Influence of UDP-Glucuronosyltransferase Polymorphisms on Mycophenolic Acid Metabolism in Renal Transplant Patients.

This study aimed to evaluate the effects of UDP-glucuronosyltransferase (UGT) polymorphisms on mycophenolic acid (MPA) metabolism in renal transplant patients. A total of 11 single nucleotide polymorphisms (SNPs) of UGT1A1, UGT1A7, UGT1A8, UGT1A9, UGT1A10, and UGT2B7 were genotyped in 79 renal transplant patients. The associations of SNPs and clinical factors with dose-adjusted MPA area under the plasma concentration-time curve (AUC/D), the dose-adjusted plasma concentration (C0/D) of 7-O-MPA-glucuronide (MPAG), and the dose-adjusted plasma concentration (C0/D) of acyl MPAG (AcMPAG) were analyzed. In the univariate analysis, UGT1A1 rs4148323, age, and anion gap were associated with MPA AUC/D. MPA AUC/D was higher in patients with the GA genotype of UGT1A1 rs4148323 compared to patients with the GG genotype. UGT1A1 rs4148323, UGT1A9 rs2741049 and clinical factors, including age, serum total bilirubin, adenosine deaminase, anion gap, urea, and creatinine, were associated with MPAG C0/D. UGT2B7 rs7438135, UGT2B7 rs7439366, and UGT2B7 rs7662029 also were associated with AcMPAG C0/D. Multiple linear regression analysis showed that UGT1A9 rs2741049 and indirect bilirubin were negatively correlated with MPAG C0/D (P = .001; P = .039), and UGT2B7 rs7662029 was positively correlated with AcMPAG C0/D (P = .008). This study demonstrates a significant influence of UGT1A9 rs2741049 and UGT2B7 rs7662029 polymorphisms on the metabolism of MPA in vivo.

Effect of rt-PA intravenous thrombolysis on the prognosis of patients with minor ischemic stroke.

AIMS: The evidence of rt-PA intravenous thrombolysis in patients with minor ischemic stroke (MIS) is still controversial. This study aims to investigate the effect of rt-PA intravenous thrombolysis on the prognosis of patients with MIS. METHODS: We continuously enrolled and analyzed patients with MIS admitted into our hospital within 24 h after symptom onset between January 2016 and December 2018, including 96 patients received intravenous thrombolysis within 4.5 h after symptom onset and 84 patients not received intravenous thrombolysis. A favorable long-term outcome was a 90-day mRS score of 0-1. Good short-term outcome was a 7-day NIHSS score of 0 or less than NIHSS onset. RESULTS: There were no statistical differences between two groups of patients' age, gender, history of hypertension, coronary heart disease, atrial fibrillation, smoking, drinking, and baseline NIHSS score. Patients with history of stroke (22.62% vs. 10.42%, p < 0.05) and diabetes (46.43% vs. 22.92%, p = 0.01) were higher in group of non-thrombolysis. The difference of NIHSS score after 7 days was statistically different between the two groups (p < 0.05), while there was no significant difference in 90-day mRS score. Logistic regression analysis indicated that the prognosis of patients was correlated with neutrophil ratio and CRP at admission. CONCLUSION: Patients with MIS received intravenous thrombolysis may be associated with earlier neurological improvement, but might has no significant effect on long-term prognosis. The level of neutrophil ratio and CRP at admission are risk factors determining the prognosis, which requires further research.

Clinical and laboratory factors related to acute isolated vertigo or dizziness and cerebral infarction.

