Identification of prognostic biomarkers for cholangiocarcinoma by combined analysis of molecular characteristics of clinical MVI subtypes and molecular subtypes.

Cholangiocarcinoma (CCA) is widely noted for its high degree of malignancy, rapid progression, and limited therapeutic options. This study was carried out on transcriptome data of 417 CCA samples from different anatomical locations. The effects of lipid metabolism related genes and immune related genes as CCA classifiers were compared. Key genes were derived from MVI subtypes and better molecular subtypes. Pathways such as epithelial mesenchymal transition (EMT) and cell cycle were significantly activated in MVI-positive group. CCA patients were classified into three (four) subtypes based on lipid metabolism (immune) related genes, with better prognosis observed in lipid metabolism-C1, immune-C2, and immune-C4. IPTW analysis found that the prognosis of lipid metabolism-C1 was significantly better than that of lipid metabolism-C2 + C3 before and after correction. KRT16 was finally selected as the key gene. And knockdown of KRT16 inhibited proliferation, migration and invasion of CCA cells.

Development of a PHMB hydrogel-modified wound scaffold dressing with antibacterial activity.

Current wound scaffold dressing constructs can facilitate wound healing but do not exhibit antibacterial activity, resulting in high infection rates. We aimed to endow wound scaffold dressing with anti-infective ability by polyhexamethylenebiguanide (PHMB). We prepared PHMB hydrogel at varying concentrations (0.25%, 0.5%, 1%, 2%) and assessed release and cytotoxicity. PHMB hydrogel was added to the wound scaffold dressing to generate a PHMB hydrogel-modified wound scaffold dressing. Wound healing and infection prevention were evaluated using a full-thickness skin defect model in rats. In vitro, the hydrogel PHMB release time positively correlated with PHMB concentration, with 1% allowing sufficiently long release time to encompass the high-incidence period (3-5 days) of infection following wound scaffold dressing implantation. Implantation of 1% PHMB hydrogel into the skin did not cause adverse responses. in vitro cytotoxicity assays showed the PHMB hydrogel-modified wound scaffold dressing did not significantly affect proliferation of fibroblasts or vascular endothelial cells, 99.90% vs 99.84% for fibroblasts and 100.21% vs 99.28% for vascular endothelial cells at 21 days. Transplantation of PHMB hydrogel-modified wound scaffold dressing/unmodified wound scaffold dressing on the non-infected wounds of rats yielded no significant difference in relative vascularization rate, 47.40 vs 50.87 per view at 21 days, whereas bacterial content of the wound tissue in the PHMB hydrogel-modified wound scaffold dressing group was significantly lower than the unmodified wound scaffold dressing group, (1.80 ± 0.35) × 103 vs (9.34 ± 0.45) × 103 at 14 days. Prevalence of persistent wound infection in the rats receiving PHMB hydrogel-modified wound scaffold dressing transplantation onto infected wounds was significantly lower than the unmodified wound scaffold dressing group, 30% vs 100%. PHMB hydrogel-modified wound scaffold dressing exhibited suitable antibacterial ability, and its biological activity did not significantly differ from that of the unmodified wound scaffold dressing, thereby allowing it to effectively prevent infection following wound scaffold dressing implantation.

Deficiency of Smad3 results in enhanced inducible nitric oxide synthase-mediated hypotension in lipopolysaccharide-induced endotoxemia.

BACKGROUND: Smad3 is a principal intracellular mediator of signaling for transforming growth factor ß, a cytokine involved in pleiotropic pathophysiological processes including inflammation and immunity. The function of Smad3 in regulating inducible nitric oxide synthase (iNOS) expression and septic shock has not been characterized. METHODS: Smad3(-/-) (referred hereafter as KO) and wild-type (WT) mice were injected intraperitoneally with lipopolysaccharide (LPS) to induce the septic hypotension. Mortality, blood pressure, and plasma levels of nitrite were measured. The iNOS messenger RNA and protein levels in lung, kidney, and spleen were also analyzed. RESULTS: Mice lacking functional Smad3 respond to LPS with greater mortality than their WT littermates. The high mortality of KO mice is accompanied by enhanced hypotension after intraperitoneal injection of LPS. Both KO and WT mice displayed an increase in plasma nitrite during the experimental period; however, LPS administration caused more dramatic changes in KO mice than WT mice. Likewise, the iNOS messenger RNA and protein levels in lung, kidney, and spleen were more strongly increased in KO mice than in WT mice after LPS administration. CONCLUSIONS: Defects in the Smad3 gene may increase susceptibility to the development of septic hypotension because of enhanced iNOS production.

