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A blind carrier synchronization scheme based on minimum mean square error (MMSE) phase error detector and digital 2nd order loop is proposed for the demodulation of APSK in a low-SNR environment. In contrast to the existing decision-directed (DD) algorithm, which is widely adopted for blind synchronization, MMSE phase error detector does not involve the hard symbol decision mechanism, thus it is not sensitive to the false judgment of symbol. Obviously, the detection range is broadened and the degree of phase ambiguity is decreased. To reduce the computational overhead of MMSE detector, a look-up table methodology is used in the scheme. Implementation considerations concerning the key parameters of the carrier phase synchronization loop, the sample-rate adaptive strategy and the fixed point design are summarized. [ABSTRACT FROM AUTHOR]

In this paper, a method to generate high-precision Gaussian white noise is proposed, which takes advantage of the mapping relationship between uniform distribution and Gaussian distribution. The highlight of this method is to increase the accuracy of the generated white Gaussian noise waveform by dividing the mapping curve into different partitions with unequal quantized precision. Experimental results have demonstrated that the proposed method can meet the noise precision requirement and reduce the FPGA resource consumption at the same time. [ABSTRACT FROM AUTHOR]

Previous studies have demonstrated that extracorporeal shock wave therapy (ESWT) could accelerate diabetic wound healing and that the inhibition of glycogen synthase kinase-3β (GSK-3β) is involved in epithelial differentiation during wound healing. This study investigated whether the enhancement of diabetic wound healing by ESWT is associated with the GSK-3β-mediated Wnt/β-catenin signaling pathway. A dorsal skin wounding defect model using streptozotocin-induced diabetic rodents was established. Rats were divided into 4 groups: group 1, normal controls without diabetes; group 2, diabetic controls without treatment; group 3, diabetic rats receiving ESWT; and group 4, rats receiving 6-bromoindirubin-3′oxime (BIO), a GSK-3β inhibitor, to trigger Wnt/β-catenin signaling. Tissue samples were collected and analyzed by immunohistochemical (IHC) staining and quantitative RT-PCR. The ESWT and BIO-treated groups both exhibited significant promotion of wound healing compared to the healing in controls without treatment. RT-PCR analysis of Wnt-1, -3a, -4, -5a, and -10 and β-catenin expression showed significantly increased expression in the ESWT group. The IHC staining showed that Wnt-3a and -5a and β-catenin levels were significantly increased in the ESWT and BIO treatment groups compared to the control groups. ESWT enhancement of diabetic wound healing is associated with modulation of the GSK-3β-mediated Wnt/β-catenin signaling pathway. [ABSTRACT FROM AUTHOR]

Naturally occurring CD4+CD25+ regulatory T cells (Tregs) are essential for the suppression of autoimmunity and can control the immune-mediated pathology during the early phase of sepsis. Our previous data showed that silencing interleukin-37 (IL-37) in human CD4+CD25+ Tregs obviously reduced the suppressive activity of CD4+CD25+ Tregs. Here, we found that rhIL-37 stimulation markedly enhanced the suppressive activity of CD4+CD25+ Tregs isolated from naive C57BL/6 J mice in the absence or presence of lipopolysaccharide (LPS). Treatment with rhIL-37 could significantly upregulate the expression of cytotoxic T-lymphocyte-associated antigen (CTLA)-4 and forkhead/winged helix transcription factor p3 (Foxp3) on CD4+CD25+ Tregs. Also, rhIL-37 stimulation promoted the production of transforming growth factor-β1 (TGF-β1) but not IL-10 in the supernatants of cultured CD4+CD25+ Tregs. Pretreated CD4+CD25+ Tregs with rhIL-37 in the presence or absence of LPS were cocultured with CD4+CD25− T cells, ratio of IL-4/interferon-γ in the supernatants obviously increased in IL-37-stimulated groups. In addition, early administration of IL-37 significantly improved the survival rate of septic mice induced by cecal ligation and puncture. Taken together, we concluded that rhIL-37 enhances the suppressive activity of CD4+CD25+ Tregs and might be a potential immunomodulator for the treatment of septic complications. [ABSTRACT FROM AUTHOR]

Naturally occurring CD4 + CD25 + regulatory T cells (Tregs) are essential for the suppression of autoimmunity and can control the immune-mediated pathology during the early phase of sepsis. Our previous data showed that silencing interleukin-37 (IL-37) in human CD4 + CD25 + Tregs obviously reduced the suppressive activity of CD4 + CD25 + Tregs. Here, we found that rhIL-37 stimulation markedly enhanced the suppressive activity of CD4 + CD25 + Tregs isolated from naive C57BL/6 J mice in the absence or presence of lipopolysaccharide (LPS). Treatment with rhIL-37 could significantly upregulate the expression of cytotoxic T-lymphocyte-associated antigen (CTLA)-4 and forkhead/winged helix transcription factor p3 (Foxp3) on CD4 + CD25 + Tregs. Also, rhIL-37 stimulation promoted the production of transforming growth factor-β1 (TGF-β1) but not IL-10 in the supernatants of cultured CD4 + CD25 + Tregs. Pretreated CD4 + CD25 + Tregs with rhIL-37 in the presence or absence of LPS were cocultured with CD4 + CD25 - T cells, ratio of IL-4/interferon-γ in the supernatants obviously increased in IL-37-stimulated groups. In addition, early administration of IL-37 significantly improved the survival rate of septic mice induced by cecal ligation and puncture. Taken together, we concluded that rhIL-37 enhances the suppressive activity of CD4 + CD25 + Tregs and might be a potential immunomodulator for the treatment of septic complications.