OBJECTIVE: To clarify the relationship of clinical factors with isolated vertigo or dizziness of cerebrovascular origin. METHODS: Clinical data of patients admitted in East Hospital from Jan. 2015 to Apr. 2016, whose complaint were acute vertigo or dizziness were retrospectively collected. All patients arrived at the emergency department within 24 hr of symptom onset, had no acute ischemic lesion first CT and NIHSS score of 0. Patients were divided into cerebral infarction group and noncerebral infarction group according to subsequent cerebral imaging results and clinical and laboratory factors related to cerebral infarction were analyzed. RESULT: 51.6% of patients were female (n = 141). 46 patients (16.8%) were diagnosed with acute cerebral infarction. Baseline demographic data of the two groups was not significantly different. Univariate analysis found that history of smoking (p = 0.009), headache (p = 0.028), unsteadiness (p = 0.009), neuron specific enolase (p = 0.001), and vertebral artery abnormalities found on imaging (p = 0.009) were the significant difference between two groups. Increased neuron specific enolase (p = 0.005) and an abnormal vertebral artery (p = 0.044) were significant on multivariate analysis. CONCLUSIONS: 16.8% of acute isolated vertigo or dizziness presentations were diagnosed with acute cerebral infarction. Increased serum neuron specific enolase and vertebral artery abnormalities were the strongest indicators of acute cerebral infarction.

Association of Alcohol Consumption with Markers of Prostate Health and Reproductive Hormone Profiles: A Multi-Center Study of 4,535 Men in China.

BACKGROUND: The effect of alcohol consumption on prostate health and reproductive hormone profiles has long been investigated and currently, no consensus has been reached. Additionally, large studies focusing on this topic are relatively rare in China. PURPOSE: To investigate the association of alcohol consumption with prostate measurements and reproductive hormone profiles in Chinese population; and to examine the relationship between hormone levels and prostate measurements. METHODS: This cross-sectional study included 4535 men from four representative provinces of China. Demographic details, family history of prostate disease, tobacco and alcohol consumption, as well as International Prostate Symptom Score (I-PSS) were collected through a questionnaire. Total prostate specific antingen (total PSA), free PSA, free PSA/total PSA ratio (f/tPSA), and reproductive hormones were measured in serum. Multi-variable regression models were used to test for association of alcohol consumption with markers of prostate health, used to test for association of alcohol consumption with reproductive hormones, and reproductive hormones with markers of prostate health. RESULTS: Alcohol consumption had no obvious impact on total PSA concentration and I-PSS. Current drinkers had lower level of free PSA (ß = -0.11, p = 0.02) and f/tPSA (ß = -0.03, p = 0.005), former drinkers also had lower level of free PSA (ß = -0.19, p = 0.02) when compared with never drinkers. Lower Luteinizing hormone (LH) (ß = -1.05, p = 0.01), sex hormone-binding globulin (SHBG) (ß = -4.71, p = 0.01) and higher estradiol (ß = 7.81, p = 0.01) was found in current drinkers than never drinkers, whereas higher LH (ß = 1.04, p = 0.04) and free testosterone (FT) (ß = 0.03, p = 0.02) was detected in former drinkers than never drinkers. Furthermore, LH was positively associated with f/tPSA (ß = 0.002, p = 0.006), SHBG was also positively related with free PSA (ß = 0.003, p = 0.003) and f/tPSA (ß = 0.0004, p = 0.01). Both total testosterone (TT) and FT were inversely related with I-PSS (OR = 0.97, 95% CI, 0.95-0.98; OR = 0.23, 95% CI, 0.11-0.45, respectively). CONCLUSIONS: Alcohol consumption could affect serum free PSA concentration and also f/tPSA ratio, and also acts as an endocrine disruptor on the male reproductive hormone profiles. LH and SHBG were positively related with fPSA and f/tPSA, and higher level of TT and FT may be helpful for improving participants' subjective symptoms.

Nimesulide inhibits the growth of human esophageal carcinoma cells by inactivating the JAK2/STAT3 pathway.