Laparoscopic spleen-preserving distal pancreatectomy with or without splenic vessel conservation: a retrospective study of 20 cases.

BACKGROUND/AIMS: To compare the outcomes and potential morbidities of laparoscopic spleen-preserving distal pancreatectomy with or without splenic vessel preservation in patients with benign or low-grade malignant pancreatic lesions. METHODOLOGY: Twenty patients who underwent spleen-preserving distal pancreatectomy were retrospectively analyzed. All the patients had benign or low-grade malignant pancreatic lesions that had not invaded the spleen. Twelve patients underwent Kimura's procedure and eight patients underwent Warshaw's. Perioperative data, and procedure-specific complications were compared between the two groups. RESULTS: Age, gender, and body mass index were comparable between the two groups. Operative time and intraoperative blood loss were significantly lower for patients who underwent Warshaw's procedure than for those who received Kimura's (p <0.05 for both). There were no significant differences between the two groups with regard to perioperative blood transfusions, length of postoperative hospital stays, or complication rates. Splenic infarction and gastric varices developed only in patients who underwent Warshaw's procedure (one case each). CONCLUSIONS: Our results suggest that the Kimura technique should be the first choice for patients with benign or low-grade malignant pancreatic lesions. Warshaw's technique was associated with a higher incidence of several complications. However, Warshaw's can increase the success rate of splenic preservation in some cases.

[Association between serum homocysteine and in-hospital death in patients with acute pulmonary embolism].

OBJECTIVE: To explore the association between serum homocysteine (Hcy) level and in-hospital death in patients with acute pulmonary embolism. METHODS: A total of 186 acute pulmonary embolism patients [ (66.8 ± 12.7) years, 89 male] hospitalized in our department between June 2008 and June 2011 were included in this prospective study. Patients were divided into high Hcy group (Hcy ≥ 15.2 µmol/L, n = 95) and low Hcy group (Hcy < 15.2 µmol/L, n = 91). Patients were followed-up for 1 year for the incidence rate of early death associated with acute pulmonary embolism. The Cox proportional hazard model was used to analyze the relationship between serum Hcy level and early death in acute pulmonary embolism patients. RESULTS: Patients were hospitalized for 1-37 days [(10 ± 6) days]. In-hospital death rate was 14.5% (27/186) and was significantly higher in high Hcy group than in low Hcy group [25.3% (24/95) vs. 3.3% (3/91) , P = 0.001]. Univariate Cox regression analysis indicated that admission heart rate, oxygen saturation, enlargement of right ventricle, Hcy ≥ 15.2 µmol/L, serum creatinine level, peak TnT level and deep venous thrombosis (P < 0.05) were independent risk factors for in-hospital death. Multivariate Cox regression analysis showed that Hcy ≥ 15.2 µmol/L (HR = 4.10, 95%CI:3.00-4.98, P = 0.017), admission heart rate (HR = 1.10, 95%CI:1.01-1.20, P = 0.031) , deep venous thrombosis (HR = 1.65, 95%CI:1.45-1.76, P = 0.034) and age (HR = 1.10, 95%CI:1.02-1.19, P = 0.010) were independent predictors of in-hospital death for acute pulmonary embolism patients. One-year follow up was finished in 142 patients (89.3%). There were 19 deaths ( 5 due to repeat pulmonary embolism, 4 due to decompensated respiratory and /or cardiac diseases, 6 due to malignant tumors, 2 due to fatal bleeding and 2 due to pneumonia) . Death rate was similar between the two groups during follow up. CONCLUSION: Higher serum homocysteine is an independent for in-hospital death for patients with acute pulmonary embolism.

