KCNK levels are prognostic and diagnostic markers for hepatocellular carcinoma.

Two-pore-domain (KCNK, K2P) K+ channels are transmembrane protein complexes that control the flow of ions across biofilms, which underlie many essential cellular functions. Because KCNK family members are known to contribute to tumorigenesis in various types of cancer, we hypothesized that they might be differentially expressed in hepatocellular carcinoma (HCC) cells as compared to healthy tissue and serve as diagnostic or prognostic biomarkers. We tested this hypothesis through bioinformatic analyses of publicly available data for the expression of various KCNK subunits in HCC. We observed reduced expression of KCNK2, KCNK15, and KCNK17 in liver cancer, as well as overexpression of KCNK9, all of which correlated with a better prognosis for HCC patients per survival analyses. Moreover, ROC curves indicated that KCNK2, KCNK9, KCNK15, and KCNK17 levels could be used as a diagnostic biomarker for HCC. Finally, our western blot and qRT-PCR results were consistent with those obtained from bioinformatic analyses. Taken together, these results suggest that KCNK2, KCNK9, KCNK15, and KCNK17 could serve as potential diagnostic and prognostic biomarkers of HCC.

Sex Recognition of Skulls in Chinese Uygur / 法医学杂志

Objective Logistic regression method was used to establish a multiple regression sex discriminant function to discriminate the complete skull model and the incomplete skull model without frontal bone, occipital bone and mandible of Uygur adults in Turpan, Xinjiang. Methods A total of 117 （60 male and 57 female） three-dimensional skull models were collected by CT. Sixteen cranial measurement indexes were measured and calculated by computer software. The multivariate regression sex discriminant function was established with Logistic regression method and retrospectively tested. Results Among the 16 measurement indexes, except for nose width （x7） and maximum frontal breadth （x13）, the remaining 14 indexes had statistical significance of differences between male and female （P<0.05）. For the discriminant function of complete skull established by eyebrow arch convexity （x4）, mastoid width （x6）, maximum cranial length （x12）, cranial base length （x15）, cranial circumference （x16）, the male and female discrimination accuracy was 90.0% and 94.7%, respectively. For the sex discriminant function of incomplete skull without frontal bone established by mandibular angle width （x10）, mandibular height （x11） and cranial circumference （x16）, the discrimination accuracy of male and female was 85.0% and 84.2%, respectively. For the sex discriminant function of incomplete skull without occipital bone established by the index of eyebrow arch convexity （x4）, the discrimination accuracy of male and female was 80.0% and 73.7%, respectively. For the sex discriminant function of incomplete skull without mandible established by frontal chord （x5） and occipital protrusion angle （x9）, the discrimination accuracy of male and female was 85.0% and 78.9%, respectively. Conclusion The computer software and system developed in our study can achieve sex discrimination of complete skulls and incomplete skulls without frontal bone, occipital bone or mandible.

miR19b-3p promotes the growth and metastasis of colorectal cancer via directly targeting ITGB8.

MicroRNAs (miRNAs) are widely up-regulated or down-regulated in a variety of tumors, including lung cancer, liver cancer, and colorectal cancer (CRC). Furthermore, miRNAs can function as tumor suppressors or proto-oncogenes by controlling the growth and metastasis of cancer cells. In the present study, we found a significant increase in miR19b-3p levels in CRC compared to tumor tissue and revealed the role of miR19b-3p in CRC growth and metastasis. The exogenous overexpression of miR19b-3p induced the proliferation, migration, and invasion of CRC cells in vitro. In addition, the nude mouse xenograft model showed that miR19b-3p overexpression promoted CRC growth and lung metastasis in vivo, whereas silencing miR19b-3p showed opposite results. Mechanistic studies have shown that the integrin beta-8 (ITGB8) transcript is one of the direct targets of miR19b-3p, and the expression of ITGB8 in CRC specimens was positively correlated with miR19b-3p. Finally, ectopic expression of ITGB8 rescued cell proliferation and invasion, which was inhibited by down-regulation of miR19b-3p. In addition, knockdown of ITGB8 neutralized the effects of miR19b-3p overexpression on cell growth and metastasis in CRC cells. Together, these results suggest that the miR19b-3p/ITGB8 axis plays an important role in the growth and metastasis of CRC.

