Accidental placement of venous return catheter in the superior vena cava during venovenous extracorporeal membrane oxygenation for severe pneumonia: A case report.

BACKGROUND: Venovenous extracorporeal membrane oxygenation (V-V ECMO) has become an important treatment for severe pneumonia, but there are various complications during the treatment. This article describes a case with severe pneumonia successfully treated by V-V ECMO, but during treatment, the retrovenous catheter, which was supposed to be in the right internal vein, entered the superior vena cava directly in the mediastinum. The ECMO was safely withdrawn after multidisciplinary consultation. Our experience with this case is expected to provide a reference for colleagues who will encounter similar situations. CASE SUMMARY: A 64-year-old man had severe pulmonary infection and respiratory failure. He was admitted to our hospital and was given ventilation support (fraction of inspired oxygen 100%). The respiratory failure was not improved and he was treated by V-V ECMO, during which the venous return catheter, which was supposed to be in the right internal vein, entered the superior vena cava directly in the mediastinum. There was a risk of massive mediastinal bleeding if the catheter was removed directly when the ECMO was withdrawn. Finally, the patient underwent vena cava angiography + balloon attachment + ECMO withdrawal in the operating room (prepared for conversion to thoracotomy for vascular exploration and repair at any time during surgery) after multidisciplinary consultation. ECMO was safely withdrawn, and the patient recovered and was discharged. CONCLUSION: Patients may have different vascular conditions. Multidisciplinary cooperation can ensure patient safety. Our experience will provide a reference for similar cases.

Scleral Cross-Linking in Form-Deprivation Myopic Guinea Pig Eyes Leads to Glaucomatous Changes.

Purpose: To investigate the potential glaucomatous changes caused by scleral cross-linking (CXL) in a guinea pig form-deprivation (FD) myopia model. Methods: Eighty 4-week-old tricolor guinea pigs were divided into four groups: FD only, genipin CXL only, FD plus CXL, and control. Refractive error, axial length (AL), intraocular pressure (IOP), and structural and vasculature optic disc changes in optical coherence tomography (OCT) and OCT angiography (OCTA) were measured at baseline and day 21. CXL efficacy was evaluated by scleral rigidity Young's modulus values. Histological and molecular changes in the anterior chamber angle, retina, and sclera were assessed. Results: Baseline parameters were similar among groups (P > 0.05). The FD plus CXL group at day 21 had the least increase of AL (0.14 ± 0.08 mm) and highest IOP elevation (31.5 ± 3.6 mmHg) compared with the FD-only group (AL: 0.68 ± 0.17 mm; IOP: 22.2 ± 2.6 mmHg) and the control group (AL: 0.24 ± 0.09 mm; IOP: 17.4 ± 1.8 mmHg) (all P < 0.001). OCT and OCTA parameters of the optic disc in the FD plus CXL group at day 21 showed glaucomatous changes and decreased blood flow signals. Sclera rigidity increased in the CXL and FD plus CXL groups. Advanced glycation end products deposited extensively in the retina, choroid, and sclera of FD plus CXL eyes. Conclusions: CXL causes increased IOP and subsequent optic disc, anterior segment, and scleral changes while inhibiting myopic progression and axial elongation in FD guinea pig eyes. Therefore, applying CXL to control myopia raises safety concerns.

Synthesis and Evaluation of Glycyrrhetic Acid-aromatic Hybrids as Antiinflammatory Agents.