Gancao Nourishing-Yin decoction combined with methotrexate in treatment of aging CIA mice: a study based on DIA proteomic analysis.

BACKGROUND: Elderly rheumatoid arthritis (ERA) population faces multiple treatment dilemma. Here we aim to investigate if Gancao Nourishing-Yin decoction (GCNY) added to methotrexate (MTX) exhibit better effects in an ERA mice model. METHODS: ERA mice model was established by adding D-galactose (Dgal) to collagen-induced arthritis (CIA) mice. The model was then assigned into control group (CIA + Dgal), MTX treatment group (MTX), GCNY treatment group (GCNY), and integrative treatment group (MTX + GCNY). Pathological scoring was performed to evaluate the severity between the groups. Proteomic analysis was applied to investigate the secretory phenotype of the ERA mouse model and the underlying mechanism of GCNY, MTX and their combination. Representative cytokines related to proteomic results were further validated by ELISAs. RESULTS: CIA + Dgal mice showed more aggressive joints damage than the CIA mice. Besides changes in the inflammatory pathway such as Pi3k-Akt signaling pathway in both model, differential expressed proteins (DEPs) indicated metabolism-related pathways were more obvious in CIA + Dgal mice. Low-dose MTX failed to show pathological improvement in CIA + Dgal mice, while GCNY improved joints damage significantly. Besides down-regulated inflammation-related targets, GCNY-regulated DEPs (such as Apoc1 ~ 3, Grk2 and Creb3l3) were broadly enriched in metabolism-related pathways. MTX + GCNY showed the best therapeutic effect, and the DEPs enriched in a variety of inflammatory,metabolism and osteoclast differentiation signaling pathway. Notably, MTX + GCNY treatment up-regulated Dhfr, Cbr1, Shmt1 involved in folic acid biosynthesis and anti-folate resistance pathways indicated a coincidence synergic action. ELISAs confirmed CPR and Akt that elevated in CIA + Dgal mice were significantly ameliorated by treatments, and adding on GCNY elevated folic acid levels and its regulator Dhfr. CONCLUSION: Aging aggravated joints damage in CIA, which probably due to metabolic changes rather than more severe inflammation. GCNY showed significant effects in the ERA mice model especially when integrated with MTX to obtain a synergic action.

Role of inhibition of p38 mitogen-activated protein kinase in liver dysfunction after hemorrhagic shock and resuscitation.

BACKGROUND: The liver is one of the organs most frequently affected by trauma and hemorrhagic shock; the exact role of p38 mitogen-activated protein kinase (MAPK) activation in response to hepatic hemorrhagic shock/resuscitation (HS/R) remains unclear. MATERIALS AND METHODS: C57Bl/6 mice were divided into four groups: sham-operated group, SB-only group, control group, and SB + HS/R group. Hepatocellular injury (aspartate aminotransferase [AST] and alanine aminotransferase [ALT]) and tumor necrosis factor (TNF-α) and interleukin (IL-1ß) messenger ribonucleic acid (mRNA) expression in the liver were assessed 6 h after resuscitation, p38 MAPK activation in the liver was assessed at 30 min after resuscitation. RESULTS: p38 MAPK activation was higher in the control group than other groups 30 min after resuscitation. p38 MAPK activation level in the SB + HS/R group did not change significantly compared with that of sham and SB-only groups, but was significantly lower than that in the control group. The TNF-α mRNA expression in the control group was significantly higher than that in the sham group. The TNF-α mRNA levels after HS/R in the SB + HS/R group were significantly lower than those in the control group and were roughly the same as those in the sham and SB-only groups. IL-1ß mRNA expression showed similar changes in the four groups. Serum ALT and AST levels in the control group were significantly higher than those in the sham group. The increase in serum ALT and AST levels after HS/R in the SB + HS/R group was significantly less pronounced than that in the control group and markedly higher than that in the sham group. CONCLUSIONS: p38 MAPK was phosphorylated during the HS/R process. Inhibiting the activation of p38 MAPK may attenuate HS/R injury to the liver.

Effects of STAT3 gene silencing and rapamycin on apoptosis in hepatocarcinoma cells.

