is of help for further research of the mechanisms of silicosis.</p>
Subject(s)
Animals , Male , Rats , Disease Models, Animal , Electrophoresis, Gel, Two-Dimensional , Environmental Exposure , Lung , Metabolism , Pathology , Proteomics , Rats, Wistar , Silicon Dioxide , Toxicity , Silicosis , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To seek differentially expressed serum proteins in recovered SARS patients complicating avascular necrosis of femoral head (AVNFH).</p><p><b>METHODS</b>2-DE and MALDI-TOF MS were used to study the comparative serum proteomics among female SARS AVNFH group, female SARS non-AVNFH group and female healthy group. ELISA method was used to detect serum amyloid P component in individual serum; specificity and sensitivity of serum amyloid P component were analyzed.</p><p><b>RESULTS</b>Average protein points on 2-DE of 3 groups were 632 +/- 28, 671 +/- 55, 688 +/- 42 respectively, and the matching rate of protein points was ranged from 85% to 95%; eighteen differentially expressed proteins were discovered including transthyretin, serpin peptidase inhibitor, alpha-1-antitrypsin precursor, serum amyloid P components, etc. Compared to healthy group and SARS non-AVNFH group, transthyretin, C4B3, fibrinogen gamma, apolipoprotein L, apolipoprotein A-IV precursor, albumin and prealbumin showed lower expression, inversely serpin peptidase inhibitor, alpha-1-antitrypsin precursor and serum amyloid P components showed higher expression in serum in the SARS AVNFH necrosis group. The serum amyloid P component in 3 groups were 0.54 +/- 0.30 ng/ml, 0.83 +/- 0.39 ng/ml, 1.21 +/- 0.29 ng/ml respectively. The areas under the ROC curve on serum amyloid P component was 0.854, the specificity was 77.8% and the sensitivity was 85.2%.</p><p><b>CONCLUSION</b>There were differentially expressed serum proteins in three groups. Serum amyloid P components might be one of the potential biomarkers in serum of recovered SARS patients complicating avascular necrosis of femoral head.</p>
Subject(s)
Adult , Female , Humans , Blood Proteins , Case-Control Studies , Electrophoresis, Gel, Two-Dimensional , Femur Head Necrosis , Blood , Proteomics , Severe Acute Respiratory Syndrome , Blood , Spectrometry, Mass, Matrix-Assisted Laser Desorption-IonizationABSTRACT
<p><b>OBJECTIVE</b>To assess the feasibility of gray-scale contrast-enhanced ultrasound for characterizing thyroid nodules.</p><p><b>METHODS</b>Forty thyroid nodules from 35 patients were studied both by conventional techniques and gray-scale contrast-enhanced ultrasound. Of the nodules examined, 15 were benign and 25 malignant. The enhancement of echogenicity was evaluated. The diagnosis was confirmed by surgical biopsy and histopathological examination.</p><p><b>RESULTS</b>The study using gray-scale contrast-enhanced ultrasound showed absent contrast-enhancement in 9 of 25 malignant nodules and 1 of 11 benign solitary nodules; intense enhancement in 6 of 25 malignant nodules, with perfusion defect in the center; diffuse faint enhancement in 10 of 25 malignant nodules and 10 of 11 benign solitary nodules. Benign cystic nodules all showed absent enhancement in the cystic components and 2 of 4 intense enhancement in the solitary components.</p><p><b>CONCLUSIONS</b>Gray-scale contrast-enhanced ultrasonography imaging may be a useful tool for evaluating the perfusion of thyroid nodules. Solitary nodules showing absent enhancement or intense enhancement with absent enhancement in the nodular center may suggest malignant.</p>
Subject(s)
Female , Humans , Male , Contrast Media , Diagnosis, Differential , Image Enhancement , Methods , Thyroid Neoplasms , Diagnostic Imaging , Pathology , Thyroid Nodule , Diagnostic Imaging , Pathology , Ultrasonography , MethodsABSTRACT
<p><b>OBJECTIVE</b>1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is a potent inducer of differentiation in myeloid leukemia cells, but its clinical use is limited due to its hypercalcaemic effect and resistance. Carnosic acid is a plant-derived polyphenol food preservative with chemoprotective effects against carcinogens. Recent research has shown that carnosic acid potentiates the effects of 1,25(OH)2D3 on differentiation of human leukemia cells. This study examined the effects of 1,25(OH)2D3 in combination with carnosic acid on monocytic differentiation as well as intracellular reactive oxygen species (ROS) and Ca2+ levels in human leukemia HL-60 cells.</p><p><b>METHODS</b>HL-60 cells were randomly treated with 1 nmol/L 1,25(OH)2D3, 100 nmol/L 1,25(OH)2D3, 10 micromol/L carnosic acid, a combination of 1 nmol/L 1,25(OH)2D3 and 10 micromol/L carnosic acid or placebo. Cell growth was observed by MTT assay for 72 hrs at an interval of 24 hrs. Cells were harvested after 72 hrs of culture. Morphologic features of the cells were observed by microscopy. Flow cytometry was used to detect cell cycle, monocytic differentiation marker CD14 expression, and intracellular ROS and Ca2+ levels.</p><p><b>RESULTS</b>A combination of 1 nmol/L 1,25(OH)2D3 and 10 micromol/L carnosic acid resulted in greater proliferation inhibition (Ab: 0.56 0+/- 0.020 vs 1.482 +/- 0.327; P <0.01), mature monocytic features, G0 /G1 cell arrest, higher CD14 expression (57.62 +/- 0.817% vs 2.76 +/- 0.828%; P <0.01), lower intracellular ROS levels (52.67 +/- 10.76% vs 86.46 +/- 40.52%; P <0.01 and similar intracellular Ca2+ levels in HL-60 cells when compared with the placebo group. The ability of a combination of 1 nmol/L 1,25(OH)2D3 and 10 micromol/L carnosic acid to inhibit the proliferation and induce the differentiation of HL-60 cells was similar to that of 100 nmol/L 1,25(OH)2D3, while the intracellular Ca2+ level (115.64 +/- 17.74 nmol/L vs 185.75 +/- 27.38 nmol/L) was significantly lower than that in the 100 nmol/L 1,25(OH)2D3 group.</p><p><b>CONCLUSIONS</b>Low concentration of 1,25(OH)2D3 combined with 10 micromol/L carnosic acid can produce enhanced differentiation, proliferation inhibition and antioxidant effects of HL-60 cells. The combination of the two inducers dose not increases intracellular Ca2+ levels.</p>