BACKGROUND: Inflammation is a biological response of body tissues to harmful stimuli, so it is desirable to search for novel anti-inflammatory agents with improved pharmaceutical profiles and reduced adverse effects. OBJECTIVE: This study was to explore natural anti-inflammatory agents and improve therapeutic application of glycyrrhetic acid (GA) through molecular hybridization with active aromatics. METHODS: Fourteen novel GA-aromatic hybrids were synthesized and evaluated for their antiinflammatory activities by inhibiting LPS-induced nitric oxide (NO) release in RAW264.7 cells. The synthesized compounds were characterized by single-crystal X-ray diffraction, 1H NMR, 13C NMR and HRMS. RESULTS: The structure-activity relationship (SAR) study indicated that compounds with styryl displayed better NO inhibitory activity. Among them, compounds 2a and 3c exhibited the most promising activity with IC50 values of 9.93 µM and 12.25 µM, respectively. In addition, X-ray singlecrystal diffraction data for compounds 2e and 3c showed that the absolute configuration of GA skeleton was consistent with that of natural 18 ß-glycyrrhetic acid. CONCLUSION: The results showed that GA-aromatic hybrids were a new class of anti-inflammatory agents and this study provided useful information on further optimization.

[Angelica Polysaccharide Resists Platelets Apoptosis Induced by LY294002].

OBJECTIVE: To investigate the anti-apoptotic effect of Angelica polysaccharide (APS) on cryopreservated platelets and its mechanism. METHODS: The platelets were divided into 4 group: control group(4 ℃ stored platelets),APS group (APS-treated platelets stored at 4 ℃), LY294002 group (LY294002-treated platelets stored at 4 ℃) and LY294002+APS group(LY294002+APS treated platelets stored at 4 ℃ ). The expression of platelet membrane glycoprotein CD41 and CD61, as well as the platelet apoptotic rate, Caspase 3 expression and mitochondrial membrane potential (MMP) were detected by flow cytometry; the anti-apoptotic mechanism of APS by PI3K /AKT signaling pathway was analyzed by Western blot assay. RESULTS: The apoptosis rate of platelets in LY294002 group obviously increased, the activity of CD41 and CD61 expression gradually decreased along with the enhancement of LY294002 concentrations (r=-0.953)； compared with control group, the apoptosis rate of platelets in LY294002 group was enhanced significantly(P<0.05)，while the apoptosis rate of platelets in LY294002+APS group significantly was reduced(P<0.05) as compare with LY294002 group, which suggest that APS has an anti-apoptotic effect on the cryopreserved platelets. APS decreased the expression of Caspase-3 and inhibited the reduction of mitochondrial membrane potential induced by LY294002, moreover, APS could increase the activation of PI3K /AKT pathway in Plt. CONCLUSION: APS has an anti-apoptotic effect on the cryopreserved platelets through activating the PI3K /AKT pathway, decreasing the expression of apoptosis protease Caspase-3 and inhibiting the reduction of MMP.

Design, synthesis and biological evaluation of tricyclic pyrazolo[1,5-c][1,3]benzoxazin-5(5H)-one scaffolds as selective BuChE inhibitors.

Based on the structural analysis of tricyclic scaffolds as butyrylcholinesterase (BuChE) inhibitors, a series of pyrazolo[1,5-c][1,3]benzoxazin-5(5H)-one derivatives were designed, synthesized and evaluated for their acetylcholinesterase (AChE) and BuChE inhibitory activity. Compounds with 5-carbonyl and 7- or/and 9-halogen substitutions showed potential BuChE inhibitory activity, among which compounds 6a, 6c and 6g showed the best BuChE inhibition (IC50 = 1.06, 1.63 and 1.63 µM, respectively). The structure-activity relationship showed that the 5-carbonyl and halogen substituents significantly influenced BuChE activity. Compounds 6a and 6g were found nontoxic, lipophilic and exhibited remarkable neuroprotective activity and mixed-type inhibition against BuChE (Ki = 7.46 and 3.09 µM, respectively). Docking studies revealed that compound 6a can be accommodated into BuChE via five hydrogen bonds, one Pi-Sigma interaction and three Pi-Alkyl interactions.

Tricyclic pyrazolo[1,5-d][1,4]benzoxazepin-5(6H)-one scaffold derivatives: Synthesis and biological evaluation as selective BuChE inhibitors.