The PI3K/Akt/mTOR and JAK/STAT3 signaling pathways are important for regulating apoptosis, and are frequently activated in cancers. In this study, we targeted STAT3 and mTOR in human hepatocellular carcinoma Bel-7402 cells and examined the subsequent alterations in cellular apoptosis. The expression of STAT3 was silenced with small interfering RNA (siRNA)-expressing plasmid. The activity of mTOR was inhibited using rapamycin. Following treatment, Annexin V/propidium iodide staining followed by flow cytometry and Hoechst33258 immunofluorescence staining was used to examine cellular apoptosis. JC-1 staining was used to monitor depolarization of mitochondrial membrane (ΔΨm). Furthermore, the expression of activated caspase 3 protein was analyzed by Western blotting. Compared to non-treated or control siRNA-transfected cells, significantly higher levels of apoptosis were detected in siSTAT3-transfected or rapamycin-treated cells (P < 0.05), which was further enhanced in cells targeted for both molecules (P < 0.05). The pro-apoptotic effects were accompanied with concomitant depolarization of mitochondrial membrane and up-regulation of activated caspase 3. Combined treatments using rapamycin and STAT3 gene silencing significantly increases apoptosis in Bel-7402 cells, displaying more dramatic effect than any single treatment. This study provides evidence for targeting multiple molecules in cancer therapy.

[Clinical analysis of 150 patients with idiopathic pulmonary arterial hypertension].

OBJECTIVE: To explore the demographic characteristics and clinical features of patients with idiopathic pulmonary arterial hypertension (IPAH) in China. METHODS: Between March 2007 and September 2010, IPAH diagnosis was confirmed by right heart catheterization in 150 adult patients from 31 clinical centers in China. Clinical and hemodynamic data were analyzed and patients were divided into WHO functional class I/II and WHO functional class III/IV group. RESULTS: The mean age of 150 patients were 36 ± 13 years with female patient/male patient ratio of 2:1, and mean BMI was (21.3 ± 3.5) kg/m(2). Fatigue (n = 123, 82.0%) and dyspnea (n = 112, 74.7%) are the most common symptoms. Accentuated pulmonic second sound (P(2)) was detected in 92.0% (n = 138) of patients during physical examination, which was also the most common sign. About 49.0% (n = 73) patients were WHO functional class I/II patients and 46.0% (n = 68) patients were WHO functional class III/IV patients. Six minutes walking distance (6MWD) and Borg dyspnea score was (337 ± 101) m and 2.0 (2.0, 4.0), respectively. Right ventricular hypertrophy was suggested by ECG in 93.1% (n = 140) patients. Right atrial pressure was (10 ± 6) mm Hg, mean pulmonary artery pressure was (61 ± 16) mm Hg, cardiac index was (2.3 ± 0.8) L×min(-1)×m(-2) and pulmonary vascular resistance (1484 ± 699) dyn×s(-1)×cm(-5) in this cohort. 6 MWD (305 m ± 89 m vs. 377 m ± 88 m) was significantly shorter while Borg dyspnea score [3.0 (3.0, 5.0) vs. 2.0 (2.0, 3.0)] was significantly higher in WHO functional class III/IV patients than in WHO functional class I/II patients. Similarly hemodynamic parameters were also worse in WHO functional class III/IV patients than in WHO functional class I/II patients (all P < 0.05). CONCLUSION: Idiopathic pulmonary arterial hypertension patients in this cohort affect mostly young adults, dominated by female gender and lower body mass index. Fatigue and dyspnea are the most common symptoms and accentuated pulmonic second sound (P(2)) is the most common sign. IPAH patients are often displaying severe functional and hemodynamic disturbance at first visit to hospitals. Dyspnea and hemodynamic impairment are related to 6MWD and WHO functional class.

Synergistic Effects of Quercetin-Modified Silicone Gel Sheet in Scar Treatment.