Downregulation of phosphorylated MKK4 is associated with a poor prognosis in colorectal cancer patients.

Mitogen-activated protein kinase kinase 4 (MKK4) is a key mediator of Jun N-terminal kinase signaling and influences malignant metastasis. Here, we used immunohistochemistry to assess phosphorylated MMK4 (pMKK4) levels and examine their association with the clinicopathological features of a pilot set of patient samples consisting of normal colonic mucosa (NCM), colorectal adenoma (CA), and colorectal cancer (CRC) tissues. pMKK4 levels were also assessed in a validation set of CRC cases with accompanying follow-up data to confirm their clinicopathological and prognostic significance. pMKK4 levels, which were high in 79.17% of NCM samples, were downregulated in 33.33% of CA and 63.54% of CRC samples. pMKK4 downregulation was associated with metastasis, especially to the liver. In the validation set, pMKK4 downregulation was associated with increases in invasive depth, lymph node metastasis, distant metastasis, and TNM stage. Univariate analysis indicated that pMKK4 score, tumor differentiation, and TNM stage were correlated with disease-free survival and overall survival. Multivariate analysis indicated that decreased pMKK4 expression was an independent risk factor for disease-free survival in CRC patients. These results suggest that CRC patients with low pMKK4 immunochemistry scores should be monitored carefully for early detection of possible recurrences, especially liver metastasis.

Accuracy of ultrasound-measured inferior vena cava diameter in assessment of preoperative blood volume in elderly patients / 中华麻醉学杂志

Objective To evaluate the accuracy of ultrasound-measured inferior vena cava (IVC) diameter in assessment of the preoperative blood volume in elderly patients.Methods Sixty patients,aged 60-75 yr,with body mass index 20-25 kg/m2,of ASA physical status Ⅰ-Ⅲ,scheduled for elective transurethral resection of prostate,were randomly divided into 3 groups (n =20 each):control group (group C),lactated Ringer's solution group (group RL),and hydroxyethyl starch group (group H).Lactated Ringer's solution 8 ml/kg was infused intravenously in group RL.Hydroxyethyl starch 130/0.4 8 ml/kg was infused intravenously in group H.SpO2,mean arterial pressure (MAP),HR and central venous pressure (CVP) were monitored before and after fluid therapy.The IVC diameters,both during expiration (IVCe) and inspiration (IVCi),were measured using ultrasound.IVC collapsibility index (IVC-CI) was calculated.Results The IVCeand IVCi were significantly increased,and IVC-CI was decreased after fluid therapy as compared with those before fluid therapy in RL and H groups.Compared with group C,the IVCe and IVCi were significantly increased,and IVC-CI was decreased after fluid therapy in RL and H groups.IVCe and IVCi were positively correlatedwith CVP (r=0.746 and 0.697,respectively).IVC-CI was negatively correlated with CVP (r =-0.547).Conclusion Ultrasoundmeasured IVC diameter provides better accuracy in assessing the preoperative blood volume in elderly patients.

Upregulation of microRNA-106b is associated with poor prognosis in hepatocellular carcinoma.

BACKGROUND: MicroRNA-106b (miR-106b) is a member of the miR-106b ~ 25 cluster. It has been reported that miR-106b acts as an oncogene and is upregulated in many human cancers. However, the prognostic value of miR-106b in hepatocellular carcinoma (HCC) remains unclear. The aim of this study was to investigate the clinical significance of miR-106b expression in HCC. METHODS: We determined the expression level of miR-106b in 104 cases of paired HCC and adjacent non-tumor tissues by quantitative real-time PCR (qRT-PCR). The correlation between miR-106b expression and prognosis of HCC was studied by univariate and multivariate analysis. Multivariate analysis of the prognostic factors was performed with Cox proportional hazards model. RESULTS: MiR-106b expression was significantly upregulated in as high as 76.0% of HCC tissues, compared with their non-tumor counterparts (P < 0.001). High miR-106b expression was significantly associated with large tumor size (P = 0.019) and vascular invasion (P = 0.016). Kaplan-Meier analysis showed that patients with high miR-106b expression had a worse overall survival than patients with low miR-106b expression (log-rank P = 0.004). The multivariate Cox regression analysis indicated that miR-106b expression was an independent prognostic factor for overall survival (HR, 2.002; 95% CI, 1.130-6.977; P = 0.027). CONCLUSION: Our data indicated that miR-106b expression was significantly upregulated in HCC and could serve as a potential unfavorable prognostic biomarker. VIRTUAL SLIDES: The virtual slide(s) for this article can be found here: http://www.diagnosticpathology.diagnomx.eu/vs/13000_2014_226.