Objective: To investigate the expression of high mobility group protein B1 (HMGB1) and its receptor, receptor for advanced glycation end-product (RAGE), in renal cancer tissue and surrounding normal tissue and to analyze the relationship between the expression level of the protein and receptor as well as the clinical pathological characteristics and prognosis in renal cancer patients., Methods: A total of 80 renal carcinoma patients who were surgically treated in our hospital from February 2004 to December 2012 were included in this study. Normal paratumoral tissues were collected as a control. All diagnoses were confirmed with a postoperative pathological examination. All patients had complete pathological data. The expression of HMGB1/RAGE proteins in renal cancer tissue and paratumoral tissue was examined using immunohistochemical methods., Results: The positive expression rate of HMGB1 was 71% in renal cancer tissue, which was significantly higher than that in the paratumoral normal tissue (25%). The positive expression rate of RAGE was 72% in renal cancer tissue, which was significantly higher than that in the paratumoral normal tissue (27%). Further analysis did not indicate a correlation between the positive expression of HMGB1 and RAGE proteins and gender, age and tumor size (P > 0.05), whereas the expression patterns were shown to correlate with tumor differentiation, clinical stage and lymph node metastasis (P < 0.05). The expression of HMGB1 exhibited a significant positive correlation with RAGE level (P < 0.05), the expression of HMGB1/RAGE proteins exhibited a negative correlation with the prognosis of patients, and the five-year survival rate of patients with positive expression was significantly lower than that of patients with negative expression (P < 0.05)., Conclusion: HMGB1/RAGE exhibited significantly elevated expression in renal cancer tissues that was closely related to the clinical prognosis of patients; thus, the expression levels may become a new target in the treatment of renal carcinoma.

Objective: This study aims to identify morphological changes in the lung parenchyma of acute respiratory distress syndrome (ARDS) survivors after extracorporeal membrane oxygenation  (ECMO) by high-resolution computed tomography (HRCT) follow-up. Factors influencing these changes are also examined., Methods: Information and lung HRCT scans were collected and studied 1, 3, 6, and 12 months after the withdrawal of severe ARDS survivors rescued by ECMO in the Respiratory Care Unit of Beijing Chaoyang Hospital from November 2009 to August 2012. The observation endpoint was set as the time when the lung lesions were basically absorbed or 12 months after withdrawal., Results: Among nine survivors, one survivor was lost to follow-up. The lesions of two patients, which were attributed to bacterial pneumonia and pneumocystis pneumonia, were basically absorbed 1 month after surgery. Six patients completed the 12 month follow-up. Although initial morphological changes varied, different degrees of absorption improvement were observed in later stages of treatment. Lung HRCT analysis on the sixth month indicated that the degree of involvement of the ventral region was greater than that of the dorsal area. No significant difference was observed in patients in terms of ECMO support time, pre-ECMO Murray score, and APACHE II score, among others., Conclusion: Lung HRCT of severe ARDS survivors after ECMO treatment showed various degrees of morphological changes in the lung parenchyma. The severity of these changes may be associated with the disease duration.

Objective: To investigate the effect of continuous high-volume hemofiltration (CHVHF) in patients with severe acute respiratory distress syndrome (ARDS)., Methods: A prospective randomized controlled trial was conducted. Sixty-five patients with severe ARDS admitted to intensive care unit (ICU) from June 2007 to June 2011 were divided into control group (n=28) and treatment group (n=37). Patients in treatment group were treated with CHVHF and other routine treatments. Patients in control group received routine treatments only. The oxygenation index (PaO2/FiO2), extravascular lung water index (EVLWI), arterial partial pressure of carbon dioxide (PaCO2), heart rate (HR), mean arterial pressure (MAP) were compared between control group and treatment group before and 6, 24, 48, 72 hours after treatment. The duration of mechanical ventilation (MV), ICU stay time, percentage of weaning from MV, and 28-day survival rate were also compared., Results: The indexes of pulmonary function were improved after treatment in both groups. With prolonged time of treatment, PaO2/FiO2 was elevated, and EVLWI, PaCO2 were lowered, and the improvements were more marked in treatment group compared with control group (6-hour PaO2/FiO2: 92.6±7.2 mm Hg vs. 83.8±11.4 mm Hg, 24-hour EVLWI: 10.8±3.7 ml/kg vs. 12.6±4.5 ml/kg, 24-hour PaCO2: 47.2±8.5 mm Hg vs. 51.4±4.8 mm Hg, all P<0.05). HR and MAP were improved after the treatment in both groups, and there was no significant difference between groups. Compared with control group, the duration of MV and ICU stay were shortened in treatment group (duration of MV: 12±4 days vs. 19±6 days, ICU stay time: 21±4 days vs. 33±8 days, both P<0.05), and percentage of successful weaning from MV and 28-day survival rate were higher in treatment group (percentage of successful weaning from MV: 81.1% vs. 64.3%, 28-day survival rate: 86.5% vs. 71.4%, both P<0.05)., Conclusions: CHVHF is an effective adjuvant treatment for severe ARDS. It can improve the lung function, shorten the duration of MV, improve the percentage of successful weaning from MV, and the survival rate, and it lowers the mortality, but it imparts no obvious influence to hemodynamics in patients.