Both silicone gel and quercetin are effective in scar treatment but have different action mechanisms. Quercetin is mainly applied in the gel form and can lead to poor adhesion of silicone gel sheet; therefore, they cannot be combined in clinical use. In this study, a silicone gel sheet that releases quercetin in a sustained manner for 48 hours was successfully developed. Four round scars (Ø: 1 cm) were made in the ears of New Zealand albino rabbits (n = 10). After scar healing, the rabbits were divided into four groups: blank control group with no treatment, silicone gel sheet group with dressing change every 2 days, quercetin group with dressing change three times daily, and combination treatment group with dressing change every 2 days. Scar assessment was performed 3 months later. Transepidermal water loss showed no difference between the combination treatment group and the silicone gel sheet group, but was lower than that in the quercetin group and the blank control group. Immunohistochemistry of CD 31 and proliferating cell nuclear antigen showed the following results: combination treatment group < silicone gel sheet group = quercetin group < blank control group. Polymerase chain reaction results showed that the expression of type-I and type-III collagen in the combination treatment group and the quercetin group was significantly lower than that in the other two groups. Thus, quercetin-modified silicone gel sheet combines the advantages of the two treatments and is more effective at inhibiting cell proliferation in scar tissue than either of the two treatments alone.

Successful treatment of a patient with an extraordinarily large deep burn.

BACKGROUND: Treatment of extraordinarily large deep burns remains a huge clinical challenge. CASE REPORT: This article is a summary of our experience with the treatment of a patient with an extraordinarily large deep burn (99.5% TBSA and 23% fourth degree burn) by using the "microskin autografting and alloskin repeated grafting" method to close the deep burn wound because of scarcity of skin sources of the patient. CONCLUSIONS: The patient has been observed for 2 years, and is able to face the reality of life peacefully with the support of his family.

[Effect of RhoC on hepatocellular carcinoma cell growth and related molecular mechanisms].

OBJECTIVE: To clarify the role of RhoC in the growth of hepatocellular carcinoma cells and its molecular mechanism, so as to explore the molecular target of tumor cell growth. METHODS: siRNA-RhoC plasmid was constructed and RhoC gene silencing the cell-line of hepatocellular carcinoma was setup. Cell growth was assessed by MTT assay. AgNORs staining was applied to determine cell proliferation. Plate cell clone test was conducted to examine the capacity of cell clone formation. FACS was adopted to measure the course of cell cycle and semi-quantitative RT-PCR was used to determine the expression of cell cycle proteins. In order to further determine the effect of RhoC expression on cell growth, a RhoC over-expression human hepatocellular cell line was setup by PcDNA3-RhoC plasmid transfection. RESULTS: The inhibition rate of RhoC was 82.3%. From the fourth day of cell culture, the growth of cells in RNAi group was significantly slower than that in parental Bel7402 and negative control groups (0.41 ± 0.10 vs. 0.73 ± 0.11 and 0.71 ± 0.07 respectively, P < 0.05). AgNORs staining showed that average cell stained particles in RNAi group was significantly lower than that in parental Bel7402 and negative control(1.23 ± 0.35 vs. 3.47 ± 0.93 and 3.17 ± 0.78, P < 0.01). Plate clone formation test showed that clone formation efficiency in the RNAi group was notably lower than that in the control group [(20.33 ± 5.42)% vs. (70.58 ± 10.10)% and (69.83 ± 14.77)%, respectively, P < 0.01]. Cell cycle analysis by FACS showed that G(0)/G(1) cell percentage in the RNAi group was significantly higher than that in the control group [(73.14 ± 5.93)% vs. (57.05 ± 5.97)% and (52.99 ± 4.80)%, P < 0.05]. Compared with Bel7402 and negative control groups, the expression of following growth associated genes was significantly decreased: cyclin D1(0.45 ± 0.21 vs. 1.25 ± 0.24 and 1.12 ± 0.15, respectively, P < 0.05)and CDK4 (0.55 ± 0.08 vs. 1.18 ± 0.32 and 1.10 ± 0.29, respectively, P < 0.05); the following genes were notably increased: p16(1.07 ± 0.23 vs. 0.36 ± 0.12 and 0.35 ± 0.13, respectively, P < 0.01)and p21(0.42 ± 0.12 vs. 0.17 ± 0.06 and 0.19 ± 0.08, respectively, P < 0.05). RhoC was highly expressed in PcDNA3-RhoC transfected hepatocellular cell line. From the third day on of the cell culture, cell growth in PcDNA3-RhoC group was remarkably higher than that in the HL7702 and PcDNA3 groups (0.83 ± 0.10 vs. 0.54 ± 0.11 and 0.58 ± 0.55, respectively, P < 0.05). CONCLUSIONS: RhoC is the key molecule in promoting hepatocellular cell growth, and is a promising target for tumor cell growth controlling.