BuChE inhibitors play important roles in treatment of patients with advanced Alzheimer's disease (AD). A series of tricyclic pyrazolo[1,5-d][1,4]benzoxazepin-5(6H)-one derivatives were synthesized and evaluated as acetylcholinesterase (AChE) and butyrylcholinesterase (BuChE) inhibitors. Some derivatives showed selective BuChE inhibitory activity, which was influenced by the volumes of the substituted groups at the C6 position and halogen substituents at the benzene ring of tricyclic scaffold. Among them, compounds 3f and 3o with dihalogen and 6-ethyl substituent showed the most potent activity (IC50 = 2.95, 2.04 µM, and mixed-type, non-competitive inhibition against BuChE, respectively). Eutomer (6R)-3o exhibited better BuChE inhibitory activity than (6S)-3o. Compound 3o exhibited nontoxic, good ADMET properties, and remarkable neuroprotective activity. Docking studies revealed the same binding orientation within the active site of target enzyme. Compound 3o was nicely bound to BuChE via three hydrogen bonds, one Alkyl interaction and three Pi-Alkyl interactions. The selective BuChE inhibitors had a potential use in progressive neurodegenerative disorder.

Novel unsaturated glycyrrhetic acids derivatives: Design, synthesis and anti-inflammatory activity.

To develop novel anti-inflammatory agents, a series of unsaturated glycyrrhetic acids were designed, synthesized and evaluated for anti-inflammatory activity using RAW264.7 cells. The structure-activity relationship (SAR) of NO inhibitory activity was analyzed. α,ß-Unsaturated glycyrrhetic acids showed better activity, among them, compounds 6k and 6l with piperazine unit exhibited the most potent nitric oxide (NO) and interleukin-6 (IL-6) inhibitory activity (IC50 = 13.3 and 15.5 µM respectively). Furthermore, compound 6k could also significantly suppress LPS-induced iNOS and COX-2 expression and IL-6 production through MAPKs and NF-kB signaling pathway.

Bisoprolol reverses epinephrine-mediated inhibition of cell emigration through increases in the expression of ß-arrestin 2 and CCR7 and PI3K phosphorylation, in dendritic cells loaded with cholesterol.

The effect of bisoprolol on dendritic cell (DC) migration was investigated, including the analysis of protein expression, cytokine secretion and activation of the PI3K-pathway. The chemotactic cell numbers in cholesterol-loaded DCs treated with epinephrine were significantly decreased by 26.66±6.29% (6h), 35.67±2.91% (12h) and 29.33±1.09% (24h). This effect was significantly reversed by 46.00±10.65% (6h), 64.25±6.77% (12h) and 55.74±5.51% (24h) when bisoprolol and epinephrine were both present. In cholesterol-loaded DCs, treatment with epinephrine significantly increased AR-ß1 protein expression by 56.99±4.87%, but inhibited ß-arrestin 2 and CCR7 protein expression by 30.51±4.22% and 25.31±0.04%, respectively. These effects were reversed by bisoprolol by 36.87±4.40%, 41.47±3.95% and 30.14±0.54%, respectively. TNF-α and MMP9 levels were decreased by 68.33±4.00% and 39.57±9.21% in cholesterol-loaded DCs treated with epinephrine. In contrast, when bisoprolol and epinephrine were administered together, the secretion of these proteins was significantly increased by 233.81±37.06 % and 76.66±14.21%, respectively. Treatment with epinephrine decreased PI3K-phosphorylation by 31.88±2.79%, 40.24±5.69% and 30.93±4.66% at 15, 30 and 60min, respectively, whereas the effect of epinephrine on the expression of phosphorylated PI3K was reversed by 49.49±12.12%, 70.93±16.14% and 47.62±6.00%, respectively, when cells were treated with both bisoprolol and epinephrine. Wortmannin inhibited the effects of bisoprolol on PI3K-phosphorylation (38.63±6.12%), the expression of CCR7 (23.4±2.72%), the secretion of TNF-α (69.46±4.48%) and MMP9 (43.15±4.63%), and the number of chemotactic cells (36.84±5.22%). This is the first study to establish a signaling pathway, epinephrine-AR-ß1-ß-arrestin2-PI3K-MMP9/CCR7, which plays a critical role in the migration of DCs.