Although selective COX-2 inhibitors have cancer-preventive effects and induce apoptosis, the mechanisms underlying these effects are not fully understood. This study investigated the effects of nimesulide, a selective COX-2 inhibitor, on apoptosis and on the JAK/STAT signaling pathway in Eca-109 human esophageal squamous carcinoma cells. The effects and mechanisms of nimesulide on Eca-109 cell growth were studied in culture and in nude mice with Eca-109 xenografts. Cells were cultured with or without nimesulide and/or the JAK2 inhibitor AG490. Cell proliferation was evaluated using the MTT assay, and apoptosis was investigated. COX-2 mRNA expression was measured using reverse transcription polymerase chain reaction, and protein expression was detected by Western blot analysis, immunohistochemistry, and flow cytometry. Nimesulide significantly inhibited Eca-109 cell viability in vitro in a dose- and time-dependent manner (P<0.05). Nimesulide also induced apoptosis, which was accompanied by a significant decrease in the expression of COX-2 and survivin and an increase in caspase-3 expression. Nimesulide downregulated the phosphorylation levels of JAK2 and STAT3, and JAK2 inhibition by AG490 significantly augmented both nimesulide-induced apoptosis and the downregulation of COX-2 and survivin (P<0.05). In vivo, nimesulide inhibited the growth of Eca-109 tumors and the expression of p-JAK2 and p-STAT3. Thus, nimesulide downregulates COX-2 and survivin expression and upregulates caspase-3 expression in Eca-109 cells, by inactivating the JAK2/STAT3 pathway. These effects may mediate nimesulide-induced apoptosis and growth inhibition in Eca-109 cells in vitro and in vivo.

Artesunate induces apoptosis and inhibits growth of Eca109 and Ec9706 human esophageal cancer cell lines in vitro and in vivo.

Esophageal cancer is a common malignant tumor worldwide with a high incidence rate in China and it is a great threat to human health. Combined modality therapy is used for chemotherapeutic treatment of esophageal cancer; however, drug resistance and side effects of the drugs is a major barrier to the success of chemotherapy. As chemotherapy with common drugs is far from providing satisfactory clinical outcomes for patients with esophageal cancer, more efficient drugs are urgently required. Artesunate (Art) is the first-line treatment option for malaria; however, it was recently revealed that Art has remarkable anti-tumor activity, making it a novel candidate for cancer chemotherapy. Although the anti-cancer effects of Art have been well documented, its potential against esophageal cancer has rarely been explored. The present study aimed to investigate the significance and mechanism of the anti-proliferative activity of Art on esophageal cancer cells in vitro and in vivo. In the in vitro experiments, Art inhibited the growth as well as induced cell apoptosis and cell cycle arrest of esophageal cancer cell lines (Eca109 and Ec9706) in a concentration-dependent manner. Furthermore, downregulation of mitochondrial membrane potential, B-cell lymphoma-2 (BCL-2) and CDC25A, as well as upregulation of BCL-2­associated X protein (Bax) and caspase-3 expression in Art-treated cells were identified. In addition, an in vivo study showed that Art produced a dose-dependent tumor regression in nude mice, while side effects were low. The anti-tumor activity of 200 mg/kg Art was similar to that of 3 mg/kg cisplatin. In conclusion, Art exerted concentration-dependent inhibitory activity against esophageal cancer in vivo and in vitro by inducing cell apoptosis and cell cycle arrest through affecting mitochondrial membrane potential, BCL-2, Bax, caspase-3 and CDC25A.

A retrospective analysis of negative diffusion-weighted image results in patients with acute cerebral infarction.

We aimed to investigate the clinicoradiologic determinants of negative diffusion-weighted image (DWI) results in patients with acute cerebral infarction (ACI). The medical records were reviewed of ACI patients. Patients were divided to the DWI positive and negative group. Positive DWI was used as independent variable and patients' clinicoradiologic factors were used as co-variables for multivariate logistic regression analysis. 349 patients received initial cerebral MRI within 72 hours of admission. Lacunar infarction was most common (42.1%) followed by posterior circulation infarction (30.1%) and partial anterior circulation infarction (18.1%). The majority of the patients (72.2%) had an NIHSS score of less than 5 at admission. 316 patients (90.54%) were positive on initial DWI. Patients with smoking, initial SBP ≥ 140 or DBP ≥ 90 mmHg, initial fasting plasma glucose (FPG) ≥ 7.0 mmol/L, initial MRI from onset of disease >1 d and anterior circulation infarction were liable to show positive DWI. Furthermore, DWI negative patients had significantly lower NIHSS scores (IQR 0,1,2) than DWI positive patients (IQR 1,2,4) (P = 0.000) at two weeks post onset of acute cerebral infarction. In conclusion, multiple clinicoradiologic factors are associated with negative and positive DWI and further delineation of these factors is required in future prospective studies.