[Effect of mild hypothermia in combination of acupuncture on Bcl-2 and Bax protein expressions of local cerebral ischemia/reperfusion rats].

OBJECTIVE: To observe and compare the effects of acupuncture in combination of mild hypothermia therapy on the expressions of Bcl-2 and Bax. METHODS: Fifty rats were randomly divided into 5 groups, i.e., the control group, the model group, the acupuncture group, the mild hypothermia group, and the combination group (acupuncture in combination of mild hypothermia therapy), 10 in each group. Focal cerebral ischemia model was prepared with modified Longa's suture method. The effects of acupuncture, the mild hypothermia, and the acupuncture in combination of mild hypothermia therapy on the expressions of Bcl-2 and Bax were observed. The indices were detected using immunohistochemical assay. RESULTS: Compared with the control group, the expression of Bcl-2 in the model groups significantly decreased (P < 0.01), and the expression of Bax significantly increased (P < 0.01). Compared with the model group, the expression of Bcl-2 in the three treatment groups significantly increased (P < 0.01) and the expression of Bax significantly decreased (P < 0.01). Compared with the acupuncture group and the mild hypothermia group, the expression of Bcl-2 significantly increased and the expression of Bax significantly decreased in the combination group (both P < 0.01). Compared with the mild hypothermia group, increased expression of Bcl-2 and decreased expression of Bax in the acupuncture group were not obvious (P > 0.05). There was insignificant difference in the therapeutic efficacy between the acupuncture group and the mild hypothermia group. CONCLUSIONS: Both acupuncture and the mild hypothermia therapy could increase the expression of Bcl-2 and decrease the expression of Bax, intervene cerebral ischemia, and protect neurons. In the early cerebral ischemia/reperfusion stage, it is necessary to perform acupuncture and mild hypothermia therapy as soon as possible.

[The efficacy and safety of bosentan therapy for Chinese patients with idiopathic pulmonary arterial hypertension: an open-label, prospective multicenter study].

OBJECTIVE: To investigate the efficacy, safety and tolerance of bosentan, a dual endothelin receptor antagonist, in Chinese patients with idiopathic pulmonary arterial hypertension (IPAH). METHODS: Totally 79 IPAH patients (hemodynamic criteria confirmed by right heart catheterization) were included in this open-label, prospective multicenter study. Patients received 62.5 mg of bosentan twice daily for the first 4 weeks, and then up-titrated to 125 mg twice daily for another 12 weeks. The primary end point was the change in exercise capacity showed by six-minute walk distance (6MWD) from baseline to 16 weeks. Secondary end points included the change in World Health Organization (WHO) functional class, Borg dyspnoea scale and systolic pulmonary artery pressure measured by echocardiography. RESULTS: The 6MWD increased from (343.7 ± 93.7) meters at baseline to (397.5 ± 104.4) meters after 16 weeks (P < 0.01), WHO functional class and Borg dyspnoea scale were also significantly improved after 16 weeks therapy compared to baseline levels (all P < 0.01). Furthermore, the systolic pulmonary artery pressure was significantly decreased from (97.8 ± 25.2) mm Hg (1 mm Hg = 0.133 kPa) to (92.8 ± 29.5) mm Hg (P < 0.05) after 16 weeks bosentan treatment. There was no patient withdrawal from this study for safety consideration. CONCLUSION: Bosentan therapy is well tolerated and can improve the exercise capacity and WHO functional class in Chinese IPAH patients.

High concentrations of environmental ammonia induced changes in large-scale loach (Paramisgurnus dabryanus) immunity.