LIN28 expression and prognostic value in hepatocellular carcinoma patients who meet the Milan criteria and undergo hepatectomy.

Stem cell marker LIN28, related closely with SOX2 and OCT4, has been studied as a biomarker for the maintainance of pluripotent cells in several malignancies. Our previous study showed that SOX2 and OCT4 were negative predictors for hepatocellular carcinoma (HCC). However, the predictive value of LIN28 in HCC outcome is still undetermined. We hypothesized that LIN28 may also play a role as a biomarker for HCC. To test this hypothesis, we examined the expression of LIN28 in 129 radically resected HCC tissues using reverse transcription-polymerase chain reaction and analyzed the association of LIN28 expression with clinicopathologic features and prognosis. Our study showed that LIN28 was expressed at a higher frequency in tumor tissues than in non-HCC tissues (45.0% vs. 21.7%, P = 0.020). Moreover, LIN28 expression was significantly increased in cases with large tumor size (P = 0.010). Univariate analysis did not reveal a significant correlation between LIN28 expression and overall survival or recurrence-free survival. For HCC patients who met the Milan criteria, stratified analysis revealed shorter overall survival (P = 0.007) and recurrence-free survival (P < 0.001) in those with detectable LIN28 expression compared to those with no detectable LIN28 expression. Furthermore, multivariate analysis revealed that LIN28 was a negative independent predictor for both overall survival (hazard ratio= 7.093, P = 0.017) and recurrence-free survival (hazard ratio=5.518, P = 0.004) in patients who met the Milan criteria. Taken together, our results suggest that LIN28 identifies low-risk and high-risk subsets of HCC patients meeting the Milan criteria who undergo hepatectomy.

LIN28 expression and prognostic value in hepatocellular carcinoma patients who meet the Milan criteria and undergo hepatectomy / 癌症

Stem cell marker LIN28, related closely with SOX2 and OCT4, has been studied as a biomarker for the maintainance of pluripotent cells in several malignancies. Our previous study showed that SOX2 and OCT4 were negative predictors for hepatocellular carcinoma (HCC). However, the predictive value of LIN28 in HCC outcome is still undetermined. We hypothesized that LIN28 may also play a role as a biomarker for HCC. To test this hypothesis, we examined the expression of LIN28 in 129 radically resected HCC tissues using reverse transcription-polymerase chain reaction and analyzed the association of LIN28 expression with clinicopathologic features and prognosis. Our study showed that LIN28 was expressed at a higher frequency in tumor tissues than in non-HCC tissues (45.0% vs. 21.7%, P = 0.020). Moreover, LIN28 expression was significantly increased in cases with large tumor size (P = 0.010). Univariate analysis did not reveal a significant correlation between LIN28 expression and overall survival or recurrence-free survival. For HCC patients who met the Milan criteria, stratified analysis revealed shorter overall survival (P = 0.007) and recurrence-free survival (P < 0.001) in those with detectable LIN28 expression compared to those with no detectable LIN28 expression. Furthermore, multivariate analysis revealed that LIN28 was a negative independent predictor for both overall survival (hazard ratio= 7.093, P = 0.017) and recurrence-free survival (hazard ratio=5.518, P = 0.004) in patients who met the Milan criteria. Taken together, our results suggest that LIN28 identifies low-risk and high-risk subsets of HCC patients meeting the Milan criteria who undergo hepatectomy.

Determination for Enterobacter cloacae based on a europium ternary complex labeled DNA probe.