Objective: To summarize the experience of extracorporeal membrane oxygenation (ECMO) for patients with severe acute respiratory failure in adults and to investigate the factors associated with death., Methods: The clinical data of patients with severe acute respiratory failure supported with ECMO in respiratory intensive care unit of Beijing Chaoyang Hospital from November 2009 to December 2011 were prospectively collected and analyzed. The data included general condition before EMCO, blood gas analysis, hemodynamics, ventilator settings of mechanical ventilation and complications during ECMO. The primary outcome was death or severe disability within 3 months. Statistical software of SPSS (version 16.0) was used for data analysis., Results: Twenty-five patients with severe respiratory failure received ECMO treatment, of which 16 patients were analyzed. The mean age was (45 ± 14) years old (range, 22 - 64 years old). Thirteen patients were male. Before ECMO, all of the patients were treated with invasive positive pressure ventilation for (72 ± 64) hours. Eight patients had been treated with noninvasive ventilation for a median of 55(10-114) hours. Patients had severe respiratory failure despite advanced mechanical ventilator support. The mean PaO2/fraction of inspired oxygenation (FiO2) ratio was (54 ± 18) mm Hg (1 mm Hg = 0.133 kPa), positive end-expiratory pressure (PEEP) was (11 ± 6) cm H2O(1 cm H2O = 0.098 kPa), Murray lung injury score was 3.6 ± 0.5, serum lactate was (2.5 ± 2.0) mmol/L, serum white blood cell count was (16 ± 6)×10(9)/L, and APACHEII score was 17 ± 8. All of the patients were treated with venous-venous ECMO (VV-ECMO). The change of mechanical ventilation settings were (pre-ECMO vs 2 hours post-ECMO): FiO2 1.0 vs 0.55 ± 0.21, PEEP (11 ± 6) vs (9 ± 6) cm H2O, V(T) (6.8 ± 2.2) vs (4.4 ± 2.0) ml/kg PBW, peak airway pressure (27 ± 8) vs (24 ± 7) cm H2O, respiratory rate (37 ± 10) vs (23 ± 10) breaths/min. Arterial blood gas, including pH, PaO2 and PaCO2 were significantly improved after ECMO running 24 and 48 hours (P < 0.05). The mean VV-ECMO support interval was (9.7 ± 9.6) days (range, 2 - 41 days). Ten patients were successfully weaned from ECMO, of whom 2 died in ICU. Three patients died during ECMO, while another 3 patients died after withdrawal of further treatment. Eight patients survived to 3 months without severe disability. In a multi-variate Cox regression model, pre-ECMO factors including lower PaO2/FiO2 and increasing white blood cell count were associated with increased risks of death (RR was 0.733, 1.701 respectively, both P values < 0.05)., Conclusions: VV-ECMO is a potentially effective approach for severe acute respiratory failure. PaO2/FiO2 and white blood cell count pre-ECMO may be the risk factors for poor outcome.

Objective: To investigate the role of small G-protein RhoA in neointimal formation following rat carotid artery balloon injury and related mechanisms., Methods: Male 3-4-month-old Sprague-Dawley rats were used in the present study (10 rats per group). Group A: control; Group B: carotid artery balloon injury; Group C: injury + Ad-CMV-eGFP + Pluronic F-127; Group D: injury + Ad-CMV-N19RhoA-eGFP + Pluronic F-127; Group E: non injury + Ad-CMV-eGFP + Pluronic F-127. Perivascular gene transfer of an adenovirus co-expressing N19RhoA was performed to rat carotid artery following balloon injury and the effect on neointimal formation and the expressions of PCNA and α-SM-actin examined. Rats were killed after 14 days., Results: The protein expression of RhoA in group B was significantly higher than in group A (P = 0.001), and the positive cells rate of PCNA and α-SM-actin which were assessed by immunohistochemistry in group C (45.2% and 75.6%) was significantly higher than in group D (28.4% and 51.9%, all P < 0.01). The area of neointima was significantly smaller [(0.14 ± 0.08) mm(2) vs. (0.23 ± 0.10) mm(2), P < 0.01], the luminal area was significantly larger [(0.47 ± 0.11) mm(2) vs. (0.31 ± 0.06) mm(2), P < 0.01] in group D than in group C., Conclusion: Gene transfer of N19RhoA attenuates neointimal formation after balloon injury in rat carotid arteries possibly related to the modulating capacities of small G-protein RhoA on the proliferation, phenotypic differentiation and migration of vascular adventitial fibroblasts.

The present study investigated the expression of the novel gene hBiot2 in colorectal cancer (CRC) and its relationships with clinicopathological variables in CRC patients. The expression of hBiot2 in 163 primary CRCs together with the corresponding normal mucosa, 36 liver metastases and 5 colon cancer cell lines was examined using real-time PCR. In situ hybridization (ISH) was performed to evaluate the localization of hBiot2 expression in CRC and normal mucosa. hBiot2 expression at the RNA level was localized in the nucleus of tumor cells and normal epithelial cells. The mean expression of hBiot2 in the CRCs (243.571±564.569) was higher compared to the normal mucosa (107.252±413.635, P<0.0001) and liver metastasis samples (42.002±40.809, P=0.0002). hBiot2 expression was increased from stages I+II to III (P=0.047), and no difference in the expression was found in stages III and IV (P=0.452). A high value of hBiot2 was associated with a poorer prognosis compared with a low value independently of gender, age, tumor site, stage and differentiation (P=0.007, RR 7.519, 95% CI 1.729-32.704). Liver metastasis, smaller tumors, non-local recurrence and primary liver surgery alone were associated with a higher value of hBiot2 compared to larger tumors, local recurrence and repeated liver surgery (P=0.003, 0.044 and 0.026, respectively). An inverse relationship was found between hBiot2 expression and the metastatic potential of the colon cancer cell lines. Thus, increased expression of hBiot2 may be an early and interim event in the development of CRC. A higher expression of hBiot2 in primary CRC patients independently indicates a poorer prognosis.