High concentrations of environmental ammonia can cause reduced immunity and death in fish, causing enormous economic losses. Air-breathing fish usually have a high ammonia tolerance and are very suitable for high-density fish farming. However, research on the effects of environmental ammonia on air-breathing fish immunity is lacking. Therefore, this study investigated the effects of environmental ammonia on the immunity of large-scale loach (Paramisgurnus dabryanus) by exposing fish to 30 mmol/L NH4Cl solution and subsequently analyzing the changes in serum and liver immune indicators, including total protein, albumin, globulin, immunoglobulin (Ig) M, lysozyme, complement component (C) 3 and C4, heat shock protein (HSP) 70, HSP90, tumor necrosis factor (TNF)-α, interleukin (IL)-1ß, IL-6, and IL-12. Results revealed that ammonia exposure significantly affected the total protein, albumin, globulin, IgM, complement C3 and C4, HSP70, HSP90, and inflammatory cytokine contents in the body, indicating that ammonia exposure induced a significant immune response and lowered bodily immunity. However, most of the immune indicators significantly decreased in the later stages of the experiment, suggesting a weakened immune response, which may be due to the species-specific ammonia detoxification ability of large-scale loach that reduces ammonia toxicity in the body.

Low-dose contrast-enhanced magnetic resonance imaging of brain metastases at 3.0 T using high-relaxivity contrast agents.

BACKGROUND: Lesion-to-brain contrast after gadolinium administration is significantly higher at 3.0 Tesla (T) compared to 1.5 T. The high in vivo relaxivity of gadobenate dimeglumine (Gd-BOPTA) may permit the use of lower-dose contrast agents. PURPOSE: To investigate whether low-dose contrast-enhanced MRI at 3.0 T using a high-relaxivity contrast agent (Gd-BOPTA) can achieve a comparable or improved contrast-to-noise ratio (CNR) for the detection of brain metastases compared with examination of the same patient at 1.5 T using a standard dose of gadopentetate dimeglumine (Gd-DTPA). MATERIAL AND METHODS: A total of 18 patients with known brain metastases were first imaged at 1.5 T with 0.1 mmol/kg Gd-DTPA. Patients returned at least 24 hours later for imaging at 3.0 T with Gd-BOPTA at cumulative doses of 0.025 mmol/kg, 0.05 mmol/kg, 0.075 mmol/kg, and 0.1 mmol/kg (0.1 mmol/kg body weight overall). The CNR of enhancing brain lesions compared to the normal contralateral white matter was calculated. For the 3.0 T study using different cumulative doses of Gd-BOPTA, the CNR of lesions was compared with CNR of the same lesions imaged at 1.5 T using 0.1 mmol/kg Gd-DTPA, by using the Wilcoxon matched-pairs signed-rank test. RESULTS: At 1.5 T with 0.1 mmol/kg Gd-DTPA, the mean CNR between enhanced lesions and cerebral white matter was 12.01 +/- 2.53. With 3.0 T imaging using different cumulative doses of Gd-BOPTA, the mean CNRs were 7.19 +/- 4.06, 15.31 +/- 6.37, 25.44 +/- 11.02, and 31.88 +/- 13.21. At 3.0 T with 0.05 mmol/kg Gd-BOPTA, CNR was 1.34-fold higher compared to CNR at 1.5 T with 0.1 mmol/kg Gd-DTPA (P <0.01). CONCLUSION: Comparable contrast enhancement of brain metastases can be achieved with a 0.05-mmol/kg dose of Gd-BOPTA at 3.0 T compared to imaging at 1.5 T using 0.1 mmol/kg Gd-DTPA.

CD47 blockade alleviates acute rejection of allogeneic mouse liver transplantation by reducing ischemia/reperfusion injury.