The fast detection and accurate diagnosis of the prevalent pathogenic bacteria is very important for the treatment of disease. Nowadays, fluorescence techniques are important tools for diagnosis. A two-probe tandem DNA hybridization assay was designed for the detection of Enterobacter cloacae based on time-resolved fluorescence. In this work, the authors synthesized a novel europium ternary complex Eu(TTA)(3)(5-NH(2)-phen) with intense luminescence, high fluorescence quantum yield and long lifetime before. We developed a method based on this europium complex for the specific detection of original extracted DNA from E. cloacae. In the hybridization assay format, the reporter probe was labeled with Eu(TTA)(3)(5-NH(2)-phen) on the 5'-terminus, and the capture probe capture probe was covalent immobilized on the surface of the glutaraldehyde treated glass slides. The original extracted DNA of samples was directly used without any DNA purification and amplification. The detection was conducted by monitoring the fluorescence intensity from the glass surface after DNA hybridization. The detection limit of the DNA was 5×10(-10) mol L(-1). The results of the present work proved that this new approach was easy to operate with high sensitivity and specificity. It could be conducted as a powerful tool for the detection of pathogen microorganisms in the environment.

NADPH oxidase DUOX1 and DUOX2 but not NOX4 are independent predictors in hepatocellular carcinoma after hepatectomy.

NADPH oxidase DUOX1, DUOX2, and NOX4 have recently been gained considerable concerns, owing to the fact that they involve in reactive oxygen species-induced genetic and epigenetic alternations of human carcinogenesis and serve as biomarkers in several cancers. Whether they predict survival in hepatocellular carcinoma (HCC) is still uncertain. Here, we detected the expressions of DUOX1, DUOX2, and NOX4 in one normal liver cell line, seven HCC cell lines, 30 non-cirrhotic normal liver tissues, and 107 paired HCC tissues using reverse transcription-polymerase chain reaction. The correlations of genes expression with prognoses were analyzed. DUOX1 was expressed at high levels in MHCC-97H and MHCC-97L, but at low levels in Bel-7402. In contrast to low expression level at SMMC-7721, DUOX2 was expressed at considerably high levels in MHCC-97H and MHCC-97L. The transcript of NOX4 was only detected in SMMC-7721. All the 30 normal liver tissues failed to express the three candidate markers. Compared with adjacent non-neoplastic tissues, DUOX1, DUOX2, and NOX4 were expressed at higher frequencies in tumor specimens. Both univariate and multivariate analyses revealed that elevated expression of DUOX1 or DUOX2 predicted poorer recurrence-free survival and overall survival. No such significance trend regarding NOX4 predictive value in survival, however, was seen in univariate analysis. These results suggested DUOX1 and DUOX2, but not NOX4, could predict HCC prognoses after hepatectomy.

A novel bifunctional europium chelate applied in quantitative determination of human immunoglobin G using time-resolved fluoroimmunoassay.

The authors demonstrate herein a novel time-resolved fluoroimmunoassay (TRFIA) protocol for quantification of human IgG with the new bifunctional chelate Eu(TTA)3(5-NH2-phen) (ETNP) labeling the goat anti-human IgG. The immunoassay was conducted by following the typical procedure for sandwich-type immunoreactions. Goat anti-human IgG was immobilized on aldehyde-modified glass slides. The human IgG analyte was first captured by the primary antibody and then sandwiched by a secondary antibody labeled with the chelate ETNP. The experimental procedure was simple to follow and gave desirable levels of sensitivity and low limits of detection. To the best of our knowledge, this is the first application of the new chelate, ETNP, in an immunoassay. In comparison to typical organic, fluorescent compounds and other lanthanide fluorescent chelates used in immunoassay, the detection sensitivity of our method using ETNP chelate in the solid phase was greatly improved and a concentration of human IgG about 5 µg/L could be detected under optimal conditions. The main result of this work shows that the new chelate ETNP can be applied as a powerful fluorescent labeling material for constructing ultrasensitive TRFIAs. The detection of human IgG, using ETNP as the chelate, is a model example of the effectiveness of this immunoassay. Many other types of antigen-antibody immunoassays should be possible using the protocol described herein.

