Insertion/deletion polymorphism at angiotensin-converting enzyme gene in PTSD individuals and their reciprocal effects on blood pressure.

OBJECTIVES: The aim of the present study was to investigate relationships between insertion/deletion (I/D) polymorphism at angiotensin-converting enzyme gene (ACE) and post-traumatic stress disorder (PTSD), as well as their interactions on blood pressure. METHODS: Variants of ACE I/D were identified by polymerase chain reaction method and verified by DNA sequencing. PTSD symptoms were assessed by the PTSD Checklist-Civilian Version (PCL-C) based on DSM-IV-TR criteria among high school students at 6 months after the 2008 Wenchuan earthquake. RESULTS: Female subjects were found to have higher prevalence of PTSD and PCL-C scores than male counterparts in the II homozygotes (p = .038 for PTSD and p = .003 for PCL-C scores) and the ID heterozygotes (p = .000 for PTSD and p = .000 for PCL-C scores), but not in the DD homozygotes. Male subjects with the ID (p = .046) or the DD genotype (p = .039) had lower pulse pressure (PP) than the male II homozygotes, while the female II homozygotes had lower diastolic blood pressure (DBP) than the female DD homozygotes (p = .036). ACE I/D, PTSD, or PCL-C scores, as well as gender and BMI, were found to be the predictors of PP. CONCLUSIONS: These results indicate that there are interactions of ACE I/D and PTSD, together with gender and BMI, on PP. This finding may be the additional explanation for the heterogeneous relationships between PTSD and blood pressure, and suggest psychiatry care and different medication strategies for patients with comorbidities of PTSD and hypertension and with different genotypes of ACE I/D.

Iatrogenic Pseudoaneurysm with Acute Arterial Thrombosis in Multiple Branches of the Lower Limbs Treated by Ultrasound-guided Thrombin Injection: A Case Report.

INTRODUCTION: With the development of vascular intervention, pseudoaneurysm complications are increasing. Ultrasound-guided thrombin injection (UGTI) is currently the treatment of choice for pseudoaneurysm, but the pharmacological properties of thrombin may trigger acute thrombosis within the vessel lumen. Despite a very low incidence, this type of primary arterial thrombosis is a serious complication of UGTI, and cases involving multiple branches of the lower limb arteries are particularly rare. CASE PRESENTATION: Here, we report a case of a 65-year-old male who underwent UGTI for the treatment of an iatrogenic pseudoaneurysm of the femoral artery complicated by acute thrombosis of multiple arteries in the lower limbs, and the patient ultimately underwent a successful thrombectomy. CONCLUSION: We reviewed the case and analyzed the possible etiologic causes, providing a reference for future clinical work.

Reduced hepatic AdipoR2 by increased glucocorticoid mediates effect of psychosocial stress to elevate serum cholesterol.

Understanding the effects of psychosocial stress on serum cholesterol may offer valuable insights into the relationship between psychological disorders and endocrine diseases. However, these effects and their underlying mechanisms have not been elucidated yet. Here we show that serum corticosterone, total cholesterol and low-density lipoprotein cholesterol (LDL-C) are elevated in a mouse model of psychosocial stress. Furthermore, alterations occur in AdipoR2-mediated AMPK and PPARα signaling pathways in liver, accompanied by a decrease in LDL-C clearance and an increase in cholesterol synthesis. These changes are further verified in wild-type and AdipoR2 overexpression HepG2 cells incubated with cortisol and AdipoR agonist, and are finally confirmed by treating wild-type and hepatic-specific AdipoR2 overexpression mice with corticosterone. We conclude that increased glucocorticoid mediates the effects of psychosocial stress to elevate serum cholesterol by inhibiting AdipoR2-mediated AMPK and PPARα signaling to decrease LDL-C clearance and increase cholesterol synthesis in liver.

Combining Serum Procalcitonin Level, Thromboelastography, and Platelet Count to Predict Short-term Development of Septic Shock in Intensive Care Unit.