Despite advances in immunosuppressive therapies, acute rejection response is still a serious concern especially in the early phase after liver transplantation. This study aimed to evaluate whether blocking the TSP1-CD47 signaling pathway could attenuate the acute rejection after liver transplantation. An allogeneic mouse orthotopic liver transplantation model (Balb/c→C3H) with prolonged cold ischemic phase was used to induce severe IRI and lethal acute rejection. CD47mAb or isotype matched-control IgG2a was administered to donor liver during graft perfusion. Recipients were sacrificed at 1d, 3d, 5d and 7d after reperfusion. Blood samples were collected to evaluate serum alanine aminotransferase, total bilirubin, HMGB-1,TNF-α, IL-2 and INF-γ level. Flow cytometric analysis was used to detect the strength of innate and adaptive immune response. Liver tissue was obtained for HE, TUNEL staining and F4/80 immumohistochemical staining. Moreover, we conducted a mixed lymphocyte reaction treated with IgG2a or CD47mAb. Mice in CD47mAb-treated group demonstrated improved survival and significantly lower increase in Suzuki score, apoptosis index, acute rejection index, serum alanine aminotransferase, total bilirubin, HMGB-1, TNF-α, IL-2, INF-γ level and the degree of Kupffer cells' activation especially in the early phase of acute rejection. In addition, Pearson's correlation analysis confirmed significant correlation between Suzuki score/ALT and acute rejection index. The in vitro inhibition assay showed that CD47 blockade couldn't directly inhibit recipient lymphocyte proliferation. Based on the evidence that TSP1-CD47 signaling blockade with CD47mAb could alleviate acute rejection by reducing the extent of IRI after liver transplantation indirectly, this study provided a basis for new interventions and management methods to support better transplant outcomes.

Herpotrichones A and B, Two Intermolecular [4 + 2] Adducts with Anti-Neuroinflammatory Activity from a Herpotrichia Species.

Herpotrichones A and B (1 and 2), two intermolecular [4 + 2] adducts with an unprecedented pentacyclic 6/6/6/6/3 skeleton, were isolated from Herpotrichia sp. SF09, an isopod-associated fungus, along with a new shunt product protrichone (3). Their structures were elucidated by the analysis of spectroscopic data, residual dipolar coupling (RDC)-based computer-assisted 3D structure elucidation (CASE-3D), and single-crystal X-ray diffraction in combination with electronic circular dichroism (ECD) calculations. Compounds 1 and 2 were assessed to be potent anti-neuroinflammatory agents in lipopolysaccharide (LPS)-induced BV-2 microglial cells with the half maximal inhibitory concentration (IC50) values of 0.41 and 0.11 µM, respectively.

In vitro labeling of mesenchymal stem cells with superparamagnetic iron oxide by means of microbubble-enhanced US exposure: initial experience.

PURPOSE: To investigate the feasibility of magnetically labeling stem cells with superparamagnetic iron oxide (SPIO) by means of microbubble-enhanced ultrasonographic (US) exposure (MUE) and to study the effects of this approach--without secondary transfection agents--on the viability, proliferation activity, and differentiation capability of MUE-labeled stem cells. MATERIALS AND METHODS: Institutional review board approval was obtained for this study. Human mesenchymal stem cells (MSCs) ([1 to 2] x 10(6)/mL) were studied in four experiment groups: sham exposure to US with microbubbles and SPIO (group A), exposure to US with SPIO but without microbubbles (group B), exposure to US with microbubbles and SPIO (group C), and sham exposure to US without SPIO or microbubbles (group D). Intracellular iron uptake was analyzed qualitatively at light and electron microscopy. The viability and proliferation activity of MSCs were evaluated. The adipogenic and osteogenic differentiation capability of the labeled MSCs was also evaluated. Ninety-five percent confidence intervals were derived for assessment of differences in cell viability and proliferation activity between groups C and D. RESULTS: Light and electron microscopy revealed intracytoplasmic iron uptake and nearly 100% cell labeling efficiency. The MUE-labeled MSCs had unaltered viability and uncompromised proliferation activity compared with the nonlabeled MSCs. Similar to the nonlabeled MSCs, the MUE-labeled MSCs differentiated into adipogenic and osteogenic lineages. CONCLUSION: Initial study results show that stem cells can be effectively labeled with SPIO by using MUE without secondary transfection agents and thus that MUE labeling is an appealing alternative cell-labeling approach that warrants investigation for intracellular magnetic labeling of stem cells. SUPPLEMENTAL MATERIAL: http://radiology.rsna.org/lookup/suppl/doi:10.1148/radiol.2531081974/-/DC1.