Role of 5-HT1A receptor in distal cerebrospinal fluid-contacting neurons in neuropathic pain in rats / 中华麻醉学杂志

Objective To evaluate the role of 5-HT1A receptors in distal cerebrospinal fluid (CSF)-contacting neurons in neuropathic pain (NP) in rats. Methods Forty male SD rats weighing 230-270 g were randomly divided into 4 groups (n = 8 each): sham operation group (group S); NP group; dimethyl sulfoxide (DMSO) group and 8-OH-DPAT (a specific 5-HT1A receptor agonist) group. NP was induced by chronic constrictive injury (CCI) in groups NP, DMSO and 8-OH-DPAT. Four silk ligatures were placed on the sciatic nerve at 1 mm intervals . In group S, the sciatic nerve was exposed but not ligated. 8-OH-DPAT and DMSO 1 μl were injected into the region where most of CSF-contacting neurons are present over 5 min on 7th day after CCI in groups 8-OH-DPAT and DMSO respectively. Paw withdrawal latency (PWL) and paw withdrawal threshold (PWT) were measured before CCI, on 7th day after CCI, and at 3 and 6 h after administration. The rats were sacrificed 6 h after administration, and the brain tissues removed for determination of the expression of 5-HT1A receptors in the distal CSF-contacting neurons by immunofluorescence. Results Compared with group S, PWL was significantly shorten and PWT decreased at T, in groups NP, DMSO and 8-OH- DPAT (P ＜ 0.01) . Compared with group DMSO, PWL was significantly prolonged and PWT increased at T2 and T3 in group 8-OH-DPAT ( P ＜ 0.01). The 5-HT1A receptor expression was significantly down-regulated in groups NP and DMSO compared with group S, while up-regulated in group 8-OH-DPAT compared with groups NP and DMSO ( P ＜ 0.01). There was no significant difference in 5-HT1A receptor expression between groups NP and DMSO ( P ＞ 0.05). Conclusion 5-HT1A receptors in distal CSF-contacting neurons are involved in the regulation of NP in rats.

Time-resolved fluorescence aptamer-based sandwich assay for thrombin detection.

In the present study, the authors report a novel sensitive method for the detection of thrombin using time-resolved fluorescence sensing platform based on two different thrombin aptamers. The thrombin 15-mer aptamer as a capture probe was covalently attached to the surface of glass slide, and the thrombin 29-mer aptamer was fluorescently labeled as a detection probe. A bifunctional europium complex was used as the fluorescent label. The introduction of thrombin triggers the two different thrombin aptamers and thrombin to form a sandwich structure. The fluorescence intensity is proportional to the thrombin concentration. The present sensing system could provide both a wide linear dynamic range and a low detection limit. The proposed sensing system also presented satisfactory specificity and selectivity. Results showed that thrombin was retained at the aptamer-modified glass surface while nonspecific proteins were removed by rinsing with buffer solution. This approach successfully showed the suitability of aptamers as low molecular weight receptors on glass slides for sensitive and specific protein detection.

A novel bifunctional europium complex as a potential fluorescent label for DNA detection.

A bifunctional europium complex of Eu(TTA)(3)(5-NH(2)-phen) using 2-thenoyltrifluoroacetonate (TTA) and 5-amino-1,10-phenanthroline (5-NH(2)-phen) as ligand reagents was applied in DNA detection assays for the first time. The complex has a long fluorescence lifetime, high fluorescence quantum yield, and is easy to label oligonucleotides for time-resolved fluorescence bioanalysis. A two-probe tandem DNA hybridization assay including capture DNA(1), probe DNA(2), and target DNA(3) was employed to detect microbial pathogens. The DNA sequences in the assay were designed using software Primer Premier 5.0 based on published specific nucleotide sequences of Staphylococcus aureus and Escherichia coli. 3'-Amino-modified capture DNA(1) was covalently immobilized on the common glass slide surface and the 5'-amino-modified probe DNA(2) was combined with the functionalized Eu(TTA)(3)(5-NH(2)-phen) via glutaraldehyde. The detection was done by monitoring the fluorescence intensity from the glass surface after the hybridization reaction with complementary target DNA(3). The optimal concentration of capture DNA(1) of 1.0 x 10(-6) mol l(-1) dropped onto the glass slides and optimal hybridization temperatures of 48 degrees C and 39 degrees C respectively for Staphylococcus aureus and Escherichia coli were obtained. The proposed DNA detection system showed higher sensitivity than such a complex doped nanoparticle-based detection system in our previous study for the better uniformity and dispersity of monomolecular labels. The sensing system presented a short hybridization time of 2 h, satisfactory stability, and high selectivity. The results demonstrate that this complex might be a potentially excellent dye in area of biochemical analysis.

Sensitive and simple detection of Escherichia coli strain based on time-resolved fluorescence DNA hybridization assay.