Metastasis is the most lethal attribute of human malignancy. High-level expression of survivin is involved in both carcinogenesis and angiogenesis in cancer. Previous studies indicate that a mutation of the threonine residue at position 34 (Thr34Ala) of survivin generates a dominant-negative mutant that induces apoptosis, inhibits angiogenesis, and suppresses highly metastatic breast carcinoma in mouse models. We investigated the efficacy of gene therapy with a survivin dominant-negative mutant and possible factors related to lymph node metastasis. The metastasis rate was compared between each group in order to find a survivin-targeted therapy against lymphangiogenesis in its earliest stages. We established lymph node metastasis models and treated animals with H22 tumors with Lip-mSurvivinT34A (Lip-mS), Lip-plasmid (Lip-P), or normal saline (NS). Eight days after the last dose, five randomly chosen mice from each group were sacrificed. We detected the apoptotic index, microvessel density (MVD), lymphatic microvessel density (LMVD), and the expression of VEGF-D with immunohistochemistry. After the remaining animals were sacrificed, we compared the tumor-infiltrated lymph nodes in each group. Administration of mSurvivinT34A plasmid complexed with cationic liposome (DOTAP/chol) resulted in the efficacious inhibition of tumor growth and lymph node metastasis within the mouse H22 tumor model. These responses were associated with tumor cell apoptosis, and angiogenesis and lymphangiogenesis inhibition. Our results suggested that Lip-mSurvivinT34A induced apoptosis and inhibited tumor angiogenesis and lymphangiogenesis, thus suppressing tumor growth and lymphatic metastasis. The mSurvivinT34A survivin mutant is a promising strategy of gene therapy to inhibit lymphatic metastasis.

Objective: To evaluate the effects of administration of 6% hydroxyethyl starch (6% HES 130/0.4, voluven) in combination with high volume hemofiltration (HVHF) in patients with ALI and AKI., Methods: One hundred and eight patients with acute lung injury (ALI) and acute kidney injury (AKI) were enrolled from Department of Intensive Care Unit (ICU) of the provincial Hospital Affiliated to Shandong University between August 2006 and May 2011. The patients were randomly divided into two groups A (n = 68) and B (n = 40) to receive voluven (i.v., for volume resuscitation) and voluven+HVHF for 72 hours. The arterial blood lactate concentration (Lac), high sensitivity C-reactive protein (hs-CRP) serum concentration, pulmonary function index alveolar-arterial oxygen pressure difference [P(A-a)DO2] and oxygenation index (OI), as well as kidney function index serum cystatin C (Cyst C) and serum creatinine clearance rate (CCr) were measured at the time of admission and 72 hours after the treatment for statistical analysis., Results: In comparison with group A, group B had significantly (all P < 0.01) lower mean value in the level of arterial Lac (mmol/L: 1.7 ± 0.7 vs. 2.7 ± 1.5), serum hs-CRP (mg/L: 35.8 ± 18.8 vs. 99.5 ± 20.4), P(A-a)DO2 (mm Hg, 1 mm Hg=0.133 kPa: 115.5 ± 23.1 vs. 155.4 ± 27.4), Cyst C (mg/L: 2.06 ± 1.12 vs. 3.95 ± 2.06) and significantly higher (both P < 0.01) mean value of OI (mm Hg: 295.2 ± 38.8 vs. 239.5 ± 32.7) and CCr (ml/min: 108.71 ± 31.33 vs. 90.21 ± 30.35) 72 hours after treatment. The mortality rate of group B was significantly lower than group A [10.00%(4/40) vs. 29.41%(20/68), P < 0.05] 7 days after the admission., Conclusion: 6% HES 130/0.4 in combination with HVHF could improve the lung and kidney function of the patients with ALI and AKI, prevent the development of multiple organ dysfunction syndrome (MODS), therefore improve the survival rate of these patients.

Objective: To study the antitumor effects and mechanism of cationic liposome-mediated SurvivinT34A mutants in mice with pulmonary metastases of melanoma B16., Methods: The pulmonary metastases model of B16, a mouse melanoma cell line, was established by the injection of B16 cell suspension via tail vein of C57BL/6 mouse. 3 days later, 18 tumor-bearing mice were randomly divided into 3 groups, which were intravenously administrated with normal saline (100 microL), vector (5 microg DNA, 100 microL) and recombinant plasmid SurvivinT34A mutants (5 microg DNA, 100 microL) every other day for five doses, respectively. After 28 days, the mice were sacrificed, the lung tissue was weighted, and the number of metastasis foci on lung surface was counted and measured. Then, pathological change of lung tissue was studied with HE stainning. The number of apoptotic cells in tumor tissues was assessed by TUNEL assay., Results: Compared with the control, mice treated with mSurvivnT34A had an intact structure of lung, with significant reduction in the number of metastasis foci on lung surface (P < 0.05), and high level of apoptosis in tumor tissue (P < 0. 05)., Conclusion: Recombinant plasmid SurvivinT34A mutants (pORF9-mSurvivinT34A) may inhibit the formation of pulmonary metastases of melanoma.