The PTEN/PI3K/Akt signaling pathway mediates HMGB1-induced cell proliferation by regulating the NF-κB/cyclin D1 pathway in mouse mesangial cells.

Our previous experiment confirmed that high-mobility group box chromosomal protein 1 (HMGB1) was involved in the pathogenesis of Lupus nephritis (LN) by upregulating the proliferation of the mouse mesangial cell line (MMC) through the cyclin D1/CDK4/p16 system, but the precise mechanism is still unknown. Therefore, in the present study, we demonstrated that HMGB1 induced the proliferation of MMC cells in a time- and concentration-dependent manner, downregulated phosphatase and tensin homolog deleted on chromosome ten (PTEN) expression, increased the level of Akt serine 473 phosphorylation, and induced p65 subunit nuclear translocation. The overexpression of PTEN prevented the upregulation of HMGB1-induced proliferation by blocking the activation of Akt. The knockdown of Akt by siRNA technology and blocking the nuclear factor-κB (NF-κB) pathway using pyrrolidine dithiocarbamate (PDTC) and SN50, inhibitors of NF-κB, both attenuated the HMGB1-induced proliferation by counteracting the activation of the cyclin D1. In addition, while sh-Akt partly blocked the nuclear translocation of the p65 subunit, PDTC did not affect the activation of the Akt induced by HMGB1 in MMC cells. These findings indicate that HMGB1 induced the proliferation of MMC cells by activating the PTEN/phosphoinositide-3-kinase (PI3K)/Akt/NF-κB signaling pathway.

ABCG2 gene amplification and expression in esophageal cancer cells with acquired adriamycin resistance.

Resistance to chemotherapeutic agents is the main reason for treatment failure in patients with cancer. The primary mechanism of multidrug resistance (MDR) is the overexpression of drug efflux transporters, including ATP­binding cassette transporter G2 (ABCG2). To the best of our knowledge, the MDR mechanisms of esophageal cancer have not been described. An adriamycin (ADM)-resistant subline, Eca109/ADM, was generated from the Eca109 esophageal cancer cell line by a stepwise selection in ADM from 0.002 to 0.02 ng/µl. The resulting subline, designated Eca109/ADM, revealed a 3.29-fold resistance against ADM compared with the Eca109 cell line. The ABCG2 gene expression in the Eca109/ADM cells was increased compared with that of the Eca109 cells. The cellular properties of the Eca109/ADM cells were detected by reverse transcription polymerase chain reaction (RT-PCR), flow cytometry and western blotting. The ABCG2 expression levels were detected by RT-PCR and flow cytometry, and the drug efflux effect was detected by flow cytometry. The present study detected the correlation between ABCG2 and the multidrug resistance of esophageal cancer. ABCG2 gene expression and the drug efflux effect of the Eca109/ADM cells were increased compared with those of the Eca109 cells. Collectively, the results of this study indicated that the overexpression of ABCG2 in the Eca109/ADM cells resulted in drug efflux, which may be responsible for the development of esophageal cancer MDR.

Reversal of multidrug resistance by the anti-malaria drug artesunate in the esophageal cancer Eca109/ABCG2 cell line.

The overexpression of ATP-binding cassette (ABC) transporters confers multidrug resistance (MDR) to tumor cells. ABCG2 is a member of the ABC superfamily. The present study aimed to investigate the correlation between ABCG2 expression and the MDR of esophageal cancer and to estimate the therapeutic benefit of downregulating ABCG2 expression and reversing chemoresistance in esophageal cells using artesunate (Art). The Eca109/ABCG2 cell line was established by transfecting the ABCG2 gene into Eca109 cells. The Eca109/ABCG2 esophageal cancer cells with ABCG2 gene overexpression were resistant to adriamycin (ADM), daunorubicin (DNR) and mitoxantrone (MIT), which indicated that ABCG2 may be associated with drug resistance in esophageal cancer. Art is a noteworthy antimalarial agent, particularly in severe and drug-resistant cancer cases, as Art is able to reverse drug resistance. In the present study, Art also exerted profound anticancer activity. The mechanism for the reversal of multidrug resistance by Art in esophageal carcinoma was analyzed using cellular experiments, but still remains largely unknown.