OBJECTIVE: Despite the recent advances in diagnosis and treatment, sepsis continues to lead to high morbidity and mortality. Early diagnosis and prompt treatment are essential to save lives. However, most biomarkers can only help to diagnose sepsis, but cannot predict the development of septic shock in high-risk patients. The present study determined whether the combined measurement of procalcitonin (PCT), thromboelastography (TEG) and platelet (PLT) count can predict the development of septic shock. METHODS: A retrospective study was conducted on 175 septic patients who were admitted to the intensive care unit between January 2017 and February 2021. These patients were divided into two groups: 73 patients who developed septic shock were assigned to the septic shock group, while the remaining 102 patients were assigned to the sepsis group. Then, the demographic, clinical and laboratory data were recorded, and the predictive values of PCT, TEG and PLT count for the development of septic shock were analyzed. RESULTS: Compared to the sepsis group, the septic shock group had statistically lower PLT count and TEG measurements in the R value, K value, α angle, maximum amplitude, and coagulation index, but had longer prothrombin time (DT), longer activated partial thromboplastin time (APTT), and higher PCT levels. Furthermore, the Sequential Organ Failure Assessment (SOFA) score was higher in the septic shock group. The multivariate logistic regression analysis revealed that PCT, TEG and PLT count were associated with the development of septic shock. The area under the curve analysis revealed that the combined measurement of PCT, TEG and PLT count can be used to predict the development of septic shock with higher accuracy, when compared to individual measurements. CONCLUSION: The combined measurement of PCT, TEG and PLT count is a novel approach to predict the development of septic shock in high-risk patients.

Anlotinib in combination with pembrolizumab for low-grade myofibroblastic sarcoma of the pancreas: A case report.

BACKGROUND: Low-grade myofibroblastic sarcoma (LGMS) is a rare spindle cell sarcoma especially in the pancreas, with myofibroblastic differentiation. Hitherto, only a few cases have been reported. CASE SUMMARY: Herein, we report a case involving the discovery of a pancreatic mass detected during a routine physical examination. Subsequent imaging and pathological tests of the patient led to the diagnosis of LGMS of the pancreas. Following surgical intervention, the patient experienced recurrence and metastasis. Conventional treatment is not effective for postoperative recurrent pancreatic LGMS with multiple metastases. After communicating with the patients and their families, informed consent was obtained for the treatment of anlotinib combined with pembrolizumab. Evaluation of imaging and clinical symptoms post-treatment revealed a relatively favorable response to the combination of anlotinib and pembrolizumab. CONCLUSION: Based on the comprehensive literature review, our report aimed to provide evidence for a better understanding of the disease characteristics, diagnostic criteria, imaging findings, and identification of LGMS. And explore novel treatment strategies for this disease.

The Waist-Hip Ratio is a Mediator Between Serum Levels of Brain-Derived Neurotrophic Factor and Its Val66Met Polymorphism in Adolescents.

Aim: To explore anthropometric, metabolic and dietary factors associated with and their interplays with the Val66Met polymorphism at brain-derived neurotrophic factor (BDNF) gene (Bdnf) on serum BDNF levels in adolescents. Methods: Serum BDNF levels were quantified using an enzyme-linked immunosorbent assay in 644 high school students (278 males/366 females). A polymerase chain reaction and restriction fragment length polymorphism assay were utilized for Bdnf Val66Met genotyping followed by verification using DNA sequencing. Serum levels of metabolic characteristics were assayed by routine methods. The intake of macro and micronutrients was collected by a three-day food record. Results: Serum BDNF levels were found to be significantly different in the subjects with different genotypes of Bdnf Val66Met (Val/Val homozygotes, 60.05 ± 28.07 ng/mL vs Val/Met heterozygotes, 56.37 ± 29.34 ng/mL vs Met/Met homozygotes, 51.32 ± 24.54 ng/mL, p = 0.022). Among the 36 tested variables, waist-hip ratio (WHR) (ß = -0.163, p < 0.001), iodine intake (ß = 0.132, p = 0.001), heart rate (ß = 0.108, p = 0.005), high-density lipoprotein cholesterol (HDL-C) (ß = 0.098, p = 0.011) and dietary fiber intake (ß = 0.082, p = 0.084) were the predictor of serum BDNF levels, while SBP (ß = 0.097, p = 0.013) and WHR (ß = 0.091, p = 0.021) were related with Bdnf Val66Met. Moreover, WHR was observed to play a partial mediating role in the relationship between Bdnf Val66Met and serum BDNF levels (95% CI [-1.161, -0.087]) and contribute 13.05% of its total effect on serum BDNF levels. Conclusion: There are interplays between WHR and Bdnf Val66Met on serum BDNF levels, which may be among the explanations for the previous heterogeneous reports and provide novel insights into the regulation of serum BDNF levels.