Effects of total dissolved gas supersaturated water on lethality and catalase activity of Chinese sucker (Myxocyprinus asiaticus Bleeker).

Total dissolved gas (TDG) supersaturation caused by dam sluicing can result in gas bubble trauma (GBT) in fish and threaten their survival. In the present study, Chinese suckers (Myxocyprinus asiaticus Bleeker) were exposed to TDG supersaturated water at levels ranging from 120% to 145% for 48 h. The median lethal concentration (LC(50)) and the median lethal time (LT(50)) were determined to evaluate acute lethal effects on Chinese suckers. The results showed that the LC(50) values of 4, 6, 8, and 10 h were 142%, 137%, 135%, and 130%, respectively. The LT(50) values were 3.2, 4.7, 7.8, 9.2, and 43.4 h, respectively, when TDG supersaturated levels were 145%, 140%, 135%, 130%, and 125%. Furthermore, the biological responses in Chinese suckers were studied by assaying the catalase (CAT) activities in gills and muscles at the supersaturation level of 140% within LT(50). The CAT activities in the gills and muscle tissues exhibited a regularity of a decrease after an increase. CAT activities in the muscles were increased significantly at 3/5LT(50) (P<0.05) and then came back to the normal level. However, there were no significant differences between the treatment group (TDG level of 140%) and the control group (TDG level of 100%) on CAT activities in the gills before 3/5LT(50) (P>0.05), but the activities were significantly lower than the normal level at 4/5LT(50) and LT(50) (P<0.05).

Effect of overexpression of human SR-AI on oxLDL uptake and apoptosis in 293T cells.

Type I class A macrophage scavenger receptor (SR)-AI plays an important role in foam cell formation and in apoptosis in atherosclerosis, however the mechanism remains unclear. Therefore, we generated a pEGFP-C1-SR-AI plasmid construct for transient transfection of 293T human embryonic kidney cells and observed if SR-AI expression led: (i) to foam cell formation or apoptosis; and (ii) to expression of apoptosis-related genes Bcl-2 and Bak-1 in cells treated with oxidized low-density lipoprotein (oxLDL). The pEGFP-C1 (empty vector) transfected cell line was used as a control. Transfection efficiency of each group was >90% and transfected cells expressed functional SR-AI protein. Binding and uptake of 3,3'-dioctadecylindocarbocyanine-labeled oxLDL (DiI-oxLDL) were verified by flow cytometry; increases in the rate of oxLDL binding and uptake were observed in pEGFP-C1-SR-AI transfected 293T cells and incubation with oxLDL also led to increased apoptosis (≈50%) compared with controls. A decrease in Bcl-2 and an increase in Bak-1 mRNA and protein expression were observed in pEGFP-C1-SR-AI transfected cells compared with controls. We conclude that transient over-expression of SR-AI leads to an increase in oxLDL uptake and binding in a non-macrophage cell line. In addition, over-expression of SR-AI induced non-macrophage cell apoptosis via downregulation of Bcl-2 and upregulation of Bak-1 expression. We conclude that the 293T cell expression described here is a model for foam cell formation. These results may form the basis of further research into SR-AI structure and function (including lipoprotein uptake, apoptosis modulation and adhesion), which may give an insight into the progression of atherosclerosis in vivo.

The effect of interleukin-10 on apoptosis in macrophages stimulated by oxLDL.