PLC-ε1 gene polymorphisms significantly enhance the risk of esophageal squamous cell carcinoma in individuals with a family history of upper gastrointestinal cancers.

BACKGROUND AND AIMS: Phospholipase C epsilon 1 (PLCε1) may regulate cell growth, differentiation, apoptosis and angiogenesis and play an important role in carcinogenesis and the progression of several cancers. This study was designed to validate the association of the PLCε1 rs2274223 single nucleotide polymorphism (SNP) with esophageal squamous cell carcinoma (ESCC) as identified by genome-wide association studies (GWAS) and further assess whether the rs11599672 SNP could affect an individual's susceptibility to ESCC. METHODS: These two SNPs were genotyped by polymerase chain reaction ligase detection reaction (PCR-LDR) in 527 ESCC patients and 527 controls. RESULTS: Compared with the rs2274223 SNP AA genotype, other genotypes or combined genotypes all enhanced the risk of ESCC. Further analyses showed that AG/GG genotype carriers with a family history of upper gastrointestinal cancers (UGIC) had an increased risk of ESCC than those AA genotype carriers without UGIC family history (OR = 2.10, 95% CI = 1.46-3.10). Overall, rs11599672 SNP had no influence on ESCC susceptibility. However, UGIC family history elevated the risk of ESCC for subjects with the TT genotype (OR = 1.59, 95% CI = 1.13-2.24). CONCLUSIONS: These results highlighted the role of a genetic factor in ESCC and suggested that the PLCε1 rs2274223 SNP might be an effective genetic marker to assess the risk of ESCC in individuals with a UGIC family history from a region of high incidence in northern China.

Mild-to-moderate neurogenic pyrexia in acute cerebral infarction.

BACKGROUND: Pyrexia is often associated with unfavorable stroke outcomes. However, limited information is available on the relationship between the causes of poststroke hyperthermia and stroke prognosis, especially for mild-to-moderate neurogenic pyrexia in acute cerebral infarction. AIMS: To compare the differences in the clinical features and characteristics of pyrexia as well as its prognosis among acute cerebral infarction patients with mild-to-moderate neurogenic pyrexia, with infectious pyrexia, and without pyrexia. The focus was on mild-to-moderate neurogenic pyrexia. METHODS: A total of 709 patients with acute cerebral infarction were prospectively recruited and their clinical data were analyzed. RESULTS: No significant difference was detected in age, gender, history of smoking, hypertension, or diabetes among the 3 groups (p > 0.05). Patients with mild-to-moderate neurogenic pyrexia and those with infectious pyrexia had higher baseline National Institutes of Health Stroke Scale (NIHSS) scores (15.1 ± 6.7, p = 0.003; 14.3 ± 8.1, p = 0.002, respectively), lower 3-month Barthel index (BI) values (64.2 ± 40.7, p < 0.001; 61.9 ± 49.3, p < 0.001, respectively) and higher 3-month mortality rates (13%, p = 0.026; 16%, p < 0.001, respectively) than patients without pyrexia (NIHSS score 11.4 ± 7.9; BI 82.6 ± 39.8, and mortality rate 6%, respectively). No difference existed in these parameters between the 2 pyrexia groups (p > 0.05), but mild-to-moderate neurogenic pyrexia had an earlier onset and a shorter duration than infectious pyrexia (p < 0.001). CONCLUSIONS: Acute cerebral infarction patients with mild-to-moderate neurogenic pyrexia had a similar prognosis compared to those with infectious pyrexia. Mild-to-moderate neurogenic pyrexia is possibly associated with stroke severity.

Progesterone reverses the mesenchymal phenotypes of basal phenotype breast cancer cells via a membrane progesterone receptor mediated pathway.