The novel gene human Biot2 (hBiot2) was first reported by our laboratory. Previously, we indicated its function of proliferation and carcinogenesis in human endometrial cancer. In the present study, we aimed to investigate whether hBiot2 played a similar role in human cervical cancer. We tested hBiot2 expression profile in cervical cancer, the corresponding adjacent normal tissues, normal cervix and the cervical cancer cell lines by RT-PCR and compared the mean value of hBiot2 expression between cervical cancer and normal cervix, and cervical cancer with or without lymphatic metastasis by real-time PCR. The location of hBiot2 in normal cervix and cancer tissues together with the corresponding adjacent normal tissues was determined by RNA-RNA in situ hybridization (ISH). hBiot2 expression in the cervical cancer (20/25), the corresponding adjacent normal tissues (3/12), normal cervix (17/18) and the cervical cell lines (2/3) was shown by RT-PCR. The mean value of hBiot2 expression was higher in the cervical cancer than in the normal cervix (0.478±1.612 vs. 0.091±0.107, P=0.0004), higher in the lymphatic metastasis than in the non-lymphatic metastasis in the cervical cancer (1.117±2.483 vs. 0.052±0.071, P=0.014). hBiot2 expression location was mainly in the parenchymal cells of the cervical cancer and normal cervix rather than in the stromal cells. Overexpression of hBiot2 is associated with early and interim development of human cervical cancer.

Background: High-volume hemofiltration (HVHF) is technically possible in severe acute pancreatitis (SAP) patients complicated with multiple organ dysfunction syndrome (MODS). Continuous HVHF is expected to become a beneficial adjunct therapy for SAP complicated with MODS. In this study, we aimed to explore the effects of fluid resuscitation and HVHF on alveolar-arterial oxygen exchange, the Acute Physiology and Chronic Health Evaluation II (APACHE II) score in patients with refractory septic shock., Methods: A total of 89 refractory septic shock patients, who were admitted to ICU, the Provincial Hospital affiliated to Shandong University from August 2006 to December 2009, were enrolled in this retrospective study. The patients were randomly divided into two groups: fluid resuscitation (group A, n=41), and fluid resuscitation plus high-volume hemofiltration (group B, n=48). The levels of O2 content of central venous blood (CcvO2), arterial oxygen content (CaO2), alveolar-arterial oxygen pressure difference P(A-a)DO2, ratio of arterial oxygen pressure/alveolar oxygen pressure (PaO2/PaO2), respiratory index (RI) and oxygenation index (OI) were determined. The oxygen exchange levels of the two groups were examined based on the arterial blood gas analysis at different times (0, 24, 72 hours and 7 days of treatment) in the two groups. The APACHE II score was calculated before and after 7-day treatment in the two groups., Results: The levels of CcvO2, CaO2 on day 7 in group A were significantly lower than those in group B (CcvO2: 0.60±0.24 vs. 0.72±0.28, P<0.05; CaO2: 0.84±0.43 vs. 0.94±0.46, P<0.05). The level of oxygen extraction rate (O2ER) in group A on the 7th day was significantly higher than that in group B (28.7±2.4 vs. 21.7±3.4, P<0.01). The levels of P(A-a)DO2 and RI in group B on the 7th day were significantly lower than those in group A. The levels of PaO2/PaO2 and OI in group B on 7th day were significantly higher than those in group A (P<0.05 or P<0.01). The APACHE II score in the two groups reduced gradually after 7-day treatment, and the APACHE II score on the 7th day in group B was significantly lower than that in group A (8.2±3.8 vs. 17.2±6.8, P<0.01)., Conclusion: HVHF combined with fluid resuscitation can improve alveolar-arterial-oxygen exchange, decrease the APACHE II score in patients with refractory septic shock, and thus it increases the survival rate of patients.

Objective: To investigate the regulation mechanism of p38 mitogen-activated protein kinase (p38MAPK) in interleukin-6 (IL-6) expression of vascular smooth muscle cell (VSMC) induced by lipopolysaccharide (LPS)., Methods: Rat VSMCs were divided into LPS group, SB203580+LPS group, SB203580 group and control group. LPS group was treated with 100 microg/L LPS for 0, 3, 6, 12, 24 hours, SB203580+LPS group was first treated with 10 micromol/L SB203580 for 2 hours and then exposed to 100 microg/L LPS for 0, 3, 6, 12, 24 hours, SB203580 group was pretreated with 10 micromol/L SB203580 for 2 hours. The level of IL-6 mRNA was determined by real-time polymerase chain reaction (PCR) and IL-6 secretion in the culture medium was measured by enzyme linked immunosorbent assay (ELISA) at different time points., Results: The expression of IL-6 mRNA and the release of IL-6 were increased significantly in VSMC as early as 3 hours after being treated with LPS [mRNA: (21.3+/-3.2)x10(4), protein: (296.2+/-19.6) ng/L], peaked in 12 hours [mRNA: (131.4+/-11.2)x10(4), protein: (897.7+/-34.0) ng/L], and the elevation persisted up to 24 hours after treatment [mRNA: (15.3+/-4.7)x10(4), protein: (194.3+/-24.0) ng/L] compared with control group [mRNA: (9.4+/-1.9)x10(4), protein: (29.4+/-4.4) ng/L, all P<0.05]. On the other hand, the expression of IL-6 was significantly suppressed by p38MAPK inhibitor SB203580 at 3, 6, 12 hours [mRNA: (15.4+/-3.6)x10(4), (43.2+/-6.6)x10(4), (56.2+/-5.5)x10(4), protein: (180.3+/-23.6), (432.2+/-56.8), (546.2+/-57.9) ng/L, all P<0.05]., Conclusion: The release of IL-6 and the expression of IL-6 mRNA was increased significantly in LPS-challenged VSMC; however, the induction of IL-6 was significantly suppressed by p38MAPK inhibitor. p38MAPK may play an important role in the release of IL-6 induced by LPS.