Associations of TNF-RII rs1061622 With Post-Traumatic Stress Disorder and Their Interplays on Serum Lipids Levels in Adolescents.

OBJECTIVE: To verify effects of rs1061622 at tumor necrosis factor-α receptor II (TNF-RII) gene (TNF-RII) on post-traumatic stress disorder (PTSD) and its interactive effects with PTSD on serum lipids levels in adolescents. METHODS: PTSD was measured by PTSD Checklist-Civilian Version (PCL-C) in 699 adolescent survivors at 6 months after Wenchuan earthquake in China. A polymerase chain reaction and restriction fragment length polymorphism assay were utilized for TNF-RII rs1061622 genotyping followed by verification using DNA sequencing. Serum triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol were tested using routine methods. RESULTS: G (deoxyguanine) allele carriers had higher PCL-C scores than TT (deoxythymidine) homozygotes in female subjects. Female adolescents had higher PCL-C scores than male subjects in TT homozygotes. Predictors of PTSD prevalence and severity were different between G allele carriers and TT homozygotes. Subjects with PTSD had lower TG, TG/HDL-C, TC/HDL-C, and higher HDL-C than adolescents without PTSD in male G allele carriers. G allele carriers had higher TG/HDL-C and TC/HDL-C than TT homozygotes in male adolescents without PTSD, and lower TG and TG/HDL-C in male PTSD patients. G allele carriers had higher TG than TT homozygotes only in female adolescents without PTSD. CONCLUSION: These results suggest reciprocal actions of TNF-RII rs1061622 with other factors on PTSD severity, interplays of TNF-RII rs1061622 with PTSD on serum lipid levels, and novel treatment strategies for PTSD and comorbidities of PTSD with hyperlipidemia among adolescents with different genetic backgrounds of TNF-RII rs1061622 after experiencing traumatic events.

Effectiveness of combining plasma exchange with continuous hemodiafiltration on acute Fatty liver of pregnancy complicated by multiple organ dysfunction.

Acute fatty liver of pregnancy (AFLP) is a rare disease of progressive hepatic insufficiency and secondary systemic complications that induce significant maternal risk. The application of combining plasma exchange (PE) and continuous hemodiafiltration (CHDF) is a novel concept for patients with AFLP. Since 2002, we have utilized the combination of PE with CHDF as adjunctive medical therapy for 11 AFLP patients with multiple organ dysfunction. Before PE and CHDF initiation, four patients had signs and symptoms of encephalopathy, four required ventilatory support, and all 11 were developing liver failure, significant renal compromise, and coagulopathy. PE combined with CHDF for patients was initiated a mean of 2 days postpartum (range, days 0-3). Daily or every other day PE combined with CHDF was undertaken on two to eight occasions for each of the 11 patients. Ten patients responded with composite clinical and laboratory improvement and were discharged to the ward, then cured and discharged from hospital; one patient died of septic shock. Average duration of hospitalization was 17 days (range, days 9-38) from time of admission to discharge; the average duration of intensive care unit was 10 days (range, days 4-23). No significant PE- and CHDF-related complications occurred. These results indicate that combing PE and CHDF in a series-parallel circuit is an effective and safe treatment for patients with severe AFLP. This finding may have important implications for the development of an effective treatment for patients with AFLP suffering multiple organ dysfunction.

Comparison of the Diagnostic Accuracy of Percutaneous Fistula Contrast-Enhanced Ultrasound Combined with Transrectal 360° 3-D Imaging and Conventional Transrectal Ultrasound in Complex Anal Fistula.