INTRODUCTION: Basal phenotype breast cancers (BPBC) are often associated with apparent epithelial to mesenchymal transition (EMT). The role of progesterone (P4) in regulating EMT of BPBC has not been reported. METHODS: The EMT relevant biology was investigated in vitro using human BPBC cell models (MDA-MB468 and MDA-MB231) with P4, PR agonist (RU486), and PR antagonist (R5020) treatments. The essential role of membrane progesterone receptor alpha (mPRalpha) in the P4-regulated EMT was demonstrated by knocking down the endogenous gene and/or stably transfecting exogenous mPRalpha gene in the BPBC cell models. RESULTS: The expression of snail and down-stream EMT proteins such as occludin, fibronectin, and E-cadherin was significantly regulated by P4 incubation, which was accompanied by cell morphological reversion from mesenchymal to epithelial phenotypes. In searching for the cell mediator of P4' action in the MDA-MB468 (MB468) cells, it was found that mPRalpha but not the nuclear PR has an essential role in the P4 mediated EMT inhibition. Knocking down the expression of mPRalpha with specific siRNA blocked the P4's effects on expression of the EMT proteins. In another BPBC cell line--MDA-MB231 (MB231), which is mPRalpha negative by Western blotting--P4 treatment did not alter cell proliferation and EMT protein expressions. Introduction of the exogenous mPRalpha cDNA into these cells caused cell proliferation, but not EMT, to become responsive to P4 treatment. In further studies, it was found that activation of the PI3K/Akt pathway is necessary for the P4-induced EMT reversion. To define the potential inter-mediate steps between mPRalpha and PI3K, we demonstrated that mPRalpha, caveolin-1 (Cav-1), and epidermal growth factor receptor (EGFR) are colocalized in the membrane of caveolar vesicle and the P4-repressed EMT in MB468 cells can be blocked by EGFR inhibitor (AG1478) and PI3K inhibitor (wortmannin). CONCLUSIONS: Our data suggest that the signaling cascade of P4 induced mesenchymal repression is mediated through mPRalpha and other caveolae bound signaling molecules namely Cav-1, EGFR, and PI3K. This novel finding may have great impact on fully understanding the pathogenesis of BPBC and provide an essential clue for developing a targeted therapeutic strategy for treatment of BPBC.

Optimizing the time of Doxil injection to increase the drug retention in transplanted murine mammary tumors.

Sex hormonal milieus during the female fertility cycle modulate the tumor vascular permeability of breast cancer. It has been proposed that the liposomal formulated doxorubicin (ie, Doxil), given at the menstrual/estrous stage with the predicted highest tumor vascular permeability, allows significantly increased drug retention in the breast tumor. In the current study, syngeneic murine 4T1 mammary tumors were established on the backs of female BALB/c mice and Doxil was administered at particular mouse estrous cycle stages. The results indicated that Doxil administration during certain times in the mouse estrous cycle was crucial for drug retention in 4T1 tumor tissues. Significantly higher drug concentrations were detected in the tumor tissues when Doxil was administered during the diestrus stage, as compared to when the drug injection was given at all other estrous stages. Our study also showed that the tumor-bearing mice exhibited nearly normal rhythmicity of the estrous cycle post drug injection, indicating the feasibility of continual injection of Doxil at the same estrous cycle stage. By using 4T1 cells cultured in vitro, we showed that progesterone (P4) significantly inhibited cell proliferation and the production of six tumor-derived cytokines, eg, sTNF-RI, CXCL-16, GM-CSF, MIP-1alpha, MIP-1gamma, and Flt3-L. Some of these factors have been shown to be vascular modulators in diverse tissues. In this report, we demonstrated that the concentration of P4 in the plasma and/or estrous cycle stage of 4T1 tumor-bearing mice can be used to select the best time for administrating the liposomal anticancer drugs.

[Expression of EphA2 and EphrinA1 in human renal cell carcinoma and its relationship with angiogenesis].