Objective: To investigate the effect of Survivin-T34A mutant on murine prostate cancer and its apoptosis-inducing efficacy in vivo and in vitro., Methods: In vitro, prostate cancer cells TRAMP-C1 were transfected with Survivin-T34A plasmid encapsulated by cationic liposome. The apoptosis of TRAMP-C1 was evaluated with flow cytometry. C57BL/6 mice model with TRAMP-C1 prostate cancer was established. Twenty four male mice with TRAMP-C1 prostate cancers were divided randomly into three groups, which were intravenously injected with normal saline, empty vector PORF-9-null encapsulated by cationic liposome and Survivin-T34A plasmid encapsulated by cationic liposome respectively twice a week for eight doses. The size of tumors was measured and the tumor sections of each group were stained with TUNEL reagent for apoptosis detection., Results: An apoptotic index of 46% of TRAMP-C1 transfected with Survivin-T34A plasmid encapsulated by cationic liposome was observed. The tumor volume of Survivin-T34A group of C57BL/6 mice with TRAMP-C1 prostate cancer was far smaller than those in the control groups (P < 0.05) and the tumors treated with Survivin-T34A showed significant increase of apoptosis compared with those of control groups (P < 0.05)., Conclusion: Survivin-T34A mutant efficiently inhibits the growth of prostate cancer, which is based on the mechanism of Survivin-T34A mutant inducing apoptosis of tumor cells.

Objective: To investigate the effects of high volume hemofiltration (HVHF) on the expression of Toll-like receptor 4 (TLR4) mRNA in myocardium in endotoxin (lipopolysaccharide, LPS) induced shock in dogs., Methods: Sixteen healthy male dogs were injected with LPS 650 microg/kg via central vein to reproduce the model of endotoxin shock. All dogs were divided randomly into two groups: control group and therapy group, with 8 dogs in each group. Contents of tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), IL-10 in circulation were measured by radioimmunological method. The expression levels of TLR4 mRNA in all group were measured by reverse transcription-polymerase chain reaction (RT-PCR). Change in myocardial histopathology was observed and analyzed with the aid of electron microscope., Results: The contents of TNF-alpha (microg/L: 0.59+/-0.15, 0.51+/-0.12, 0.41+/-0.10), IL-6 (ng/L: 11.08+/-2.83, 9.82+/-2.58, 8.25+/-2.05), IL-10 (microg/L: 57.28+/-5.93, 53.81+/-5.83, 50.67+/-6.33) in therapy group were found to have decreased significantly at 1, 2, and 4 hours after HVHF compared with those when the model was completed [(0.84+/-0.16) microg/L, (16.97+/-2.50) ng/L, (70.86+/-5.43) microg/L], showing a continuous trend of lowering (all P<0.01). The contents of TNF-alpha, IL-6, IL-10 in therapy group were lower than those in control group significantly at any time point [TNF-alpha (microg/L): 0.75+/-0.14, 0.74+/-0.11, 0.72+/-0.11, IL-6 (ng/L): 15.33+/-3.20, 14.66+/-3.24, 14.20+/-3.33, IL-10 (microg/L): 71.54+/-4.73, 70.71+/-4.34, 69.35+/-4.60, all P<0.01]. Compared with control group, HVHF treatment group could down-regulate mRNA expression of TLR4 in myocardium (t=3.58, P<0.01). Correlation analysis revealed significant positive-correlation between tissue TLR4 mRNA expression and contents of TNF-alpha, IL-6, IL-10 in circulation (r(1)=0.785, r(2)=0.569, r(3)=0.653, all P<0.05). Injury to the myocardium was significantly ameliorated in therapy group compared with control group as shown by electron microscopic observation., Conclusion: HVHF can down-regulate mRNA expression of TLR4 in myocardium in LPS induced shock in dogs, and myocardial inflammatory response was alleviated resulting in amelioration of myocardial injury.

Radiosensitivity of tumors is due to a complex interaction of various factors, it has been reported that survivin also acts as a constitutive and inducible radioresistance factor in a panel of tumor cells and approaches designed to inhibit survivin expression or function may lead to tumor sensitisation to chemical and physical agents. Previously, we found that the plasmid encoding the phosphorylation-defective mouse survivin threonine 34-->alanine mutant complexed to DOTAP-chol liposome (Lip-mS) can suppress murine primary breast carcinoma. However, little is known regarding the biological effect of Lip-mS combined with radiation. The present study was designed to determine whether Lip-mS could enhance the anti-tumor activity of radiation. The Lewis Lung Carcinoma (LLC) cells treated with a combination of Lip-mS and radiation displayed apparently increased apoptosis compared with those treated with Lip-mS or radiation alone. Mice bearing LLC tumors were treated with intravenous injections of Lip-mS and radiation, the combined treatment significantly reduced mean tumor volume compared with either treatment alone. Moreover, the anti-tumor effect of Lip-mS combined with radiation was greater than their additive effect when compared with the expected effect of the combined treatment. These data suggest that inhibition of survivin using a dominant-negative mutant, survivin T34A, could sensitize LLC cells to radiation efficiently and the synergistic anti-tumor activity may in part result from increasing the apoptosis of tumor cells, inhibiting tumor angiogenesis and inducing a tumor-protective immune response in the combined treatment.