This study compared the diagnostic accuracy of percutaneous fistula contrast-enhanced ultrasound (CEUS) combined with 360° 3-D transrectal ultrasound (TRUS) imaging (CEUS + 360°-TRUS) with that of conventional transrectal ultrasound in the diagnosis of complex anal fistulas. A total of 156 patients clinically diagnosed with complex anal fistula from January 2020 to December 2021 were studied and randomly divided into an experimental group (n = 82) and a control group (n = 74). Patients in the experimental group were examined by percutaneous fistula CEUS combined with CEUS + 360°-TRUS, while patients in the control group were examined using TRUS. The detection of fistulas (main tract, branch and internal orifice) and the accuracy of Parks classification were compared between the two groups. Recurrences were followed up at 1, 3 and 6 mo after the surgery. A total of 156 patients were included, aged 23-68 y (average: 37.7 ± 18.2 y). In both groups, the course of disease was <1 mo in 128 cases, 1-2 mo in 22 cases and >3 mo in 6 cases. A total of 474 fistulas were confirmed by surgery in the aforementioned patients, including 224 main fistulas, 250 branch pipes and 254 internal orifices. The CEUS + 360°-TRUS group had 96.87%, 90.41% and 90.14% diagnostic accuracy for the main tract, branch and internal orifice, which was statistically significant (p < 0.001) compared with the 85.00%, 70.00% and 72.46% for the TRUS group, respectively. The overall accuracy of Parks classification of anal fistula in the CEUS + 360°-TRUS group was significantly higher than that in the TRUS group (90.24% vs. 78.38%, p < 0.001). After 6 mo of follow-up, the recurrence rate in the CEUS + 360°-TRUS group was 4.87%, and the recurrence rate in the TRUS group was 18.91%. Percutaneous fistula CEUS combined with transrectal 360° 3-D imaging has significantly higher accuracy than conventional TRUS in the diagnosis of complex anal fistula, especially for anal fistula branches, internal openings and Parks classification and is beneficial in reducing post-operative occurrence of complex anal fistulas.

Facile and efficient approach to speed up layer-by-layer assembly: dipping in agitated solutions.

A facile and efficient approach has been developed to speed up the fabrication of LBL films through sequential dipping in vigorously agitated solutions. By this agitated-dipping (AD) LBL technique, the multilayer films of PAH and PSS were fabricated. The resulting films were explored by UV-vis spectroscopy, X-ray reflectivity, and AFM. Meanwhile, the comparison of the AD and conventional LBL films was made, which demonstrated that AD LBL can decrease dipping time by more than 15 times without reducing film quality remarkably. In addition, to verify the generality of AD LBL, we studied the AD LBL films of PDDA/PSS and PAH/PAA preliminarily as well. AD LBL promotes the efficiency of conventional LBL greatly while preserving its most advantages, such as simplicity, cheapness, precise control, universality in substrates, recycling use of sample solutions, and so on. It would be a promising alternative to build up LBL films rapidly.

[The effects of joint administration of 6% hydroxyethyl starch 130/0.4 and high-volume hemofiltration on patients with acute lung injury and acute kidney injury].

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.

[The role of p38 mitogen-activated protein kinase in interleukin-6 induction by lipopolysaccharide in vascular smooth muscle cells].

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.

Effects of earthquake and related environmental factors on relationship of posttraumatic stress disorder with LDLR rs5925.

To test the relationship not yet explored before among earthquake and related environmental factors, low-density lipoprotein receptor (LDLR) and posttraumatic stress disorder (PTSD), the genetic variation of LDLR rs5925 was selected and PTSD was examined by PTSD Checklist-Civilian Version (PCLC) in adolescents with different genotypes of LDLR rs5925 longitudinally at 6, 12 and 18 months after the 2008 Wenchuan earthquake. The C allele carriers were observed to have higher PTSD prevalence than the TT homozygotes in the male subjects, and higher PTSD prevalence and PCL-C scores in the female subjects only at 6 months. When compared to that at 12 months, decreased PTSD prevalence was observed at 18 months only in the female C allele carriers, but not in the female TT homozygotes or the male subjects. The potential risk factors of PTSD and predictors of PCL-C scores were different during the follow-up. LDLR rs5925 was one of the predictors for PCL-C scores at 6 and 12 months, and one of the potential factors for PTSD prevalence at 6 months. These results suggest that interactions may occur between earthquakes and other related environmental factors, which could affect the relationship of LDLR rs5925 with PTSD and be considered for individualized treatment.

[Study the effects of high-volume hemofiltration and fluid resuscitation on removing blood lactic acid and pro-inflammatory cytokines in patients with refractory septic shock].

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.

The Leptin Gene rs7799039 Modulates the Prevalence of Posttraumatic Stress Disorder After an Earthquake in Han Chinese Adolescents.