OBJECTIVE: To investigate the expression of EphA2 and EphrinA1 and its relationship with angiogenesis in renal cell carcinoma and its relevance to clinicopathologic features. METHODS: The expression of the EphA2 and EphrinA1 was detected by immunohistochemistry (IHC) in the tissues samples from 68 renal cell carcinomas and 24 normal kidneys, and quantitatively analyzed. The microvessel density (MVD) was determined by CD34 immunostaining of microvascular endothelial cells. Statistical analysis was performed using the software SPSS (version 13.0). RESULTS: The expression of EphA2, EphrinA1 and MND in the cancerous tissues were significantly higher (P<0.01) than that in the normal ones. Significantly increased expression of EphA2, EphrinA1 and MVD (P<0.01) was detected in cancer tissues with higher grade differentiation, more advanced stage and more lymph node metastasis, respectively (P<0.05 for each group). Expression of the EphA2 and EphrinA1 protein was shown to be positively associated with the MVD assessed by Spearman's correlation and factor analysis (r=0.555, r=0.485, P<0.01). The MVD was also significantly correlated with the diameter of the tumor (P<0.01). CONCLUSION: EphA2 and EphrinA1 are highly expressed in renal cell carcinoma, and positively correlated with histological differentiation, clinical stage and angiogenesis in the cancer.

The regulatory effect of the p38 signaling pathway on valdecoxib-induced apoptosis of the Eca109 cell line.

Valdecoxib is a second generation selective COX-2 inhibitor that can induce cell apoptosis in a variety of cell types, but its precise regulatory mechanism is unknown. Apoptosis of Eca109 cells and p38 mRNA expression were investigted. The expression of p-p38MAPK, Fas and FasL proteins were detected by immunohistochemical staining and FCM. Valdecoxib increased the apoptosis rate of Eca109 cells. Fas and FasL protein expression was up-regulated in the valdecoxib groups, while SB203580 partly inhibited the valdecoxib-induced overexpression. Valdecoxib increased p38MAPK expression, while SB203580 inhibited the overexpression of this protein and the apoptosis rate decreased. The expression of Fas, FasL and p38MAPK protein were positively correlated with the apoptotic rate. In conclusion, valdecoxib activates the p38MAPK pathway, thus up-regulating expression of the Fas and FasL proteins, which may be one of the mechanisms through which valdecoxib induces apoptosis.

Mammaglobin as a potential molecular target for breast cancer drug delivery.

BACKGROUND: Mammaglobin (MAM) has been used as a specific molecular marker for breast cancer diagnosis. Recently, several groups of researchers proposed a number of therapeutic strategies targeting this molecule. Some of the strategies are based upon an essential but not demonstrated hypothesis - mammaglobin is associated with the surface of breast cancer cells, which strongly disputes the therapeutic strategies. RESULTS: We conducted a computer-based predictive analysis and identified a small fragment at the N-end of MAM as a potential transmembrane domain. We provided several evidences to demonstrate the presence of the membrane-associated MAM. We isolated the membrane protein components from known MAM positive breast cancer cells (MDA-MB361 and MDA-MB415). We showed that about 22-64% of MAM proteins, depending upon the types of the cancer cells, directly attached on the membrane of breast cancer cells, by Western blotting assays. To directly visualize the presence of the membrane-bound MAM protein, we incubated the MAM positive cancer cells with FITC labeled anti-MAM antibody, and observed clear fluorescent signals on the surface of the cells. In studying the MAM protein distribution in human breast cancer tissues, we first identified two immunostain patterns that are associated with the membrane-bound MAM: the membrane stain pattern and luminary surface stain pattern. To test whether the membrane-associated MAM can serve as a molecular target for drug delivery, we conjugated anti-MAM antibody to human low-density lipoprotein (LDL) and loaded doxorubicin (Dox) in the core of LDL. Specific binding and cytotoxicity of the MAM targeted and Dox loaded LDL was tested in the MAM positive breast cancer cells in vitro. CONCLUSION: We first showed that some of MAM protein directly associated with the surface of breast cancer cells. The membrane-associated MAM protein may be utilized as a useful molecular marker for breast cancer targeted drug delivery.

[The functional mechanism of CIK cells on ovarian carcinoma cell lines SKOV3/CDDP].