Background: The widespread use of gastrointestinal bleeding prophylaxis in critically ill patients was one of the most controversial issues. Since few studies reported the incidence of gastrointestinal bleeding in mechanically ventilated patients, this study aimed to identify the incidence and risk factors related to gastrointestinal bleeding in patients undergoing mechanical ventilation for more than 48 hours., Methods: A total of 283 ICU patients who had received mechanical ventilation for longer than 48 hours at a provincial hospital affiliated to Shandong University from January 1, 2007 to December 31, 2009 were analyzed retrospectively. Those were excluded from the study if they had a history of gastrointestinal bleeding or ulceration, recent gastrointestinal surgery, brain death and active bleeding from the nose or throat. Demographic data of the patients included patient age, diagnosis on admission, duration of ICU stay, duration of ventilation, patterns and parameters of ventilation, ICU mortality, APACHE II score, multiple organ dysfunction, and indexes of biochemistry, kidney function, liver function and coagulation function. Risk factors of gastrointestinal bleeding were analyzed by univariate analysis and multiple logistic regression analysis., Results: In the 242 patients who were given mechanical ventilation longer than 48 hours, the incidence of gastrointestinal bleeding was 46.7%. The bleeding in 3.3% of the patients was clinically significant. Significant risk factors were peak inspiratory pressure ≥30cmH2O, renal failure, liver failure, PLT count<50×10(9)/L and prolonged APTT. Enteral nutrition had a beneficial effect on gastrointestinal bleeding. However, the multiple logistic regression analysis revealed that the independent risk factors of gastrointestinal bleeding were as follows: high pressure ventilator setting ≥ 30cmH2O(RR=3.478, 95%CI=2.208-10.733), renal failure(RR=1.687, 95%CI = 1.098-3.482), PLT count<50×1 0(9)/L (RR=3.762, 95%CI=2.346-14.685), and prolonged APTT(RR=5.368, 95%CI=2.487-11.266). Enteral nutrition(RR=0.436, 95%CI= 0.346-0.764) was the independent protective factor., Conclusions: The incidence of gastrointestinal bleeding was high in the patients who received mechanical ventilation, and bleeding usually occurred within the first 48 hours. High pressure ventilator setting, renal failure, decreased PLT count and prolonged APTT were the significant risk factors of gastrointestinal bleeding. However, enteral nutrition was the independent protective factor.

Down-regulated in renal cell carcinoma gene (DRR1) is one of the candidate tumor suppressor genes (TSGs) on human 3p21.1. This study was performed to validate the expression status of DRR1 gene in cancer cells and the expression pattern of the protein in clinical specimens of human lung cancer and to examine its potential as a molecular target for treatment of lung cancer in vivo. DRR1 expression was analyzed in 7 human lung cancer cell lines. DRR1 protein expression was also examined in clinical non-small cell lung cancer (NSCLC) specimens. Furthermore, effects of DRR1 re-expression on A549 cells in vitro and A549 xenograft tumors in nude mice were evaluated. Loss of DRR1 mRNA expression was detected in 6 of the 7 human cancer cell lines, the exception was the renal cancer cell line OS-RC-2. DRR1 protein expression was absent in 15 of 20 (75%) human NSCLC specimens by immunostaining. Transfection of DRR1 gene into DRR1-negative-expressing A549 cells resulted in significant cell growth suppression and apoptosis. Plasmids containing DRR1 cDNA complexed with DOTAP:Chol liposomes were administered intravenously via tail vein to nude mice bearing A549 xenograft tumors resulting in tumor growth inhibition and elevation of apoptosis compared with the controls. DRR1 is a potent growth suppressor of NSCLC, acting through apoptosis pathway in vivo and it may be a potential therapeutic gene for human lung cancer.

Objective: To study on the expression pattern of a novel gene, Biot2-L, in mice and its primary bio-function on the development of testis., Methods: Bioinformatics analysis were applied to identify the structure characteristics and homologene of Biot2-L. RT-PCR and Real-time RT-PCR were used to analyze its expression profile in normal tissues and the time-dependent expression pattern during the testes sexual maturation., Results: The complete sequence of Biot2-L was obtained. A conserved domain-CCDC7 was detected and there were homolog proteins in advanced mammals with about 50% identity by bioinformatics analysis. The highly expression of Biot2-L in adult testis but low or absent expressed in other tissues was observed. A regular change of Biot2-L expression level during the testis development and spermatogonia was demonstrated by the results of Real-time RT-PCR. The mRNA transcript appeared from the fifth week of postnatal development, and the expression level increased up to adulthood, but decreased when get older., Conclusion: Biot2-L was highly expressed in adult testis and regular changed during the testis development, which suggested that the Biot2-L would be involved in the testis development and spermatogonia.