Marked anti-atheromatous effects of the anti-inflammatory cytokine interleukin-10 (IL-10) were observed in several lipid-driven animal models of arteriosclerosis. We have previously demonstrated that IL-10 significantly inhibited lipid uptake in macrophages induced by oxLDL (Wang et al., 2008; Yang et al., 2008b). In this study, we investigated whether IL-10 affects the apoptosis related gene BCL2L11 and BMF expression in macrophages treated with oxLDL from THP-1 cells, which served as macrophage models. Cell apoptosis assays were performed by flow cytometry. Expression of the apoptosis related genes BCL2L11 and BMF mRNA was quantified by real-time RT-PCR (mRNA expression) and Western blotting (protein expression). IL-10 markedly blocked oxLDL induced cells undergoing early stage apoptosis. In the foam cell group, as compared with the macrophage group, the percentage of apoptosis increased by 100%. Here the expression of BCL2L11 was 45% (mRNA) and 41% (protein) elevated, while the expression of BMF was 54% (mRNA) and 44% (protein) elevated. When macrophages were co-stimulated by 100mg/l oxLDL and 20 µg/l IL-10 for 24h, compared with the foam cell group, the percentage of the apoptosis decreased by 21%, the expression of apoptosis related gene BMF was inhibited, the expression of mRNA and protein was both depressed by 23% and 20%, respectively, but the BCL2L11 expression was unchanged. These results may explain why decrement of early stage apoptosis cells was observed during co-stimulation and raise the possibility that IL-10 reduces foam cell undergoing apoptosis partly through down-regulating the expression of BMF, which demonstrates a critical role of IL-10 in anti-atherogenesis.

[Expression of Twist in papillary thyroid carcinomas and its roles in differential diagnosis].

OBJECTIVE: To study Twist expression in thyroid papillary carcinoma (PTC) by immunohistochemistry and to assess its usefulness as marker in the differential diagnosis of PTC, follicular adenomas (FA) and benign papillary lesions (BPL). METHODS: Fifty cases of PTC, 48 cases of FA and 47 cases of BPL were evaluated using manual tissue chip and SP immunohistochemical stain to detect the expression of Twist and HBME-1, and comparing the staining to that of cytokeratin 19 (CK19). RESULTS: In PTC, positive rates of Twist, HBME-1 and CK19 were 100% (48/48), 94.0% (47/50) and 78.0% (39/ 50) respectively; in FA, positive rates were 0, 6.7% (3/45) and 0 respectively; in BPL, positive rates were 7.0% (3/34), 2.1% (1/47) and 0, respectively. The differences between PTC and FA and between PTC and BPL were both statistically significant (P = 0. 000). The sensitivity of Twist, HBME-1 and CK19 was 100%, 94.0% and 78.0%; the specifity was 96.4%, 95.7% and 100%; overall accurary was 97.7%, 95.1% and 91.9%, respectively. CONCLUSIONS: Positive rates of Twist is higher than the other markers in PTC. Immunohistochemical staining of Twist has important significance in the differential diagnosis of thyroid lesions. Twist immunohistochemistry maybe helpful in diagnosis and differential diagnosis of PTC.

Preliminary identification of Citrus changshanhuyou elite genotypes by molecular markers.

Random amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) analyses were used in elite genotypes identification of Citrus changshan-huyou and analysis of its origin. 12 out of 100 RAPD primers and 11 out of 105 ISSR primers could generate reproducible polymorphic fragments. The RAPD-PCR and ISSR-PCR assays revealed that 64 bands out of 117 (the percentage of polymorphic bands, PPB=54.7%) and 58 bands out of 94 (PPB = 61.7%) were polymorphic, respectively. ISSR and RAPD produced 15 and 12 genotype-specific and species-specific molecular markers. Analysis of molecular variance (AMOVA) was used to calculate the similarity values according to these polymorphic bands, and a dendrogram was constructed using NTSYS-pc software. Each genotype and species in this study could be distinguished from others, suggesting that DNA profiles based on ISSR and RAPD markers have produced potential diagnostic fingerprints for various species, and also for genotypes. The molecular phylogenetic tree shows that C. changshan-huyou and C. sinensis formed a subcluster, so we can conclude that C. sinensis is an assured parent of C. changshan-huyou. However the largest genetic distance was found between C. grandis and C. changshan-huyou, it might be explained that C. changshan-huyou is the origin of multitude of natural hybrids from C. sinensis, C. grandis and other species of Citrus.