OBJECTIVES: The study's aim was to examine the prevalence and severity of posttraumatic stress disorder (PTSD) longitudinally among high school students with different genotypes of the leptin gene (LEP) rs7799039 after the 2008 Wenchuan earthquake in China. METHODS: The symptoms of PTSD were measured by the PTSD Checklist-Civilian Version (PCL-C) based on DSM-IV-TR criteria in 462 students at 6, 12, and 18 months after the earthquake. The genotypes of LEP rs7799039 were identified by polymerase chain reaction-restriction fragment length polymorphism analyses in 2018 using genomic DNA prepared in 2008 and stored at -80°C and verified by DNA sequencing. The association of LEP genotypes with PTSD was then analyzed by various statistical methods. RESULTS: The AA homozygotes had higher prevalence of PTSD than the G allele carriers at 12 months (22.30% vs 10.53%, P = .013) and higher median (interquartile range [IQR]) PCL-C scores at 12 (27.00 [24.00-35.75] vs 26.00 [22.00-31.25], P = .010) and 18 months (27.00 [21.00-32.00] vs 24.00 [19.00-29.00], P = .003) post-earthquake among female subjects. Female students had higher PCL-C scores than male subjects at 6 and 12 months regardless of the genotypes but only among the AA homozygotes at 18 months (27.00 [21.00-32.00] vs 22.00 [18.00-26.00], P = .000). The potential risk factors for and predictors of PTSD severity differed at different time points during follow-up. LEP rs7799039 was a potential factor for PTSD at 12 months and a predictor of PTSD severity at 18 months post-earthquake. CONCLUSIONS: An association of LEP rs7799039 with the prevalence and severity of PTSD in Chinese adolescents was observed. These results indicate that females with the LEP rs7799039 AA genotype had more severe PTSD characteristics compared to female G allele carriers, suggesting that psychosocial or pharmacologic managements may particularly be needed by these female subjects.

Growth inhibition and gene induction in human hepatocellular carcinoma cell exposed to sodium 4-phenylbutanoate.

BACKGROUND: Sodium 4-phenylbutanoate (NaPB) can induce cellular differentiation and cell cycle arrest. However, its potential anticancer properties in hepatocellular carcinoma and influence on normal liver cell are still unclear. We observed the effects of NaPB on growth inhibition, including differentiation and phase growth arrest in normal liver cell line L-02 and hepatocellular carcinoma cell line Bel-7402. Furthermore, we investigated its mechanism in Bel-7402. METHODS; Hepatocellular carcinoma cells Bel-7402 and normal liver cell line L-02 were treated with NaPB at different concentrations. Light microscopy was used to find morphological change in cells. Cell cycle was detected by flow cytometry. Expression of acetylating histone H4 and of histones deacetylase 4 (HDAC4) were determined by Western blot. The expression of P21WAF1/CIP1 and E-cadherin were observed through immunocytochemistry. RESULTS: NaPB treatment led to time dependent growth inhibition in hepatocellular carcinoma cells Bel-7402. NaPB treatment caused a significant decline in the fraction of S phase cells and a significant increase in G0/G1 cells. NaPB increased the expression of P21(WAF1/CIP1) and E-cadherin in Bel-7402 and significantly decreased the level of HDAC4 in Bel-7402. NaPB significantly improved the level of acetylating histone H4. The normal liver cell line L-02 showed no distinct changes under treatment with NaPB. CONCLUSIONS: NaPB inhibited the growth of hepatocellular carcinoma cells Bel-7402 and induced partial differentiation through enhancing the acetylating histones. In Bel-7402, the expressions of P21(WAF1/CIP1) and E-cadherin may be related to level of acetylating histones and inhibition of cellular growth. NaPB showed no significant effect on normal liver cells.

Expression of angiostatin cDNA in human gallbladder carcinoma cell line GBC-SD and its effect on endothelial proliferation and growth.