Objective: To evaluate the effects of high-volume hemofiltration (HVHF) and fluid resuscitation on the levels of arterial lactate and inflammatory cytokines, the acute physiology and chronic health evaluation II (APACHE II) score, and sepsis-related organ failure assessment (SOFA) score in patients with refractory septic shock and multiple organ dysfunction syndrome (MODS)., Methods: Eighty-nine patients with refractory septic shock and MODS undergoing HVHF, fluid resuscitation were included in the trial, and they were randomly divided into fluid resuscitation group (group A, 41 cases) and HVHF and fluid resuscitation group (group B, 48 cases). The changes in arterial lactate, lactate clearance rate, interleukin-6 (IL-6), procalcitonin (PCT), high sensitivity C-reactive protein (hs-CRP), APACHE II score and SOFA score in patients with refractory septic shock and MODS were determined before and after the treatment., Results: (1) The arterial lactate clearance rate after treatment in both groups was elevated gradually. The level of lactate clearance rate in group B at 6, 12, 24 hours [(18.8+/-10.3)%, (31.6+/-11.4)%, (39.2+/-16.4)%] were higher than that of group A, respectively [(10.7+/-7.5)%, (14.7+/-10.3)%, (16.5+/-10.2)%, P < 0.05 or P < 0.01]. (2) The levels of pro-inflammatory cytokine IL-6, PCT and hs-CRP were reduced gradually after treatment in both groups. After treatment, the serum concentration of IL-6, PCT and hs-CRP levels in group B were reduced significantly more than that in group A at 1 day and 3 days (P < 0.05 or P < 0.01). (3) The APACHE II and SOFA scores in both groups were reduced gradually on 3 days and 7 days after treatment, but the APACHE II and SOFA scores on 7 days in group B were lower compared with group A (P < 0.05 and P < 0.01)., Conclusion: HVHF and fluid resuscitation could reduce arterial lactate and cytokines contents, at the same time lower the APACHE II score and SOFA score in patients with refractory septic shock and MODS, thus it could improve the survival rate of MODS patients.

Objective: To explore the feasibility of constructing a dendritic cell targeted adenovirus as a possible universal tumor vaccine., Methods: The cDNA of human telomerase reverse transcriptase (hTRT) from pBABE-PURO-hTERT was subcloned into pAdTrack-CMV. Recombinant adenovirus for hTRT was constructed in AdEasy system. The confirmed recombinant adenovirus plasmid was transfected into 293 cells to generate the recombinant adenovirus. Later the recombinant adenovirus was multiplied, purified and modified with mannan as a vaccine to immunize mice., Results: An obvious cytopathic effect was observed with the rescue of recombinant adenovirus. The expression of hTRT was confirmed by PCR and Western blot analysis. Mannan modified adenovirus stained positively by periodic acid Schiff stain, and expression of the adenovirus was found mainly restricted in dendritic cells (DC) in spleen of mice., Conclusion: A DC-targeted adenovirus was constructed. This might be helpful (or the development of universal tumor vaccine in future.

We have identified a novel Xenopus gene (xVAP019) encoding a DUF1208 domain containing protein. Using whole-mount in situ hybridization and RT-PCR, we found abundant xVAP019 maternal transcripts in the animal hemisphere during the cleavage stages and blastula stages. During gastrulation xVAP019 is differentially expressed with higher levels in the animal helf and the highest in marginal zone, then further expressed widely at neuronal stages with strongest signals in the prospective CNS regions and the epidermal ectoderm. Subsequently xVAP019 was expressed predominantly in the head, the eyes, the otic vesicle, branchial arches, spinal cord, notochord, somites, and tailbud. It is absent or very weak in the endoderm. Injecting a morpholino oligo complementary to xVAP019 mRNA or injecting a caped xVAP019 mRNA caused most of embryos to die during gastrulation and neurulation. Overexpression of xVAP019 mRNA also led to eye defect, shorten interocular distance, small body size and abnormal pigment formation in parts of the survival embryos. Similar effects were induced by injecting the xVAP019 human homologous gene FAM92A1. Our results suggest that xVAP019 is essential for the normal ectoderm and axis mesoderm differentiation and embryos survival. This investigation is for the first time in vivo study examining the role of this novel gene and reveals an important role of xVAP019 in embryonic development., ((c) 2007 Wiley-Liss, Inc.)

Aim: To study the biological effects of the novel tumor/testis antigen Biot2 on proliferation of NIH3T3 cell and to identify the preliminary biological functions of it., Methods: The Biot2 gene was amplified by RT-PCR and cloned into the eukaryotic expression vector pcDNA3.1(+) to construct the pcDNA3.1-Biot2 plasmid. The recombinant plasmid pcDNA3.1-Biot2 was transfected into NIH3T3 cell by lipofectamine 2000, and then the positive clones were screened by G418. The expression of pcDNA3.1(+) and Biot2 mRNA in positive clones were detected by RT-PCR, Realtime RT-PCR and Western blot, respectively. The cell growth curves were measured by MTT assay to study the changes of proliferation of the NIH3T3 cells stably transfected by pcDNA3.1-Biot2 plasmid and the controls., Results: The eukaryotic expression vecter pcDNA3.1-Biot2 was successfully constructed. The expression of pcDNA3.1(+) vecter was found by RT-PCR. Biot2 mRNA transcription and protein were detected by Realtime RT-PCR and Western blot respectively. The growth rate of NIH3T3 cells stably transfected by the recombinant plasmid was faster than that of the controls., Conclusion: The stable pcDNA3.1-Biot2 transfected NIH3T3 cell line is successfully established, and the proliferation of the cells stadly transfected is promoted, which will provide experimental basis for further study on biotherapy using Biot2 target antigen.