A two-probe tandem DNA hybridization assay based on time-resolved fluorescence was employed to detect Escherichia coli strain. The amino modified capture probe was covalently immobilized on the common glass slide surface. The Eu(TTA)(3)(5-NH(2)-phen) with the characteristics of long lifetime and intense luminescence was labeled with reporter probe. The original extracted DNA samples without the purification and amplification process were directly used in the hybridization assay. The concentration of capture probe, hybridization temperature, hybridization and washing time were optimized. The detection limit is about 1.49x10(3) CFU mL(-1) E. coli cells, which is comparable to the value of most microbiology methods. The proposed method has the advantages of easy operation, satisfactory sensitivity and specificity, which can provide a promising technique for monitoring the microorganisms.

[OCT4 expression in hepatocellular carcinoma and its clinical significance].

BACKGROUND AND OBJECTIVE: Recently, many studies have focused on stem cells in hepatocellular carcinoma (HCC) and found some stem cell markers in HCC, which are associated with the prognosis. OCT4, as a member of the POU transcription factor family, is a key factor to maintain self-renewal and pluripotency of embryonic stem cells (ESCs). This study was to explore the expression of the ESCs marker OCT4A in HCC, and its correlations with clinicopathologic features and prognosis of HCC. METHODS: OCT4A mRNA expression was detected in five liver cancer cell lines (SMMC-7721, BEL-7402, Hep-G2, MHCC97-L, and MHCC97-H), one immortalized liver cell line L-O2, tumor tissues with matched non-neoplastic liver tissues in 107 HCC patients, and normal liver tissues of 20 cases using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The correlations between OCT4A mRNA and clinicopathologic features and prognosis of HCC were analyzed. RESULTS: OCT4A mRNA was detected in SMMC-7721, BEL-7402, Hep-G2, MHCC-97L, and MHCC-97H cells, but not in L-O2 cells. The positive rate of OCT4A mRNA expression was significantly higher in the HCC tissues than in the non-neoplastic liver tissues (72.0% vs. 30.8%, P<0.001). No OCT4A mRNA expression was found in the normal liver tissues. OCT4A mRNA expression was correlated with the tumor size, vascular invasion, and TNM stage (P<0.05). Kaplan-Meier survival curves showed that patients with positive expression of OCT4A mRNA had lower overall survival and disease-free survival rates. CONCLUSIONS: OCT4A mRNA, which is highly expressed in a subset of liver cancer cell lines and HCC tissues, may be involved in the carcinogenesis of HCC. OCT4A mRNA may be a valuable biomarker for assessing the prognosis of HCC.

OCT4 expression in hepatocellular carcinoma and its clinical significance / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Recently, many studies have focused on stem cells in hepatocellular carcinoma (HCC) and found some stem cell markers in HCC, which are associated with the prognosis. OCT4, as a member of the POU transcription factor family, is a key factor to maintain self-renewal and pluripotency of embryonic stem cells (ESCs). This study was to explore the expression of the ESCs marker OCT4A in HCC, and its correlations with clinicopathologic features and prognosis of HCC.</p><p><b>METHODS</b>OCT4A mRNA expression was detected in five liver cancer cell lines (SMMC-7721, BEL-7402, Hep-G2, MHCC97-L, and MHCC97-H), one immortalized liver cell line L-O2, tumor tissues with matched non-neoplastic liver tissues in 107 HCC patients, and normal liver tissues of 20 cases using semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). The correlations between OCT4A mRNA and clinicopathologic features and prognosis of HCC were analyzed.</p><p><b>RESULTS</b>OCT4A mRNA was detected in SMMC-7721, BEL-7402, Hep-G2, MHCC-97L, and MHCC-97H cells, but not in L-O2 cells. The positive rate of OCT4A mRNA expression was significantly higher in the HCC tissues than in the non-neoplastic liver tissues (72.0% vs. 30.8%, P<0.001). No OCT4A mRNA expression was found in the normal liver tissues. OCT4A mRNA expression was correlated with the tumor size, vascular invasion, and TNM stage (P<0.05). Kaplan-Meier survival curves showed that patients with positive expression of OCT4A mRNA had lower overall survival and disease-free survival rates.</p><p><b>CONCLUSIONS</b>OCT4A mRNA, which is highly expressed in a subset of liver cancer cell lines and HCC tissues, may be involved in the carcinogenesis of HCC. OCT4A mRNA may be a valuable biomarker for assessing the prognosis of HCC.</p>

Time-resolved fluorescence based DNA detection using novel europium ternary complex doped silica nanoparticles.