AIM: To explore the influence of angiostatin up-regulation on the biologic behavior of gallbladder carcinoma cells in vitro and in vitro, and the potential value of angiostatin gene therapy for gallbladder carcinoma. METHODS: A eukaryotic expression vector of pcDNA3.1(+) containing murine angiostatin was constructed and identified by restriction endonuclease digestion and sequencing. The recombinant vector pcDNA3.1-angiostatin was transfected into human gallbladder carcinoma cell line GBC-SD with Lipofectamine 2000, and paralleled with the vector and mock control. The resistant clone was screened by G418 filtration. Angiostatin transcription and protein expression were examined by RT-PCR, immunofluorescence and Western-blot. The supernatant was collected to treat endothelial cells. Cell proliferation and growth in vitro were observed under microscope. RESULTS: Murine angiostatin cDNA was successfully cloned into the eukaryotic expression vector pcDNA3.1 (+). After 14 d of transfection and selection with G418, macroscopic resistant cell cloning was formed in the experimental group transfected with pcDNA 3.1(+)-angiostatin and vector control. But untreated cells died in the mock control. Angiostatin was detected by RT-PCR and protein expression was detected in the experimental group by immunofluorescence and Western-blot. Cell proliferation and growth in vitro in the three groups were observed respectively under microscope. No significant difference was observed in the growth speed of GBC-SD cells between groups that were transfected with and without angiostatin. After treatment with supernatant, significant differences were observed in endothelial cell (ECV-304) growth in vitro. The cell proliferation and growth were inhibited. CONCLUSION: Angiostatin does not directly inhibit human gallbladder carcinoma cell proliferation and growth in vitro, but the secretion of angiostatin inhabits endothelial cell proliferation and growth.

Effects of exogenous p53 transfection on the gene expression in the human brain glioma cell line U251.

The p53 gene is activated in response to several malignancy-associated stress signals by transactivation of downstream genes and by transcription-independent mechanisms. In order to identify new p53 downstream genes, we established a new system of p53 gene inducible expression, U251-pTet-p53 cell line, with the Tet-On Gene Expression System, in which exogenous p53 gene could overexpress in doxycycline (Dox) medium but not in the medium without Dox. By comparing their random primer RT-PCR products, it was proved that exogenous p53 gene expression could lead to many genes differential expression, some up-expressed and others down-expressed. All of these differential expressed genes may be p53 downstream genes. We can gain the magnitude of p53 downstream genes, which provides the basis of directly cloning of novel p53 downstream genes and further studying of p53 regulatory network.

Fe-Promoted Chlorobenzylation of Terminal Alkynes through Benzylic C(sp(3))-H Bond Functionalization.

A chlorobenzylation of terminal alkynes through Fe(II)-promoted benzylic C(sp(3))-H bond functionalization in the presence of NCS as a chloride source was developed. Compared with previous methods to prepare polysubstituted alkenyl halides, the presented procedure provides an efficient alternative with high atom and step economy under mild conditions. The transformation was established to proceed through a single-electron transfer (SET) process with benzyl cations as key intermediates.

Effect of probiotics on the incidence of ventilator-associated pneumonia in critically ill patients: a randomized controlled multicenter trial.

PURPOSE: To evaluate the potential preventive effect of probiotics on ventilator-associated pneumonia (VAP). METHODS: This was an open-label, randomized, controlled multicenter trial involving 235 critically ill adult patients who were expected to receive mechanical ventilation for ≥48 h. The patients were randomized to receive (1) a probiotics capsule containing live Bacillus subtilis and Enterococcus faecalis (Medilac-S) 0.5 g three times daily through a nasogastric feeding tube plus standard preventive strategies or (2) standard preventive strategies alone, for a maximum of 14 days. The development of VAP was evaluated daily, and throat swabs and gastric aspirate were cultured at baseline and once or twice weekly thereafter. RESULTS: The incidence of microbiologically confirmed VAP in the probiotics group was significantly lower than that in the control patients (36.4 vs. 50.4 %, respectively; P = 0.031). The mean time to develop VAP was significantly longer in the probiotics group than in the control group (10.4 vs. 7.5 days, respectively; P = 0.022). The proportion of patients with acquisition of gastric colonization of potentially pathogenic microorganisms (PPMOs) was lower in the probiotics group (24 %) than the control group (44 %) (P = 0.004). However, the proportion of patients with eradication PPMO colonization on both sites of the oropharynx and stomach were not significantly different between the two groups. The administration of probiotics did not result in any improvement in the incidence of clinically suspected VAP, antimicrobial consumption, duration of mechanical ventilation, mortality and length of hospital stay. CONCLUSION: Therapy with the probiotic bacteria B. Subtilis and E. faecalis are an effective and safe means for preventing VAP and the acquisition of PPMO colonization in the stomach.