AIM: To investigate the functional mechanism of CIK cells or ovarian carcinoma cell lines SKOV3/CDDP. METHODS: The changes of ultramicrostructure, cell cycle, apoptosis, expression of multidrug resistance-associated protein (MDR1, Topo-IIbeta) and other molecules (hB7-1, hB7-2, MHCIb, HLA-DR) of SKOV3/CDDP cells treated with or without CIK cells were detected by electron microscope, MTT and FCM. The changes of cytokine (IL-2, TNF-alpha, IFN-gamma, GM-CSF) in sera of SCID mice bearing SKOV3/CDDP cells were detected by radioimmunit and ELISA. RESULTS: CIK cells could induce apoptosis of the SKOV3/CDDP cells by electronmicroscopic observations. The apoptosis rate in CIK group was 9.07%, and its cell cycle was arrested at S and G2/M phase (P<0.05). Compared with NS Group, the co-expression of MDR-1 and Topo-IIbeta were decreased significantly in the CIK treated group(P<0.05), and the expression of MHCIb, HLA-DR, hB7-1 and hB7-2 antigen were increased significantly (P<0.01). Compared with NS Group, the contents of IL-2, TNF-alpha, INF-gamma, GM-CSF were increased significantly (P<0.01) in SCID mice of CIK group. CONCLUSION: CIK cells have several important biological effects on the ovarian carcinoma cell line SKOV3/CDDP, which may lay the foundation for further research on anti-tumor therapy.

New technology and clinical applications of nanomedicine.

Nanomedicine is the use of nanotechnology to achieve innovative medical breakthroughs. Nanomedicine, with its broad range of ideas, hypotheses, concepts, and undeveloped clinical devices, is still in its early stage. This article outlines present developments and future prospects for the use of nanotechnology techniques in experimental in vivo and in vitro studies and in engineering nanodevices and biosensors for clinical and investigative use n diagnosis and therapy in the fields of genetics, oncology, cardiology, and dermatology. Toxicologic considerations also are discussed.

[Expression and clinical significance of COX-2, p-Stat3, and p-Stat5 in esophageal carcinoma].

BACKGROUND & OBJECTIVE: Cyclooxygenase-2 (COX-2) and signal transducers and activators of transcription (STAT) are closely correlated to the genesis of tumors. This study was to investigate the expression and clinical significance of COX-2, p-Stat3 and p-Stat5 (the activated forms of Stat3 and Stat5) in various lesions of esophageal tissues, and to analyze their correlations to clinicopathologic features of esophageal squamous cell carcinoma (ESCC). METHODS: The expression of COX-2, p-Stat3, and p-Stat5 in 59 specimens of ESCC, 24 specimens of squamous dysplasia, and 18 specimens of normal squamous epithelium was examined by SP immunohistochemistry. Their correlations to clinicopathologic features of ESCC were analyzed. RESULTS: The protein level of COX-2 was significantly higher in ESCC and squamous dysplasia than in normal squamous epithelium (2.10+/-1.77 and 1.85+/-1.24 vs. 0.83+/-0.46, P<0.05). The protein level of p-Stat3 was 0 in normal squamous epithelium, 0.76+/-0.59 in squamous dysplasia, and 2.83+/-1.27 in ESCC. The protein level of p-Stat5 was 1.98+/-0.78 in normal squamous epithelium, 3.92+/-0.41 in squamous dysplasia, and 5.02+/-0.34 in ESCC. There were significant differences among the 3 groups (P<0.05). In ESCC, COX-2 expression was correlated to lymph node metastasis and differentiation (P<0.05); p-Stat3 expression was correlated to tumor invasion depth (P<0.05); but p-Stat5 expression had no correlation to clinicopathologic features. COX-2 expression was positively correlated to both p-Stat3 expression and p-Stat5 expression in ESCC. CONCLUSIONS: The up-regulation of COX-2, p-Stat3, and p-Stat5 may be correlated to the carcinogenesis of ESCC. The activation of Stat3 is correlated to the aggressive behavior of ESCC.