A two-probe tandem DNA hybridization assay including capture DNA(1), probe DNA(2), and target DNA(3) was prepared. The long-lived luminescent europium complex doped nanoparticles (NPs) were used as the biomarker. The complex included in the particle was Eu(TTA)(3)(5-NH(2)-phen)-IgG (ETN-IgG), the europium complex Eu(TTA)(3)(5-NH(2)-phen) linking an IgG molecule. Silica NPs containing ETN-IgG were prepared by the reverse microemulsion method, and were easy to label oligonucleotide for time-resolved fluorescence assays. The luminophores were well-protected from the environmental interference when they were doped inside the silica network. The sequences of Staphylococcus aureus and Escherichia coli genes were designed using software Primer Premier 5.0. Amino-modified capture DNA(1) was covalently immobilized on the common glass slides surface. The detection was done by monitoring the fluorescence intensity from the glass surface after the hybridization reaction with the NPs labeled probe DNA(2) and complementary target DNA(3). The sensing system presented short hybridization time, satisfactory stability, sensitivity, and selectivity. This approach was successfully employed for preliminary application in the detection of pure cultured E. coli, it might be an effective tool for pathogen DNA monitoring.

Fluorescence sensor for water in organic solvents prepared from covalent immobilization of 4-morpholinyl-1, 8-naphthalimide.

A new fluorescent dye, N-allyl-4-morpholinyl-1,8-naphthalimide (AMN), was synthesized as a fluorescence indicator in the fabrication of a sensor for determining water content in organic solvents. To prevent leakage of the fluorophore, AMN was photo-copolymerized with acrylamide, (2-hydroxyethyl)methacrylate, and triethylene glycol dimethacrylate on a glass surface treated with a silanizing agent. The sensing mechanism is based on the solvatochromic feature of the covalently immobilized AMN. The fluorescence intensity of AMN decreased with increasing water contents when it was excited at 400 nm. In the range of ca. 0.00-4.40% (v/v), the fluorescence intensity of AMN changed as a linear function of water content. The sensor exhibited satisfactory reproducibility, reversibility, and a response time (t (99)) of the order of 50 s. The detection limit was solvent-dependent, when acetonitrile was used as the solvent, and the detection limit could be as low as 0.006% (v/v) of water. Additionally, the prepared sensor is pH-insensitive and possesses a relatively long lifetime of at least one month.

Peripheral blood stem cell mobilization with low dose rhG-CSF in 56 unrelated healthy donors / 中国实验血液学杂志

The study was aimed to observe the effect of recombinant human granulocyte-colony stimulating factors (rhG-CSF) in low dose on peripheral blood stem cell (PBSC) mobilization in unrelated healthy normal donors. G-CSF was administered at 5 microg/(kg x d) subcutaneously for successive 5 or 6 days to 56 unrelated donors. Stem cells were harvested on the fourth and fifth days or on the fifth and sixth days. The numbers of mononuclear cells (MNC), CD34(+) cells and Hb, Plt, and CD3(+), CD4(+), CD8(+) and CD20(+) cells were determined during the mobilization. The results showed that most common adverse events were bone pain (17.9%, 10/56), agrypnia (8.9%, 5/56) and lassitude (4.5%, 3/56) during rhG-CSF mobilization, but all donors were suffered less than grade II according to the WHO criteria, and did not need to stop the mobilization and not need to give special treatment. In harvest on day 4 - 5 and 5 - 6, MNC count was (5.95 +/- 1.52) x 10(8)/kg and (7.19 +/- 2.12) x 10(8)/kg; CD34(+) cells count was (3.03 +/- 1.09) x 10(6)/kg and (7.92 +/- 2.50) x 10(6)/kg. There were no significant differences in hemoglobin level and platelet count, the percentage of CD3(+) cells, CD4(+) cells, CD8(+) cells and CD20(+) cells between pre-mobilization and post-mobilization of rhG-CSF. It is concluded that the low dose of rhG-CSF 5 microg/(kg x d) for peripheral blood stem cell mobilization in unrelated healthy normal donors is safe and effective.